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Sample records for irradiated babesia bovis

  1. Transient transfection of purified Babesia bovis merozoites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transient transfection of intraerythrocytic Babesia bovis parasites has been previously reported. In this study, we describe the development and optimization of methods for transfection of purified B. bovis merozoites using either nucleofection (Amaxa) or conventional electroporation (Gene Pulser II...

  2. Babesiosis (Babesia bovis) stability in unstable environments.

    PubMed

    Smith, R D; Evans, D E; Martins, J R; Ceresér, V H; Correa, B L; Petraccia, C; Cardozo, H; Solari, M A; Nari, A

    2000-01-01

    Enzootic stability (herd immunity) in bovine babesiosis occurs when the rate of transmission (inoculation rate) of Babesia spp by the tick vector is sufficient to immunize a majority of susceptible calves before the loss of calfhood resistance. The effect of three tick (Boophilus microplus) control strategies (none, threshold, and strategic) on enzootic stability and the likelihood of babesiosis (Babesia bovis) outbreaks was studied using a spreadsheet age-class computer simulation model. The model was driven by weekly bovine tick counts from Brazil and Uruguay. The Eldorado do Sul, RS, Brazil bovine population (30 degrees 05' South latitude) was found to be in a naturally occurring state of enzootic stability, corresponding to an inoculation rate exceeding 0.005 throughout the year. Threshold dipping strategies should not increase the risk of babesiosis in cattle so managed. Strategic dipping resulted in an extended period of enzootic instability lasting 30 weeks, which requires protection of the herd through immunization. Because of the more prolonged low winter temperature conditions, the Tacuarembó, Uruguay bovine population (31 degrees 40' South latitude) was found to be in a naturally occurring state of enzootic instability, characterized by a 28 week period in which the inoculation rate was below 0.005. Strategic dipping should lead to eradication of the babesial parasite from tick and bovine populations, but would not result in eradication of the tick vector. This could lead to subsequent outbreaks if Babesia carrier animals were to be introduced into the herd. In both populations, strategic tick control could be accompanied by concurrent babesiosis vaccination. PMID:11193666

  3. Tick passage results in enhanced attenuation of babesia bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Serial blood passage of virulent Babesia bovis in splenectomized cattle results in attenuated derivatives that do not cause neurologic disease. Tick transmissibility can be lost with attenuation, and has been reported to result in a reversion to virulence following tick passage. This study provides ...

  4. An impedance spectroscopy method for the detection and evaluation of Babesia bovis antibodies in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An immunosensor method for diagnosis of Babesia bovis in cattle based on impedance measurement is presented in this study. The method probes the interaction between serum antibodies against B. bovis infected cattle and recombinant protein, RAP-1, with C-terminal obtained from a Portuguese B. bovis s...

  5. Impact of Babesia bovis and Babesia bigemina on the production of beef cattle in Uruguay.

    PubMed

    Solari, M A; Nari, A; Cardozo, H

    1992-01-01

    Uruguay is situated in a marginal area for the development of Boophilus microplus (30 degrees 35 degrees South Lat.) with important areas of enzootic instability for Babesia bovis and B. bigemina. The livestock products represent 70% of our exports, for which reason it is fundamental to evaluate the losses in the production that these haemoparasites cause as basic information to take future decisions. In the period 1988-1990, several works were carried out by our laboratory to know the incidence of babesiosis in the reduction of liveweight gains. The results are shown and discussed in the work. Experiment I: the weight increase of the control group (x = 0.248 kg/day), was 23% higher than that of the infected group with Babesia spp (from Uruguay), but significant statistical differences were not found (P < 0.05). These animals were kept in boxes and the food was controlled for 76 days. Experiment II: the incidence of Babesia spp (same strain) was studied for 140 days on Hereford heifers (n = 14) on natural pastures. The control group obtained x = 25.29 kg of liveweight gain and it was 45% higher than that of the infected group, significant statistical difference were found (P < 0.05). Experiments with attenuated strains III: four studies were carried out inoculating B. bovis and B. bigemina in bovines about one year old, in different growth systems, searching for the limit of application. Significant statistical differences between those groups were not found during the experiment (about 180 days) (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1343684

  6. Expression of 6-Cys gene superfamily defines babesia bovis sexual stage development within rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia bovis, an intra-erythrocytic tick-borne apicomplexan protozoan, is one of the agents of bovine babesiosis. Its life cycle includes sexual reproduction within cattle fever ticks, Rhipicephalus spp. Six B. bovis 6-Cys gene superfamily members were previously identified (A, B, C, D, E, F) and t...

  7. Water Buffalos as carriers of Babesia bovis in Argentina.

    PubMed

    Ferreri, Lucas; Benitez, Daniel; Dominguez, Mariana; Rodriguez, Anabel; Asenzo, Gustavo; Mesplet, Maria; Florin-Christensen, Monica; Schnittger, Leonhard

    2008-12-01

    The tick-transmitted hemoprotozoan Babesia bovis is a major causative agent of bovine babesiosis, an often fatal disease of cattle. The disease is widespread in the northeastern region of Argentina, where an increasing part of the livestock is composed of water buffalos. Although clinical cases of buffalo babesiosis have not been reported so far, the pathogen-transmitting tick vector has been occasionally observed by us to be feeding on water buffalos. We therefore set out to examine whether buffalos may constitute a reservoir of the parasite. Competitive enzyme-linked immunosorbent assay (cELISA) detected B. bovis-specific antibodies in 20% of investigated buffalos (21/103), while direct detection of the pathogen by nested PCR was demonstrated in 34% of the animals (35/103). In one field, more than 60% of investigated animals (22/36) tested positive by nested PCR. These results are discussed in the context of buffalo babesiosis reported in other countries and in view of the currently effected control measures against bovine babesiosis in the region.

  8. Acute and persistent infection by a transfected Mo7 strain of Babesia bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Stable transfection of the Mo7 strain of Babesia bovis and expression of an exogenous gene has been demonstrated in long term culture. However, the use of transfected parasites as marker vaccines or vehicles for expressing exogenous antigens in vivo requires demonstration of acute and persistent inf...

  9. A virulent babesia bovis strain failed to infect white-tailed deer (Odocoileus virginianus)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus...

  10. The glycosylphosphatidylinositol-anchored protein repertoire of babesia bovis and its significance for erythrocyte invasion

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Glycosylphosphatidyl-anchored proteins are particularly abundant on the surface of pathogenic protozoans and might play an important role for parasite survival. In the present work the relevance of GPI-anchored proteins for erythrocyte invasion of Babesia bovis, one of the tick-transmitted causative...

  11. Babesia bovis expresses a neutralization-sensitive antigen that contains a microneme adhesive repeat (MAR) domain

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A gene coding for a protein with sequence similarity to the Toxoplasma gondii micronemal 1 (MIC1) protein that contains a copy of a domain described as a sialic acid-binding micronemal adhesive repeat was identified in the Babesia bovis genome. The single copy gene, located in chromosome 3, contains...

  12. Genome Sequence of Babesia bovis and Camparative Analysis of Apicomplexan Hemoprotozoa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia bovis is an apicomplexan tick-transmitted pathogen of cattle imposing a global risk and severe constraints to livestock health and economic development. The complete genome sequence was undertaken to facilitate vaccine antigen discovery, and to allow for comparative analysis with the related...

  13. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the northeast region of Thailand.

    PubMed

    Terkawi, Mohamad Alaa; Huyen, Nguyen Xuan; Shinuo, Cao; Inpankaew, Tawin; Maklon, Khuanwalai; Aboulaila, Mahmoud; Ueno, Akio; Goo, Youn-Kyoung; Yokoyama, Naoaki; Jittapalapong, Sathaporn; Xuan, Xuenan; Igarashi, Ikuo

    2011-06-10

    Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and Babesia bigemina and is characterized by significant morbidity and mortality worldwide. The disease is widespread in the northeastern region of Thailand, where an increasingly large part of the livestock is composed of water buffaloes. The present study was therefore conducted to investigate the epidemiological distribution of B. bovis and B. bigemina in water buffaloes in the northeastern region of Thailand. A total of 305 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the nested PCR (nPCR) assay, ELISA, and IFAT techniques. The overall prevalence of B. bovis and B. bigemina was 11.2% and 3.6% by nPCR, 14.7% and 5.9% by ELISA, and 16.8% and 5.6% by IFAT, respectively. The high concordance between the molecular and the serological detection tests revealed the specificity and sensitivity of the diagnostic assays used for the detection of infection as well as the endemic stability status of the parasites in the surveyed areas. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and location but not gender. Our data provide valuable information regarding the epidemiology of B. bovis and B. bigemina infection in water buffaloes in the northeastern region of Thailand which will likely be very beneficial for management and control programs of this disease.

  14. Molecular and serological prevalence of Babesia bovis and Babesia bigemina in water buffaloes in the north region of Brazil.

    PubMed

    da Silva, Jenevaldo Barbosa; André, Marcos Rogério; da Fonseca, Adivaldo Henrique; de Albuquerque Lopes, Cinthia Távora; da Silva Lima, Danillo Henrique; de Andrade, Stefano Juliano Tavares; Oliveira, Carlos Magno Chaves; Barbosa, José Diomedes

    2013-11-01

    Bovine babesiosis is a tick-borne disease caused mainly by Babesia bovis and Babesia bigemina, which are associated to considerable economic losses in cattle herds worldwide. Approximately 60% of buffalo herds in South America are located in Northern Brazil. Little is known about the impact of babesiosis on buffalo herds in Brazil. The present work aimed to verify the occurrence of B. bovis and B. bigemina in 542 water buffaloes in the state of Pará, Northern Brazil, using molecular and serological techniques. The percentage of seropositive animals for B. bovis and B. bigemina was 41.2% and 19.0%, respectively, by ELISA. B. bovis and B. bigemina DNA were detected in 15 and 16% of sampled buffaloes, respectively. A high correlation (Kappa index of 0.9) between serological and molecular tests suggests that the combination of the utilized techniques in the present study is suitable for babesiosis diagnosis in an endemic unstable area. Significantly difference of positivity for serological and molecular assays was verified to localities and reproductive status of sampled animals, but not between buffalo breeds. The immune status of sampled buffaloes associated to the circulation of babesiosis agents in sampled population suggests that the studied area is at risk to clinical babesiosis outbreaks. Furthermore, this study demonstrated that this region can be classified as endemically unstable.

  15. Expression of 6-Cys Gene Superfamily Defines Babesia bovis Sexual Stage Development within Rhipicephalus microplus

    PubMed Central

    Alzan, Heba F.; Herndon, David R.; Ueti, Massaro W.; Scoles, Glen A.; Kappmeyer, Lowell S.; Suarez, Carlos E.

    2016-01-01

    Babesia bovis, an intra-erythrocytic tick-borne apicomplexan protozoan, is one of the causative agents of bovine babesiosis. Its life cycle includes sexual reproduction within cattle fever ticks, Rhipicephalus spp. Six B. bovis 6-Cys gene superfamily members were previously identified (A, B, C, D, E, F) where their orthologues in Plasmodium parasite have been shown to encode for proteins required for the development of sexual stages. The current study identified four additional 6-Cys genes (G, H, I, J) in the B. bovis genome. These four genes are described in the context of the complete ten 6-Cys gene superfamily. The proteins expressed by this gene family are predicted to be secreted or surface membrane directed. Genetic analysis comparing the 6-Cys superfamily among five distinct B. bovis strains shows limited sequence variation. Additionally, A, B, E, H, I and J genes were transcribed in B. bovis infected tick midgut while genes A, B and E were also transcribed in the subsequent B. bovis kinete stage. Transcription of gene C was found exclusively in the kinete. In contrast, transcription of genes D, F and G in either B. bovis infected midguts or kinetes was not detected. None of the 6-Cys transcripts were detected in B. bovis blood stages. Subsequent protein analysis of 6-Cys A and B is concordant with their transcript profile. The collective data indicate as in Plasmodium parasite, certain B. bovis 6-Cys family members are uniquely expressed during sexual stages and therefore, they are likely required for parasite reproduction. Within B. bovis specifically, proteins encoded by 6-Cys genes A and B are markers for sexual stages and candidate antigens for developing novel vaccines able to interfere with the development of B. bovis within the tick vector. PMID:27668751

  16. Targeted Surface Expression of an Exogenous Antigen in Stably Transfected Babesia bovis

    PubMed Central

    Laughery, Jacob M.; Knowles, Donald P.; Schneider, David A.; Bastos, Reginaldo G.; McElwain, Terry F.; Suarez, Carlos E.

    2014-01-01

    Babesia bovis is a tick-borne intraerythocytic protozoan responsible for acute disease in cattle which can be controlled by vaccination with attenuated B. bovis strains. Emerging B. bovis transfection technologies may increase the usefulness of these live vaccines. One use of transfected B. bovis parasites may be as a vaccine delivery platform. Previous transfection methods for B. bovis were limited by single expression sites and intracellular expression of transfected antigens. This study describes a novel transfection system in which two exogenous genes are expressed: one for selection and the other for a selected antigen designed to be delivered to the surface of the parasites. The strategy for duplicating the number of transfected genes was based on the use of the putative bidirectional promoter of the B. bovis 1.4 Kb ef-1α intergenic region. The ability of this region to regulate two independent expression sites was demonstrated using a luciferase assay on transiently transfected B. bovis parasites and then incorporated into a stable transfection plasmid to control independent expression of the selectable marker GFP-BSD and another gene of interest. A chimeric gene was synthetized using sequences from the protective B-cell epitopes of Rhipicephalus microplus tick antigen Bm86 along with sequences from the surface exposed B. bovis major surface antigen-1. This chimeric gene was then cloned into the additional expression site of the transfection plasmid. Transfection of the B. bovis Mo7 strain with this plasmid resulted in stable insertion into the ef-1α locus and simultaneous expression of both exogenous genes. Expression of the Bm86 epitopes on the surface of transfected merozoites was demonstrated using immunofluorescence analyses. The ability to independently express multiple genes by the inclusion of a bidirectional promoter and the achievement of surface expression of foreign epitopes advances the potential of transfected B. bovis as a future vaccine

  17. Molecular and serological detection of Babesia bovis- and Babesiabigemina-infection in bovines and water buffaloes raised jointly in anendemic field

    Technology Transfer Automated Retrieval System (TEKTRAN)

    tBabesia bovis and Babesia bigemina are causative agents of bovine babesiosis, a tick-borne disease of cattlein tropical and subtropical regions. Babesia spp. infection adversely affects cattle health and can be fatalresulting in considerable economic loss worldwide. Under endemic stability conditio...

  18. First survey for Babesia bovis and Babesia bigemina infection in cattle from Central and Southern regions of Portugal using serological and DNA detection methods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Incidence of bovine babesiosis in Portugal is currently unknown. In this study, a first survey of Babesia bovis and B. bigemina infection in cattle was carried out using blood samples from 406 clinically healthy individuals from different districts from Central and Southern regions of Portugal and a...

  19. A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus)

    PubMed Central

    Freeman, Jeanne M.; Johnson, Wendell C.; Scoles, Glen A.

    2015-01-01

    Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus spp., that can cause up to 90% mortality in naive adult cattle. While cattle are the primary host for cattle fever ticks, wild and exotic ungulates, including white-tailed deer (WTD), are known to be viable alternative hosts. The presence of cattle fever tick populations resistant to acaricides raises concerns regarding the possibility of these alternative hosts introducing tick-borne babesial parasites into areas free of infection. Understanding the B. bovis reservoir competence of these alternative hosts is critical to mitigating the risk of introduction. In this study, we tested the hypothesis that WTD are susceptible to infection with a B. bovis strain lethal to cattle. Two groups of deer were inoculated intravenously with either B. bovis blood stabilate or a larval extract supernatant containing sporozoites from infected R. microplus larvae. The collective data demonstrated that WTD are neither a transient host nor reservoir of B. bovis. This conclusion is supported by the failure of B. bovis to establish an infection in deer regardless of inoculum. Although specific antibody was detected for a short period in the WTD, the PCR results were consistently negative at multiple time points throughout the experiment and blood from WTD that had been exposed to parasite, transferred into naïve recipient susceptible calves, failed to establish infection. In contrast, naïve steers inoculated intravenously with either B. bovis blood stabilate or the larval extract supernatant containing sporozoites rapidly succumbed to disease. These findings provide evidence that WTD are not an epidemiological component in the maintenance of B. bovis infectivity to livestock. PMID:26083429

  20. A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus).

    PubMed

    Ueti, Massaro W; Olafson, Pia U; Freeman, Jeanne M; Johnson, Wendell C; Scoles, Glen A

    2015-01-01

    Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus spp., that can cause up to 90% mortality in naive adult cattle. While cattle are the primary host for cattle fever ticks, wild and exotic ungulates, including white-tailed deer (WTD), are known to be viable alternative hosts. The presence of cattle fever tick populations resistant to acaricides raises concerns regarding the possibility of these alternative hosts introducing tick-borne babesial parasites into areas free of infection. Understanding the B. bovis reservoir competence of these alternative hosts is critical to mitigating the risk of introduction. In this study, we tested the hypothesis that WTD are susceptible to infection with a B. bovis strain lethal to cattle. Two groups of deer were inoculated intravenously with either B. bovis blood stabilate or a larval extract supernatant containing sporozoites from infected R. microplus larvae. The collective data demonstrated that WTD are neither a transient host nor reservoir of B. bovis. This conclusion is supported by the failure of B. bovis to establish an infection in deer regardless of inoculum. Although specific antibody was detected for a short period in the WTD, the PCR results were consistently negative at multiple time points throughout the experiment and blood from WTD that had been exposed to parasite, transferred into naïve recipient susceptible calves, failed to establish infection. In contrast, naïve steers inoculated intravenously with either B. bovis blood stabilate or the larval extract supernatant containing sporozoites rapidly succumbed to disease. These findings provide evidence that WTD are not an epidemiological component in the maintenance of B. bovis infectivity to livestock.

  1. A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus).

    PubMed

    Ueti, Massaro W; Olafson, Pia U; Freeman, Jeanne M; Johnson, Wendell C; Scoles, Glen A

    2015-01-01

    Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus spp., that can cause up to 90% mortality in naive adult cattle. While cattle are the primary host for cattle fever ticks, wild and exotic ungulates, including white-tailed deer (WTD), are known to be viable alternative hosts. The presence of cattle fever tick populations resistant to acaricides raises concerns regarding the possibility of these alternative hosts introducing tick-borne babesial parasites into areas free of infection. Understanding the B. bovis reservoir competence of these alternative hosts is critical to mitigating the risk of introduction. In this study, we tested the hypothesis that WTD are susceptible to infection with a B. bovis strain lethal to cattle. Two groups of deer were inoculated intravenously with either B. bovis blood stabilate or a larval extract supernatant containing sporozoites from infected R. microplus larvae. The collective data demonstrated that WTD are neither a transient host nor reservoir of B. bovis. This conclusion is supported by the failure of B. bovis to establish an infection in deer regardless of inoculum. Although specific antibody was detected for a short period in the WTD, the PCR results were consistently negative at multiple time points throughout the experiment and blood from WTD that had been exposed to parasite, transferred into naïve recipient susceptible calves, failed to establish infection. In contrast, naïve steers inoculated intravenously with either B. bovis blood stabilate or the larval extract supernatant containing sporozoites rapidly succumbed to disease. These findings provide evidence that WTD are not an epidemiological component in the maintenance of B. bovis infectivity to livestock. PMID:26083429

  2. Phylogenetic analysis of Mexican Babesia bovis isolates using msa and ssrRNA gene sequences.

    PubMed

    Genis, Alma D; Mosqueda, Juan J; Borgonio, Verónica M; Falcón, Alfonso; Alvarez, Antonio; Camacho, Minerva; de Lourdes Muñoz, Maria; Figueroa, Julio V

    2008-12-01

    Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2), and msa-2b). Small subunit ribosomal (ssr)RNA gene is subject to evolutive pressure and has been used in phylogenetic studies. To determine the phylogenetic relationship among B. bovis Mexican isolates using different genetic markers, PCR amplicons, corresponding to msa-1, msa-2c, msa-2b, and ssrRNA genes, were cloned and plasmids carrying the corresponding inserts were sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 12 geographically different B. bovis isolates and a reference strain. Overall sequence identities of 47.7%, 72.3%, 87.7%, and 94% were determined for msa-1, msa-2b, msa-2c, and ssrRNA, respectively. A robust phylogenetic tree was obtained with msa-2b sequences. The phylogenetic analysis suggests that Mexican B. bovis isolates group in clades not concordant with the Mexican geography. However, the Mexican isolates group together in an American clade separated from the Australian clade. Sequence heterogeneity in msa-1, msa-2b, and msa-2c coding regions of Mexican B. bovis isolates present in different geographical regions can be a result of either differential evolutive pressure or cattle movement from commercial trade.

  3. Natural Babesia bovis Infection in Water Buffaloes (Bubalus bubalis) and Crossbred Cattle under Field Conditions in Egypt: a Preliminary Study

    PubMed Central

    Mahmmod, Yasser

    2014-01-01

    Background There is a little or no data available on the natural Babesia bovis (B. bovis) infection in water buffaloes (Bubalus bubalis) comparing to the available one for cattle. This study was conducted to investigate the natural B. bovis infection in water buffaloes in comparison to crossbred cattle under field conditions in Egypt. Methods: A total of 35 buffaloes and cattle were clinically and laboratory investigated from March to June 2008. Twenty-nine buffaloes and cattle out of 35 were naturally infected with B. bovis and showed signs of bovine babesiosis. Three cows and three buffaloes showed no clinical signs and were free from external, internal, and blood parasites served as control group. Results: Babesia bovis-infected cattle showed typical signs of bovine babesiosis while B. bovis-infected buffaloes showed a milder form (less severe) of the clinical signs. Advanced cases of cattle showed dark brown to dark red (coffee-color) urine, hemoglobinuria and nervous manifestations while these manifestations were not detected in the infected buffaloes. Hematological changes in both species however, these changes were less significant in buffaloes than those reported in cattle. Conclusion: This paper documents the first description of natural B. bovis infection in water buffaloes which were found to be more likely to be tolerant than cattle to the natural clinical infection with B. bovis and its subsequent haematological changes. Our finding may lead to a better understanding of the disease pattern of B. bovis infection under field conditions in buffaloes. PMID:25629060

  4. Sequence conservation of the 12D3 gene in Mexican isolates of Babesia bovis.

    PubMed

    Perez, J; Javier Perez, J; Vargas, P; Antonio Alvarez, J; Rojas, C; Figueroa, J V

    2010-04-01

    The 12D3 antigen present in Babesia bovis has been evaluated as a recombinant vaccine candidate and the 12d3 coding sequence has been reported for an Australian and an USA (Texas) isolate of B. bovis. However, no approach has been conducted to perform analysis of 12d3 sequence conservation on a larger number of B. bovis isolates. This could provide important information to determine whether a recombinant vaccine containing this antigen could be widely used. This study reports the cloning and sequencing analysis of the 12d3 coding region in 20 different B. bovis isolates collected from various geographical regions in the tropics and subtropics of Mexico. Comparative analysis of the consensus nucleotide sequences obtained for each isolate revealed a high degree of conservation (94-99% sequence identity) among the 12d3 alleles present in the Mexican isolates when compared with the 12d3 ORF sequences from the Texan (T2Bo) B. bovis isolate. Similarly, BLASTX sequence homology search showed a high percent identity (93-99%) of the deduced amino acid 12D3 sequence as compared with the T2Bo isolate sequence. The high level of sequence conservation in 12d3 among the 20 B. bovis isolates collected from geographically distant locations in Mexico suggests that there exists a minimal bovine-host immunological pressure which could be translated into antigenic diversity or variation, and most probably this is reflected in the non-inmunodominant characteristic of the 12D3 antigen as it has been previously described in the literature. 12D3 antigen can be considered as a viable candidate for inclusion in a recombinant vaccine for cattle babesiosis caused by B. bovis in Mexico.

  5. Dielectrophoretic sorting on a microfabricated flow cytometer: label free separation of Babesia bovis infected erythrocytes.

    PubMed

    Nascimento, Elisabete M; Nogueira, Nuno; Silva, Tiago; Braschler, Thomas; Demierre, Nicolas; Renaud, Philippe; Oliva, Abel G

    2008-08-01

    Dielectrophoresis is a method that has demonstrated great potential in cell discrimination and isolation. In this study, the dielectrophoretic sorting of normal and Babesia bovis infected erythrocytes was performed using a microfabricated flow cytometer. Separation was possible through exploitation of the dielectric differences between normal and infected erythrocytes, essentially due to the higher ionic membrane permeability of B. bovis infected cells. Sorting experiments were performed inside a microchip made from Pt microelectrodes and SU-8 channels patterned on a glass substrate. Optimum cell separation was achieved at 4 MHz using an in vitro culture of B. bovis suspended in 63 mS/m phosphate buffer and applying a sinusoidal voltage of 15 V peak-to-peak. Normal erythrocytes experienced stronger positive dielectrophoresis (pDEP) than B. bovis infected cells, moving them closer to the microelectrodes. Under these conditions it was possible to enrich the fraction of infected cells from 7 to 50% without the need of extensive sample preparation or labelling. Throughout the experiments very few microliters of sample were used, suggesting that this system may be considered suitable for integration in a low-cost automated device to be used in the in situ diagnostic of babesiosis. PMID:18511353

  6. Evaluation of the growth-inhibitory effect of trifluralin analogues on in vitro cultured babesia bovis parasites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia bovis caused bovine babesiosis is a world tick borne hemoprotozoan disease leading to fever, anemia, weight losses and ultimately death. Several babesicidal drugs that have been in use in cattle for years have proven to be partially ineffective and the development of alternative highly speci...

  7. Expressed gene sequences from larval genes over-expressed upon Babesia bovis infection of Rhipecephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  8. Expression and strain variation of the novel “Small Open Reading Frame” 3 (smorf) multigene family in Babesia bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Small open reading frame (smorf) genes comprise the second largest Babesia bovis multigene family. All known 44 variant smorf genes are located in close chromosomal proximity to ves1 genes, which encode proteins that mediate cytoadhesion and contribute to immune evasion. In this study, we characte...

  9. A novel neutralization sensitive and subdominant RAP-1-related antigen (RRA) is expressed by babesia bovis merozoites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objective: The Babesia bovis genome encodes a rap-1 related gene denominated RAP-1 related antigen (RRA). In this study, we analyzed the pattern of expression, immunogenicity and functional relevance of RRA. Methods: Phylogenetic analysis was performed using the program Phylip. Expression of rra wa...

  10. Differential response of splenic monocytes and DC from cattle to microbial stimulation with Mycobacterium bovis BCG and Babesia bovis merozoites.

    PubMed

    Bastos, R G; Johnson, W C; Brown, W C; Goff, W L

    2007-02-15

    Both bovine peripheral blood monocyte-derived dendritic cells (DC) and myeloid DC from afferent lymph have been described, but resident DC from other bovine tissues have not been fully characterized. The spleen as a secondary lymphoid organ is central to the innate and acquired immune response to various diseases particularly hemoprotozoan infections like babesiosis. Therefore, we developed methods to demonstrate the presence of myeloid DC from the spleen of cattle and have partially characterized a DC population as well as another myeloid cell population with monocyte characteristics. The phenotypic profile of each population was CD13+CD172a+/-CD14-CD11a-CD11b+/-CD11c+ and CD172a+CD13+/-CD14+CD11a-CD11b+/-CD11c+, respectively. The CD13+ population was found exclusively in the spleen whereas the CD172a+ population was present at the same percentage in the spleen and peripheral blood. CD13+ cells developed a typical veiled appearance when in culture for 96 h. The two cell populations differed in their ability to produce nitric oxide and had a different pattern of cytokine mRNA when stimulated with Mycobacterium bovis BCG or Babesia bovis merozoites. The data demonstrate the presence of a myeloid splenic DC with attributes consistent with an immature status.

  11. Babesia bovis and Babesia bigemina infection levels estimated by qPCR in Angus cattle from an endemic area of São Paulo state, Brazil.

    PubMed

    Giglioti, R; Oliveira, H N; Santana, C H; Ibelli, A M G; Néo, T A; Bilhassi, T B; Rabelo, M D; Machado, R Z; Brito, L G; Oliveira, M C S

    2016-07-01

    The levels of infection by Babesia bovis and Babesia bigemina were estimated by absolute quantification through the quantitative PCR technique (qPCR). Fifty-one contemporaneous Angus cattle were evaluated on two occasions. The number of standard female Rhipicephalus microplus ticks present on the left side of the body was counted and blood samples were drawn from the tail vein into tubes containing the anticoagulant EDTA. The blood samples were submitted to DNA extraction and used to quantify the number of copies (NC) of DNA from B. bovis and B. bigemina by qPCR. The data on tick count and number of DNA copies were transformed for normalization and analyzed by a mixed model method. A multivariate model with repeated measures of the same animal, including the effects of collection, parasite species and their interaction, was used. The repeatability values were obtained from the matrix of (co)variances and were expressed for each species. The correlations between the counts of different species on the same animal, in the same collection or different collections, were also estimated. The results showed the qPCR could distinguish the two between infection by the two Babesia species. Infection levels by B. bovis and B. bigemina were detected in 100% and 98% of the animals, respectively. Significant differences were found (P<0.05) between the NC of the two Babesia species, B. bovis 1.49±0.07 vs. B. bigemina 0.82±0.06. Low repeatabilities were found for the counts of R. microplus and NC of B. bovis and B. bigemina: 0.05, 0.10 and 0.02, respectively. The correlations between R. microplus count and NC of B. bovis and B. bigemina were both very near zero. However, an association was observed between the NC of the two species, with a correlation coefficient of 0.30 for measures from the same collection. The absence of associations between the quantity of DNA from B. bovis and B. bigemina and the tick counts suggests that the variation of parasitemia by the hemoparasites did

  12. Babesia bovis and Babesia bigemina infection levels estimated by qPCR in Angus cattle from an endemic area of São Paulo state, Brazil.

    PubMed

    Giglioti, R; Oliveira, H N; Santana, C H; Ibelli, A M G; Néo, T A; Bilhassi, T B; Rabelo, M D; Machado, R Z; Brito, L G; Oliveira, M C S

    2016-07-01

    The levels of infection by Babesia bovis and Babesia bigemina were estimated by absolute quantification through the quantitative PCR technique (qPCR). Fifty-one contemporaneous Angus cattle were evaluated on two occasions. The number of standard female Rhipicephalus microplus ticks present on the left side of the body was counted and blood samples were drawn from the tail vein into tubes containing the anticoagulant EDTA. The blood samples were submitted to DNA extraction and used to quantify the number of copies (NC) of DNA from B. bovis and B. bigemina by qPCR. The data on tick count and number of DNA copies were transformed for normalization and analyzed by a mixed model method. A multivariate model with repeated measures of the same animal, including the effects of collection, parasite species and their interaction, was used. The repeatability values were obtained from the matrix of (co)variances and were expressed for each species. The correlations between the counts of different species on the same animal, in the same collection or different collections, were also estimated. The results showed the qPCR could distinguish the two between infection by the two Babesia species. Infection levels by B. bovis and B. bigemina were detected in 100% and 98% of the animals, respectively. Significant differences were found (P<0.05) between the NC of the two Babesia species, B. bovis 1.49±0.07 vs. B. bigemina 0.82±0.06. Low repeatabilities were found for the counts of R. microplus and NC of B. bovis and B. bigemina: 0.05, 0.10 and 0.02, respectively. The correlations between R. microplus count and NC of B. bovis and B. bigemina were both very near zero. However, an association was observed between the NC of the two species, with a correlation coefficient of 0.30 for measures from the same collection. The absence of associations between the quantity of DNA from B. bovis and B. bigemina and the tick counts suggests that the variation of parasitemia by the hemoparasites did

  13. Genome Sequence of Babesia bovis and Comparative Analysis of Apicomplexan Hemoprotozoa

    PubMed Central

    Brayton, Kelly A; Lau, Audrey O. T; Herndon, David R; Hannick, Linda; Kappmeyer, Lowell S; Berens, Shawn J; Bidwell, Shelby L; Brown, Wendy C; Crabtree, Jonathan; Fadrosh, Doug; Feldblum, Tamara; Forberger, Heather A; Haas, Brian J; Howell, Jeanne M; Khouri, Hoda; Koo, Hean; Mann, David J; Norimine, Junzo; Paulsen, Ian T; Radune, Diana; Ren, Qinghu; Smith, Roger K; Suarez, Carlos E; White, Owen; Wortman, Jennifer R; Knowles, Donald P; McElwain, Terry F; Nene, Vishvanath M

    2007-01-01

    Babesia bovis is an apicomplexan tick-transmitted pathogen of cattle imposing a global risk and severe constraints to livestock health and economic development. The complete genome sequence was undertaken to facilitate vaccine antigen discovery, and to allow for comparative analysis with the related apicomplexan hemoprotozoa Theileria parva and Plasmodium falciparum. At 8.2 Mbp, the B. bovis genome is similar in size to that of Theileria spp. Structural features of the B. bovis and T. parva genomes are remarkably similar, and extensive synteny is present despite several chromosomal rearrangements. In contrast, B. bovis and P. falciparum, which have similar clinical and pathological features, have major differences in genome size, chromosome number, and gene complement. Chromosomal synteny with P. falciparum is limited to microregions. The B. bovis genome sequence has allowed wide scale analyses of the polymorphic variant erythrocyte surface antigen protein (ves1 gene) family that, similar to the P. falciparum var genes, is postulated to play a role in cytoadhesion, sequestration, and immune evasion. The ∼150 ves1 genes are found in clusters that are distributed throughout each chromosome, with an increased concentration adjacent to a physical gap on chromosome 1 that contains multiple ves1-like sequences. ves1 clusters are frequently linked to a novel family of variant genes termed smorfs that may themselves contribute to immune evasion, may play a role in variant erythrocyte surface antigen protein biology, or both. Initial expression analysis of ves1 and smorf genes indicates coincident transcription of multiple variants. B. bovis displays a limited metabolic potential, with numerous missing pathways, including two pathways previously described for the P. falciparum apicoplast. This reduced metabolic potential is reflected in the B. bovis apicoplast, which appears to have fewer nuclear genes targeted to it than other apicoplast containing organisms. Finally

  14. Diversity of Babesia bovis merozoite surface antigen genes in the Philippines.

    PubMed

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Ybanez, Adrian Patalinghug; Ybanez, Rochelle Haidee Daclan; Perez, Zandro Obligado; Guswanto, Azirwan; Igarashi, Ikuo; Yokoyama, Naoaki

    2014-02-01

    Babesia bovis is the causative agent of fatal babesiosis in cattle. In the present study, we investigated the genetic diversity of B. bovis among Philippine cattle, based on the genes that encode merozoite surface antigens (MSAs). Forty-one B. bovis-positive blood DNA samples from cattle were used to amplify the msa-1, msa-2b, and msa-2c genes. In phylogenetic analyses, the msa-1, msa-2b, and msa-2c gene sequences generated from Philippine B. bovis-positive DNA samples were found in six, three, and four different clades, respectively. All of the msa-1 and most of the msa-2b sequences were found in clades that were formed only by Philippine msa sequences in the respective phylograms. While all the msa-1 sequences from the Philippines showed similarity to those formed by Australian msa-1 sequences, the msa-2b sequences showed similarity to either Australian or Mexican msa-2b sequences. In contrast, msa-2c sequences from the Philippines were distributed across all the clades of the phylogram, although one clade was formed exclusively by Philippine msa-2c sequences. Similarities among the deduced amino acid sequences of MSA-1, MSA-2b, and MSA-2c from the Philippines were 62.2-100, 73.1-100, and 67.3-100%, respectively. The present findings demonstrate that B. bovis populations are genetically diverse in the Philippines. This information will provide a good foundation for the future design and implementation of improved immunological preventive methodologies against bovine babesiosis in the Philippines. The study has also generated a set of data that will be useful for futher understanding of the global genetic diversity of this important parasite.

  15. Using msa-2b as a molecular marker for genotyping Mexican isolates of Babesia bovis.

    PubMed

    Genis, Alma D; Perez, Jocelin; Mosqueda, Juan J; Alvarez, Antonio; Camacho, Minerva; Muñoz, Maria de Lourdes; Rojas, Carmen; Figueroa, Julio V

    2009-12-01

    Variable merozoite surface antigens of Babesia bovis are exposed glycoproteins having a role in erythrocyte invasion. Members of this gene family include msa-1 and msa-2 (msa-2c, msa-2a(1), msa-2a(2) and msa-2b). To determine the sequence variation among B. bovis Mexican isolates using msa-2b as a genetic marker, PCR amplicons corresponding to msa-2b were cloned and plasmids carrying the corresponding inserts were purified and sequenced. Comparative analysis of nucleotide and deduced amino acid sequences revealed distinct degrees of variability and identity among the coding gene sequences obtained from 16 geographically different Mexican B. bovis isolates and a reference strain. Clustal-W multiple alignments of the MSA-2b deduced amino acid sequences performed with the 17 B. bovis Mexican isolates, revealed the identification of three genotypes with a distinct set each of amino acid residues present at the variable region: Genotype I represented by the MO7 strain (in vitro culture-derived from the Mexico isolate) as well as RAD, Chiapas-1, Tabasco and Veracruz-3 isolates; Genotype II, represented by the Jalisco, Mexico and Veracruz-2 isolates; and Genotype III comprising the sequences from most of the isolates studied, Tamaulipas-1, Chiapas-2, Guerrero-1, Nayarit, Quintana Roo, Nuevo Leon, Tamaulipas-2, Yucatan and Guerrero-2. Moreover, these three genotypes could be discriminated against each other by using a PCR-RFLP approach. The results suggest that occurrence of indels within the variable region of msa-2b sequences can be useful markers for identifying a particular genotype present in field populations of B. bovis isolated from infected cattle in Mexico.

  16. Rapid and sensitive detection of Babesia bovis and Babesia bigemina by loop-mediated isothermal amplification combined with a lateral flow dipstick.

    PubMed

    Yang, Yimin; Li, Qun; Wang, Suhua; Chen, Xueqiu; Du, Aifang

    2016-03-30

    Babesia spp. are apicomplexan protozoan parasites of the red blood cells of mammals and are transmitted by ticks. Bovine babesiosis mainly caused by Babesia bovis and Babesia bigemina occurs worldwide, which is a great threat to animal health. Microscopy examination is a gold standard for the diagnosis of babesiosis. However, its sensitivity is too low. This study was conducted to establish a simple, efficient and fast LAMP-LFP method used for early diagnosis of animal babesiosis. LAMP was developed with a set of four primers targeting and amplifying six distinct regions of cytochrome b gene of Babesia spp. under isothermal conditions. Afterwards, a chromatographic lateral-flow dipstick (LFD) was used to detect LAMP products that were labeled with FITC at the 5' end, avoiding gel electrophoresis. The LAMP-LFD method was very specific, yielding no positive results with DNA templates of Theileria sergenti, Thenileria ovis, Theileria equi and Toxoplasma gondii. The LAMP-LFP was highly sensitive and could detect 0.85 fg B. bigemina DNA and 0.14 fg B. bovis DNA, 100-fold higher than a conventional PCR assay. This method could be adapted for quick and accurate diagnosis of bovine babesiosis in the fields in case the whole blood could be directly used, especially for identifying carrier animals with very low parasitaemia. PMID:26921043

  17. Studies on the pathogenicity of Babesia bovis in water buffaloes after cryopreservation and resuscitation.

    PubMed

    Yao, B; Zhao, J; Ma, L; Liu, Z

    1997-11-01

    Packed erythrocytes infected with Babesia bovis were mixed with an equal volume of 16% dimethyl sulphoxide (DMSO) in Alsever's solution and dispensed into 1.5 or 5 ml cryotubes. The vials were kept in liquid nitrogen (-196 degrees C) for 26, 78, 142 or 149 days. The samples were removed from the liquid nitrogen container and rapidly thawed in a 40 degrees C water bath. The thawed blood successfully infected splenectomised buffalo calves by injection via subcutaneous or intravenous or via intravenous and subcutaneous routes. The parasites, typical B. bovis, were discovered in red blood cells 5, 8 or 9 days after inoculation. The highest percentage of parasitised erythrocytes (PPE) was 15%. The babesiosis resulting from cryopreserved parasites was the same as that resulting from fresh parasites inoculated by ticks. Typical clinical signs were found, such as continuous fever (the highest temperature was 41.3 degrees C), anaemia, icterus and haemoglobinuria. Infected calves, which were not treated, died. Cryopreservation is a simple and reliable method for longterm preservation of B. bovis of water buffaloes.

  18. Molecular and serological prevalence of Babesia bigemina and Babesia bovis in cattle and water buffalos under small-scale dairy farming in Beheira and Faiyum Provinces, Egypt.

    PubMed

    Ibrahim, Hany M; Adjou Moumouni, Paul F; Mohammed-Geba, Khaled; Sheir, Sherin K; Hashem, Ihab S Y; Cao, Shinuo; Terkawi, Mohamad A; Kamyingkird, Ketsarin; Nishikawa, Yoshifumi; Suzuki, Hiroshi; Xuan, Xuenan

    2013-11-15

    In order to determine the molecular and serological prevalence of Babesia bigemina and Babesia bovis, a total of 247 blood samples were collected from cattle and water buffalos in Beheira and Faiyum Provinces in Egypt and examined by nested polymerase chain reaction (nPCR) and enzyme-linked immunosorbent assay (ELISA). In cattle, the prevalence of B. bigemina and B. bovis was 5.30% and 3.97% by nPCR and 10.60% and 9.27% by ELISA, respectively, whereas those of water buffalos were 10.42% and 4.17% by nPCR and 15.63% and 11.46% by ELISA, respectively. Statistically significant differences in the prevalence of the two infections were observed on the basis of age and health status. Sequencing analysis revealed two genotypes for B. bovis spherical body protein-4. In conclusion, the current data provide valuable information regarding the epidemiology of B. bigemina and B. bovis infections in cattle and water buffalos from Egypt, which can be employed in developing future strategies for disease management and control.

  19. Molecular and serological detection of Babesia bovis- and Babesia bigemina-infection in bovines and water buffaloes raised jointly in an endemic field.

    PubMed

    Romero-Salas, Dora; Mira, Anabela; Mosqueda, Juan; García-Vázquez, Zeferino; Hidalgo-Ruiz, Mario; Vela, Noot Aditya Ortiz; de León, Adalberto Angel Perez; Florin-Christensen, Monica; Schnittger, Leonhard

    2016-02-15

    Babesia bovis and Babesia bigemina are causative agents of bovine babesiosis, a tick-borne disease of cattle in tropical and subtropical regions. Babesia spp. infection adversely affects cattle health and can be fatal resulting in considerable economic loss worldwide. Under endemic stability conditions, herds contain high numbers of chronically infected, asymptomatic carrier animals, in which no parasitemia is detected by microscopic blood smear examination. In addition to bovines, also water buffaloes are infected by both Babesia spp. commonly leading to a subclinical infection. The infection rate (by nPCR) and herd exposure (by IFAT) of bovines and water buffaloes reared under similar field conditions in an area of endemic stability were determined and compared. In order to optimize direct parasite detection, highly sensitive nPCR assays were developed and applied, allowing the detection of as little as 0.1 fg DNA of each Babesia pathogen. Significantly lower percentages (p<0.001) of seropositive water buffaloes compared to bovines were observed for B. bovis (71.4% vs. 98%) and B. bigemina (85% vs. 100%). Interestingly, in comparison, differences noticed between water buffaloes and bovines were considerably larger with direct parasite detection by nPCR (16.2% vs. 82.3% and 24% vs. 94.1% for B. bovis and B. bigemina, respectively). As expected, bovines subjected to monthly acaricide applications exhibited a significant lower infection rate as determined by nPCR than bovines not subjected to these measures (B. bovis 33.3% vs. 90.7%, p<0.001; B. bigemina 80% vs. 96.5%, p<0.001, for treated vs. untreated animals). Interestingly no differences between these groups were observed with respect to seropositivity, suggesting similar rates of parasite exposure (B. bovis 100% vs. 97.7%, p<0.001; B. bigemina 100% vs. 100%, p<0.001). Importantly, a significantly higher number of water buffaloes as determined by nPCR were infected when reared jointly with bovines not subjected

  20. Molecular and serological detection of Babesia bovis- and Babesia bigemina-infection in bovines and water buffaloes raised jointly in an endemic field.

    PubMed

    Romero-Salas, Dora; Mira, Anabela; Mosqueda, Juan; García-Vázquez, Zeferino; Hidalgo-Ruiz, Mario; Vela, Noot Aditya Ortiz; de León, Adalberto Angel Perez; Florin-Christensen, Monica; Schnittger, Leonhard

    2016-02-15

    Babesia bovis and Babesia bigemina are causative agents of bovine babesiosis, a tick-borne disease of cattle in tropical and subtropical regions. Babesia spp. infection adversely affects cattle health and can be fatal resulting in considerable economic loss worldwide. Under endemic stability conditions, herds contain high numbers of chronically infected, asymptomatic carrier animals, in which no parasitemia is detected by microscopic blood smear examination. In addition to bovines, also water buffaloes are infected by both Babesia spp. commonly leading to a subclinical infection. The infection rate (by nPCR) and herd exposure (by IFAT) of bovines and water buffaloes reared under similar field conditions in an area of endemic stability were determined and compared. In order to optimize direct parasite detection, highly sensitive nPCR assays were developed and applied, allowing the detection of as little as 0.1 fg DNA of each Babesia pathogen. Significantly lower percentages (p<0.001) of seropositive water buffaloes compared to bovines were observed for B. bovis (71.4% vs. 98%) and B. bigemina (85% vs. 100%). Interestingly, in comparison, differences noticed between water buffaloes and bovines were considerably larger with direct parasite detection by nPCR (16.2% vs. 82.3% and 24% vs. 94.1% for B. bovis and B. bigemina, respectively). As expected, bovines subjected to monthly acaricide applications exhibited a significant lower infection rate as determined by nPCR than bovines not subjected to these measures (B. bovis 33.3% vs. 90.7%, p<0.001; B. bigemina 80% vs. 96.5%, p<0.001, for treated vs. untreated animals). Interestingly no differences between these groups were observed with respect to seropositivity, suggesting similar rates of parasite exposure (B. bovis 100% vs. 97.7%, p<0.001; B. bigemina 100% vs. 100%, p<0.001). Importantly, a significantly higher number of water buffaloes as determined by nPCR were infected when reared jointly with bovines not subjected

  1. Babesia bovis: a bipartite signal directs the glutamyl-tRNA synthetase to the apicoplast.

    PubMed

    Pedroni, Monica J; Luu, Tracy N K; Lau, Audrey O T

    2012-06-01

    Babesia bovis contains a prokaryotic derived organelle known as the apicoplast. Many participants of the metabolic pathways within the apicoplast are encoded in the nuclear genome and post-translationally imported with the help of a bipartite signal. Recently, an all encompassing algorithm was derived to predict apicoplast targeted proteins for many non-Plasmodium apicomplexans in which it reported the presence of 260 apicoplast targeted proteins in Babesia. One of these proteins is glutamyl tRNA synthetase (GltX). This study investigates if the putative bipartite signal of GltX alone is sufficient to direct proteins into the apicoplast. Using a transient transfection system consisting of a green fluorescent protein as the reporter, we tested the signal and transit portions of the bipartite signal in apicoplastic transport. We first identified the transcript of gltX to be expressed during the asexual blood stages and subsequently confirmed that the complete bipartite signal is responsible for directing the reporter protein into a compartment distinct from the nucleus and the mitochondrion. As GltX bipartite signal successfully guided the reporter protein into the apicoplast, our finding implies that it also directs native GltX into the same organelle.

  2. Transfection of babesia bovis by double selection with WR99210 and blasticidin-S and its application for functional analysis of thioredoxin peroxidase-1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Genetic manipulation is an essential technique to analyze gene function; however, limited methods are available for Babesia bovis, a causative pathogen of the globally important cattle disease, bovine babesiosis. To date, two stable transfection systems have been developed for B. bovis, using select...

  3. Molecular and seroepidemiological survey of Babesia bovis and Babesia bigemina infections in cattle and water buffaloes in the central region of Vietnam.

    PubMed

    Li, Yan; Luo, Yuzi; Cao, Shinuo; Terkawi, Mohamad Alaa; Lan, Dinh Thi Bich; Long, Phung Thang; Yu, Longzheng; Zhou, Mo; Gong, Haiyan; Zhang, Houshuang; Zhou, Jinlin; Yokoyama, Naoaki; Suzuki, Hiroshi; Xuan, Xuenan

    2014-09-01

    In the present study, a total of 137 blood samples were collected from cattle and water buffaloes in central region of Vietnam and tested using nested polymerase chain reaction (nPCR), enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT) to determine the molecular and serological prevalence of Babesia bovis and Babesia bigemina. In cattle, the prevalence of B. bovis and B. bigemina was 21.3% and 16.0% by nPCR, 73.4% and 42.6% by ELISA and 60.6% and 59.6% by IFAT, respectively, whereas those of water buffalos were 23.3% and 0% by nPCR, 37.2% and 9.3% by ELISA and 27.9% and 18.6% by IFAT, respectively. IFAT and ELISA detected a higher number of infected cattle and water buffaloes than nPCR totally. Statistically significant differences in the prevalence of the two infections were observed on the basis of age. Overall, the current data suggest high incidence of B. bovis and B. bigemina infections in the central region of Vietnam, which is needed to develop comprehensive approach to the modern surveillance, diagnosis and control of bovine babesiosis.

  4. Genotypic diversity in Babesia bovis field isolates and vaccine strains from South Africa.

    PubMed

    Combrink, M P; Troskie, P C; Pienaar, R; Latif, A A; Mans, B J

    2014-01-31

    Genotypic diversity in Babesia bovis (cause of Asiatic redwater in cattle) vaccine strains and field isolates from South Africa were investigated using the Bv80 gene as well as microsatellites. The S11 vaccine strain possessed both A and B alleles of the Bv80 gene, as well as genotypic diversity within each allele type as defined by repeat variation resulting in different amplicon sizes. Rapid serial passage of vaccine strain from passage S10 to S24 resulted in loss of genotypic diversity that yielded a single allele A genotype with an amplicon size of 558 bp. This suggested that clonal selection occurred during rapid passaging. Extensive genotypic diversity exists in 44 field isolates characterized with both Bv80 A and B alleles, but can be readily distinguished from the S24 vaccine strain using either the Bv80 allele specific PCR assays or using multi-locus micro-satellite typing. This indicated that no recent documented clinical cases of Asiatic redwater were caused by the reversion to virulence of the current vaccine strain.

  5. The molecular prevalence and MSA-2b gene-based genetic diversity of Babesia bovis in dairy cattle in Thailand.

    PubMed

    Simking, Pacharathon; Saengow, Sinsamuth; Bangphoomi, Kunan; Sarataphan, Nachai; Wongnarkpet, Sirichai; Inpankaew, Tawin; Jittapalapong, Sathaporn; Munkhjargal, Tserendorj; Sivakumar, Thillaiampalam; Yokoyama, Naoaki; Igarashi, Ikuo

    2013-11-01

    Bovine babesiosis is an economically significant disease that affects dairy farming operations in Thailand. In the present study, 1824 blood-DNA samples prepared from cattle bred in 4 different regions of the country (North, Northeast, Central, and South) were screened using a nested PCR for the specific detection of Babesia bovis. While the overall prevalence of B. bovis was 8.8%, the Central region of Thailand was found to be a high-risk area of the country, as the prevalence of the parasite was 15.0%. The positive rate was relatively higher among the animals of 1-5 years of age. The genetic diversity among the B. bovis parasites was also studied based on their MSA-2b gene, and the findings showed that the Thai sequences were dispersed across 8 of 13 total clades observed in the phylogram. Three of these clades were formed only of Thai sequences. Similarity among the deduced MSA-2b amino acid sequences determined in the present study was 68.3-100%. In conclusion, the present study found that all the locations surveyed were infected with B. bovis and that the parasite populations in Thailand were genetically diverse. Our findings highlight the need for further studies in Thailand to generate more information before a sound control strategy could be implemented against B. bovis.

  6. Unusual chromatin structure associated with monoparalogous transcription of the Babesia bovis ves multigene family.

    PubMed

    Huang, Yingling; Xiao, Yu-Ping; Allred, David R

    2013-02-01

    Rapid antigenic variation in Babesia bovis involves the variant erythrocyte surface antigen-1 (VESA1), a heterodimeric protein with subunits encoded by two branches of the ves multigene family. The ves1α and ves1β gene pair encoding VESA1a and 1b, respectively, are transcribed in a monoparalogous manner from a single locus of active ves transcription (LAT), just one of many quasi-palindromic ves loci. To determine whether this organization plays a role in transcriptional regulation, chromatin structure was first assessed. Limited treatment of isolated nuclei with micrococcal nuclease to assay nucleosomal patterning revealed a periodicity of 156-159 bp in both bulk chromatin and specific gene coding regions. This pattern also was maintained in the intergenic regions (IGr) of non-transcribed ves genes. In contrast, the LAT IGr adopts a unique pattern, yielding an apparent cluster of five closely-spaced hypersensitive sites flanked by regions of reduced nucleosomal occupancy. ves loci fall into three patterns of overall sensitivity to micrococcal nuclease or DNase I digestion, with only the LAT being consistently very sensitive. Non-transcribed ves genes are inconsistent in their sensitivity to the two enzymatic probes. Non-linear DNA structure in chromatin was investigated to determine whether unique structure arising as a result of the quasi-palindromic nature of the LAT may effect transcriptional control. The in vitro capacity of ves IGr sequences to adopt stable higher-order DNA structure is demonstrated here, but the presence of such structure in vivo was not supported. Based upon these results a working model is proposed for the chromatin structural remodeling responsible for the sequential expression of ves multigene family members from divergently-organized loci.

  7. The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

    PubMed Central

    2013-01-01

    Background Cattle babesiosis is a tick-borne disease of cattle with the most severe form of the disease caused by the apicomplexan, Babesia bovis. Babesiosis is transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus. The most prevalent species is Rhipicephalus (Boophilus) microplus, which is distributed throughout the tropical and subtropical countries of the world. The transmission of B. bovis is transovarian and a previous study of the R. microplus ovarian proteome identified several R. microplus proteins that were differentially expressed in response to infection. Through various approaches, we studied the reaction of the R. microplus ovarian transcriptome in response to infection by B. bovis. Methods A group of ticks were allowed to feed on a B. bovis-infected splenectomized calf while a second group fed on an uninfected splenectomized control calf. RNA was purified from dissected adult female ovaries of both infected and uninfected ticks and a subtracted B. bovis-infected cDNA library was synthesized, subtracting with the uninfected ovarian RNA. Four thousand ESTs were sequenced from the ovary subtracted library and annotated. Results The subtracted library dataset assembled into 727 unique contigs and 2,161 singletons for a total of 2,888 unigenes, Microarray experiments designed to detect B. bovis-induced gene expression changes indicated at least 15 transcripts were expressed at a higher level in ovaries from ticks feeding upon the B. bovis-infected calf as compared with ovaries from ticks feeding on an uninfected calf. We did not detect any transcripts from these microarray experiments that were expressed at a lower level in the infected ovaries compared with the uninfected ovaries. Using the technique called serial analysis of gene expression, 41 ovarian transcripts from infected ticks were differentially expressed when compared with transcripts of controls. Conclusion Collectively, our experimental approaches provide

  8. msa-1 and msa-2c gene analysis and common epitopes assessment in Mexican Babesia bovis isolates.

    PubMed

    Borgonio, Veronica; Mosqueda, Juan; Genis, Alma D; Falcon, Alfonso; Alvarez, J Antonio; Camacho, Minerva; Figueroa, Julio V

    2008-12-01

    Babesia bovis msa-1 and msa-2c genes belong to the variable merozoite surface antigen gene family. These genes code for antigenic proteins present on the merozoite surface (MSA) and are involved in the parasite invasion to the bovine erythrocyte. Previous studies carried out on MSA-1 have evidenced antigen allelic variation in B. bovis isolates from similar endemic regions, as well as in isolates from different geographic regions of the world (Argentina, Australia, Israel). Studies conducted on MSA-2c, however, have shown that this antigen is widely conserved on isolates from distinct geographic regions. In this study, it was hypothesized that MSA-1 and MSA-2c antigens would contain common epitopes despite the presence of nucleotide sequence differences found in 13 B. bovis isolates and strains collected in geographically distant regions of Mexico. Bioinformatics analysis of the primary structure from DNA fragments derived from PCR amplification, cloning, and sequencing of msa-1 and msa-2c genes from the 13 B. bovis populations revealed that the msa-1 gene product present in the various isolates tested is less conserved among isolates obtained within a similar geographic region in Mexico (51-99.7% sequence identity). Results obtained by immunoblot analysis of B. bovis protein extracts reacted with a monoclonal antibody to MSA-1 42-kDa antigen, conclusively showed cross-reactive common epitopes only in Mexican isolates having high sequence identity (>/=99%, eight isolates). Sequence analysis and multiple alignment of deduced MSA-2c demonstrated a high degree of sequence identity (90-100%) among the Mexican B. bovis isolates and strains. Immunoblot results using a polyclonal antibody to MSA-2c reacted against the protein extracts recognized conserved epitopes in at least nine of the B. bovis isolates. The results obtained in this study agree with those previously reported by other researchers and confirm that, based in sequence identity conservation in Mexican B

  9. Genetic variations in merozoite surface antigen genes of Babesia bovis detected in Vietnamese cattle and water buffaloes.

    PubMed

    Yokoyama, Naoaki; Sivakumar, Thillaiampalam; Tuvshintulga, Bumduuren; Hayashida, Kyoko; Igarashi, Ikuo; Inoue, Noboru; Long, Phung Thang; Lan, Dinh Thi Bich

    2015-03-01

    The genes that encode merozoite surface antigens (MSAs) in Babesia bovis are genetically diverse. In this study, we analyzed the genetic diversity of B. bovis MSA-1, MSA-2b, and MSA-2c genes in Vietnamese cattle and water buffaloes. Blood DNA samples from 258 cattle and 49 water buffaloes reared in the Thua Thien Hue province of Vietnam were screened with a B. bovis-specific diagnostic PCR assay. The B. bovis-positive DNA samples (23 cattle and 16 water buffaloes) were then subjected to PCR assays to amplify the MSA-1, MSA-2b, and MSA-2c genes. Sequencing analyses showed that the Vietnamese MSA-1 and MSA-2b sequences are genetically diverse, whereas MSA-2c is relatively conserved. The nucleotide identity values for these MSA gene sequences were similar in the cattle and water buffaloes. Consistent with the sequencing data, the Vietnamese MSA-1 and MSA-2b sequences were dispersed across several clades in the corresponding phylogenetic trees, whereas the MSA-2c sequences occurred in a single clade. Cattle- and water-buffalo-derived sequences also often clustered together on the phylogenetic trees. The Vietnamese MSA-1, MSA-2b, and MSA-2c sequences were then screened for recombination with automated methods. Of the seven recombination events detected, five and two were associated with the MSA-2b and MSA-2c recombinant sequences, respectively, whereas no MSA-1 recombinants were detected among the sequences analyzed. Recombination between the sequences derived from cattle and water buffaloes was very common, and the resultant recombinant sequences were found in both host animals. These data indicate that the genetic diversity of the MSA sequences does not differ between cattle and water buffaloes in Vietnam. They also suggest that recombination between the B. bovis MSA sequences in both cattle and water buffaloes might contribute to the genetic variation in these genes in Vietnam.

  10. Validation and Field Evaluation of a Competitive Enzyme-Linked Immunosorbent Assay for Diagnosis of Babesia bovis Infections in Argentina

    PubMed Central

    Dominguez, Mariana; Echaide, Ignacio; de Echaide, Susana Torioni; Wilkowsky, Silvina; Zabal, Osvaldo; Mosqueda, Juan J.; Schnittger, Leonhard

    2012-01-01

    Infections by Babesia bovis limit cattle production and cause important economic losses in tropical and subtropical areas around the world. Monitoring of calf sera can be used to detect unprotected cattle herds and to decide on strategic control measures, as well as for epidemiological studies. Merozoite surface antigen 2c (MSA-2c) is an immunodominant surface protein expressed in B. bovis merozoites and sporozoites and contains B-cell epitopes that are conserved among geographic isolates. A monoclonal antibody against recombinant MSA-2c (rMSA-2c) was previously shown to inhibit the binding of anti-B. bovis antibodies to a parasite B-cell epitope in a competitive enzyme-linked immunosorbent assay (cELISA) format. In the work at hand, the parameters of this cELISA were reevaluated and adjusted when necessary, and a cutoff value was determined by receiver operator characteristic (ROC) curve analysis of a total of 357 bovine sera of known reactivity, as assessed by indirect immunofluorescence assay (IFAT). The established rMSA-2c cELISA demonstrated a specificity of 98% and a sensitivity of 96.2%. An additional set of 303 field bovine sera from regions where ticks are endemic and tick-free regions of Argentina was tested by both rMSA-2c cELISA and IFAT, and the results were shown to be in very good agreement (kappa index, 0.8325). The performance shown by rMSA-2c cELISA in the detection of B. bovis-specific antibodies and its suitability for standardization and large-scale production, as well as the possibility of its application in most veterinary diagnostic laboratories, make the assay a powerful tool for the surveillance of herd immunity as a strategic measure for the control of bovine babesiosis. PMID:22492742

  11. Type-specific PCR assays for Babesia bovis msa-1 genotypes in Asia: Revisiting the genetic diversity in Sri Lanka, Mongolia, and Vietnam.

    PubMed

    Liyanagunawardena, Nilukshi; Sivakumar, Thillaiampalam; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Battsetseg, Badgar; Lan, Dinh Thi Bich; Inoue, Noboru; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-01-01

    Babesia bovis is the most virulent Babesia organism, resulting in a high mortality rate in cattle. The genetic diversity of B. bovis merozoite surface antigens (MSAs), such as MSA-1, MSA-2b, and MSA-2c, might be linked to altered immune profiles in the host animals. The present study aimed to develop type-specific PCR assays for Asian msa-1 genotypes, thereby re-analyzing the genetic diversity of msa-1 in Sri Lanka, Mongolia, and Vietnam. Specific primers were designed for nine Asian msa-1 genotypes, which had been detected based on the phylogeny constructed using msa-1 gene sequences retrieved from the GenBank database. Specificity of the type-specific PCR assays was confirmed using plasmids containing the inserts of msa-1 gene fragments that represent Asian genotypes. Furthermore, no amplicons were observed by these PCR assays when DNA samples of Babesia bigemina, Babesia ovata, Theileria annulata, Theileria orientalis, Trypanosoma evansi, Trypanosoma theileri, Anaplasma marginale, and Anaplasma bovis, and non-infected bovine blood were analyzed. In total, 109 B. bovis-positive blood DNA samples sourced from Sri Lanka (44 cattle), Mongolia (26 cattle), and Vietnam (23 cattle and 16 water buffaloes) were then screened by the type-specific PCR assays. The sequences derived from all of the PCR amplicons were phylogenetically analyzed. Out of 109 DNA samples, 23 (20 from cattle and 3 from water buffaloes) were positive for at least one genotype. In agreement with previous studies, five and four different genotypes were detected among the DNA samples from Sri Lanka and Vietnam, respectively. In contrast, four genotypes, including three novel genotypes, were detected from Mongolia. Five DNA samples were found to be co-infected with multiple genotypes. The sequences of the PCR amplicons clustered phylogenetically within the corresponding clades. These findings indicated that the type-specific PCR assays described herein are useful for the determination of genotypic

  12. Type-specific PCR assays for Babesia bovis msa-1 genotypes in Asia: Revisiting the genetic diversity in Sri Lanka, Mongolia, and Vietnam.

    PubMed

    Liyanagunawardena, Nilukshi; Sivakumar, Thillaiampalam; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Battsetseg, Badgar; Lan, Dinh Thi Bich; Inoue, Noboru; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-01-01

    Babesia bovis is the most virulent Babesia organism, resulting in a high mortality rate in cattle. The genetic diversity of B. bovis merozoite surface antigens (MSAs), such as MSA-1, MSA-2b, and MSA-2c, might be linked to altered immune profiles in the host animals. The present study aimed to develop type-specific PCR assays for Asian msa-1 genotypes, thereby re-analyzing the genetic diversity of msa-1 in Sri Lanka, Mongolia, and Vietnam. Specific primers were designed for nine Asian msa-1 genotypes, which had been detected based on the phylogeny constructed using msa-1 gene sequences retrieved from the GenBank database. Specificity of the type-specific PCR assays was confirmed using plasmids containing the inserts of msa-1 gene fragments that represent Asian genotypes. Furthermore, no amplicons were observed by these PCR assays when DNA samples of Babesia bigemina, Babesia ovata, Theileria annulata, Theileria orientalis, Trypanosoma evansi, Trypanosoma theileri, Anaplasma marginale, and Anaplasma bovis, and non-infected bovine blood were analyzed. In total, 109 B. bovis-positive blood DNA samples sourced from Sri Lanka (44 cattle), Mongolia (26 cattle), and Vietnam (23 cattle and 16 water buffaloes) were then screened by the type-specific PCR assays. The sequences derived from all of the PCR amplicons were phylogenetically analyzed. Out of 109 DNA samples, 23 (20 from cattle and 3 from water buffaloes) were positive for at least one genotype. In agreement with previous studies, five and four different genotypes were detected among the DNA samples from Sri Lanka and Vietnam, respectively. In contrast, four genotypes, including three novel genotypes, were detected from Mongolia. Five DNA samples were found to be co-infected with multiple genotypes. The sequences of the PCR amplicons clustered phylogenetically within the corresponding clades. These findings indicated that the type-specific PCR assays described herein are useful for the determination of genotypic

  13. Delivery of recombinant vaccines against bovine herpesvirus type 1 gD and Babesia bovis MSA-2c to mice using liposomes derived from egg yolk lipids.

    PubMed

    Rodriguez, Anabel E; Zamorano, Patricia; Wilkowsky, Silvina; Torrá, Florencia; Ferreri, Lucas; Dominguez, Mariana; Florin-Christensen, Mónica

    2013-06-01

    Liposomes prepared from total egg yolk lipid extracts were used to deliver experimental DNA vaccines to mice consisting of pCI-neo plasmids encoding bovine herpesvirus type 1 (BoHV-1) gD or Babesia bovis MSA-2c. A significantly higher proportion of mice in the B. bovis MSA-2c group, but not those in the BoHV-1 gD group, developed detectable immunoglobulin G responses when vaccinated with liposome encapsulated DNA in comparison with mice vaccinated with naked DNA. In both groups, antibody titres were similar between mice vaccinated with liposome encapsulated DNA and naked DNA. PMID:23183017

  14. Characterization of the unusual bidirectional ves promoters driving VESA1 expression and associated with antigenic variation in Babesia bovis.

    PubMed

    Wang, Xinyi; Xiao, Yu-Ping; Bouchut, Anne; Al-Khedery, Basima; Wang, Hongbin; Allred, David R

    2012-03-01

    Rapid clonal antigenic variation in Babesia bovis involves the variant erythrocyte surface antigen-1 (VESA1) protein expressed on the infected-erythrocyte surface. Because of the significance of this heterodimeric protein for demonstrated mechanisms of parasite survival and virulence, there is a need to understand how expression of the ves multigene family encoding this protein is controlled. As an initial step toward this goal, we present here initial characterization of the ves promoter driving transcription of VESA1a and -1b subunits. A series of transfection constructs containing various sequence elements from the in vivo locus of active ves transcription (LAT) were used to drive expression of the firefly luciferase gene in a dual luciferase-normalized assay. The results of this approach reveal the presence of two bidirectional promoter activities within the 434-bp intergenic region (IGr), influenced by putative regulatory sequences embedded within the flanking ves1α and ves1β genes. Repressor-like effects on the apposing gene were observed for intron 1 of both ves1α and ves1β. This effect is apparently not dependent upon intronic promoter activity and acts only in cis. The expression of genes within the ves family is likely modulated by local elements embedded within ves coding sequences outside the intergenic promoter region in concert with chromatin modifications. These results provide a framework to help us begin to understand gene regulation during antigenic variation in B. bovis.

  15. Cell-mediated immune responses to Babesia bovis merozoite antigens in cattle following infection with tick-derived or cultured parasites.

    PubMed Central

    Brown, W C; Logan, K S; Wagner, G G; Tetzlaff, C L

    1991-01-01

    Peripheral blood mononuclear cells from cattle experimentally infected with Babesia bovis were examined for parasite-specific cell-mediated immune responses. Unfractionated merozoites and soluble and membrane fractions derived from merozoites were all antigenic for immune cattle, although the membrane fraction was the most stimulatory. Cattle responded to different antigenic fractions in a differential manner, and only that animal immunized with autologous cultured parasites responded to parasitized erythrocyte culture supernatants. Plastic-adherent cells (presumably monocytes/macrophages) were required for a proliferative response to babesial antigens but not to the T-cell mitogen concanavalin A, suggesting that babesial proteins are not simply mitogenic for T cells. Lymphocyte responses directed against a different hemoparasite from Mexico, Babesia bigemina, indicate that this parasite shares cross-reactive T-cell epitopes with B. bovis. These studies define a system whereby T lymphocytes from babesia-immune cattle can be used in proliferation assays to identify babesial merozoite antigens which are immunogenic for T cells. Because identification of helper T-cell epitopes is important for the design of a babesial subunit vaccine which will evoke anamnestic responses, the studies described here provide a basis for such experiments. Images PMID:2050406

  16. Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab

    PubMed Central

    Zulfiqar, Samreen; Shahnawaz, Sadia; Ali, Muhammad; Bhutta, Arif Mahmood; Iqbal, Shahid; Hayat, Sikandar; Qadir, Shazia; Latif, Muhammad; Kiran, Nazia; Saeed, Ali; Ali, Muhammad; Iqbal, Furhan

    2012-01-01

    Objective To determine the presence of Babesia bovis (B. bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals. Methods Blood samples were collected from 144 large ruminants, including 105 cattle and 39 buffaloes, from six districts in southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar and Vehari. Data on the characteristics of animals and herds were collected through questionnaires. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B. bovis on the blood and serological profile of infected animals. Results 27 out of 144 animals, from 5 out of 6 sampling districts, produced the 541-bp fragment specific for B. bovis. Age of animals (P=0.02), presence of ticks on animals (P=0.04) and presence of ticks on dogs associated with herds (P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area. ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle. Conclusions : This study has reported for the first time the presence of B. bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output. PMID:23569878

  17. The genetic diversity of merozoite surface antigen 1 (MSA-1) among Babesia bovis detected from cattle populations in Thailand, Brazil and Ghana.

    PubMed

    Nagano, Daisuke; Sivakumar, Thillaiampalam; De De Macedo, Alane Caine Costa; Inpankaew, Tawin; Alhassan, Andy; Igarashi, Ikuo; Yokoyama, Naoaki

    2013-11-01

    In the present study, we screened blood DNA samples obtained from cattle bred in Brazil (n=164) and Ghana (n=80) for Babesia bovis using a diagnostic PCR assay and found prevalences of 14.6% and 46.3%, respectively. Subsequently, the genetic diversity of B. bovis in Thailand, Brazil and Ghana was analyzed, based on the DNA sequence of merozoite surface antigen-1 (MSA-1). In Thailand, MSA-1 sequences were relatively conserved and found in a single clade of the phylogram, while Brazilian MSA-1 sequences showed high genetic diversity and were dispersed across three different clades. In contrast, the sequences from Ghanaian samples were detected in two different clades, one of which contained only a single Ghanaian sequence. The identities among the MSA-1 sequences from Thailand, Brazil and Ghana were 99.0-100%, 57.5-99.4% and 60.3-100%, respectively, while the similarities among the deduced MSA-1 amino acid sequences within the respective countries were 98.4-100%, 59.4-99.7% and 58.7-100%, respectively. These observations suggested that the genetic diversity of B. bovis based on MSA-1 sequences was higher in Brazil and Ghana than in Thailand. The current data highlight the importance of conducting extensive studies on the genetic diversity of B. bovis before designing immune control strategies in each surveyed country.

  18. Evaluation of different heterologous prime-boost immunization strategies against Babesia bovis using viral vectored and protein-adjuvant vaccines based on a chimeric multi-antigen.

    PubMed

    Jaramillo Ortiz, José Manuel; Molinari, María Paula; Gravisaco, María José; Paoletta, Martina Soledad; Montenegro, Valeria Noely; Wilkowsky, Silvina Elizabeth

    2016-07-19

    Protection against the intraerythrocytic bovine parasite Babesia bovis requires both humoral and cellular immune responses. Therefore, tailored combinations of immunogens targeted at both arms of the immune system are strategies of choice to pursue sterilizing immunity. In this study, different heterologous prime-boost vaccination schemes were evaluated in mice to compare the immunogenicity induced by a recombinant adenovirus, a modified vaccinia Ankara vector or a subunit vaccine all expressing a chimeric multi-antigen. This multi-antigen includes the immunodominant B and T cell epitopes of three B. bovis proteins: Merozoite Surface Antigen - 2c (MSA-2c), Rhoptry Associated Protein - 1 (RAP-1) and Heat Shock Protein 20 (HSP20). Both priming with the adenovirus or recombinant multi-antigen and boosting with the modified vaccinia Ankara vector achieved a high degree of activation of TNFα and IFNγ-secreting CD4(+) and CD8(+) specific T cells 60days after the first immunization. High titers of specific IgG antibodies were also detected at the same time point and lasted up to day 120 of the first immunization. Only the adenovirus - MVA combination triggered a marked isotype skew for the IgG2a antibody subclass meanwhile for the other immune traits analyzed here, both vaccination schemes showed similar performances. The immunological characterization in the murine model of these rationally designed immunogens led us to propose that adenoviruses as well as the bacterially expressed multi-antigen are highly reliable primer candidates to be considered in future experiments in cattle to test protection against bovine babesiosis. PMID:27269058

  19. DNA from Protozoan Parasites Babesia bovis, Trypanosoma cruzi, and T. brucei Is Mitogenic for B Lymphocytes and Stimulates Macrophage Expression of Interleukin-12, Tumor Necrosis Factor Alpha, and Nitric Oxide

    PubMed Central

    Shoda, Lisl K. M.; Kegerreis, Kimberly A.; Suarez, Carlos E.; Roditi, Isabel; Corral, Ricardo S.; Bertot, Gustavo M.; Norimine, Junzo; Brown, Wendy C.

    2001-01-01

    The activation of innate immune responses by genomic DNA from bacteria and several nonvertebrate organisms represents a novel mechanism of pathogen recognition. We recently demonstrated the CpG-dependent mitogenic activity of DNA from the protozoan parasite Babesia bovis for bovine B lymphocytes (W. C. Brown, D. M. Estes, S. E. Chantler, K. A. Kegerreis, and C. E. Suarez, Infect. Immun. 66:5423–5432, 1998). However, activation of macrophages by DNA from protozoan parasites has not been demonstrated. The present study was therefore conducted to determine whether DNA from the protozan parasites B. bovis, Trypanosoma cruzi, and T. brucei activates macrophages to secrete inflammatory mediators associated with protective immunity. DNA from Escherichia coli and all three parasites stimulated B-lymphocyte proliferation and increased macrophage production of interleukin-12 (IL-12), tumor necrosis factor alpha (TNF-α), and nitric oxide (NO). Regulation of IL-12 and NO production occurred at the level of transcription. The amounts of IL-12, TNF-α, and NO induced by E. coli and protozoal DNA were strongly correlated (r2 > 0.9) with the frequency of CG dinucleotides in the genome, and immunostimulation by DNA occurred in the order E. coli ≥ T. cruzi > T. brucei > B. bovis. Induction of inflammatory mediators by E. coli, T. brucei, and B. bovis DNA was dependent on the presence of unmethylated CpG dinucleotides. However, at high concentrations, E. coli and T. cruzi DNA-mediated macrophage activation was not inhibited following methylation. The recognition of protozoal DNA by B lymphocytes and macrophages may provide an important innate defense mechanism to control parasite replication and promote persistent infection. PMID:11254571

  20. Involvement of TLR6 in the induction of COX-2, PGE2 and IL-10 in macrophages by lipids from virulent S2P and attenuated R1A Babesia bovis strains.

    PubMed

    Gimenez, G; Belaunzarán, M L; Magalhães, K G; Poncini, C V; Lammel, E M; González Cappa, S M; Bozza, P T; Isola, E L D

    2016-06-15

    Toll like receptors (TLRs) are involved in the modulation of diverse host genes expression through a complex network of signalling events that allow for an appropriate response to a microbial pathogen. In the present work we used TLR6KO mice in order to study the role of TLR6 in the immune discrimination of lipids from two Babesia bovis strains, attenuated R1A (LA) and virulent S2P (LV), and the consequent macrophage activation. We demonstrated that TLR6 is required for lipid body induction in murine peritoneal macrophages by both LA and LV. Interestingly, as regards IL-10 and COX-2/PGE2 pathway induction by LA and LV, we observed differences in the biological effects produced by these lipid extracts. Our results indicate a role of TLR6 in the down-modulation of these immunoregulators only in the case of LA, whereas this receptor was not implicated in pro-inflammatory TNFα, IL-6 and KC release induced by LA. Remarkably, LV did not exert the down-modulatory effect observed for LA, supporting the notion that LA and LV possess different lipid composition that could correlate with the polar pathogenic effect of both B. bovis strains.

  1. Involvement of TLR6 in the induction of COX-2, PGE2 and IL-10 in macrophages by lipids from virulent S2P and attenuated R1A Babesia bovis strains.

    PubMed

    Gimenez, G; Belaunzarán, M L; Magalhães, K G; Poncini, C V; Lammel, E M; González Cappa, S M; Bozza, P T; Isola, E L D

    2016-06-15

    Toll like receptors (TLRs) are involved in the modulation of diverse host genes expression through a complex network of signalling events that allow for an appropriate response to a microbial pathogen. In the present work we used TLR6KO mice in order to study the role of TLR6 in the immune discrimination of lipids from two Babesia bovis strains, attenuated R1A (LA) and virulent S2P (LV), and the consequent macrophage activation. We demonstrated that TLR6 is required for lipid body induction in murine peritoneal macrophages by both LA and LV. Interestingly, as regards IL-10 and COX-2/PGE2 pathway induction by LA and LV, we observed differences in the biological effects produced by these lipid extracts. Our results indicate a role of TLR6 in the down-modulation of these immunoregulators only in the case of LA, whereas this receptor was not implicated in pro-inflammatory TNFα, IL-6 and KC release induced by LA. Remarkably, LV did not exert the down-modulatory effect observed for LA, supporting the notion that LA and LV possess different lipid composition that could correlate with the polar pathogenic effect of both B. bovis strains. PMID:27198789

  2. Observations on cattle schistosomiasis in the Sudan, a study in comparative medicine. III. Field testing of an irradiated Schistosoma bovis vaccine

    SciTech Connect

    Majid, A.A.; Bushera, H.O.; Saad, A.M.; Hussein, M.F.; Taylor, M.G.; Dargie, J.D.; Marshall, T.F.; Nelson, G.S.

    1980-05-29

    Previous work has shown that cattle can acquire a strong resistance to Schistosoma bovis infection following repeated natural exposure. Partial resistance to a laboratory challenge with S. bovis has also been demonstrated in calves after immunization with an irradiated schistosomular or cercarial vaccine. The aim of the present study was to see whether this type of caccine could protect calves under the very different conditions of natural exposure to S. bovis in the field. Thirty 6- to 9-month-old calves were each immunized with 10,000 irradiated S. bovis schistosomula by intramuscular injection and 8 weeks later were released into an enzootic area along with 30 unvaccinated animals. The calves were followed up for 10 months, during which period protection was evidenced by a lower mortality rate, a slower rate of acquisition of infection, and lower fecal egg counts in the vaccinated calves. Necropsy of the survivors showed 60 to 70% reductions in worm and tissue egg counts of the vaccinated calves as compared to those not vaccinated.

  3. Molecular detection of Babesia spp. and other haemoparasitic infections of cattle in Maputo Province, Mozambique.

    PubMed

    Martins, Tiago M; Neves, Luís; Pedro, Olívia C; Fafetine, José M; Do Rosário, Virgílio E; Domingos, Ana

    2010-05-01

    Molecular detection of Babesia species in apparently healthy cattle within an endemic region was carried out in order to determine the prevalence of carriers and the geographical distribution of Babesia bigemina and Babesia bovis in Maputo Province, Mozambique. Samples from 477 animals at 5 localities were analysed using 2 techniques, the semi-nested hot-start PCR and the reverse line blot (RLB) assay. With the semi-nested hot-start PCR, detection of B. bigemina ranged between 30% and 89%, and of B. bovis between 27% and 83%. The RLB assay was comparatively less sensitive in this study and detection of B. bovis ranged from 0% to 17%, and B. bigemina was not detected at all by this technique. Analysis of new sequences of the 18S rRNA gene revealed that the current B. bigemina RLB probe is not specific for the identification of isolates in Mozambique. The RLB assay, however, resulted in the detection of 8 other haemoparasite species belonging to the genera Babesia, Theileria, Anaplasma and Ehrlichia. 18S rRNA gene sequences from the Theileria spp. were identified, and a phylogenic tree constructed with these sequences yielded a heterogeneous T. mutans-like group. In conclusion, infection with B. bigemina and B. bovis is endemic in Maputo Province, but rates of transmission vary. Furthermore, mixed infections with the haemoparasites responsible for several tick-borne diseases in cattle are common in Mozambique. PMID:20128941

  4. Identification and characterization of profilin antigen among Babesia species as a common vaccine candidate against babesiosis.

    PubMed

    Munkhjargal, Tserendorj; Aboge, Gabriel Oluga; Ueno, Akio; Aboulaila, Mahmoud; Yokoyama, Naoaki; Igarashi, Ikuo

    2016-07-01

    We have characterized a member of the profilin (PROF) family protein as a common antigen in three pathogens-Babesia bovis (B. bovis), Babesia bigemina (B. bigemina), and Babesia microti (B. microti)-and evaluated its immunogenic and cross-protective properties against a challenge infection with B. microti in BALB/c mice. The recombinant PROF proteins of B. bovis, B. bigemina, and B. microti were successfully expressed in Escherichia coli (E. coli) as soluble GST fusion proteins (rBboPROF, rBbigPROF, and rBmPROF, respectively), and they were found to be antigenic. On probing with mouse anti-rPROF serum, green fluorescence was observed on the parasites' cytosols by confocal laser microscopy. Immunization regimes in BALB/c mice using rPROFs induced cross-protective immunity against B. microti infection based on high levels of cytokines and immunoglobulin (IgG) titers, a reduction in peak parasitemia levels, and earlier clearance of the parasite as compared with control mice. The findings of the present study indicate that PROF is a common antigen among bovine and murine Babesia parasites, and it might be used as a common vaccine candidate against babesiosis.

  5. Serial analysis of gene expression associated with Babesoa bovis infection of Rhipecephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized Serial Analysis of Gene Expression (SAGE) to quant...

  6. Genetic characterization of Babesia and Theileria parasites in water buffaloes in Sri Lanka.

    PubMed

    Sivakumar, Thillaiampalam; Tattiyapong, Muncharee; Fukushi, Shintaro; Hayashida, Kyoko; Kothalawala, Hemal; Silva, Seekkuge Susil Priyantha; Vimalakumar, Singarayar Caniciyas; Kanagaratnam, Ratnam; Meewewa, Asela Sanjeewa; Suthaharan, Kalpana; Puvirajan, Thamotharampillai; de Silva, Weligodage Kumarawansa; Igarashi, Ikuo; Yokoyama, Naoaki

    2014-02-24

    Water buffaloes are thought to be the reservoir hosts for several hemoprotozoan parasites that infect cattle. In the present study, we surveyed Sri Lankan bred water buffaloes for infections with Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis using parasite-specific PCR assays. When 320 blood-derived DNA samples from water buffaloes reared in three different districts (Polonnaruwa, Mannar, and Mullaitivu) of Sri Lanka were PCR screened, B. bovis, B. bigemina, and T. orientalis were detected. While T. orientalis was the predominant parasite (82.5%), low PCR-positive rates were observed for B. bovis (1.9%) and B. bigemina (1.6%). Amplicons of the gene sequences of the Rhoptry Associated Protein-1 (RAP-1) of B. bovis, the Apical Membrane Antigen-1 (AMA-1) of B. bigemina, and the Major Piroplasm Surface Protein (MPSP) of T. orientalis were compared with those characterized previously in Sri Lankan cattle. While the B. bigemina AMA-1 sequences from water buffaloes shared high identity values with those from cattle, B. bovis RAP-1 sequences from water buffaloes diverged genetically from those of cattle. For T. orientalis, none of the MPSP sequence types reported previously in Sri Lankan cattle (types 1, 3, 5, and 7) were detected in the water buffaloes, and the MPSP sequences analyzed in the present study belonged to types N1 or N2. In summary, in addition to reporting the first PCR-based survey of Babesia and Theileria parasites in water buffaloes in Sri Lanka, the present study found that the predominant variants of water buffalo-derived B. bovis RAP-1 and T. orientalis MPSP sequences were different from those previously described from cattle in this country.

  7. Repeated Aerosolized-Boosting with Gamma-Irradiated Mycobacterium bovis BCG Confers Improved Pulmonary Protection against the Hypervirulent Mycobacterium tuberculosis Strain HN878 in Mice

    PubMed Central

    Kim, Jong-Seok; Kim, Hongmin; Kwon, Kee Woong; Han, Seung Jung; Eum, Seok-Yong; Cho, Sang-Nae; Shin, Sung Jae

    2015-01-01

    Mycobacterium bovis bacillus Calmette-Guerin (BCG), the only licensed vaccine, shows limited protection efficacy against pulmonary tuberculosis (TB), particularly hypervirulent Mycobacterium tuberculosis (Mtb) strains, suggesting that a logistical and practical vaccination strategy is urgently required. Boosting the BCG-induced immunity may offer a potentially advantageous strategy for advancing TB vaccine development, instead of replacing BCG completely. Despite the improved protection of the airway immunization by using live BCG, the use of live BCG as an airway boosting agent may evoke safety concerns. Here, we analyzed the protective efficacy of γ-irradiated BCG as a BCG-prime boosting agent for airway immunization against a hypervirulent clinical strain challenge with Mycobacterium tuberculosis HN878 in a mouse TB model. After the aerosol challenge with the HN878 strain, the mice vaccinated with BCG via the parenteral route exhibited only mild and transient protection, whereas BCG vaccination followed by multiple aerosolized boosting with γ-irradiated BCG efficiently maintained long-lasting control of Mtb in terms of bacterial reduction and pathological findings. Further immunological investigation revealed that this approach resulted in a significant increase in the cellular responses in terms of a robust expansion of antigen (PPD and Ag85A)-specific CD4+ T cells concomitantly producing IFN-γ, TNF-α, and IL-2, as well as a high level of IFN-γ-producing recall response via both the local and systemic immune systems upon further boosting. Collectively, aerosolized boosting of γ-irradiated BCG is able to elicit strong Th1-biased immune responses and confer enhanced protection against a hypervirulent Mycobacterium tuberculosis HN878 infection in a boosting number-dependent manner. PMID:26509812

  8. An epidemiological survey of bovine Babesia and Theileria parasites in cattle, buffaloes, and sheep in Egypt.

    PubMed

    Elsify, Ahmed; Sivakumar, Thillaiampalam; Nayel, Mohammed; Salama, Akram; Elkhtam, Ahmed; Rizk, Mohamed; Mosaab, Omar; Sultan, Khaled; Elsayed, Shimaa; Igarashi, Ikuo; Yokoyama, Naoaki

    2015-02-01

    Cattle, buffaloes, and sheep are the main sources of meat and milk in Egypt, but their productivity is thought to be greatly reduced by hemoprotozoan parasitic diseases. In this study, we analyzed the infection rates of Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis, using parasite-specific PCR assays in blood-DNA samples sourced from cattle (n=439), buffaloes (n=50), and sheep (n=105) reared in Menoufia, Behera, Giza, and Sohag provinces of Egypt. In cattle, the positive rates of B. bovis, B. bigemina, T. annulata, and T. orientalis were 3.18%, 7.97%, 9.56%, and 0.68%, respectively. On the other hand, B. bovis and T. orientalis were the only parasites detected in buffaloes and each of these parasites was only found in two individual DNA samples (both 2%), while one (0.95%) and two (1.90%) of the sheep samples were positive for B. bovis and B. bigemina, respectively. Sequence analysis showed that the B. bovis Rhoptry Associated Protein-1 and the B. bigemina Apical Membrane Antigen-1 genes were highly conserved among the samples, with 99.3-100% and 95.3-100% sequence identity values, respectively. In contrast, the Egyptian T. annulata merozoite surface antigen-1 gene sequences were relatively diverse (87.8-100% identity values), dispersing themselves across several clades in the phylogenetic tree containing sequences from other countries. Additionally, the T. orientalis Major Piroplasm Surface Protein (MPSP) gene sequences were classified as types 1 and 2. This is the first report of T. orientalis in Egypt, and of type 2 MPSP in buffaloes. Detection of MPSP type 2, which is considered a relatively virulent genotype, suggests that T. orientalis infection may have veterinary and economic significance in Egypt. In conclusion, the present study, which analyzed multiple species of Babesia and Theileria parasites in different livestock animals, may shed an additional light on the epidemiology of hemoprotozoan parasites in Egypt.

  9. Molecular epidemiology of bovine Babesia spp. and Theileria orientalis parasites in beef cattle from northern and northeastern Thailand.

    PubMed

    Jirapattharasate, Charoonluk; Adjou Moumouni, Paul Franck; Cao, Shinuo; Iguchi, Aiko; Liu, Mingming; Wang, Guanbo; Zhou, Mo; Vudriko, Patrick; Changbunjong, Tanasak; Sungpradit, Sivapong; Ratanakorn, Parntep; Moonarmart, Walasinee; Sedwisai, Poonyapat; Weluwanarak, Thekhawet; Wongsawang, Witsanu; Suzuki, Hiroshi; Xuan, Xuenan

    2016-02-01

    Beef cattle production represents the largest cattle population in Thailand. Their productivity is constrained by tick-borne diseases such as babesiosis and theileriosis. In this study, we determined the prevalence of Babesia bigemina, Babesia bovis and Theileria orientalis using polymerase chain reaction (PCR). The genetic markers that were used for detection of the above parasites were sequenced to determine identities and similarity for Babesia spp. and genetic diversity of T. orientalis. Furthermore the risk factors for the occurrence of the above protozoan parasites in beef cattle from northern and northeastern parts of Thailand were assessed. A total of 329 blood samples were collected from beef cattle in 6 provinces. The study revealed that T. orientalis was the most prevalent (30.1%) parasite in beef cattle followed by B. bigemina (13.1%) and B. bovis (5.5%). Overall, 78.7% of the cattle screened were infected with at least one of the above parasites. Co-infection with Babesia spp. and T. orientalis was 30.1%. B. bigemina and T. orientalis were the most prevalent (15.1%) co-infection although triple infection with the three parasites was observed in 3.0% of the samples. Sequencing analysis revealed that B. bigemina RAP1 gene and B. bovis SBP2 gene were conserved among the parasites from different cattle samples. Phylogenetic analysis showed that the T. orientalis MPSP gene from parasites isolated from cattle in north and northeast Thailand was classified into types 5 and 7 as reported previously. Lack of tick control program was the universal risk factor of the occurrence of Babesia spp. and T. orientalis infection in beef cattle in northern and northeastern Thailand. We therefore recommend training of farmers on appropriate tick control strategies and further research on potential vectors for T. orientalis and elucidate the effect of co-infection with Babesia spp. on the pathogenicity of T. orientalis infection on beef in northern and northeastern Thailand.

  10. Detection of Babesia and Theileria species infection in cattle from Portugal using a reverse line blotting method.

    PubMed

    Silva, M G; Marques, P X; Oliva, A

    2010-12-15

    Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR-reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases.

  11. Evaluation of in vitro inhibitory effect of enoxacin on Babesia and Theileria parasites.

    PubMed

    Omar, Mosaab A; Salama, Akram; Elsify, Ahmed; Rizk, Mohamed Abdo; Al-Aboody, Mohammad Saleh; AbouLaila, Mahmoud; El-Sayed, Shimaa Abd El-Salam; Igarashi, Ikuo

    2016-02-01

    Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P < 0.05) by micro molar concentrations of enoxacin (IC50 values = 33.5, 15.2, 7.5 and 23.2 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or incase of animal sensitivity against diminazin toxicity. PMID:26724376

  12. Evaluation of in vitro inhibitory effect of enoxacin on Babesia and Theileria parasites.

    PubMed

    Omar, Mosaab A; Salama, Akram; Elsify, Ahmed; Rizk, Mohamed Abdo; Al-Aboody, Mohammad Saleh; AbouLaila, Mahmoud; El-Sayed, Shimaa Abd El-Salam; Igarashi, Ikuo

    2016-02-01

    Enoxacin is a broad-spectrum 6-fluoronaphthyridinone antibacterial agent (fluoroquinolones) structurally related to nalidixic acid used mainly in the treatment of urinary tract infections and gonorrhea. Also it has been shown recently that it may have cancer inhibiting effect. The primary antibabesial effect of Enoxacin is due to inhibition of DNA gyrase subunit A, and DNA topoisomerase. In the present study, enoxacin was tested as a potent inhibitor against the in vitro growth of bovine and equine Piroplasms. The in vitro growth of five Babesia species that were tested was significantly inhibited (P < 0.05) by micro molar concentrations of enoxacin (IC50 values = 33.5, 15.2, 7.5 and 23.2 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Enoxacin IC50 values for Babesia and Theileria parasites were satisfactory as the drug is potent antibacterial drug with minimum side effects. Therefore, enoxacin might be used for treatment of Babesiosis and Theileriosis especially in case of mixed infections with bacterial diseases or incase of animal sensitivity against diminazin toxicity.

  13. Molecular cloning and characterization of Babesia orientalis rhoptry-associated protein 1.

    PubMed

    Yu, Qian; He, Lan; Zhang, Wen-Jie; Cheng, Jian-Xi; Hu, Jin-Fang; Miao, Xiao-Yan; Huang, Yuan; Fan, Li-Zhe; Khan, Muhammad Kasib; Zhou, Yan-Qin; Hu, Min; Zhao, Jun-Long

    2014-10-15

    The rhoptry-associated protein 1 (RAP-1) gene of Babesia orientalis was obtained from a cDNA expression library by immunoscreening with B. orientalis-infected water buffalo sera. The nucleotide sequence of the cDNA was 1732 bp with an open reading frame (ORF) of 1434 bp, encoding a polypeptide of 478 amino acid residues with a predicted size of 52.5 kDa. The ORF was cloned into a pGEX-KG plasmid and subsequently expressed as a GST-fusion protein. The recombinant RAP-1 of B. orientalis (rBoRAP-1) was purified and evaluated as an antigen using Western blotting. The native BoRAP-1 was recognized by the antibodies raised in rabbits against rBoRAP-1. Strong immunofluorescence signals were observed in erythrocytes infected with B. orientalis. Phylogentic analysis revealed that B. orientalis fell into a Babesia clade and most closely related to Babesia bovis and Babesia ovis, which was similar to the previous reported trees based on 18S rRNA and HSP70 genes. The present study suggests that the BoRAP-1 might be a potential diagnostic antigen, and the RAP-1 genes can aid in the classification of Babesia and Theileria species.

  14. Plasmids containing small subunit ribosomal RNA gene fragments from Babesia bovis and Babesia bigemina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BEI Resources was developed by NIAID as a centralized biological resource center for research reagents to the scientific community (http://www.beiresources.org/). They have a considerable amount of reagents and isolates for parasitologists working with Entamoeba histolytica, Giardia, Toxoplasma, and...

  15. In silico predicted conserved B-cell epitopes in the Merozoite Surface Antigen -2 family of B. bovis are neutralization-sensitive

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Merozoite Surface Antigens-2 of Babesia bovis conform a family of GPI-anchored glycoproteins located at the parasite cell surface, that contain neutralization-sensitive B-cell epitopes, thus constituting putative vaccine candidates for bovine babesiosis. It was previously shown that (i) the MSA-...

  16. Molecular cloning, phylogenetic analysis and heat shock response of Babesia gibsoni heat shock protein 90.

    PubMed

    Yamasaki, Masahiro; Tsuboi, Yoshihiro; Taniyama, Yusuke; Uchida, Naohiro; Sato, Reeko; Nakamura, Kensuke; Ohta, Hiroshi; Takiguchi, Mitsuyoshi

    2016-09-01

    The Babesia gibsoni heat shock protein 90 (BgHSP90) gene was cloned and sequenced. The length of the gene was 2,610 bp with two introns. This gene was amplified from cDNA corresponding to full length coding sequence (CDS) with an open reading frame of 2,148 bp. A phylogenetic analysis of the CDS of HSP90 gene showed that B. gibsoni was most closely related to B. bovis and Babesia sp. BQ1/Lintan and lies within a phylogenetic cluster of protozoa. Moreover, mRNA transcription profile for BgHSP90 exposed to high temperature were examined by quantitative real-time reverse transcription-polymerase chain reaction. BgHSP90 levels were elevated when the parasites were incubated at 43°C for 1 hr. PMID:27149891

  17. Molecular cloning, phylogenetic analysis and heat shock response of Babesia gibsoni heat shock protein 90

    PubMed Central

    YAMASAKI, Masahiro; TSUBOI, Yoshihiro; TANIYAMA, Yusuke; UCHIDA, Naohiro; SATO, Reeko; NAKAMURA, Kensuke; OHTA, Hiroshi; TAKIGUCHI, Mitsuyoshi

    2016-01-01

    The Babesia gibsoni heat shock protein 90 (BgHSP90) gene was cloned and sequenced. The length of the gene was 2,610 bp with two introns. This gene was amplified from cDNA corresponding to full length coding sequence (CDS) with an open reading frame of 2,148 bp. A phylogenetic analysis of the CDS of HSP90 gene showed that B. gibsoni was most closely related to B. bovis and Babesia sp. BQ1/Lintan and lies within a phylogenetic cluster of protozoa. Moreover, mRNA transcription profile for BgHSP90 exposed to high temperature were examined by quantitative real-time reverse transcription-polymerase chain reaction. BgHSP90 levels were elevated when the parasites were incubated at 43°C for 1 hr. PMID:27149891

  18. Babesia microti, upstate New York.

    PubMed

    Kogut, Sarah J; Thill, Charles D; Prusinski, Melissa A; Lee, Joon-Hak; Backerson, P Bryon; Coleman, James L; Anand, Madhu; White, Dennis J

    2005-03-01

    Five cases of human babesiosis were reported in the Lower Hudson Valley Region of New York State in 2001. An investigation to determine if Babesia microti was present in local Ixodes scapularis ticks yielded 5 positive pools in 123 pools tested, the first detection of B. microti from field-collected I. scapularis in upstate New York.

  19. Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.

    PubMed

    El-Ashker, Maged; Hotzel, Helmut; Gwida, Mayada; El-Beskawy, Mohamed; Silaghi, Cornelia; Tomaso, Herbert

    2015-01-30

    In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement.

  20. Immunoprophylactic control of bovine babesiosis: role of exoantigens of Babesia.

    PubMed

    Montenegro-James, S

    1989-01-01

    A major breakthrough in babesiosis vaccine research was achieved with the development of methods for continuous propagation in vitro of Babesia parasites. The development of a soluble, exoantigen-containing immunogen, free of erythrocytic antigens, has been proposed as a practical and realistic means of immunoprophylactic control of babesiosis. Such immunogens have been tested for immunogenicity of different strains, minimal protective dose, and protective efficacy of a combined B. bovis-B. bigemina immunogen. During the last 7 years, most of these aspects have been studied in a co-operative research programme between the University of Illinois and the Veterinary Research Institute of Venezuela. The Babesia exoantigen vaccine prevents clinical disease, is effective against different parasite strains, induces protective immunity for at least 13 months, does not immunize against host blood groups, is antigenically stable for at least 3 years at 4 degrees C, and is available in large quantities. Continued improvement of currently available culture-derived immunogens will best guarantee the successful production of food-producing animals in the tropics. Since July 1984, 16 field vaccination trials have been conducted in 7 states of Venezuela in areas where large-scale dairy and beef production is crucial for the livestock industry. These ranches encompassed a cattle production of 14,000, of which 3000 have been vaccinated with a combined B. bovis-B. bigemina immunogen in 2 subcutaneous inoculations with a 4-week interval. In the first 5 trials, a 1-year monitoring period following vaccination and natural tick exposure indicated that the culture-derived vaccine has effectively controlled the clinical manifestation of infection. Studies on antigen characterization are shifting the emphasis from crude antigenic preparations to purified polypeptides, isolated and identified using new biochemical techniques. Current research efforts are directed toward isolation of

  1. Mycobacterium bovis in Panama, 2013

    PubMed Central

    Acosta, Fermín; Chernyaeva, Ekatherina; Mendoza, Libardo; Sambrano, Dilcia; Correa, Ricardo; Rotkevich, Mikhail; Tarté, Miroslava; Hernández, Humberto; Velazco, Bredio; de Escobar, Cecilia; de Waard, Jacobus H.

    2015-01-01

    Panama remains free of zoonotic tuberculosis caused by Mycobacterium bovis. However, DNA fingerprinting of 7 M. bovis isolates from a 2013 bovine tuberculosis outbreak indicated minimal homology with strains previously circulating in Panama. M. bovis dispersion into Panama highlights the need for enhanced genotype testing to track zoonotic infections. PMID:25988479

  2. Identification of papain-like cysteine proteases from the bovine piroplasm Babesia bigemina and evolutionary relationship of piroplasms C1 family of cysteine proteases.

    PubMed

    Martins, Tiago M; do Rosário, Virgílio E; Domingos, Ana

    2011-01-01

    Papain-like cysteine proteases have been shown to have essential roles in parasitic protozoa and are under study as promising drug targets. Five genes were identified by sequence similarity search to be homologous to the cysteine protease family in the ongoing Babesia bigemina genome sequencing project database and were compared with the annotated genes from the complete bovine piroplasm genomes of Babesia bovis, Theileria annulata, and Theileria parva. Multiple genome alignments and sequence analysis were used to evaluate the molecular evolution events that occurred in the C1 family of cysteine proteases in these piroplasms of veterinary importance. BbiCPL1, one of the newly identified cysteine protease genes in the B. bigemina genome was expressed in Escherichia coli and shows activity against peptide substrates. Considerable differences were observed in the cysteine protease family between Babesia and Theileria genera, and this may partially explain the diverse infection mechanisms of these tick-borne diseases. PMID:20655912

  3. Molecular diagnosis of Theileria and Babesia species infecting cattle in Northern Spain using reverse line blot macroarrays

    PubMed Central

    García-Sanmartín, Josune; Nagore, Daniel; García-Pérez, Ana L; Juste, Ramón A; Hurtado, Ana

    2006-01-01

    Background Piroplasmosis in cattle is caused by tick-borne haemoprotozoan parasites of the genera Theileria and Babesia. Molecular detection techniques offer higher sensitivity and specificity than microscopy examination methods and serological tests. A reverse line blot (RLB) macroarray that included generic and species-specific probes for Theileria annulata, Theileria buffeli, Babesia bovis, Babesia bigemina, Babesia divergens and Babesia major was used to study the presence and identity of the piroplasm species infecting 263 bovine blood samples from 79 farms, most of them in Northern Spain. Microscopy examination of blood smears and haematology were also performed whenever possible to identify animals with parasitaemia. Results RLB hybridisation identified infection in 54.0% of the samples, whereas only 28.8% were positive by microscopy examination. The most frequently found species was T. buffeli, present in 42.6% of the samples. T. annulata was found in 22 samples (8.4%) from 12 farms, including 9 farms (14 samples) located in Northern Spain where presence of the vector is not very common. Babesia infections were less frequently detected: B. major was found in 3.0% of the samples, B. bigemina in 2.7%, B. bovis in 2.3% and B. divergens in 1.1%. Mixed infections were detected in 14 samples, accounting for six different combinations of species. Conclusion This is the first report in which B. major and B. divergens have been detected in Spain using molecular identification techniques and the first time that B. bovis has been detected in Northern Spain. The detection of T. annulata in Northern Spain suggests that the distribution of Mediterranean theileriosis might be changing. Samples with positive RLB hybridisation but negative microscopy had haematology values within the normal ranges suggesting that they corresponded to chronic carriers that may serve as reservoirs of the infection. In this sense, sensitive and specific laboratorial tests like RLB that clearly

  4. A PCR-based survey of selected Babesia and Theileria parasites in cattle in Sri Lanka.

    PubMed

    Sivakumar, Thillaiampalam; Kothalawala, Hemal; Abeyratne, Sembukutti Arachchige Eranga; Vimalakumar, Singarayar Caniciyas; Meewewa, Asela Sanjeewa; Hadirampela, Dilhani Thilanka; Puvirajan, Thamotharampillai; Sukumar, Subramaniyam; Kuleswarakumar, Kulanayagam; Chandrasiri, Alawattage Don Nimal; Igarashi, Ikuo; Yokoyama, Naoaki

    2012-11-23

    Hemoprotozoan parasites are responsible for significant economic losses in cattle. We screened Sri Lankan cattle populations for the presence of Babesia bovis, Babesia bigemina, Theileria annulata, and Theileria orientalis, using species-specific PCR assays. Out of 316 samples collected from animals in four different districts of Sri Lanka (Nuwara Eliya, Polonnaruwa, Ampara, and Jaffna), 231 (73.1%) were positive for at least one parasite species. All four parasite species were detected among the study groups from all of the districts surveyed. The first and second commonest hemoprotozoan parasites identified were T. orientalis (53.5%) and B. bigemina (30.1%), respectively. We found that the dry zones (Polonnaruwa, Ampara, and Jaffna) had more Babesia-positive animals than the hill country wet zone (Nuwara Eliya). In contrast, T. orientalis was the predominant species detected in Nuwara Eliya, while infection with T. annulata was more common in the dry zones. In addition, 81 (35.1%) of the 231 positive samples were infected with more than one parasite species. The presence of multiple parasite species among the different cattle populations is of clinical and economic significance. Therefore, island-wide control and prevention programs against bovine babesiosis and theileriosis are needed to minimize the financial burden caused by these parasites.

  5. A promising new ELISA diagnostic test for cattle babesiosis based on Babesia bigemina Apical Membrane Antigen-1.

    PubMed

    Torina, Alessandra; Cordaro, Antonio; Blanda, Valeria; D'Agostino, Rosalia; Scimeca, Salvatore; Scariano, Maria E; Sireci, Guido; Lelli, Rossella

    2016-01-01

    Babesiosis due to Babesia bigemina is a relevant tick-borne disease, affecting cattle worldwide. Many surface proteins of the pathogen including the Apical Membrane Antigen 1 (AMA-1) - have been analysed for vaccine and diagnostic purposes. This study focused on B. bigemina AMA-1 and on its use for the assessment of diagnostic tests. After bioinformatic analyses, AMA-1 codifying region was amplified and cloned into an expression vector used to induce protein synthesis in Escherichia coli cells. AMA-1 was purified by affinity chromatography and used to set up the best condition for an ELISA protocol. Bovine field sera positive to B. bigemina were used to evaluate the presence of anti-AMA-1 antibodies. In order to verify the assay specificity, sera positive to Babesia bovis or to the piroplasm Theileria annulata were also included. Significant differences were obtained between sera negative to both B. bigemina and B. bovis and samples positive to B. bigemina, to B. bovis or to both pathogens. No significant reaction was observed with T. annulata positive sera. The results showed that AMA-1 protein is suitable to be used as antigen in diagnostic assays for babesiosis diagnosis in cattle, as it does not show any cross reaction with anti-T. annulata antibodies. PMID:27033532

  6. Babesia species in questing Ixodes ricinus, Sweden.

    PubMed

    Karlsson, Maria E; Andersson, Martin O

    2016-02-01

    Babesiosis is an emerging tick-transmitted zoonosis in large parts of the world. In Sweden, the occurrence and diversity of Babesia species is largely unknown. In order to estimate the exposure to Babesia from infected ticks, we collected questing Ixodes ricinus from several sites across southern Sweden during two consecutive field seasons and investigated the occurrence of Babesia species. We report for the first time the occurrence of the zoonotic species Babesia venatorum in Swedish ticks, with a prevalence of 1%. We also detected B. microti (prevalence 3.2%) and B. divergens (prevalence 0.2%). The incidence of Babesia in questing ticks is substantially lower than that of several other tick-borne diseases in Sweden. Nevertheless, babesiosis should not be neglected as a possible diagnosis following tick bites in humans and animals in Sweden.

  7. Occurrence of Theileria and Babesia species in water buffalo (Bubalus babalis, Linnaeus, 1758) in the Hubei province, South China.

    PubMed

    He, Lan; Feng, Hui-Hui; Zhang, Wen-Jie; Zhang, Qing-Li; Fang, Rui; Wang, Li-Xia; Tu, Pan; Zhou, Yan-Qin; Zhao, Jun-Long; Oosthuizen, Marinda C

    2012-05-25

    The presence and prevalence of tick-borne haemoparasites in water buffalo from the Hubei province, south China was investigated using the reverse line blot (RLB) hybridization assay and phylogenetic analysis of the parasite 18S rRNA gene. Theileria buffeli (19.1%) was the most frequently found species in all of the locations, followed by Babesia orientalis (8.9%), Babesia bovis (1.0%) and Babesia bigemina (0.7%). Only 12 (3.9%) of the samples had mixed infections. Eleven samples with single infections were selected for further characterization using 18S rRNA gene sequence analysis. Phylogenetic analysis showed that the eight T. buffeli 18S rRNA gene sequences obtained grouped into four clusters, of which three grouped with the known T. buffeli types B and D. The remaining five grouped separately from the previously describe T. buffeli types, constituting new T. buffeli types. The two B. bigemina 18S rRNA gene sequences obtained grouped closely with B. bigemina Kunming; this serves as the first report of B. bigemina in the Hubei province. The B. orientalis Daye 18S rRNA gene sequence obtained grouped closely with the previously reported B. orientalis Wuhan strain and with Babesia sp. Kashi 1 and Kashi 2.

  8. Clofazimine Inhibits the Growth of Babesia and Theileria Parasites In Vitro and In Vivo.

    PubMed

    Tuvshintulga, Bumduuren; AbouLaila, Mahmoud; Davaasuren, Batdorj; Ishiyama, Aki; Sivakumar, Thillaiampalam; Yokoyama, Naoaki; Iwatsuki, Masato; Otoguro, Kazuhiko; Ōmura, Satoshi; Igarashi, Ikuo

    2016-05-01

    The present study evaluated the growth-inhibitory effects of clofazimine, currently used for treating leprosy, against Babesia bovis, B. bigemina, B. caballi, and Theileria equi in in vitro culture and against Babesia microti in mice. The 50% inhibitory concentrations (IC50s) of clofazimine against the in vitro growth of B. bovis, B. bigemina, B. caballi, and T. equi were 4.5, 3, 4.3, and 0.29 μM, respectively. In mice infected with B. microti, treatment with 20 mg/kg of body weight of clofazimine administered orally resulted in a significantly lower peak parasitemia (5.3%) than that in the control group (45.9%), which was comparable to the subcutaneous administration of 25 mg/kg diminazene aceturate, the most widely used treatment for animal piroplasmosis. Although slight anemia was observed in both clofazimine- and diminazene aceturate-treated infected mice, the level and duration of anemia were lower and shorter, respectively, than those in untreated infected mice. Using blood transfusions and PCR, we also examined whether clofazimine completely killed B. microti On day 40 postinfection, when blood analysis was performed, parasites were not found in blood smears; however, the DNA of B. microti was detected in the blood of clofazimine-treated animals and in several tissues of clofazimine- and diminazene aceturate-treated mice by PCR. The growth of parasites was observed in mice after blood transfusions from clofazimine-treated mice. In conclusion, clofazimine showed excellent inhibitory effects against Babesia and Theileria in vitro and in vivo, and further study on clofazimine is required for the future development of a novel chemotherapy with high efficacy and safety against animal piroplasmosis and, possibly, human babesiosis. PMID:26883713

  9. Clofazimine Inhibits the Growth of Babesia and Theileria Parasites In Vitro and In Vivo

    PubMed Central

    Tuvshintulga, Bumduuren; AbouLaila, Mahmoud; Davaasuren, Batdorj; Ishiyama, Aki; Sivakumar, Thillaiampalam; Yokoyama, Naoaki; Iwatsuki, Masato; Otoguro, Kazuhiko; Ōmura, Satoshi

    2016-01-01

    The present study evaluated the growth-inhibitory effects of clofazimine, currently used for treating leprosy, against Babesia bovis, B. bigemina, B. caballi, and Theileria equi in in vitro culture and against Babesia microti in mice. The 50% inhibitory concentrations (IC50s) of clofazimine against the in vitro growth of B. bovis, B. bigemina, B. caballi, and T. equi were 4.5, 3, 4.3, and 0.29 μM, respectively. In mice infected with B. microti, treatment with 20 mg/kg of body weight of clofazimine administered orally resulted in a significantly lower peak parasitemia (5.3%) than that in the control group (45.9%), which was comparable to the subcutaneous administration of 25 mg/kg diminazene aceturate, the most widely used treatment for animal piroplasmosis. Although slight anemia was observed in both clofazimine- and diminazene aceturate-treated infected mice, the level and duration of anemia were lower and shorter, respectively, than those in untreated infected mice. Using blood transfusions and PCR, we also examined whether clofazimine completely killed B. microti. On day 40 postinfection, when blood analysis was performed, parasites were not found in blood smears; however, the DNA of B. microti was detected in the blood of clofazimine-treated animals and in several tissues of clofazimine- and diminazene aceturate-treated mice by PCR. The growth of parasites was observed in mice after blood transfusions from clofazimine-treated mice. In conclusion, clofazimine showed excellent inhibitory effects against Babesia and Theileria in vitro and in vivo, and further study on clofazimine is required for the future development of a novel chemotherapy with high efficacy and safety against animal piroplasmosis and, possibly, human babesiosis. PMID:26883713

  10. Mycoplasma bovis research update

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis in bison is a newly emerging and potentially devastating threat to the bison industry. This bacterium is increasingly being identified, both in the United States and Canada, as the cause of severe respiratory disease outbreaks with devastating consequences for the health of the ani...

  11. Mycoplasma bovis research update

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Research conducted at the USDA/ARS/National Animal Disease Center in Ames, Iowa, reveals that ELISAs designed for detection of M. bovis-specific serum IgG in cattle may not be optimal for identification of seropositive bison, particularly those with low to moderate levels of antibody. In a study so...

  12. Molecular cloning and characterization of a novel heat shock protein 20 of Babesia orientalis.

    PubMed

    He, Lan; Yu, Qian; Zhang, Wen-Jie; Zhang, Qing-Li; Fan, Li-Zhe; Miao, Xiao-Yan; Khan, Muhammad Kasib; Hu, Min; Zhou, Yan-Qin; Zhao, Jun-Long

    2014-08-29

    The heat shock protein 20 (HSP20) gene of Babesia orientalis (BoHSP20) was identified from both genomic DNA and cDNA. The full-length BoHSP20 gene was 690bp with one intron from position 88-243bp. The amplicon obtained from cDNA corresponded to a full-length open reading frame (ORF) with a length of 534bp, encoding a polypeptide of 178 amino acid residues with a predicted size of 20kDa. The ORF was cloned into a pET-28a plasmid and subsequently expressed as a His-fusion protein. The recombinant HSP20 of B. orientalis (rBoHSP20) was purified and evaluated as an antigen using Western blotting. Anti-B. orientalis water buffalo serum reacted with rBoHSP20, indicating that this protein was an immunodominant antigen and could be a useful diagnostic reagent to detect antibodies against B. orientalis in water buffalo. The native BoHSP20 was recognized by polyclonal antibody from the serum of rabbit immunized with rBoHSP20. Strong immunofluorescence signals were observed from B. orientalis in blood smears by fluorescence microscopy. Bacterial survival experiments indicated that HSP20 can significantly increase the viability of bacteria when the culture is exposed to thermal stress. The results suggest that BoHSP20 might play an important role during B. orientalis transmission from tick to host animal, given the sudden shifts in temperature involved. Phylogenetic analysis revealed that B. orientalis is in the Babesia clade and most closely related to Babesia bovis. Similar topologies were obtained from trees based on 18S rRNA and the HSP70 gene. The present study suggests that BoHSP20 might be a potential diagnostic antigen and that the HSP20 genes can aid in the classification of Babesia and Theileria species.

  13. Mycobacterium bovis Infection, Lyon, France

    PubMed Central

    Pichat, Catherine; Carret, Gerard

    2006-01-01

    In a 5-year retrospective study, we used spoligotyping and mycobacterial interspersed repetitive units to type 13 strains of Mycobacterium bovis isolated from human sources. Despite the relatively high incidence of human tuberculosis caused by M. bovis (2%), these tools showed no clonal evolution and no relationships between the isolates. PMID:17073096

  14. Streptococcus bovis meningitis and hemorrhoids.

    PubMed

    Smith, Adam Hewitt; Sra, Harminder K; Bawa, Sandeep; Stevens, Richard

    2010-07-01

    We report a case of Streptococcus bovis (Streptococcus gallolyticus subsp. pasteurianus) meningitis, a rare cause of central nervous system (CNS) infection in an adult, and comment on the importance of investigation of the lower gastrointestinal tract to identify a portal of entry in cases of systemic Streptococcus bovis infection. PMID:20421434

  15. Canine babesiosis in Romania due to Babesia canis and Babesia vogeli: a molecular approach.

    PubMed

    Ionita, Mariana; Mitrea, Ioan Liviu; Pfister, Kurt; Hamel, Dietmar; Buzatu, Catalin Marius; Silaghi, Cornelia

    2012-05-01

    Canine babesiosis is a tick-borne disease caused by the protozoa Babesia spp. that affects dogs worldwide. In Romania, canine babesiosis has become quite frequent in the last few years, with a wide variety of clinical signs, ranging from mild, nonspecific illness to peracute collapse, and even death. Traditionally, a Babesia infection in dogs is diagnosed based on the morphologic appearance of the intraerythrocytic piroplasms observed in peripheral blood smears. To date, no data on genetic characterization of Babesia species in dogs has been documented for Romania. Therefore, a molecular survey on natural Babesia infections of dogs in Romania using polymerase chain reaction and genetic sequence analysis of a fragment of the ssRNA gene was performed. A total number of 16 blood samples were tested for the presence of Babesia DNA. Blood samples were collected from 11 dogs with symptoms of babesiosis and microscopically proven positive for Babesia and from a group of five asymptomatic dogs, not tested microscopically for Babesia, which were included in the study for comparative analysis. The piroplasm-specific PCR amplifying the partial 18S rRNA gene confirmed Babesia spp. infection in all 11 samples from dogs with clinical babesiosis, and in one of the clinically normal dogs. Sequence analysis revealed the presence of Babesia canis in all clinically affected dogs and Babesia vogeli in one clinically normal dog. This is the first molecular evidence of B. canis and B. vogeli in dogs from Romania. The results of the study provide basic information toward a better understanding of the epidemiology of canine babesiosis in Romania and will help to promote an effective control program.

  16. Supernatants from ultraviolet-irradiated keratinocytes decrease the resistance and delayed-type hypersensitivity response to Mycobacterium bovis bacillus Calmette-Guerin in mice and impair the phagocytic ability of macrophages.

    PubMed

    Jeevan, A; Ullrich, S E; Dizon, V V; Kripke, M L

    1991-12-01

    We recently demonstrated that exposure of mice to a single high dose or multiple smaller doses of ultraviolet (UV) radiation decreased the induction of the delayed-type hypersensitivity (DTH) response to bacillus Calmette-Guerin (BCG) from Mycobacterium bovis injected into unexposed sites. In view of the limited ability of UV radiation to penetrate beyond the epidermis and upper layers of the dermis, it is not entirely clear how exposing the dorsal skin of mice to UV radiation causes systemic impairment of the immune response to BCG. In this study we report that mice injected with supernatants from keratinocyte cultures exposed to UV radiation in vitro impaired host resistance to BCG. Both induction and elicitation of the DTH reaction were suppressed after the intravenous injection of supernatants from UV-irradiated keratinocytes. Furthermore, these supernatants interfered with the elimination of viable bacteria from the lymphoid organs. To determine whether macrophages were the target of the UV-induced, keratinocyte-derived, suppressive cytokine, macrophages were isolated from mice injected with the suppressive cytokine or treated in vitro with the supernatants and tested for their ability to ingest and kill BCG in vitro. Injection of the suppressive factor significantly reduced the phagocytosis of BCG by the macrophages but did not alter the rate of intracellular killing. Similarly, phagocytosis was reduced when normal macrophages were treated in vitro with the suppressive factor. These findings suggest that the suppressive cytokine interferes with the elimination of bacteria in vivo by inhibiting the initial step in bacterial clearance, the uptake of the bacteria by host macrophages.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1823151

  17. Differential expression of three members of the multidomain adhesion CCp family in babesia bigemina, babesia bovis and theileria equi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Members of the CCp protein family have been previously described to be expressed on gametocytes of apicomplexan Plasmodium parasites. Knocking out Plasmodium CCp genes blocks the development of the parasite in the mosquito vector, making the CCp proteins potential targets for the development of a tr...

  18. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host–parasite interaction

    PubMed Central

    Jackson, Andrew P.; Otto, Thomas D.; Darby, Alistair; Ramaprasad, Abhinay; Xia, Dong; Echaide, Ignacio Eduardo; Farber, Marisa; Gahlot, Sunayna; Gamble, John; Gupta, Dinesh; Gupta, Yask; Jackson, Louise; Malandrin, Laurence; Malas, Tareq B.; Moussa, Ehab; Nair, Mridul; Reid, Adam J.; Sanders, Mandy; Sharma, Jyotsna; Tracey, Alan; Quail, Mike A.; Weir, William; Wastling, Jonathan M.; Hall, Neil; Willadsen, Peter; Lingelbach, Klaus; Shiels, Brian; Tait, Andy; Berriman, Matt; Allred, David R.; Pain, Arnab

    2014-01-01

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5′ ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct. PMID:24799432

  19. The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host-parasite interaction.

    PubMed

    Jackson, Andrew P; Otto, Thomas D; Darby, Alistair; Ramaprasad, Abhinay; Xia, Dong; Echaide, Ignacio Eduardo; Farber, Marisa; Gahlot, Sunayna; Gamble, John; Gupta, Dinesh; Gupta, Yask; Jackson, Louise; Malandrin, Laurence; Malas, Tareq B; Moussa, Ehab; Nair, Mridul; Reid, Adam J; Sanders, Mandy; Sharma, Jyotsna; Tracey, Alan; Quail, Mike A; Weir, William; Wastling, Jonathan M; Hall, Neil; Willadsen, Peter; Lingelbach, Klaus; Shiels, Brian; Tait, Andy; Berriman, Matt; Allred, David R; Pain, Arnab

    2014-06-01

    Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B. bigemina, B. divergens and B. bovis). Ves1 structure diverges rapidly after speciation, notably through the evolution of shortened forms (ves2) from 5' ends of canonical ves1 genes. Phylogenetic analyses show that ves1 genes are transposed between loci routinely, whereas ves2 genes are not. Similarly, analysis of sequence mosaicism shows that recombination drives variation in ves1 sequences, but less so for ves2, indicating the adoption of different mechanisms for variation of the two families. Proteomic analysis of the B. bigemina PR isolate shows that two dominant VESA1 proteins are expressed in the population, whereas numerous VESA2 proteins are co-expressed, consistent with differential transcriptional regulation of each family. Hence, VESA2 proteins are abundant and previously unrecognized elements of Babesia biology, with evolutionary dynamics consistently different to those of VESA1, suggesting that their functions are distinct.

  20. Transfusion-Transmitted Babesia microti.

    PubMed

    Fang, Deanna C; McCullough, Jeffrey

    2016-07-01

    Because testing of donors for Babesia microti has become available, it is important to determine the kinds of patients who should receive B microti-tested blood. We searched PubMed, AABB abstracts, and FDA Web site to identify all cases of transfusion-transmitted babesiosis (TTB). Cases were analyzed for underlying medical condition, age, presence of spleen, and reason for transfusion in relation to 5 classes of recipient outcome severity. Sixty-seven reports included 256 transfusion cases where donor tested positive for B microti, 165 of which resulted in TTB. Sixty recipients did not develop disease or become test positive, and test results were not known for 31 more. The 165 cases of TTB involved hematologic (19%), neonate (10%), cardiovascular (8%), and gastrointestinal (6%) patients. Thirty-two (19%) of the 165 infected patients died with death attributed to babesiosis in 25 of the cases. Nine (5%) were asymptomatic, 27 (16%) were symptomatic but had uncomplicated disease, and 16 (10%) had complicated disease. The severity of disease was mixed among many disease categories. Patients >65 years of age included the largest number of recipients (59/165, 36%) and deaths (11/32, 34%), although deaths occurred in other age groups as well. TTB cases were predominantly due to red cells (133 of 140 specified units), with red blood cell units processed in a variety of ways and at all storage duration. TTB with complicated babesiosis and/or death occurred in patients of all age groups and with a variety of underlying medical conditions. PMID:27260107

  1. Identification and characterization of a novel 34 kDa merozoite protein in Babesia orientalis.

    PubMed

    He, Lan; Fan, Lizhe; Liu, Qin; Hu, Jinfang; Miao, Xiaoyan; Huang, Yuan; He, Pei; He, Junwei; Yu, Long; Khan, Muhammad Kasib; Zhou, Yanqin; Shen, Bang; Zhao, Junlong

    2015-09-15

    A novel Babesia orientalis 34 kDa protein (designated BoP34) was obtained by immunoscreening of a cDNA expression library using B. orientalis infected water buffalo serum. The complete nucleotide sequence of the BoP34 was 1088 bp, which contained one open reading frame (ORF), two untranslated regions (UTRs) and a poly (A) tail. The length of ORF was 933 bp, encoding a polypeptide of 310 aa with a predicted size of 34 kDa. BLAST analysis showed that the nucleotide sequence of BoP34 had 71% similarity with that of the Babesia bovis gene XM_001611335, which encodes a nuclear movement family protein. This suggested that BoP34 is a homologous of the movement family protein. Structural analysis of the BoP34 protein indicated a CS domain which may interact with the ATPase domain of the heat shock protein 90. A truncated version of BoP34 was cloned into the expression vector pET-32a and subsequently expressed and purified from the Escherichia coli Rosetta™ (DE3) pLysS stain as a Trx-fusion (designated rBoP34-T). Antibodies in the serum of a B. orientalis-infected water buffalo were able to recognize this protein in immune-bloting analysis. Rabbit antibodies raised against rBoP34-T could detecte native BoP34 (34 kDa) in B. orientalis-infected water buffalo erythrocytes. These results suggested that BoP34 might be a good diagnostic antigen for the specific detection of anti-B. orientalis antibody in water buffalo. Further research is required to explore the biological function and diagnostic potential of this molecule.

  2. CD4+ T-helper lymphocyte responses against Babesia bigemina rhoptry-associated protein I.

    PubMed Central

    Rodríguez, S D; Palmer, G H; McElwain, T F; McGuire, T C; Ruef, B J; Chitko-McKown, M G; Brown, W C

    1996-01-01

    A multigene family of 58- to 60-kDa proteins, which are designated rhoptry-associated protein 1 (RAP-1) and which come from the parasites Babesia bigemina and Babesia bovis, is a target for vaccine development. The presence of multiple gene copies and conserved sequences and epitopes of RAP-1 implies that these proteins are functionally important for the survival of these parasites. Furthermore, it was previously shown that B. bigemina RAP-1 induced partial protection against challenge infection. However, the lack of correlation between protective immunity to B. bigemina infection and antibody titers against a merozoite surface-exposed, neutralization-sensitive epitope of B. bigemina RAP-1 indicated the potential importance of RAP-1-specific T helper (Th) cells in the observed protection. To begin to understand the mechanism of RAP-1-induced protective immunity, RAP-1-specific T-cell responses were characterized in cattle. Vigorous and sustained proliferative responses of peripheral blood mononuclear cells from native RAP-1-immunized cattle were observed. The anamnestic response in immunized cattle was specific for B. bigemina RAP-1 and predominantly comprised CD4+ T cells, which upon cloning expressed type 1 cytokine mRNA profiles and high levels of gamma interferon protein. The T cells responded to both native and recombinant forms of RAP-1, indicating the potential to use recombinant protein or epitopes derived therefrom as a vaccine that could evoke specific recall responses after exposure to natural infection. The differential responses of peripheral blood mononuclear cells and seven Th-cell clones derived from RAP-1-immunized cattle to different Central American strains of B. bigemina indicated the presence of at least one conserved and one variable Th-cell epitope. The lack of response to B. bovis RAP-1 indicated that a strictly conserved 14-amino-acid peptide shared by the two babesial species was not immunogenic for Th cells in these experiments. However

  3. Babesia spp. in European wild ruminant species: parasite diversity and risk factors for infection

    PubMed Central

    2014-01-01

    Babesia are tick-borne parasites that are increasingly considered as a threat to animal and public health. We aimed to assess the role of European free-ranging wild ruminants as maintenance mammalian hosts for Babesia species and to determine risk factors for infection. EDTA blood was collected from 222 roe deer (Capreolus c. capreolus), 231 red deer (Cervus e. elaphus), 267 Alpine chamois (Rupicapra r. rupicapra) and 264 Alpine ibex (Capra i. ibex) from all over Switzerland and analysed by PCR with pan-Babesia primers targeting the 18S rRNA gene, primers specific for B. capreoli and Babesia sp. EU1, and by sequencing. Babesia species, including B. divergens, B. capreoli, Babesia sp. EU1, Babesia sp. CH1 and B. motasi, were detected in 10.7% of all samples. Five individuals were co-infected with two Babesia species. Infection with specific Babesia varied widely between host species. Cervidae were significantly more infected with Babesia spp. than Caprinae. Babesia capreoli and Babesia sp. EU1 were mostly found in roe deer (prevalences 17.1% and 7.7%, respectively) and B. divergens and Babesia sp. CH1 only in red deer. Factors significantly associated with infection were low altitude and young age. Identification of Babesia sp. CH1 in red deer, co-infection with multiple Babesia species and infection of wild Caprinae with B. motasi and Babesia sp. EU1 are novel findings. We propose wild Caprinae as spillover or accidental hosts for Babesia species but wild Cervidae as mammalian reservoir hosts for B. capreoli, possibly Babesia sp. EU1 and Babesia sp. CH1, whereas their role regarding B. divergens is more elusive. PMID:24925474

  4. Babesia spp. in European wild ruminant species: parasite diversity and risk factors for infection.

    PubMed

    Michel, Adam O; Mathis, Alexander; Ryser-Degiorgis, Marie-Pierre

    2014-06-13

    Babesia are tick-borne parasites that are increasingly considered as a threat to animal and public health. We aimed to assess the role of European free-ranging wild ruminants as maintenance mammalian hosts for Babesia species and to determine risk factors for infection. EDTA blood was collected from 222 roe deer (Capreolus c. capreolus), 231 red deer (Cervus e. elaphus), 267 Alpine chamois (Rupicapra r. rupicapra) and 264 Alpine ibex (Capra i. ibex) from all over Switzerland and analysed by PCR with pan-Babesia primers targeting the 18S rRNA gene, primers specific for B. capreoli and Babesia sp. EU1, and by sequencing. Babesia species, including B. divergens, B. capreoli, Babesia sp. EU1, Babesia sp. CH1 and B. motasi, were detected in 10.7% of all samples. Five individuals were co-infected with two Babesia species. Infection with specific Babesia varied widely between host species. Cervidae were significantly more infected with Babesia spp. than Caprinae. Babesia capreoli and Babesia sp. EU1 were mostly found in roe deer (prevalences 17.1% and 7.7%, respectively) and B. divergens and Babesia sp. CH1 only in red deer. Factors significantly associated with infection were low altitude and young age. Identification of Babesia sp. CH1 in red deer, co-infection with multiple Babesia species and infection of wild Caprinae with B. motasi and Babesia sp. EU1 are novel findings. We propose wild Caprinae as spillover or accidental hosts for Babesia species but wild Cervidae as mammalian reservoir hosts for B. capreoli, possibly Babesia sp. EU1 and Babesia sp. CH1, whereas their role regarding B. divergens is more elusive.

  5. Characterization of a papain-like cysteine protease essential for the survival of Babesia ovis merozoites.

    PubMed

    Carletti, Tamara; Barreto, Carmo; Mesplet, Maria; Mira, Anabela; Weir, William; Shiels, Brian; Oliva, Abel Gonzalez; Schnittger, Leonhard; Florin-Christensen, Monica

    2016-02-01

    Babesia ovis, a tick-transmitted intraerythrocytic protozoan parasite, causes severe infections in small ruminants from Southern Europe, Middle East, and Northern Africa. With the aim of finding potential targets for the development of control methods against this parasite, sequence analysis of its genome led to the identification of four putative cysteine proteases of the C1A family. Orthology between B. ovis, B. bovis, T. annulata, and T. parva sequences showed that each B. ovis C1A peptidase sequence clustered within one of the four ortholog groups previously reported for these piroplasmids. The ortholog of bovipain-2 of B. bovis and falcipain-2 of Plasmodium falciparum, respectively, was designated "ovipain-2" and further characterized. In silico analysis showed that ovipain-2 has the typical topology of papain-like cysteine peptidases and a highly similar predicted three dimensional structure to bovipain-2 and falcipain-2, suggesting susceptibility to similar inhibitors. Immunoblotting using antibodies raised against a recombinant form of ovipain-2 (r-ovipain-2) demonstrated expression of ovipain-2 in in vitro cultured B. ovis merozoites. By immunofluorescence, these antibodies reacted with merozoites and stained the cytoplasm of infected erythrocytes. This suggests that ovipain-2 is secreted by the parasite and could be involved in intra- and extracellular digestion of hemoglobin and/or cleavage of erythrocyte proteins facilitating parasite egress. A significant reduction in the percentage of parasitized erythrocytes was obtained upon incubation of B. ovis in vitro cultures with anti-r-ovipain-2 antibodies, indicating an important functional role for ovipain-2 in the intra erythrocytic development cycle of this parasite. Finally, studies of the reactivity of sera from B. ovis-positive and negative sheep against r-ovipain-2 showed that this protease is expressed in vivo, and can be recognized by host antibodies. The results of this study suggest that ovipain-2

  6. Natural history of Zoonotic Babesia: Role of wildlife reservoirs

    PubMed Central

    Yabsley, Michael J.; Shock, Barbara C.

    2012-01-01

    Babesiosis is an emerging zoonotic disease on all inhabited continents and various wildlife species are the principal reservoir hosts for zoonotic Babesia species. The primary vectors of Babesia are Ixodid ticks, with the majority of zoonotic species being transmitted by species in the genus Ixodes. Species of Babesia vary in their infectivity, virulence and pathogenicity for people. Various factors (e.g., increased interactions between people and the environment, increased immunosuppression, changes in landscape and climate, and shifts in host and vector species abundance and community structures) have led to an increase in tick-borne diseases in people, including babesiosis. Furthermore, because babesiosis is now a reportable disease in several states in the United States, and it is the most common blood transfusion-associated parasite, recognized infections are expected to increase. Because of the zoonotic nature of these parasites, it is essential that we understand the natural history (especially reservoirs and vectors) so that appropriate control and prevention measures can be implemented. Considerable work has been conducted on the ecology of Babesia microti and Babesia divergens, the two most common causes of babesiosis in the United States and Europe, respectively. However, unfortunately, for many of the zoonotic Babesia species, the reservoir(s) and/or tick vector(s) are unknown. We review the current knowledge regarding the ecology of Babesia among their reservoir and tick hosts with an emphasis of the role on wildlife as reservoirs. We hope to encourage the molecular characterization of Babesia from potential reservoirs and vectors as well from people. These data are necessary so that informed decisions can be made regarding potential vectors and the potential role of wildlife in the ecology of a novel Babesia when it is detected in a human patient. PMID:24533312

  7. Mycobacterium bovis (Bovine Tuberculosis) in Humans

    MedlinePlus

    ... such as what might occur during slaughter or hunting, or by inhaling the bacteria in air exhaled by animals infected with M. bovis. Direct transmission from animals to humans through the air is thought to be rare, but M. bovis can be spread directly from ...

  8. Theileria, Babesia, and Anaplasma detected by PCR in ruminant herds at Bié Province, Angola

    PubMed Central

    Kubelová, M.; Mazancová, J.; Široký, P.

    2012-01-01

    Distribution of Anaplasma spp., Babesia spp., Theileria spp., and Ehrlichia ruminantium, was for the first time studied in Bié Province, central Angola. We examined 76 blood samples of cattle originated from seven farms, and 13 blood samples of goats from two farms employing molecular genetic tools (PCR). Most prevalent was A. ovis-infection in goats (100%) and A. marginale-infection in cattle (38% of examined animals, and six out of seven farms). B. bigemina-infection was detected in only one specimen at Andulo, whereas B. bovis was not detected in Bié. We did not detected T. parva, the causative agent of serious diseases in cattle; nevertheless, infection by T. velifera was quite frequent (14% of examined animals, and five out of seven farms). Causative agent of heartwater disease – E. ruminantium, was not detected. Taking into account short-term perspective of PCR methods in monitoring of epidemiological status in herds, the number of infected animals and distribution of detected pathogens should not be ignored. PMID:23193527

  9. Mycobacterium bovis: characteristics of wildlife reservoir hosts.

    PubMed

    Palmer, M V

    2013-11-01

    Mycobacterium bovis is the cause of tuberculosis in animals and sometimes humans. Many developed nations have long-standing programmes to eradicate tuberculosis in livestock, principally cattle. As disease prevalence in cattle decreases these efforts are sometimes impeded by passage of M. bovis from wildlife to cattle. In epidemiological terms, disease can persist in some wildlife species, creating disease reservoirs, if the basic reproduction rate (R0) and critical community size (CCS) thresholds are achieved. Recognized wildlife reservoir hosts of M. bovis include the brushtail possum (Trichosurus vulpecula) in New Zealand, European badger (Meles meles) in Great Britain and Ireland, African buffalo (Syncerus caffer) in South Africa, wild boar (Sus scrofa) in the Iberian Peninsula and white-tailed deer (Odocoileus virginianus) in Michigan, USA. The epidemiological concepts of R0 and CCS are related to more tangible disease/pathogen characteristics such as prevalence, pathogen-induced pathology, host behaviour and ecology. An understanding of both epidemiological and disease/pathogen characteristics is necessary to identify wildlife reservoirs of M. bovis. In some cases, there is a single wildlife reservoir host involved in transmission of M. bovis to cattle. Complexity increases, however, in multihost systems where multiple potential reservoir hosts exist. Bovine tuberculosis eradication efforts require elimination of M. bovis transmission between wildlife reservoirs and cattle. For successful eradication identification of true wildlife reservoirs is critical, as disease control efforts are most effective when directed towards true reservoirs.

  10. Mycobacterium bovis: characteristics of wildlife reservoir hosts.

    PubMed

    Palmer, M V

    2013-11-01

    Mycobacterium bovis is the cause of tuberculosis in animals and sometimes humans. Many developed nations have long-standing programmes to eradicate tuberculosis in livestock, principally cattle. As disease prevalence in cattle decreases these efforts are sometimes impeded by passage of M. bovis from wildlife to cattle. In epidemiological terms, disease can persist in some wildlife species, creating disease reservoirs, if the basic reproduction rate (R0) and critical community size (CCS) thresholds are achieved. Recognized wildlife reservoir hosts of M. bovis include the brushtail possum (Trichosurus vulpecula) in New Zealand, European badger (Meles meles) in Great Britain and Ireland, African buffalo (Syncerus caffer) in South Africa, wild boar (Sus scrofa) in the Iberian Peninsula and white-tailed deer (Odocoileus virginianus) in Michigan, USA. The epidemiological concepts of R0 and CCS are related to more tangible disease/pathogen characteristics such as prevalence, pathogen-induced pathology, host behaviour and ecology. An understanding of both epidemiological and disease/pathogen characteristics is necessary to identify wildlife reservoirs of M. bovis. In some cases, there is a single wildlife reservoir host involved in transmission of M. bovis to cattle. Complexity increases, however, in multihost systems where multiple potential reservoir hosts exist. Bovine tuberculosis eradication efforts require elimination of M. bovis transmission between wildlife reservoirs and cattle. For successful eradication identification of true wildlife reservoirs is critical, as disease control efforts are most effective when directed towards true reservoirs. PMID:24171844

  11. Development of droplet digital PCR for the detection of Babesia microti and Babesia duncani

    PubMed Central

    Wilson, Melisa; Glaser, Kathleen C.; Adams-Fish, Debra; Boley, Matthew; Mayda, Maria; Molestina, Robert E.

    2014-01-01

    Babesia spp. are obligate protozoan parasites of red blood cells. Transmission to humans occurs through bites from infected ticks or blood transfusion. Infections with B. microti account for the majority of the reported cases of human babesiosis in the USA. A lower incidence is caused by the more recently described species B. duncani. The current gold standard for detection of Babesia is microscopic examination of blood smears. Recent PCR-based assays, including real-time PCR, have been developed for B. microti. On the other hand, molecular assays that detect and distinguish between B. microti and B. duncani infections are lacking. Closely related species of Babesia can be differentiated due to sequence variation within the internal transcribed spacer (ITS) regions of nuclear ribosomal RNAs. In the present study, we targeted the ITS regions of B. microti and B. duncani to develop sensitive and species-specific droplet digital PCR (ddPCR) assays. The assays were shown to discriminate B. microti from B. duncani and resulted in limits of detection of ~10 gene copies. Moreover, ddPCR for these species were useful in DNA extracted from blood of experimentally infected hamsters, detecting infections of low parasitemia that were negative by microscopic examination. In summary, we have developed sensitive and specific quantitative ddPCR assays for the detection of B. microti and B. duncani in blood. Our methods could be used as sensitive approaches to monitor the progression of parasitemia in rodent models of infection as well as serve as suitable molecular tests in blood screening. PMID:25500215

  12. Abortion associated with Mycoplasma bovis (M. bovis) in a bison (Bison bison) herd

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) has recently emerged as a significant health threat in bison and is an increasing concern and source of economic loss for producers. Clinical manifestations of infection documented in bison include pneumonia, respiratory distress and polyarthritis. The current study des...

  13. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    PubMed

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  14. Molecular assays reveal the presence of Theileria spp. and Babesia spp. in Asian water buffaloes (Bubalus bubalis, Linnaeus, 1758) in the Amazon region of Brazil.

    PubMed

    Silveira, Júlia A G; de Oliveira, Cairo H S; Silvestre, Bruna T; Albernaz, Tatiana T; Leite, Rômulo C; Barbosa, José D; Oliveira, Carlos M C; Ribeiro, Múcio F B

    2016-07-01

    Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from

  15. Observation of a novel Babesia spp. in Eastern Grey Kangaroos (Macropus giganteus) in Australia.

    PubMed

    Dawood, Kaiser E; Morgan, Jess A T; Busfield, Frances; Srivastava, Mukesh; Fletcher, Taryn I; Sambono, Jacqueline; Jackson, Louise A; Venus, Bronwyn; Philbey, Adrian W; Lew-Tabor, Ala E

    2013-12-01

    The roles and epidemiological features of tick-borne protozoans are not well elicited in wildlife. Babesia spp. are documented in many domestic animals, including cattle, horses, pigs, dogs and cats. Three cases affecting eastern grey kangaroos are described. The kangaroos exhibited neurological signs, depression and marked anaemia, and microscopic examination of blood smears revealed intraerythrocytic piroplasms. One to seven intraerythrocytic spherical, oval, pyriform and irregularly-shaped parasites consistent with Babesia spp. were seen in the blood smears and the percentage of infected erythrocytes was estimated to be approximately 7% in each case. Data suggest that the tick vector for this kangaroo Babesia sp. is a Haemaphysalis species. For Case 2, ultrastructural examination of the erythrocytes of the renal capillaries showed parasites resembling Babesia spp. and 18 of 33 erythrocytes were infected. DNA sequencing of the amplified 18S rDNA confirmed that the observed intraerythrocytic piroplasms belong to the genus Babesia. The phylogenetic position of this new kangaroo Babesia sp. (de novo Babesia macropus), as a sister species to the new Australian woylie Babesia sp., suggests a close affinity to the described Afro-Eurasian species Babesia orientalis and Babesia occultans suggesting perhaps a common ancestor for the Babesia in kangaroos.

  16. Observation of a novel Babesia spp. in Eastern Grey Kangaroos (Macropus giganteus) in Australia

    PubMed Central

    Dawood, Kaiser E.; Morgan, Jess A.T.; Busfield, Frances; Srivastava, Mukesh; Fletcher, Taryn I.; Sambono, Jacqueline; Jackson, Louise A.; Venus, Bronwyn; Philbey, Adrian W.; Lew-Tabor, Ala E.

    2012-01-01

    The roles and epidemiological features of tick-borne protozoans are not well elicited in wildlife. Babesia spp. are documented in many domestic animals, including cattle, horses, pigs, dogs and cats. Three cases affecting eastern grey kangaroos are described. The kangaroos exhibited neurological signs, depression and marked anaemia, and microscopic examination of blood smears revealed intraerythrocytic piroplasms. One to seven intraerythrocytic spherical, oval, pyriform and irregularly-shaped parasites consistent with Babesia spp. were seen in the blood smears and the percentage of infected erythrocytes was estimated to be approximately 7% in each case. Data suggest that the tick vector for this kangaroo Babesia sp. is a Haemaphysalis species. For Case 2, ultrastructural examination of the erythrocytes of the renal capillaries showed parasites resembling Babesia spp. and 18 of 33 erythrocytes were infected. DNA sequencing of the amplified 18S rDNA confirmed that the observed intraerythrocytic piroplasms belong to the genus Babesia. The phylogenetic position of this new kangaroo Babesia sp. (de novo Babesia macropus), as a sister species to the new Australian woylie Babesia sp., suggests a close affinity to the described Afro–Eurasian species Babesia orientalis and Babesia occultans suggesting perhaps a common ancestor for the Babesia in kangaroos. PMID:24533316

  17. Clinical babesiosis and molecular identification of Babesia canis and Babesia gibsoni infections in dogs from Serbia.

    PubMed

    Davitkov, Darko; Vucicevic, Milos; Stevanovic, Jevrosima; Krstic, Vanja; Tomanovic, Snezana; Glavinic, Uros; Stanimirovic, Zoran

    2015-06-01

    Canine babesiosis is a frequent and clinically significant tick-borne disease. Sixty symptomatic dogs with clinical findings compatible with babesiosis were included in this study conducted in Serbia. After clinical examination, blood samples were taken for microscopic examination, complete blood count (CBC), Canine SNAP 4Dx Test, DNA analyses and sequencing. The main clinical signs included apathy, anorexia, fever, brown/red discoloration of urine, pale mucous membranes, icterus, splenomegaly, and vomiting. The main clinicopathological findings in Babesia infections were a slight to severe thrombocytopenia and a mild to very severe normocytic normochromic anaemia. Microscopic evaluation revealed 58 positive samples with the presence of large and small intraerythrocytic piroplasms in 57 and 1 sample(s), respectively. No co-infections were found using SNAP test. Two Babesia species, B. canis (58/60) and B. gibsoni (2/60), were differentiated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Species identification was further confirmed by sequencing PCR products of B. gibsoni samples and six randomly selected B. canis samples. All dogs were treated with imidocarb dipropionate (6.6 mg/kg of body weight), given intramuscularly twice at an interval of 14 days. This report presents the first molecular evidence of the occurrence of B. gibsoni and B. canis, confirmed by DNA sequencing, in sick dogs from Serbia. PMID:26051258

  18. Identification of Streptococcus bovis and Streptococcus salivarius in clinical laboratories.

    PubMed Central

    Ruoff, K L; Ferraro, M J; Holden, J; Kunz, L J

    1984-01-01

    Streptococci identified as Streptococcus bovis, S. bovis variant, and Streptococcus salivarius were examined with respect to physiological and serological characteristics and cellular fatty acid content. Similarities in physiological reactions and problems encountered in serological analysis were noted, suggesting that an expanded battery of physiological tests is needed to definitively identify these streptococci. Cellular fatty acid analysis provided an accurate method for distinguishing S. salivarius from S. bovis and S. bovis variant. PMID:6490816

  19. First record of Babesia sp. in Antarctic penguins.

    PubMed

    Montero, Estrella; González, Luis Miguel; Chaparro, Alberto; Benzal, Jesús; Bertellotti, Marcelo; Masero, José A; Colominas-Ciuró, Roger; Vidal, Virginia; Barbosa, Andrés

    2016-04-01

    This is the first reported case of Babesia sp. in Antarctic penguins, specifically a population of Chinstrap penguins (Pygoscelis antarctica) in the Vapour Col penguin rookery in Deception Island, South Shetlands, Antarctica. We collected peripheral blood from 50 adult and 30 chick Chinstrap penguins. Examination of the samples by microscopy showed intraerythrocytic forms morphologically similar to other avian Babesia species in 12 Chinstrap penguin adults and seven chicks. The estimated parasitaemias ranged from 0.25×10(-2)% to 0.75×10(-2)%. Despite the low number of parasites found in blood smears, semi-nested PCR assays yielded a 274 bp fragment in 12 of the 19 positive blood samples found by microscopy. Sequencing revealed that the fragment was 97% similar to Babesia sp. 18S rRNA from Australian Little Penguins (Eudyptula minor) confirming presence of the parasite. Parasite prevalence estimated by microscopy in adults and chicks was higher (24% vs. 23.3%, respectively) than found by semi-nested PCR (16% vs. 13.3% respectively). Although sampled penguins were apparently healthy, the effect of Babesia infection in these penguins is unknown. The identification of Babesia sp. in Antarctic penguins is an important finding. Ixodes uriae, as the only tick species present in the Antarctic Peninsula, is the key to understanding the natural history of this parasite. Future work should address the transmission dynamics and pathogenicity of Babesia sp. in Chinstrap penguin as well as in other penguin species, such as Gentoo penguin (Pygoscelis papua) and Adélie penguin (Pygoscelis adeliae), present within the tick distribution range in the Antarctic Peninsula.

  20. First record of Babesia sp. in Antarctic penguins.

    PubMed

    Montero, Estrella; González, Luis Miguel; Chaparro, Alberto; Benzal, Jesús; Bertellotti, Marcelo; Masero, José A; Colominas-Ciuró, Roger; Vidal, Virginia; Barbosa, Andrés

    2016-04-01

    This is the first reported case of Babesia sp. in Antarctic penguins, specifically a population of Chinstrap penguins (Pygoscelis antarctica) in the Vapour Col penguin rookery in Deception Island, South Shetlands, Antarctica. We collected peripheral blood from 50 adult and 30 chick Chinstrap penguins. Examination of the samples by microscopy showed intraerythrocytic forms morphologically similar to other avian Babesia species in 12 Chinstrap penguin adults and seven chicks. The estimated parasitaemias ranged from 0.25×10(-2)% to 0.75×10(-2)%. Despite the low number of parasites found in blood smears, semi-nested PCR assays yielded a 274 bp fragment in 12 of the 19 positive blood samples found by microscopy. Sequencing revealed that the fragment was 97% similar to Babesia sp. 18S rRNA from Australian Little Penguins (Eudyptula minor) confirming presence of the parasite. Parasite prevalence estimated by microscopy in adults and chicks was higher (24% vs. 23.3%, respectively) than found by semi-nested PCR (16% vs. 13.3% respectively). Although sampled penguins were apparently healthy, the effect of Babesia infection in these penguins is unknown. The identification of Babesia sp. in Antarctic penguins is an important finding. Ixodes uriae, as the only tick species present in the Antarctic Peninsula, is the key to understanding the natural history of this parasite. Future work should address the transmission dynamics and pathogenicity of Babesia sp. in Chinstrap penguin as well as in other penguin species, such as Gentoo penguin (Pygoscelis papua) and Adélie penguin (Pygoscelis adeliae), present within the tick distribution range in the Antarctic Peninsula. PMID:26874670

  1. Mycoplasam Bovis - an emerging pathogen of ranched bison

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) is an emerging bacterial pathogen that has caused severe disease among ranched bison (Bison bison) herds in North America. Unlike cattle, M. bovis in bison seems to be a primary pathogen, affecting animals in feedlots as well as breeding-age cows on pasture. Mortality r...

  2. Fatal Babesia canis canis infection in a splenectomized Estonian dog.

    PubMed

    Tiškina, Valentina; Capligina, Valentina; Must, Külli; Berzina, Inese; Ranka, Renate; Jokelainen, Pikka

    2016-01-25

    A previously splenectomized dog from Estonia was presented with a sudden lack of appetite and discoloration of the urine. Despite supportive therapy, its condition deteriorated dramatically during 1 day. Severe thrombocytopenia and high numbers of protozoan hemoparasites were evident in blood smears, and the hematocrit dropped from 46 to 33 %. The dog was euthanized before specific antibabesial treatment was initiated. Blood samples from the dog and from two other dogs in the same household tested positive for Babesia using molecular methods, and the sequences of partial 18S rRNA gene confirmed the causative species as Babesia canis canis. The risk of severe, rapidly progressing babesiosis in splenectomized dogs merits awareness.

  3. Efficacy and Immunogenicity of Mycobacterium bovis Delta RD1 against Aerosol M. bovis Infection in Neonatal Calves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An attenuated Mycobacterium bovis RD1 knockout (Delta RD1) vaccine administered to calves at 2 weeks of age provided similar efficacy as M. bovis bacille Calmette Guerin (BCG) against low dose, aerosol challenge with virulent M. bovis at 3.5m of age. Approximately 4.5 months after challenge, both De...

  4. Lack of transplacental transmission of Bartonella bovis.

    PubMed

    Chastant-Maillard, S; Boulouis, H-J; Reynaud, K; Thoumire, S; Gandoin, C; Bouillin, C; Cordonnier, N; Maillard, R

    2015-02-01

    Transplacental transmission of Bartonella spp. has been reported for rodents, but not for cats and has never been investigated in cattle. The objective of this study was to assess vertical transmission of Bartonella in cattle. Fifty-six cow-calf pairs were tested before (cows) and after (calves) caesarean section for Bartonella bacteremia and/or serology, and the cotyledons were checked for gross lesions and presence of the bacteria. None of the 29 (52%) bacteremic cows gave birth to bacteremic calves, and all calves were seronegative at birth. Neither placentitis nor vasculitis were observed in all collected cotyledons. Bartonella bovis was not detected in placental cotyledons. Therefore, transplacental transmission of B. bovis and multiplication of the bacteria in the placenta do not seem likely. The lack of transplacental transmission may be associated with the particular structure of the placenta in ruminants or to a poor affinity/agressiveness of B. bovis for this tissue.

  5. Wild Cervids Are Host for Tick Vectors of Babesia Species with Zoonotic Capability in Belgium

    PubMed Central

    Wirtgen, Marc; Nahayo, Adrien; Caron, Yannick; Shiels, Brian; Saegerman, Claude; Losson, Bertrand; Linden, Annick

    2012-01-01

    Abstract Babesiosis is a tick-borne disease caused by different species of intraerythrocytic protozoan parasites within the genus Babesia. Different species of Babesia are described as potentially zoonotic and cause a malaria-like disease mainly in immunocompromised humans. Interest in the zoonotic potential of Babesia is growing and babesiosis has been described by some authors as an emergent zoonotic disease. The role of cervids to maintain tick populations and act as a reservoir host for some Babesia spp. with zoonotic capability is suspected. To investigate the range and infection rate of Babesia species, ticks were collected from wild cervids in southern Belgium during 2008. DNA extraction was performed for individual ticks, and each sample was evaluated for the absence of PCR inhibition using a PCR test. A Babesia spp. genus-specific PCR based on the 18S rRNA gene was applied to validated tick DNA extracts. A total of 1044 Ixodes ricinus ticks were collected and 1023 validated samples were subsequently screened for the presence of Babesia spp. DNA. Twenty-eight tick samples were found to be positive and identified after sequencing as containing DNA representing: Babesia divergens (3), B. divergens-like (5), Babesia sp. EU1 (11), Babesia sp. EU1-like (3), B. capreoli (2), or unknown Babesia sp. (4). This study confirms the presence of potentially zoonotic species and Babesia capreoli in Belgium, with a tick infection rate of 2.7% (95% CI 1.8,3.9%). Knowledge of the most common reservoir source for transmission of zoonotic Babesia spp. will be useful for models assessing the risk potential of this infection to humans. PMID:22214270

  6. First Report of Babesia microti-Caused Babesiosis in Spain

    PubMed Central

    Arsuaga, Marta; Gonzalez, Luis M.; Lobo, Cheryl A.; de la Calle, Fernando; Bautista, Jose M.; Azcárate, Isabel G.; Puente, Sabino

    2016-01-01

    Abstract Babesiosis is an emerging zoonosis now found in several areas of the world. Using PCR and indirect immunofluorescence assay, we have diagnosed the first case of human babesiosis caused by Babesia microti in Spain. Diagnosis was delayed because of the nonspecific clinical symptoms that occurred in an immunocompetent patient. PMID:27560451

  7. First Report of Babesia microti-Caused Babesiosis in Spain.

    PubMed

    Arsuaga, Marta; Gonzalez, Luis M; Lobo, Cheryl A; de la Calle, Fernando; Bautista, Jose M; Azcárate, Isabel G; Puente, Sabino; Montero, Estrella

    2016-10-01

    Babesiosis is an emerging zoonosis now found in several areas of the world. Using PCR and indirect immunofluorescence assay, we have diagnosed the first case of human babesiosis caused by Babesia microti in Spain. Diagnosis was delayed because of the nonspecific clinical symptoms that occurred in an immunocompetent patient.

  8. Natural Transmission of Zoonotic Babesia spp. by Ixodes ricinus Ticks

    PubMed Central

    Becker, Claire A.M.; Bouju-Albert, Agnès; Jouglin, Maggy; Chauvin, Alain

    2009-01-01

    To determine characteristics of natural transmission of Babesia sp. EU1 and B. divergens by adult Ixodes ricinus ticks, we examined tick salivary gland contents. We found that I. ricinus is a competent vector for EU1 and that their sporozoites directly invade erythrocytes. We conclude that EU1 is naturally transmitted by I. ricinus. PMID:19193284

  9. Babesia microti in rodents and raccoons from northeast Florida.

    PubMed

    Clark, Kerry; Savick, Kyla; Butler, Joseph

    2012-12-01

    Human babesiosis in the United States is caused most commonly by the intraerythrocytic protozoan parasite, Babesia microti . Although a few reports have described evidence of Babesia species in animals in Florida, to date Babesia microti specifically has not been reported from Florida or most other southern states. To determine if the organism is present in vertebrates in the region, small mammals were trapped and sampled at 2 sites in northeastern Florida, and DNA extracts from blood samples were screened for B. microti DNA via PCR assays targeting portions of the nuclear small subunit rRNA (18S rDNA) and beta-tubulin genes. Amplified fragments from representative samples of PCR-positive hosts were sequenced and compared phylogenetically to reference strains of Babesia species. The B. microti strains found in cotton rats ( Sigmodon hispidus ) most closely resembles B. microti sensu stricto strains that are pathogenic to humans, and strains found in raccoons ( Procyon lotor ) most closely resembles previously described raccoon-related strains of B. microti sensu lato. The results of this study suggest that B. microti is prevalent among cotton rats and raccoons at some sites in northeast Florida and may pose a risk to humans in the region.

  10. Immunopathogenesis of Mycobacterium bovis infection of cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aerosol and intratracheal inoculation routes are commonly used for experimental biology purposes to infect cattle with virulent Mycobacterium bovis, each resulting primarily in a respiratory tract infection including lungs and lung-associated lymph nodes. Disease severity is dose and time dependent...

  11. Molecular detection and characterization of Cytauxzoon felis and a Babesia species in cougars from Florida.

    PubMed

    Yabsley, Michael J; Murphy, Staci M; Cunningham, Mark W

    2006-04-01

    Piroplasms, morphologically indistinguishable from Cytauxzoon felis, previously were detected in 36% of cougars in Florida. We utilized a nested 18S rRNA assay, which amplifies DNA from all piroplasms, to screen blood samples collected from 41 cougars from Florida (39 native Florida panthers [Puma concolor coryi] and two translocated Texas cougars [P. c. stanleyana]) from 1989-2005. Thirty-nine of the 41 cougars (95%) were positive for piroplasms; however, sequence analysis and restriction enzyme digestion revealed that only five were positive for C. felis. Samples from 32 cougars were positive for a Babesia sp. Two cougars were co-infected with both C. felis and the Babesia sp. Phylogenetic analysis of 18S rRNA gene sequence indicated that the Florida panther Babesia sp. was most closely related to a Babesia sp. reported from Ixodes ovatus from Japan, Babesia divergens, and Babesia odocoilei. This study indicates that Florida panthers harbor two distinct piroplasms, C. felis and a Babesia sp., and that some individuals are infected with both organisms. The infectivity and pathogenicity of this Babesia sp. for domestic cats is unknown. This represents the first report of a feline Babesia sp. in North America.

  12. Babesia spp. in questing ticks from eastern Poland: prevalence and species diversity.

    PubMed

    Wójcik-Fatla, Angelina; Zając, Violetta; Sawczyn, Anna; Cisak, Ewa; Dutkiewicz, Jacek

    2015-08-01

    A total of 853 questing Ixodes ricinus males, females, and nymphs and of 582 questing Dermacentor reticulatus males and females were collected from vegetation on the territory of the Lublin province (eastern Poland). The ticks were examined for the presence of Babesia by PCR detecting part of 18S ribosomal RNA (rRNA) gene and nuclear small subunit rRNA (SS-rDNA) for determining of Babesia spp. and Babesia microti, respectively. The overall incidence of Babesia strains in I. ricinus ticks was 4.6%. Three species of Babesia were identified. The prevalent species was B. microti which occurred in 2.8% of ticks, while Babesia venatorum, Babesia divergens, and unidentified Babesia species were found at the frequency of 1.2, 0.2, and 0.3%, respectively. Altogether, B. microti constituted 61.5% of the total strains detected in I. ricinus, B. venatorum-25.7%, B. divergens-5.1%, and unidentified Babesia species-7.7%. The prevalence of Babesia species in I. ricinus did not depend significantly on locality (χ(2) = 1.885, P = 0.390) nor on the tick stage (χ(2) = 4.874, P = 0.087). The incidence of Babesia strains in D. reticulatus ticks was 2.7%. Two species of Babesia were identified. Again, the prevalent species was B. microti which occurred in 2.1% of ticks, while B. canis was found in 0.7% of ticks. In one D. reticulatus female, B. canis and B. microti co-infection was found. Altogether, B. microti constituted 75% of the total strains detected in D. reticulatus while B. canis formed 25% of the total strains. The frequency of the occurrence of Babesia species in D. reticulatus did not depend significantly on locality (χ(2) = 0.463, P = 0.793). The difference between the prevalence of Babesia in males and females of D. reticulatus was insignificant (P = 0.0954); nymphs were not found. The dominance of B. microti in the species composition of tick-borne Babesia found in this study was typical for eastern Europe. In conclusion, the results

  13. Mycoplasma bovis: Mechanisms of Resistance and Trends in Antimicrobial Susceptibility

    PubMed Central

    Lysnyansky, Inna; Ayling, Roger D.

    2016-01-01

    Mycoplasma bovis is a cell-wall-less bacterium and belongs to the class Mollicutes. It is the most important etiological agent of bovine mycoplasmoses in North America and Europe, causing respiratory disease, mastitis, otitis media, arthritis, and reproductive disease. Clinical disease associated with M. bovis is often chronic, debilitating, and poorly responsive to antimicrobial therapy, resulting in significant economic loss, the full extent of which is difficult to estimate. Until M. bovis vaccines are universally available, sanitary control measures and antimicrobial treatment are the only approaches that can be used in attempts to control M. bovis infections. However, in vitro studies show that many of the current M. bovis isolates circulating in Europe have high minimum inhibitory concentrations (MIC) for many of the commercially available antimicrobials. In this review we summarize the current MIC trends indicating the development of antimicrobial resistance in M. bovis as well as the known molecular mechanisms by which resistance is acquired. PMID:27199926

  14. Dynamics of bovine spleen cell populations during the response to acute Babesia bovis infection: an immunohistological study

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The spleen is the critical organ in defense against hemoparasitic diseases like babesiosis, responding to the pathogen in blood as it passes through the organ. As a lymphoid organ, the spleen is composed of cells whose role it is to orchestrate the engagement of infected erythrocytes and free parasi...

  15. Fatal Babesia canis canis infection in a splenectomized Estonian dog.

    PubMed

    Tiškina, Valentina; Capligina, Valentina; Must, Külli; Berzina, Inese; Ranka, Renate; Jokelainen, Pikka

    2016-01-01

    A previously splenectomized dog from Estonia was presented with a sudden lack of appetite and discoloration of the urine. Despite supportive therapy, its condition deteriorated dramatically during 1 day. Severe thrombocytopenia and high numbers of protozoan hemoparasites were evident in blood smears, and the hematocrit dropped from 46 to 33 %. The dog was euthanized before specific antibabesial treatment was initiated. Blood samples from the dog and from two other dogs in the same household tested positive for Babesia using molecular methods, and the sequences of partial 18S rRNA gene confirmed the causative species as Babesia canis canis. The risk of severe, rapidly progressing babesiosis in splenectomized dogs merits awareness. PMID:26810086

  16. Prevalence of Theileria and Babesia species in Tunisian sheep.

    PubMed

    Rjeibi, Mohamed R; Darghouth, Mohamed A; Gharbi, Mohamed

    2016-01-01

    In this study, the prevalence of Theileria and Babesia species in sheep was assessed with Giemsastained blood smear examination and polymerase chain reaction to identify the different piroplasms in 270 sheep from three Tunisian bioclimatic zones (north, centre, and south). The overall infection prevalence by Babesia spp. and Theileria spp. in Giemsa-stained blood smears was 2.9% (8/270) and 4.8% (13/270) respectively. The molecular results showed that sheep were more often infected by Theileria ovis than Babesia ovis with an overall prevalence of 16.3% (44/270) and 7.8% (21/270) respectively (p = 0.01). The molecular prevalence by Babesia ovis was significantly higher in females than in males (p < 0.05). According to localities B. ovis was found exclusively in sheep from the centre of Tunisia (Kairouan) whereas Theileria ovis was found in all regions. Infections with T. ovis and B. ovis were confirmed by sequencing. The sequence of T. ovis in this study (accession numbers KM924442) falls into the same clade as T. ovis deposited in GenBank. The T. ovis amplicons (KM924442) showed 99%-100% identities with GenBank sequences. Moreover, comparison of the partial sequences of 18S rRNA gene of B. ovis described in this study (KP670199) revealed 99.4% similarity with B. ovis recently reported in northern Tunisia from sheep and goats. Three nucleotides were different at positions 73 (A/T), 417 (A/T), and 420 (G/T). It also had 99% identity with B. ovis from Spain, Turkey and Iraq. The results suggest a high T. ovis prevalence in Tunisia with a decreasing north-south gradient. This could be correlated to the vector tick distribution. PMID:27247070

  17. The zoonotic importance of Mycobacterium bovis.

    PubMed

    Moda, G; Daborn, C J; Grange, J M; Cosivi, O

    1996-04-01

    The zoonotic importance of Mycobacterium bovis has been the subject of renewed interest in the wake of the increasing incidence of tuberculosis in the human population. This paper considers some of the conditions under which transmission of M. bovis from animals to humans occurs and reviews current information on the global distribution of the disease. The paper highlights the particular threat posed by this zoonotic disease in developing countries and lists the veterinary and human public health measures that need to be adopted if the disease is to be contained. The association of tuberculosis with malnutrition and poverty has long been recognized and the need to address these basic issues are as crucial as specific measures against the disease itself.

  18. Mycobacterium bovis in coyotes from Michigan.

    PubMed

    Bruning-Fann, C S; Schmitt, S M; Fitzgerald, S D; Payeur, J B; Whipple, D L; Cooley, T M; Carlson, T; Friedrich, P

    1998-07-01

    During a survey for tuberculosis in wild carnivores and omnivores, Mycobacterium bovis was cultured from pooled lymph nodes of three adult female coyotes (Canis latrans) harvested by hunters in Michigan (USA). No gross or histologic lesions suggestive of tuberculosis were seen in these animals. One coyote was taken from Montmorency county and two coyotes from Alcona county located in the north-eastern portion of Michigan's Lower Peninsula where free-ranging white-tailed deer (Odocoileus virginianus) have been found infected with bovine tuberculosis. It is thought that these coyotes became infected with M. bovis through the consumption of tuberculous deer. Other species included in the survey were the opossum (Didelphis virginiana), raccoon (Procyon lotor), red fox (Vulpes vulpes), bobcat (Felis rufus), and badger (Taxidea taxus).

  19. Severe Babesia microti Infection in an Immunocompetent Host in Pennsylvania.

    PubMed

    Genda, Jeffrey; Negron, Elizabeth A; Lotfipour, Mona; Balabhadra, Samyuktha; Desai, Diana S; Craft, David W; Katzman, Michael

    2016-01-01

    Babesiosis, due to infection by a tick-borne protozoan (predominantly Babesia microti in North America), is an emerging health risk that is expanding into new areas and may be unfamiliar to clinicians in locations not previously considered endemic. Manifestations of infection can range from asymptomatic to life threatening, with severe disease more likely in those who have had a splenectomy, are immunocompromised, have chronic medical conditions, or are over 50 years of age. In this article, we describe an elderly but otherwise healthy man from an area not generally considered endemic for babesiosis who presented with severe hemolysis, acute renal failure, and high-level Babesia microti parasitemia; serological results suggestive of possible coinfection by Borrelia burgdorferi (the agent of Lyme disease, which is carried by the same tick as is Babesia microti) also was found. This report highlights that severe babesiosis can occur in an apparently normal host and underscores the continued geographic expansion of this pathogen and the need for early recognition and therapy. PMID:27656660

  20. Babesias of red deer (Cervus elaphus) in Ireland

    PubMed Central

    2011-01-01

    Blood samples were obtained from 38 wild red deer (Cervus elaphus) at two sites in Ireland and subjected to PCR analysis of the 18S rRNA gene followed by sequencing. Two fragments of the 18S rRNA gene were generated by two different PCR protocols and subsequent sequencing suggested that at least six of the deer were infected by a babesia that, in those loci, is indistinguishable from Babesia divergens, an important tick-borne pathogen of cattle and of zoonotic significance. Additionally, a B. odocoilei-like parasite was detected in three samples and a babesia that did not match any sequences in the GenBank database was found in five samples. Neither B. capreoli nor B. venatorum (EU1) were found. There have been several reports of B. divergens occurring in deer species, including red deer, roe deer (Capreolus capreolus) and reindeer (Rangifer tarandus). However, in view of recent re-sequencing of bovine-origin samples deposited previously in GenBank, it is unlikely that any of these sequences from deer are B. divergens. The present study describes the only deer piroplasm detected so far that shows complete identity with B. divergens, in just over half of the 18S rRNA gene. The entire gene of this deer parasite should be analysed and transmission experiments undertaken before the infectivity of B. divergens for red deer can be confirmed. PMID:21314977

  1. Severe Babesia microti Infection in an Immunocompetent Host in Pennsylvania

    PubMed Central

    Genda, Jeffrey; Negron, Elizabeth A.; Lotfipour, Mona; Balabhadra, Samyuktha; Desai, Diana S.; Craft, David W.; Katzman, Michael

    2016-01-01

    Babesiosis, due to infection by a tick-borne protozoan (predominantly Babesia microti in North America), is an emerging health risk that is expanding into new areas and may be unfamiliar to clinicians in locations not previously considered endemic. Manifestations of infection can range from asymptomatic to life threatening, with severe disease more likely in those who have had a splenectomy, are immunocompromised, have chronic medical conditions, or are over 50 years of age. In this article, we describe an elderly but otherwise healthy man from an area not generally considered endemic for babesiosis who presented with severe hemolysis, acute renal failure, and high-level Babesia microti parasitemia; serological results suggestive of possible coinfection by Borrelia burgdorferi (the agent of Lyme disease, which is carried by the same tick as is Babesia microti) also was found. This report highlights that severe babesiosis can occur in an apparently normal host and underscores the continued geographic expansion of this pathogen and the need for early recognition and therapy.

  2. Severe Babesia microti Infection in an Immunocompetent Host in Pennsylvania

    PubMed Central

    Genda, Jeffrey; Negron, Elizabeth A.; Lotfipour, Mona; Balabhadra, Samyuktha; Desai, Diana S.; Craft, David W.; Katzman, Michael

    2016-01-01

    Babesiosis, due to infection by a tick-borne protozoan (predominantly Babesia microti in North America), is an emerging health risk that is expanding into new areas and may be unfamiliar to clinicians in locations not previously considered endemic. Manifestations of infection can range from asymptomatic to life threatening, with severe disease more likely in those who have had a splenectomy, are immunocompromised, have chronic medical conditions, or are over 50 years of age. In this article, we describe an elderly but otherwise healthy man from an area not generally considered endemic for babesiosis who presented with severe hemolysis, acute renal failure, and high-level Babesia microti parasitemia; serological results suggestive of possible coinfection by Borrelia burgdorferi (the agent of Lyme disease, which is carried by the same tick as is Babesia microti) also was found. This report highlights that severe babesiosis can occur in an apparently normal host and underscores the continued geographic expansion of this pathogen and the need for early recognition and therapy. PMID:27656660

  3. The first detection of species of Babesia Starcovici, 1893 in moose, Alces alces (Linnaeus), in Norway.

    PubMed

    Puraite, Irma; Rosef, Olav; Radzijevskaja, Jana; Lipatova, Indre; Paulauskas, Algimantas

    2016-04-01

    Babesiosis is an emerging zoonotic disease and various wildlife species are reservoir hosts for zoonotic species of Babesia Starcovici, 1893. The objective of the present study was to investigate the presence and prevalence of Babesia spp. in moose Alces alces (Linnaeus) in two regions of Norway. A total of 99 spleen samples were collected from animals of various ages from an area with the occurrence of the tick Ixodes ricinus (Linnaeus, 1758), and from an area where the ticks are known to be absent. Infection was detected by the amplification of different regions of the 18S rRNA gene by using two different PCR primer sets specific of Babesia. Babesia spp. were found in the spleen samples of four moose. All Babesia-infected animals were from an area where ticks occur, with an infection rate of 6% (4 of 70). Babesia-positive samples were obtained from a five-month old moose calf and three adults. Two Babesia species, Babesia capreoli (Enigk et Friedhoff, 1962) and a B. odocoilei-like, were identified. Co-infection with Anaplasma phagocytophilum was obtained in two animals. This is the first report of the occurrence of B. capreoli and B. odocoilei-like species in moose.

  4. Babesia ugwidiensis, a new species of avian piroplasm from Phalacrocoracidae in South Africa.

    PubMed

    Peirce, M A; Parsons, N J

    2012-11-01

    A new species of haematozoa, Babesia ugwidiensis sp. nov. from a cormorant is described. This is the first species of piroplasm to be recorded from the Phalacrocoracidae and the relationship of this parasite to other Babesia spp. from marine hosts is discussed. PMID:23193522

  5. Babesia lengau sp. nov., a novel Babesia species in cheetah (Acinonyx jubatus, Schreber, 1775) populations in South Africa.

    PubMed

    Bosman, Anna-Mari; Oosthuizen, Marinda C; Peirce, Michael A; Venter, Estelle H; Penzhorn, Barend L

    2010-08-01

    In a previous paper, we reported on a large number of cheetah blood specimens that gave positive signals only for Babesia and/or Theileria genus-specific probes on the reverse line blot (RLB) assay, indicating the presence of a novel species or variant of an existing species. Some of these specimens were investigated further by microscopic, serological, sequencing, and phylogenetic analyses. The near-full-length 18S rRNA genes of 13 samples, as well as the second internal transcribed spacer (ITS2) region, were amplified, cloned, and sequenced. A species-specific RLB probe, designed to target the hypervariable V4 region of the 18S rRNA gene for detection of the novel Babesia sp., was used to screen an additional 137 cheetah blood specimens for the presence of the species. The prevalence of infection was 28.5%. Here we describe the morphology and phylogenetic relationships of the novel species, which we have named Babesia lengau sp. nov.

  6. Babesia spp. Identified by PCR in Ticks Collected from Domestic and Wild Ruminants in Southern Switzerland

    PubMed Central

    Hilpertshauser, Heidi; Deplazes, Peter; Schnyder, Manuela; Gern, Lise; Mathis, Alexander

    2006-01-01

    Concurrent infections with vector-borne pathogens affected a cattle herd in Switzerland, and one of the pathogens was identified as Babesia bigemina, which had never been observed in this country before. Therefore, a survey of the occurrence of ruminant Babesia spp. and their tick vectors in Switzerland was conducted. A total of 2,017 ticks were collected from sheep, goats, cattle, and wild ruminants (deer, roe deer, and chamois) in southern parts of Switzerland and identified morphologically. The vast majority of the ticks (99.2%) were Ixodes ricinus, but 14 ticks from sheep and goats were identified as Dermacentor marginatus and two ticks from wild ruminants were identified as Hemaphysalis punctata. PCR analyses of 700 ticks revealed the presence of Babesia divergens (n = 6), Babesia sp. genotype EU1 (n = 14), and B. major (n = 2), whose suggested occurrence was confirmed in this study by molecular analysis, and the presence of novel Babesia sp. genotype CH1 (n = 4), which is closely related to B. odocoilei and to Babesia sp. genotype RD61 reported from North America. The identification of B. divergens and B. major in ticks collected from wild ruminants cast doubt on the postulated strict host specificity of these bovine Babesia species. Furthermore, the zoonotic Babesia sp. genotype EU1 was detected in ticks collected from domestic animals but was obtained predominantly from ticks collected from wild ruminants. More than one tick containing DNA of different Babesia spp. were collected from two red deer. Hence, the role of these game animals as reservoir hosts of Babesia spp. seems to be important but requires further investigation. PMID:17021198

  7. Babesia spp. identified by PCR in ticks collected from domestic and wild ruminants in southern Switzerland.

    PubMed

    Hilpertshauser, Heidi; Deplazes, Peter; Schnyder, Manuela; Gern, Lise; Mathis, Alexander

    2006-10-01

    Concurrent infections with vector-borne pathogens affected a cattle herd in Switzerland, and one of the pathogens was identified as Babesia bigemina, which had never been observed in this country before. Therefore, a survey of the occurrence of ruminant Babesia spp. and their tick vectors in Switzerland was conducted. A total of 2,017 ticks were collected from sheep, goats, cattle, and wild ruminants (deer, roe deer, and chamois) in southern parts of Switzerland and identified morphologically. The vast majority of the ticks (99.2%) were Ixodes ricinus, but 14 ticks from sheep and goats were identified as Dermacentor marginatus and two ticks from wild ruminants were identified as Hemaphysalis punctata. PCR analyses of 700 ticks revealed the presence of Babesia divergens (n = 6), Babesia sp. genotype EU1 (n = 14), and B. major (n = 2), whose suggested occurrence was confirmed in this study by molecular analysis, and the presence of novel Babesia sp. genotype CH1 (n = 4), which is closely related to B. odocoilei and to Babesia sp. genotype RD61 reported from North America. The identification of B. divergens and B. major in ticks collected from wild ruminants cast doubt on the postulated strict host specificity of these bovine Babesia species. Furthermore, the zoonotic Babesia sp. genotype EU1 was detected in ticks collected from domestic animals but was obtained predominantly from ticks collected from wild ruminants. More than one tick containing DNA of different Babesia spp. were collected from two red deer. Hence, the role of these game animals as reservoir hosts of Babesia spp. seems to be important but requires further investigation.

  8. Isolation and characterization of Babesia pecorum sp. nov. from farmed red deer (Cervus elaphus).

    PubMed

    Jouglin, Maggy; Fernández-de-Mera, Isabel G; de la Cotte, Nathalie; Ruiz-Fons, Francisco; Gortázar, Christian; Moreau, Emmanuelle; Bastian, Suzanne; de la Fuente, José; Malandrin, Laurence

    2014-01-01

    The diversity of Babesia species infecting cervids in parts of central and southern Spain was analyzed by collecting blood from farmed red deer (Cervus elaphus). Babesia sp. was isolated in vitro from two red deer herds in Cádiz and Ciudad Real. The number of Babesia sp. carriers differed between the two herds: 36/77 in Cádiz and 1/35 in Ciudad Real. Hyalomma lusitanicum was the most prevalent tick species identified on the Cádiz farm vegetation and on sampled animals, and is therefore a candidate vector. The molecular characteristics of 21 isolates were determined by complete (8 isolates) or partial (13 isolates) 18S rRNA gene sequencing. The sequences were highly similar (over 99.4% identity) and 6 sequence types were identified at the level of one herd only, demonstrating a rather high genetic diversity. They formed a monophyletic clade, and members of the three main sequence types shared a similar morphology and the same erythrocyte susceptibility pattern. This clade also included Babesia sp. Xinjiang isolated from sheep in China and Babesia sp. identified in giraffe in South Africa, with identities higher than 98.3% and statistically relevant phylogenetic support. None of the biological properties analyzed for both Babesia from red deer and Babesia sp. Xinjiang allowed their differentiation (ability to develop in vitro in erythrocytes from cattle and sheep, as well as in erythrocytes from different cervids, unsuccessful infection of calves). We propose the Babesia isolated from red deer as a new species named B. pecorum. Whether Babesia sp. Xinjiang and the Babesia characterized in South Africa belong to the same species is debated. PMID:25155988

  9. Strong conservation of rhoptry-associated-protein-1 (RAP-1) locus organization and sequence among Babesia isolates infecting sheep from China (Babesia motasi-like phylogenetic group).

    PubMed

    Niu, Qingli; Valentin, Charlotte; Bonsergent, Claire; Malandrin, Laurence

    2014-12-01

    Rhoptry-associated-protein 1 (RAP-1) is considered as a potential vaccine candidate due to its involvement in red blood cell invasion by parasites in the genus Babesia. We examined its value as a vaccine candidate by studying RAP-1 conservation in isolates of Babesia sp. BQ1 Ningxian, Babesia sp. Tianzhu and Babesia sp. Hebei, responsible for ovine babesiosis in different regions of China. The rap-1 locus in these isolates has very similar features to those described for Babesia sp. BQ1 Lintan, another Chinese isolate also in the B. motasi-like phylogenetic group, namely the presence of three types of rap-1 genes (rap-1a, rap-1b and rap-1c), multiple conserved rap-1b copies (5) interspaced with more or less variable rap-1a copies (6), and the 3' localization of one rap-1c. The isolates Babesia sp. Tianzhu, Babesia sp. BQ1 Lintan and Ningxian were almost identical (average nucleotide identity of 99.9%) over a putative locus of about 31 Kb, including the intergenic regions. Babesia sp. Hebei showed a similar locus organization but differed in the rap-1 locus sequence, for each gene and intergenic region, with an average nucleotide identity of 78%. Our results are in agreement with 18S rDNA phylogenetic studies performed on these isolates. However, in extremely closely related isolates the rap-1 locus seems more conserved (99.9%) than the 18S rDNA (98.7%), whereas in still closely related isolates the identities are much lower (78%) compared with the 18S rDNA (97.7%). The particularities of the rap-1 locus in terms of evolution, phylogeny, diagnosis and vaccine development are discussed.

  10. Selection and application of Streptococcus bovis as a silage inoculant.

    PubMed Central

    Jones, B A; Muck, R E; Ricke, S C

    1991-01-01

    Three strains of Streptococcus bovis, a homolactic bacterium capable of utilizing starch, were evaluated for growth kinetics and ability to decrease the pH of alfalfa silage. A selected strain was evaluated for its competitiveness as an inoculant with Enterococcus faecium, an organism used in inoculants, and for its ability to enhance the effect of a commercial inoculant. Testing was completed over three studies using wilted alfalfa (28 to 34% dry matter) ensiled into laboratory silos. Treatments were control, E. faecium, E. faecium and commercial inoculant, S. bovis, and S. bovis and commercial inoculant. Replicate silos were emptied and analyzed at 0.5, 1, 2, 4, 8, and 40 days for pH, fermentation products, and nitrogen fractions. S. bovis alone lowered the pH quicker and improved silage parameters early in the fermentation compared with E. faecium, the commercial inoculant, and control treatments. When combined with a commercial inoculant, S. bovis lowered pH more quickly than the commercial inoculant alone and E. faecium plus commercial inoculant. At 40 days, S. bovis combination had lower pH and ammonia nitrogen and acetate contents than the E. faecium combination. Starch in the silage was not utilized by S. bovis as had been anticipated. Results indicate that S. bovis was more effective than E. faecium as a silage inoculant and could enhance a commercial inoculant on low-dry-matter alfalfa. PMID:1746960

  11. Mycoplasma bovis: An emerging pathogen of ranched bison

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) is an emerging, primary pathogen of ranched bison (Bison bison) in North America. It causes severe disease among animals in feedlots as well as breeding-age cows and bulls on pasture. Mortality in adult bison is as high as 25 percent, resulting in significant economic l...

  12. Pulmonary Tuberculosis Caused by Mycobacterium bovis in China

    PubMed Central

    Jiang, Guanglu; Wang, Guirong; Chen, Suting; Yu, Xia; Wang, Xiaobo; Zhao, Liping; Ma, Yifeng; Dong, Lingling; Huang, Hairong

    2015-01-01

    The epidemiology of Mycobacterium bovis infection in humans in China is unknown. In this study, pulmonary tuberculosis caused by M. bovis in China was studied. A total of 4069 clinical strains isolated from sputa during the 2007–2009 nationwide surveillance of drug-resistant tuberculosis in China were analyzed. M. bovis was identified by para-nitrobenzoic acid and thiophen-2-carboxylic acid hydrazide growth tests, spoligotyping and multiplex PCR amplification. In addition, a total of 1828 clinical specimens were recruited from Beijing Chest Hospital (Beijing, China) for Löwenstein-Jensen (LJ) culture, both on standard LJ medium and LJ medium containing 4.5 mg/ml(W/V) sodium pyruvate, the latter being the preferred medium for M. bovis growth. The isolates which demonstrated more vigorous on pyruvate containing medium than on standard LJ medium were then identified by multiplex PCR amplification. Only 1 isolate from the nationwide surveillance was confirmed as M. bovis-BCG. The isolate belonged to a predominant spoligotype SB0120 (ST482). In addition, no M. bovis isolate was acquired by the continuous screening step in Beijing Chest Hospital. M. bovis has a negligible contribution to pulmonary tuberculosis in China, so neither laboratory identification nor clinical treatment of M. bovis infection need be considered in routine work. PMID:25736338

  13. Complete Genome Sequence of Mycobacterium bovis Strain BCG-1 (Russia)

    PubMed Central

    Shitikov, Egor A.; Malakhova, Maja V.; Kostryukova, Elena S.; Ilina, Elena N.; Atrasheuskaya, Alena V.; Ignatyev, Georgy M.; Vinokurova, Nataliya V.; Gorbachyov, Vyacheslav Y.

    2016-01-01

    Mycobacterium bovis BCG (Bacille Calmette-Guérin) is a vaccine strain used for protection against tuberculosis. Here, we announce the complete genome sequence of M. bovis strain BCG-1 (Russia). Extensive use of this strain necessitates the study of its genome stability by comparative analysis. PMID:27034492

  14. Streptococcus bovis septicemia and meningitis associated with chronic radiation enterocolitis

    SciTech Connect

    Jadeja, L.; Kantarjian, H.; Bolivar, R.

    1983-12-01

    We describe the first patient with simultaneous S bovis septicemia and meningitis associated with chronic radiation enterocolitis. This case underlines the value of a thorough gastrointestinal evaluation of all patients with S bovis infection, and the need for a neurologic investigation even with minor neurologic manifestations.

  15. Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

    PubMed

    Mol, J P S; Carvalho, T F; Fonseca, A A; Sales, E B; Issa, M A; Rezende, L C; Hodon, M A; Tinoco, H P; Malta, M C C; Pessanha, A T; Pierezan, F; Mota, P M P C; Paixão, T A; Santos, R L

    2016-01-01

    Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection.

  16. Tuberculosis Caused by Mycobacterium bovis in a Capybara (Hydrochoerus hydrochaeris).

    PubMed

    Mol, J P S; Carvalho, T F; Fonseca, A A; Sales, E B; Issa, M A; Rezende, L C; Hodon, M A; Tinoco, H P; Malta, M C C; Pessanha, A T; Pierezan, F; Mota, P M P C; Paixão, T A; Santos, R L

    2016-01-01

    Tuberculosis, associated with Mycobacterium bovis, was diagnosed post mortem in an adult female capybara (Hydrochoerus hydrochaeris), kept at the Pampulha Ecological Park, Belo Horizonte, Brazil, in a large metropolitan area. On post-mortem examination, there were numerous firm white nodules scattered throughout all lobes of both lungs. Tissue samples were collected for histological and microbiological examination. Microscopically, the pulmonary nodules were multifocal to coalescing granulomas and intralesional acid-fast bacilli were evident in Ziehl-Neelsen-stained sections of the lung and spleen. Colonies with morphological features of Mycobacterium spp. were isolated from lung samples and conventional polymerase chain reaction (PCR) with genomic DNA from the isolates was positive for M. bovis; sequencing indicated 100% identity with the region of difference 4 (RD4) of M. bovis. In addition, M. bovis DNA was detected in the lung by quantitative PCR. The finding of M. bovis in a capybara indicates a potential public health risk in a zoological collection. PMID:27363904

  17. Anatomical distribution of Mycobacterium bovis genotypes in experimentally infected white-tailed deer

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium bovis (M. bovis) causes tuberculosis in white-tailed deer (WTD). Natural infection of WTD with M. bovis is most closely mimicked by instilling inoculum into palatine tonsilar crypts. One hundred fifty days after intratonsilar inoculation, M. bovis was cultured from 30 tissues originati...

  18. Susceptibility of raccoons (Procyon lotor) to infection with Mycobacterium bovis.

    PubMed

    Palmer, Mitchell V; Waters, W Ray; Whipple, Diana L

    2002-04-01

    Tuberculosis due to Mycobacterium bovis infection is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of the lower Michigan peninsula (USA). Various wild carnivores and omnivores, including raccoons (Procyon lotor), are infected with M. bovis within the endemic area. To investigate the pathogenesis of tuberculosis in raccoons and the likelihood of M. bovis transmission from infected raccoons to other susceptible hosts, we experimentally inoculated raccoons with single oral doses of M. bovis (ranging from 30 to 1.7 x 10(5) colony forming units [CFU]), five daily oral doses of M. bovis (ranging from 10 to 1 x 10(5) CFU), or a single intravenous (i.v.) dose of 1 x 10(5) CFU of M. bovis, from November 1998 through December 2000. Granulomatous lesions consistent with tuberculosis, or tissue colonization with M. bovis, were seen in one of five raccoons in the single low oral dose group, one of five raccoons in the multiple low oral dose group, two of five raccoons in the multiple medium oral dose group, five of five raccoons in the multiple high oral dose group, and five of five raccoons in the i.v. inoculated group. In oral inoculated raccoons, lesions were most common in the tracheobronchial and mesenteric lymph nodes and lung. Excretion of M. bovis in saliva or nasal secretions was noted in all i.v. inoculated raccoons and two of five multiple low oral dose raccoons. Mycobacterium bovis was not isolated from urine or feces from any experimentally inoculated raccoons. The need for multiple large oral doses to establish infection, and the low number of orally inoculated raccoons that excreted M. bovis in nasal secretions or saliva, suggest that wide-spread tuberculosis among raccoons is unlikely.

  19. Comparisons of serological tests for Babesia in British cattle.

    PubMed

    Bidwell, D E; Turp, P; Joyner, L P; Payne, R C; Purnell, R E

    1978-11-11

    A comparison was made between the microplate enzyme linked immunosorbent assay (ELISA) and the indirect fluorescent antibody (IFA) and complement fixation (CF) tests for the detection of antibodies in the serum of cattle experimentally infected with Babesia divergens and B major. Antibodies were detected using all three tests but they were detected earlier using the CF test. However CF titres were consistently lower than those obtained using the other tests. Although there was little to choose between the IFA and ELISA tests, it was suggested that the ELISA may be preferable since it is less subject to operator error and operator stress, and can be adapted readily to field use.

  20. Severe Mycoplasma bovis outbreak in an Austrian dairy herd.

    PubMed

    Pothmann, Harald; Spergser, Joachim; Elmer, Josef; Prunner, Isabella; Iwersen, Michael; Klein-Jöbstl, Daniela; Drillich, Marc

    2015-11-01

    A conventional dairy farm, housing 19 Austrian Simmental cows, experienced a spontaneous outbreak of a Mycoplasma bovis infection, showing severe clinical signs of respiratory tract disease, clinical mastitis, and tremendous drop in milk production. Despite intensive therapy, 5 cows died within 2 weeks or were euthanized. From the remaining cows, bacteriological culture and polymerase chain reaction revealed M. bovis in 10 of 14 milk samples. Mycoplasma bovis was found in 1 of 5 randomly collected nasal swabs. Autopsy of 1 cow revealed infection of the lungs and the udder with M. bovis. The 13 M. bovis isolates from milk samples, nasal swabs, lungs, and udder were genotyped by multilocus variable number of tandem-repeat analysis, and indicated that described infections were caused by a single M. bovis strain. The virulent M. bovis strain resulted in dramatic economic loss to the farmer. To control the disease, culling of all animals, including heifers and calves, was recommended, and strict hygienic measures were implemented before introducing new animals to the farm. PMID:26450838

  1. A member of the HSP90 family from ovine Babesia in China: molecular characterization, phylogenetic analysis and antigenicity.

    PubMed

    Guan, Guiquan; Liu, Junlong; Liu, Aihong; Li, Youquan; Niu, Qingli; Gao, Jinliang; Luo, Jianxun; Chauvin, Alain; Yin, Hong; Moreau, Emmanuelle

    2015-09-01

    Heat shock protein 90 (HSP90) is a key component of the molecular chaperone complex essential for activating many signalling proteins involved in the development and progression of pathogenic cellular transformation. A Hsp90 gene (BQHsp90) was cloned and characterized from Babesia sp. BQ1 (Lintan), an ovine Babesia isolate belonging to Babesia motasi-like group, by screening a cDNA expression library and performing rapid amplification of cDNA ends. The full-length cDNA of BQHsp90 is 2399 bp with an open reading frame of 2154 bp encoding a predicted 83 kDa polypeptide with 717 amino acid residues. It shows significant homology and similar structural characteristics to Hsp90 of other apicomplex organisms. Phylogenetic analysis, based on the HSP90 amino acid sequences, showed that the Babesia genus is clearly separated from other apicomplexa genera. Five Chinese ovine Babesia isolates were divided into 2 phylogenetic clusters, namely Babesia sp. Xinjiang (previously designated a new species) cluster and B. motasi-like cluster which could be further divided into 2 subclusters (Babesia sp. BQ1 (Lintan)/Babesia sp. Tianzhu and Babesia sp. BQ1 (Ningxian)/Babesia sp. Hebei). Finally, the antigenicity of rBQHSP90 protein from prokaryotic expression was also evaluated using western blot and enzyme-linked immunosorbent assay (ELISA).

  2. Effect of Sex Steroids on Babesia microti Infection in Mice

    PubMed Central

    Sasaki, Mizuki; Fujii, Yoshito; Iwamoto, Maya; Ikadai, Hiromi

    2013-01-01

    Sex-based-differences are known to affect susceptibility to protozoan infections, but their effects on parasitemia and clinical symptoms in Babesia infections remain unclear. We examined the sex-based susceptibility of various mouse strains to Babesia microti Munich strain infection. In all strains, male mice exhibited significantly higher peak parasitemia and more severe anemia than female mice. Testosterone and estradiol-17β treatment caused an increase in parasitemia and aggravation of anemia. Orchidectomized male mice receiving testosterone exhibited smaller splenic macrophage populations three days after infection, smaller B cell populations 10 days after infection, and reduced splenic tumor necrosis factor-α and interferon-γ mRNA expression than mice that did not receive testosterone. Mice receiving estradiol-17β did not exhibit immunosuppressive effects. Thus, a weakened and delayed innate immunity response may lead to acquired immunity failure. The results suggested that testosterone directly affects T or B cells, leading to delayed acquired immunity, dramatically increased parasitemia, and severe anemia. PMID:23249689

  3. Delayed-type hypersensitivity to Babesia microti-infected erythrocytes in mice

    SciTech Connect

    Ruebush, M.J.; Troutman, E.H.; Kennedy, D.A.

    1986-04-01

    Strong delayed-type hypersensitivity (DTH) to Babesia microti was elicited when intraerythrocytic parasites (IEP) were inoculated subcutaneously into the flank of normal mice 6 to 14 days before challenge in the ipsilateral footpad with 10(8) IEP. Intraperitoneal or intravenous administration of antigen did not sensitize mice for DTH. When challenge was given 21 days after immunization, the response was approximately half of the maximum and then rose again slowly over the next 3 weeks to levels that were not significantly different from those maximal values. The response was classified as a true DTH reaction on the basis of kinetics, histology, and the transfer of responsiveness with immune T lymphocytes of the Ly 1+ phenotype, but not with serum. The reaction was specific for IEP since control groups given two injections of red blood cells from uninfected syngeneic mice (NRBC) or one injection of NRBC or sheep red blood cells (SRBC) and one of IEP never developed significant footpad swelling. Freed parasites obtained by osmotic rupture, density gradient sedimentation, and lethally irradiated IEP were also effective for elicitation of DTH. Anti-IEP DTH was expressed in a dose-dependent fashion with 10(6), 10(7), or 10(8) parasites sufficing for immunizing inoculum as long as 10(8) parasites were used as the challenge dose. Mice immunized and challenged with 10(8) lethally irradiated IEP (60 krad, 60Co), were protected against subsequent intraperitoneal challenge with 10(8) viable IEP. If mice were infected intraperitoneally with 10(8) IEP at any time between 21 days before immunization to 2 hr after challenge, their ability to respond to immunization and challenge was profoundly depressed. Development of a strong anti-parasite DTH response can occur in parallel with resistance to infection, but is not a rapid sequela of bloodborne infection.

  4. Multiple co-infections of rodents with hantaviruses, Leptospira, and Babesia in Croatia.

    PubMed

    Tadin, Ante; Turk, Nenad; Korva, Miša; Margaletić, Josip; Beck, Relja; Vucelja, Marko; Habuš, Josipa; Svoboda, Petra; Zupanc, Tatjana Avšič; Henttonen, Heikki; Markotić, Alemka

    2012-05-01

    Hantaviruses, Leptospira spp., and Babesia spp. are rodent-borne pathogens present worldwide. We studied multiple co-infections of small rodents in Croatia with all three pathogens. Twenty-eight Apodemus flavicollis and 16 Myodes glareolus were tested for the presence of hantavirus RNA by real-time RT-PCR, Leptospira strains by renoculture method and Babesia DNA by PCR. Anti-hantavirus antibodies and anti-Leptospira antibodies were detected by serological methods. Very high infection rates with each pathogen were found in A. flavicollis: 20 of 28 rodents (71%) were infected with Dobrava virus, 13 rodents (46%) were infected with Leptospira, and 5 rodents (18%) were infected with Babesia. Multiple co-infections with all three pathogens were found in 3 of 28 (11%) A. flavicollis animals, suggesting that the same rodent host can be infected with several pathogens at the same time. Dual infections with both hantaviruses and Leptospira were found in 7 of 44 rodents (16%), with hantaviruses and Babesia in 2 rodents (5%), and double infection with both Leptospira and Babesia were found in 1 rodent (2%). Since hantaviruses, Leptospira, and Babesia have similar geographical distributions, it is to be expected that in other parts of the world multiple co-infections, representing a serious threat to public health, can be found.

  5. Occurrence of Babesia species in captive reindeer (Rangifer tarandus) in Germany.

    PubMed

    Wiegmann, Lisa; Silaghi, Cornelia; Obiegala, Anna; Karnath, Carolin; Langer, Sandra; Ternes, Kerstin; Kämmerling, Jens; Osmann, Christine; Pfeffer, Martin

    2015-06-30

    Two cases of acute babesiosis in captive reindeer (Rangifer tarandus) in two German zoos in 2009 and 2012 triggered this study to investigate the occurrence and species diversity of Babesia parasites infecting reindeer in different zoos and deer parks in Germany. Between June and December 2013, blood samples were taken from 123 clinically inapparent reindeer from 16 different facilities. Samples were tested for the presence of Babesia species DNA by conventional PCR and sequence analysis of part of the 18S rRNA gene. Also, Giemsa-stained smears of reindeer blood samples were examined for parasitaemia by light microscopy. The overall PCR-prevalence in blood samples was 23.6% (n=29). Comparison of sequenced amplicons with GenBank entries possibly revealed up to five different Babesia species: B. venatorum (n=19), B. capreoli (n=2) and B. capreoli-like (n=4), B. odocoilei-like (n=2) and B. divergens (n=1), while one sample turned out to be a Theileria sp. Out of the 16 facilities in the study, 12 housed at least one positive animal. In Giemsa-stained blood smears, intra-erythrocytic Babesia parasites were detected in samples of three reindeer from three locations. The high prevalence of Babesia infections implicates babesiosis to be a relevant infectious disease threat for captive reindeer in Germany. Consequently, reindeer with clinical signs compatible to those of acute babesiosis should either be tested for the presence of Babesia spp. DNA or blood smears should be examined for parasitaemia.

  6. Occurrence of Babesia species in captive reindeer (Rangifer tarandus) in Germany.

    PubMed

    Wiegmann, Lisa; Silaghi, Cornelia; Obiegala, Anna; Karnath, Carolin; Langer, Sandra; Ternes, Kerstin; Kämmerling, Jens; Osmann, Christine; Pfeffer, Martin

    2015-06-30

    Two cases of acute babesiosis in captive reindeer (Rangifer tarandus) in two German zoos in 2009 and 2012 triggered this study to investigate the occurrence and species diversity of Babesia parasites infecting reindeer in different zoos and deer parks in Germany. Between June and December 2013, blood samples were taken from 123 clinically inapparent reindeer from 16 different facilities. Samples were tested for the presence of Babesia species DNA by conventional PCR and sequence analysis of part of the 18S rRNA gene. Also, Giemsa-stained smears of reindeer blood samples were examined for parasitaemia by light microscopy. The overall PCR-prevalence in blood samples was 23.6% (n=29). Comparison of sequenced amplicons with GenBank entries possibly revealed up to five different Babesia species: B. venatorum (n=19), B. capreoli (n=2) and B. capreoli-like (n=4), B. odocoilei-like (n=2) and B. divergens (n=1), while one sample turned out to be a Theileria sp. Out of the 16 facilities in the study, 12 housed at least one positive animal. In Giemsa-stained blood smears, intra-erythrocytic Babesia parasites were detected in samples of three reindeer from three locations. The high prevalence of Babesia infections implicates babesiosis to be a relevant infectious disease threat for captive reindeer in Germany. Consequently, reindeer with clinical signs compatible to those of acute babesiosis should either be tested for the presence of Babesia spp. DNA or blood smears should be examined for parasitaemia. PMID:25986326

  7. Experimental inoculation of wild turkeys (Meleagris gallopavo) with Mycobacterium bovis.

    PubMed

    Clarke, K R; Fitzgerald, S D; Hattey, J A; Bolin, C A; Berry, D E; Church, S V; Reed, W M

    2006-03-01

    Although avian species are known to be susceptible to infection with Mycobacterium spp. organisms, much remains unknown about the susceptibility of birds to infection with M. bovis. The objective of this current study was to determine if wild turkeys (Meleagris gallopavo) can be infected with M. bovis when inoculated by the oral or intratracheal route. Six turkeys were orally inoculated and another six were inoculated via the trachea with a high dose of M. bovis, 1 x 10(5) CFU/ml. Six turkeys were sham-inoculated controls. Two turkeys from each treatment group were sacrificed on days 30, 60, and 90 postinoculation. There were no gross or microscopic lesions consistent with mycobacteriosis in the 23 inoculated turkeys over the 90-day duration of this study. Fecal cultures were also consistently negative for M. bovis when sampled before inoculation and on days 1, 30, and 60 postinoculation. Two intratracheally inoculated turkeys were positive for M. bovis in visceral tissues at 30 days postinoculation. However, this finding was only indicative of passive persistence of mycobacteria in the tissues and not of infection, as there were no attendant lesions or clinical compromise to support infection. Thus, it can be concluded that young wild turkeys are resistant to infection with M. bovis and, therefore, pose minimal threat as reservoir or spillover hosts for this organism.

  8. Tuberculosis from Mycobacterium bovis in Binational Communities, United States

    PubMed Central

    Moore, Marisa; Moser, Kathleen S.; Brodine, Stephanie K.; Strathdee, Steffanie A.

    2008-01-01

    The epidemiology of tuberculosis (TB) in the United States is changing as the incidence of disease becomes more concentrated in foreign-born persons. Mycobacterium bovis appears to be contributing substantially to the TB incidence in some binational communities with ties to Mexico. We conducted a retrospective analysis of TB case surveillance data from the San Diego, California, region from 1994 through 2005 to estimate incidence trends, identify correlates of M. bovis disease, and evaluate risk factors for deaths during treatment. M. bovis accounted for 45% (62/138) of all culture-positive TB cases in children (<15 years of age) and 6% (203/3,153) of adult cases. M. bovis incidence increased significantly (p = 0.002) while M. tuberculosis incidence declined (p<0.001). Almost all M. bovis cases from 2001 through 2005 were in persons of Hispanic ethnicity. Persons with M. bovis were 2.55× (p = 0.01) as likely to die during treatment than those with M. tuberculosis. PMID:18507901

  9. Prevalence and diversity of Babesia, Hepatozoon, Ehrlichia, and Bartonella in wild and domestic carnivores from Zambia, Africa.

    PubMed

    Williams, Brianna M; Berentsen, Are; Shock, Barbara C; Teixiera, Maria; Dunbar, Michael R; Becker, Matthew S; Yabsley, Michael J

    2014-03-01

    A molecular survey was conducted for several hemoparasites of domestic dogs and three species of wild carnivores from two sites in Zambia. Three Babesia spp. were detected including Babesia felis and Babesia leo in lions (Panthera leo) and a Babesia sp. (similar to Babesia lengau) in spotted hyenas (Crocuta crocuta) and a single lion. All wild dogs (Lycaon pictus) and domestic dogs were negative for Babesia. High prevalences for Hepatozoon were noted in all three wild carnivores (38-61%) and in domestic dogs (13%). Significantly higher prevalences were noted in hyenas and wild dogs compared with domestic dogs and lions. All carnivores were PCR negative for Ehrlichia canis, Ehrlichia ewingii, and Bartonella spp. Overall, high prevalences and diversity of Babesia and Hepatozoon were noted in wild carnivores from Zambia. This study is the first molecular characterization of Babesia from any hyena species and is the first report of a Babesia sp. closely related to B. lengau, a parasite previously only reported from cheetahs (Acinonyx jubatus), in lions and hyenas. Although usually benign in wild carnivores, these hemoparasites can be pathogenic under certain circumstances. Importantly, data on vectors for these parasites are lacking, so studies are needed to identify vectors as well as determine transmission routes, infection dynamics, and host specificity of these hemoparasites in wildlife in Africa and also the risk of transmission between domestic animals and wildlife. PMID:24363181

  10. Prevalence and diversity of Babesia, Hepatozoon, Ehrlichia, and Bartonella in wild and domestic carnivores from Zambia, Africa.

    PubMed

    Williams, Brianna M; Berentsen, Are; Shock, Barbara C; Teixiera, Maria; Dunbar, Michael R; Becker, Matthew S; Yabsley, Michael J

    2014-03-01

    A molecular survey was conducted for several hemoparasites of domestic dogs and three species of wild carnivores from two sites in Zambia. Three Babesia spp. were detected including Babesia felis and Babesia leo in lions (Panthera leo) and a Babesia sp. (similar to Babesia lengau) in spotted hyenas (Crocuta crocuta) and a single lion. All wild dogs (Lycaon pictus) and domestic dogs were negative for Babesia. High prevalences for Hepatozoon were noted in all three wild carnivores (38-61%) and in domestic dogs (13%). Significantly higher prevalences were noted in hyenas and wild dogs compared with domestic dogs and lions. All carnivores were PCR negative for Ehrlichia canis, Ehrlichia ewingii, and Bartonella spp. Overall, high prevalences and diversity of Babesia and Hepatozoon were noted in wild carnivores from Zambia. This study is the first molecular characterization of Babesia from any hyena species and is the first report of a Babesia sp. closely related to B. lengau, a parasite previously only reported from cheetahs (Acinonyx jubatus), in lions and hyenas. Although usually benign in wild carnivores, these hemoparasites can be pathogenic under certain circumstances. Importantly, data on vectors for these parasites are lacking, so studies are needed to identify vectors as well as determine transmission routes, infection dynamics, and host specificity of these hemoparasites in wildlife in Africa and also the risk of transmission between domestic animals and wildlife.

  11. Novel type of linear mitochondrial genomes with dual flip-flop inversion system in apicomplexan parasites, Babesia microti and Babesia rodhaini

    PubMed Central

    2012-01-01

    Background Mitochondrial (mt) genomes vary considerably in size, structure and gene content. The mt genomes of the phylum Apicomplexa, which includes important human pathogens such as the malaria parasite Plasmodium, also show marked diversity of structure. Plasmodium has a concatenated linear mt genome of the smallest size (6-kb); Babesia and Theileria have a linear monomeric mt genome (6.5-kb to 8.2-kb) with terminal inverted repeats; Eimeria, which is distantly related to Plasmodium and Babesia/Theileria, possesses a mt genome (6.2-kb) with a concatemeric form similar to that of Plasmodium; Cryptosporidium, the earliest branching lineage within the phylum Apicomplexa, has no mt genome. We are interested in the evolutionary origin of linear mt genomes of Babesia/Theileria, and have investigated mt genome structures in members of archaeopiroplasmid, a lineage branched off earlier from Babesia/Theileria. Results The complete mt genomes of archaeopiroplasmid parasites, Babesia microti and Babesia rodhaini, were sequenced. The mt genomes of B. microti (11.1-kb) and B. rodhaini (6.9-kb) possess two pairs of unique inverted repeats, IR-A and IR-B. Flip-flop inversions between two IR-As and between two IR-Bs appear to generate four distinct genome structures that are present at an equi-molar ratio. An individual parasite contained multiple mt genome structures, with 20 copies and 2 – 3 copies per haploid nuclear genome in B. microti and B. rodhaini, respectively. Conclusion We found a novel linear monomeric mt genome structure of B. microti and B. rhodhaini equipped with dual flip-flop inversion system, by which four distinct genome structures are readily generated. To our knowledge, this study is the first to report the presence of two pairs of distinct IR sequences within a monomeric linear mt genome. The present finding provides insight into further understanding of evolution of mt genome structure. PMID:23151128

  12. Reservoir competence of wildlife host species for Babesia microti.

    PubMed

    Hersh, Michelle H; Tibbetts, Michael; Strauss, Mia; Ostfeld, Richard S; Keesing, Felicia

    2012-12-01

    Human babesiosis is an increasing health concern in the northeastern United States, where the causal agent, Babesia microti, is spread through the bite of infected Ixodes scapularis ticks. We sampled 10 mammal and 4 bird species within a vertebrate host community in southeastern New York to quantify reservoir competence (mean percentage of ticks infected by an individual host) using real-time PCR. We found reservoir competence levels >17% in white-footed mice (Peromyscus leucopus), raccoons (Procyon lotor), short-tailed shrews (Blarina brevicauda), and eastern chipmunks (Tamias striatus), and <6% but >0% in all other species, including all 4 bird species. Data on the relative contributions of multiple host species to tick infection with B. microti and level of genetic differentiation between B. microti strains transmitted by different hosts will help advance understanding of the spread of human babesiosis. PMID:23171673

  13. Reservoir Competence of Wildlife Host Species for Babesia microti

    PubMed Central

    Tibbetts, Michael; Strauss, Mia; Ostfeld, Richard S.; Keesing, Felicia

    2012-01-01

    Human babesiosis is an increasing health concern in the northeastern United States, where the causal agent, Babesia microti, is spread through the bite of infected Ixodes scapularis ticks. We sampled 10 mammal and 4 bird species within a vertebrate host community in southeastern New York to quantify reservoir competence (mean percentage of ticks infected by an individual host) using real-time PCR. We found reservoir competence levels >17% in white-footed mice (Peromyscus leucopus), raccoons (Procyon lotor), short-tailed shrews (Blarina brevicauda), and eastern chipmunks (Tamias striatus), and <6% but >0% in all other species, including all 4 bird species. Data on the relative contributions of multiple host species to tick infection with B. microti and level of genetic differentiation between B. microti strains transmitted by different hosts will help advance understanding of the spread of human babesiosis. PMID:23171673

  14. Necrotic pharyngitis associated with Mycoplasma bovis infections in American bison (Bison bison)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) has recently emerged as a significant and costly infectious disease problem in bison, generally presenting as severe, caseonecrotic pneumonia. Here we describe three diagnostic cases in which M. bovis is strongly implicated as a causative agent of necrotic pharyngitis. ...

  15. Mycobacterium bovis infection in humans and cats in same household, Texas, USA, 2012.

    PubMed

    Ramdas, Kira E F; Lyashchenko, Konstantin P; Greenwald, Rena; Robbe-Austerman, Suelee; McManis, Cynthia; Waters, W Ray

    2015-03-01

    Mycobacterium bovis infection of cats is exceedingly rare in regions where bovine tuberculosis is not endemic. We describe the diagnosis and clinical management of pulmonary M. bovis infection in 2 indoor-housed cats and their association with at least 1 M. bovis-infected human in Texas, USA, in September 2012.

  16. Detection of Anaplasma phagocytophilum and Babesia odocoilei DNA in Ixodes scapularis (Acari: Ixodidae) collected in Indiana.

    PubMed

    Steiner, Fresia E; Pinger, Robert R; Vann, Carolyn N; Abley, Melanie J; Sullivan, Bridget; Grindle, Nate; Clay, Keith; Fuqua, Clay

    2006-03-01

    The blacklegged tick, Ixodes scapularis Say, first reported in Indiana in 1987, has now been detected in more than half of Indiana's counties. The first case of human granulocytic ehrlichiosis (human anaplasmosis) in Indiana was reported in 2002. We now report the detection of Anaplasma phagocytophilum and Babesia odocoilei (Emerson and Wright 1968) in I. scapularis ticks collected in northern Indiana. Using polymerase chain reaction analysis, 41 of 193 adult ticks (21.2%) collected from deer were positive for A. phagocytophylum, and 22 (11.4%) were positive for Babesia sp. Restriction fragment analysis of 12, and sequencing of another five of the amplified products identified these parasites as B. odocoilei. Five ticks (2.6%) were coinfected. Eight of 68 questing adult ticks (11.8%) were positive for A. phagocytophilum; seven (10.3%) were positive for Babesia sp. Six of the latter seven positive samples were determined to be B. odocoilei by restriction fragment analysis and sequencing of two samples. None of 39 pools of nymphs was positive for Babesia sp. Three of 15 ticks (20%) collected from a dog were positive for A. phagocytophilum and three ticks (20%) were positive for Babesia sp. One was confirmed as B. odocoilei. One tick was coinfected. This is the first report of the presence of these two agents in ticks in Indiana.

  17. Babesia and its hosts: adaptation to long-lasting interactions as a way to achieve efficient transmission

    PubMed Central

    Chauvin, Alain; Moreau, Emmanuelle; Bonnet, Sarah; Plantard, Olivier; Malandrin, Laurence

    2009-01-01

    Babesia, the causal agent of babesiosis, are tick-borne apicomplexan protozoa. True babesiae (Babesia genus sensu stricto) are biologically characterized by direct development in erythrocytes and by transovarial transmission in the tick. A large number of true Babesia species have been described in various vertebrate and tick hosts. This review presents the genus then discusses specific adaptations of Babesia spp. to their hosts to achieve efficient transmission. The main adaptations lead to long-lasting interactions which result in the induction of two reservoirs: in the vertebrate host during low long-term parasitemia and throughout the life cycle of the tick host as a result of transovarial and transstadial transmission. The molecular bases of these adaptations in vertebrate hosts are partially known but few of the tick-host interaction mechanisms have been elucidated. PMID:19379662

  18. Human multidrug-resistant Mycobacterium bovis infection in Mexico.

    PubMed

    Vazquez-Chacon, Carlos A; Martínez-Guarneros, Armando; Couvin, David; González-Y-Merchand, Jorge A; Rivera-Gutierrez, Sandra; Escobar-Gutierrez, Alejandro; De-la-Cruz López, Juan J; Gomez-Bustamante, Adriana; Gonzalez-Macal, Gabriela A; Gonçalves Rossi, Livia Maria; Muñiz-Salazar, Raquel; Rastogi, Nalin; Vaughan, Gilberto

    2015-12-01

    Here, we describe the molecular characterization of six human Mycobacterium bovis clinical isolates, including three multidrug resistant (MDR) strains, collected in Mexico through the National Survey on Tuberculosis Drug Resistance (ENTB-2008), a nationally representative survey conducted during 2008-2009 in nine states with a stratified cluster sampling design. The genetic background of bovine M. bovis strains identified in three different states of Mexico was studied in parallel to assess molecular relatedness of bovine and human strains. Additionally, resistance to first and second line anti-tuberculosis (TB) drugs and molecular identification of mutations conferring drug resistance was also performed. All strains were characterized by spoligotyping and 24-loci MIRU-VNTRs, and analyzed using the SITVIT2 (n = 112,000 strains) and SITVITBovis (n = 25,000 strains) proprietary databases of Institut Pasteur de la Guadeloupe. Furthermore, data from this study (n = 55 isolates), were also compared with genotypes recorded for M. bovis from USA (n = 203), Argentina (n = 726), as well as other isolates from Mexico (independent from the present study; n = 147), to determine any evidence for genetic relatedness between circulating M. bovis strains. The results showed that all human M. bovis cases were not genetically related between them or to any bovine strain. Interestingly, a high degree of genetic variability was observed among bovine strains. Several autochthonous and presumably imported strains were identified. The emergence of drug-resistant M. bovis is an important public health problem that jeopardizes the success of TB control programs in the region.

  19. A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan.

    PubMed

    Kubo, Shotaro; Tateno, Morihiro; Ichikawa, Yasuaki; Endo, Yasuyuki

    2015-10-01

    Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan.

  20. A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan.

    PubMed

    Kubo, Shotaro; Tateno, Morihiro; Ichikawa, Yasuaki; Endo, Yasuyuki

    2015-10-01

    Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan. PMID:25947226

  1. Molecular characterization of Babesia and Cytauxzoon species in wild South-African meerkats.

    PubMed

    Leclaire, Sarah; Menard, Sandie; Berry, Antoine

    2015-04-01

    Piroplasms, including Babesia, Cytauxzoon and Theileria species, frequently infect domestic and wild mammals. At present, there is no information on the occurrence and molecular identity of these tick-borne blood parasites in the meerkat, one of South Africa's most endearing wildlife celebrities. Meerkats live in territorial groups, which may occur on ranchland in close proximity to humans, pets and livestock. Blood collected from 46 healthy meerkats living in the South-African Kalahari desert was screened by microscopy and molecular methods, using PCR and DNA sequencing of 18S rRNA and ITS1 genes. We found that meerkats were infected by 2 species: one species related to Babesia sp. and one species related to Cytauxzoon sp. Ninety one percent of the meerkats were infected by the Cytauxzoon and/or the Babesia species. Co-infection occurred in 46% of meerkats. The pathogenicity and vectors of these two piroplasm species remains to be determined.

  2. A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan

    PubMed Central

    KUBO, Shotaro; TATENO, Morihiro; ICHIKAWA, Yasuaki; ENDO, Yasuyuki

    2015-01-01

    Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan. PMID:25947226

  3. Molecular characterization of Babesia and Cytauxzoon species in wild South-African meerkats.

    PubMed

    Leclaire, Sarah; Menard, Sandie; Berry, Antoine

    2015-04-01

    Piroplasms, including Babesia, Cytauxzoon and Theileria species, frequently infect domestic and wild mammals. At present, there is no information on the occurrence and molecular identity of these tick-borne blood parasites in the meerkat, one of South Africa's most endearing wildlife celebrities. Meerkats live in territorial groups, which may occur on ranchland in close proximity to humans, pets and livestock. Blood collected from 46 healthy meerkats living in the South-African Kalahari desert was screened by microscopy and molecular methods, using PCR and DNA sequencing of 18S rRNA and ITS1 genes. We found that meerkats were infected by 2 species: one species related to Babesia sp. and one species related to Cytauxzoon sp. Ninety one percent of the meerkats were infected by the Cytauxzoon and/or the Babesia species. Co-infection occurred in 46% of meerkats. The pathogenicity and vectors of these two piroplasm species remains to be determined. PMID:25374302

  4. Bacteremia with Streptococcus bovis and Streptococcus salivarius: clinical correlates of more accurate identification of isolates.

    PubMed Central

    Ruoff, K L; Miller, S I; Garner, C V; Ferraro, M J; Calderwood, S B

    1989-01-01

    Two biotypes of Streptococcus bovis can be identified by laboratory testing and can be distinguished from the phenotypically similar organism Streptococcus salivarius. We assessed the clinical relevance of careful identification of these organisms in 68 patients with streptococcal bacteremia caused by these similar species. S. bovis was more likely to be clinically significant when isolated from blood (89%) than was S. salivarius (23%). There was a striking association between S. bovis I bacteremia and underlying endocarditis (94%) compared with that of S. bovis II bacteremia (18%). Bacteremia with S. bovis I was also highly correlated with an underlying colonic neoplasm (71% of patients overall, 100% of those with thorough colonic examinations) compared with bacteremia due to S. bovis II or S. salivarius (17% overall, 25% of patients with thorough colonic examinations). We conclude that careful identification of streptococcal bacteremic isolates as S. bovis biotype I provides clinically important information and should be more widely applied. PMID:2915024

  5. First report of Babesia divergens infection in an HIV patient.

    PubMed

    González, Luis M; Castro, Emma; Lobo, Cheryl A; Richart, Alberto; Ramiro, Raquel; González-Camacho, Fernando; Luque, Daniel; Velasco, Aurelio C; Montero, Estrella

    2015-04-01

    Human babesiosis is a zoonosis primarily transmitted through Ixodes ticks and alternatively by routes such as blood transfusions from asymptomatic donors. We report the first case of human babesiosis caused by Babesia divergens in a patient with HIV. This study also focuses on elucidating the possible transmission route of infection in this patient, who received numerous blood transfusions but showed patent symptoms only after splenectomy. A battery of detection tools along with a novel Western-Blot Assay and Enzyme Linked Immunosorbent Assay using the major surface protein of B. divergens (Bd37) as a target were used to evaluate the presence of B. divergens or antibodies against the parasite in samples from the patient and the blood donors involved in this case. A retrospective study of the humoral status against the parasite revealed B. divergens IgG antibodies in one of the implicated donors, but also showed that the patient had been already exposed to the parasite before any transfusion. Thus, this analysis of natural and transfusion transmission routes suggests a pre-existing subclinical babesiosis in the patient. PMID:25686807

  6. First molecular identification of Babesia gibsoni in dogs from Slovakia, central Europe.

    PubMed

    Víchová, Bronislava; Horská, Mária; Blaňarová, Lucia; Švihran, Milan; Andersson, Martin; Peťko, Branislav

    2016-02-01

    Canine babesiosis is a severe and potentially life threatening infection. In Europe, Babesia canis is considered to be the most common species responsible for the disease. We report two cases of babesiosis caused by Babesia gibsoni. The polymerase chain reaction, restriction fragment length polymorphism analysis and further sequencing of 18S rRNA gene fragments from blood samples of both dogs revealed the identity of isolates with B. gibsoni genotypes from other dogs worldwide. This species was previously not known to infect dogs in Slovakia. It is resistant to traditional anti-babesial therapy. Therefore, correct diagnosis is crucial for the successful treatment, especially in dogs with hemolytic anemia and febrile conditions.

  7. 9 CFR 113.409 - Tuberculin-PPD Bovis, Intradermic.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Tuberculin supplied by Animal and Plant Health Inspection Service. (1) Test animals. White female guinea pigs... used in a previous test, shall be used in the specificity test. Twenty-three guinea pigs (10 sensitized... being tested, and 20 guinea pigs (10 sensitized with M. bovis and 10 sensitized with M. avium) shall...

  8. 9 CFR 113.409 - Tuberculin-PPD Bovis, Intradermic.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Tuberculin supplied by Animal and Plant Health Inspection Service. (1) Test animals. White female guinea pigs... used in a previous test, shall be used in the specificity test. Twenty-three guinea pigs (10 sensitized... being tested, and 20 guinea pigs (10 sensitized with M. bovis and 10 sensitized with M. avium) shall...

  9. [The importance of Mycoplasma bovis in bovine respiratory disease].

    PubMed

    Gevaert, D

    2006-02-15

    The annual damage caused by bovine respiratory disease is estimated at 45 up to 55 euro per calf of milking cattle and 117.50 euro per veal calf In Europe, M. bovis is responsible for at least 1/4 to 1/3 of all pneumonia cases in calves. Serology may help to identify the spreading of these bacteria in a herd.

  10. Pathology of Mycobacterium bovis infection in wild meerkats (Suricata suricatta).

    PubMed

    Drewe, J A; Foote, A K; Sutcliffe, R L; Pearce, G P

    2009-01-01

    Pathological lesions associated with Mycobacterium bovis infection (bovine tuberculosis; bTB) in free-living meerkats (Suricata suricatta) in the Kalahari Desert of South Africa are described. The pathology of bTB in meerkats was determined through detailed post-mortem examinations of 57 animals (52 meerkats showing clinical signs of bTB, and five not showing signs of disease). Lymph nodes and tissue lesions thought to be associated with bTB were cultured for mycobacteria. All 52 bTB-infected meerkats showed gross or microscopical granulomatous lesions, but M. bovis was cultured from only 42% (22/52) of these animals. The majority (96%, 50/52) of diseased meerkats had lesions in multiple sites, the pattern of which suggested haematogenous spread of M. bovis infection in this species. The histological characteristics of the tuberculous lesions, together with the gross pathology and the wide range of body systems affected, indicate that infection in meerkats is acquired principally via the respiratory and oral routes, whereas excretion is most likely via the respiratory tract and suppurating skin wounds. Urine and faeces appear to be unlikely sources of infection. The findings of this study provide information on the transmission, pathogenesis and epidemiology of bTB in meerkats that is likely to be relevant to the understanding of M. bovis infection in other social mammal species such as the European badger (Meles meles).

  11. The life and legacy of William T. Bovie.

    PubMed

    Carter, Preston L

    2013-05-01

    This Historian's Address, presented at the North Pacific Surgical Association 2012 meeting, held in Spokane, Washington, on November 9, 2012, briefly reviews the life and surgical contributions of the inventor William T. Bovie and his collaboration with Dr Harvey Cushing, which led to the widespread acceptance of surgical electrocautery for dissection and hemostasis. PMID:23592153

  12. The life and legacy of William T. Bovie.

    PubMed

    Carter, Preston L

    2013-05-01

    This Historian's Address, presented at the North Pacific Surgical Association 2012 meeting, held in Spokane, Washington, on November 9, 2012, briefly reviews the life and surgical contributions of the inventor William T. Bovie and his collaboration with Dr Harvey Cushing, which led to the widespread acceptance of surgical electrocautery for dissection and hemostasis.

  13. Pathology of Mycobacterium bovis infection in wild meerkats (Suricata suricatta).

    PubMed

    Drewe, J A; Foote, A K; Sutcliffe, R L; Pearce, G P

    2009-01-01

    Pathological lesions associated with Mycobacterium bovis infection (bovine tuberculosis; bTB) in free-living meerkats (Suricata suricatta) in the Kalahari Desert of South Africa are described. The pathology of bTB in meerkats was determined through detailed post-mortem examinations of 57 animals (52 meerkats showing clinical signs of bTB, and five not showing signs of disease). Lymph nodes and tissue lesions thought to be associated with bTB were cultured for mycobacteria. All 52 bTB-infected meerkats showed gross or microscopical granulomatous lesions, but M. bovis was cultured from only 42% (22/52) of these animals. The majority (96%, 50/52) of diseased meerkats had lesions in multiple sites, the pattern of which suggested haematogenous spread of M. bovis infection in this species. The histological characteristics of the tuberculous lesions, together with the gross pathology and the wide range of body systems affected, indicate that infection in meerkats is acquired principally via the respiratory and oral routes, whereas excretion is most likely via the respiratory tract and suppurating skin wounds. Urine and faeces appear to be unlikely sources of infection. The findings of this study provide information on the transmission, pathogenesis and epidemiology of bTB in meerkats that is likely to be relevant to the understanding of M. bovis infection in other social mammal species such as the European badger (Meles meles). PMID:19070868

  14. 9 CFR 113.409 - Tuberculin-PPD Bovis, Intradermic.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... animals. (3) Thirty-five days post-injection, the guinea pigs shall be used for tuberculin testing. (4... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Tuberculin-PPD Bovis, Intradermic. 113.409 Section 113.409 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE,...

  15. First description of Bartonella bovis in cattle herds in Israel.

    PubMed

    Rudoler, Nir; Rasis, Michal; Sharir, Benny; Novikov, Anna; Shapira, Gregory; Giladi, Michael

    2014-09-17

    Bartonella bovis has been described in beef and dairy cattle worldwide, however the reported prevalence rates are inconsistent, with large variability across studies (0-89%). This study describes the first isolation and characterization of B. bovis among cattle herds in the Middle East. Blood samples from two beef cattle herds (each sampled thrice) and one dairy herd (sampled twice) in Israel were collected during a 16-months period. Overall, 71 of 95 blood samples (75%) grew Bartonella sp., with prevalence of 78% and 59% in beef and dairy cattle, respectively. High level bacteremia (≥100,000 colony forming units/mL) was detected in 25 specimens (26%). Such high-level bacteremia has never been reported in cattle. Two dairy cows and one beef cow remained bacteremic when tested 60 or 120 days apart, respectively, suggesting that cattle may have persistent bacteremia. One third of animals were infested with ticks. Sequence analysis of a gltA fragment of 32 bacterial isolates from 32 animals revealed 100% homology to B. bovis. Species identification was confirmed by sequence analysis of the rpoB gene. Phylogenetic analysis based on the concatenated sequences of gltA and rpoB demonstrated that the isolates described herein form a monophyletic group with B. bovis strains originating from cattle worldwide. Taken together, the high prevalence of bacteremia, including high-level bacteremia, in beef and dairy cattle, the potential to develop prolonged bacteremia, the exposure of cattle to arthropod vectors, and proximity of infected animals to humans, make B. bovis a potential zoonotic agent.

  16. Compositions and characteristics of strains of Streptococcus bovis.

    PubMed

    Russell, J B; Robinson, P H

    1984-07-01

    Streptococcus bovis strains JB1, 26, 581AXY2, 21096C, and 45S1 grew on glucose, maltose, starch, sucrose, cellobiose, and lactose. None of these strains grew on xylose or ribose, but arabinose was a suitable energy source for strains 2109C and K27FF4. All strains grew at 45 degrees C, but incubation at 50 degrees C prevented growth. Growth was permitted in 2% sodium chloride, but 6.5% sodium chloride was inhibitory. Doubling times ranged from 24 to 27 min, and final pH on glucose was approximately 4.6. None of the strains had a requirement for amino acids, and growth was rapid in media containing glucose salts and B vitamins. There was no ammonia production from arginine. All strains showed aminoendopeptidase activity, but there was considerable strain variation. Strain 7H4, reported as Streptococcus bovis, was noticeably different from the other six strains. It had a doubling time that was more than four times as long, and it grew poorly on starch or in the absence of an amino acid source. Six-and-a-half percent sodium chloride was not inhibitory, and it produced ammonia from arginine. Cell morphology was coccoid rather than ovoid. Based on these criteria, classification of strain 7H4 as Streptococcus bovis seemed doubtful. Other experiments with strain 7H4 indicated that Streptococcus bovis was devoid of diaminopimelic acid. In these experiments strain 7H4 contained significant diaminopimelic acid. The six Streptococcus bovis strains all contained diaminopimelic acid as well, but concentration varied.

  17. Identification of newly isolated Babesia parasites from cattle in Korea by using the Bo-RBC-SCID mice

    PubMed Central

    Cho, Shin-Hyeong; Lee, Hyeong-Woo; Tsuji, Masayoshi; Ishihara, Chiaki; Kim, Jong-Taek; Wee, Sung-Hwan; Lee, Chung-Gil

    2002-01-01

    Attempts were made to isolate and identify Korean bovine Babesia parasite. Blood samples were collected from Holstein cows in Korea, and Babesia parasites were propagated in SCID mice with circulating bovine red blood cells for isolation. The isolate was then antigenically and genotypically compared with several Japanese isolates. The Korean parasite was found to be nearly identical to the Oshima strain isolated from Japanese cattle, which was recently designated as Babesia ovata oshimensis n. var. Haemaphysalis longicornis was the most probable tick species that transmited the parasite. PMID:11949211

  18. Corynebacterium bovis: Epizootiologic Features and Environmental Contamination in an Enzootically Infected Rodent Room

    PubMed Central

    Burr, Holly N; Wolf, Felix R; Lipman, Neil S

    2012-01-01

    Corynebacterium bovis is a common pathogen in athymic nude mouse colonies. Control and eradication of the organism are challenging because depopulation and restricted colony access are often not options within vivaria. We evaluated potential sources and dissemination routes of C. bovis in an enzootically infected colony. Immunocompetent mice and personnel were evaluated for their potential to carry C. bovis, and husbandry and sanitation methods were evaluated for their efficacy in preventing cross-contamination. C. bovis was detected in furred immunocompetent mice previously exposed to infected athymic nude mice and in the nasopharynx of humans. Microisolation cages were not effective in maintaining athymic nude mice C. bovis-free when they were housed in a room known to contain immunodeficient mice with C. bovis infections. A tunnel washer that provided a ≥180 °F final rinse provided effective elimination of C. bovis from cage components. Passive and active air sampling techniques showed airborne dispersal of C. bovis despite the use of individually ventilated caging systems and stringent operational standards. Bacterial growth was not observed in settle plates placed inside autoclaved individually ventilated microisolation cages on various ventilated racks for 24-h periods. C. bovis aerosolization was shown to be a means of spread of the bacterium during cage-change procedures inside a class II type A2 biosafety cabinet. Our findings indicate that C. bovis can be a pervasive environmental contaminant in infected rodent holding rooms and successful eradication strategies must include environmental decontamination and attention to air quality. PMID:22776119

  19. Taurine as a marker for the identification of natural Calculus Bovis and its substitutes.

    PubMed

    Shimada, Kayoko; Azuma, Yuko; Kawase, Masaya; Takahashi, Toshiharu; Schaffer, Stephen W; Takahashi, Kyoko

    2013-01-01

    Calculus Bovis (C. Bovis) is a commonly used animal-derived therapeutic preparation. To meet the increasing clinical demand for the preparation, two artificial substitutes for Bos Taurus have been introduced in China: artificial C. Bovis and in vitro cultured C. Bovis. However, information on their efficacy and safety is inadequate. Therefore, we investigated the biological differences between the commonly used natural preparation and its two substitutes, with the aim of not only identifying the differences but also providing a procedure to distinguish between the different preparations.In the study, we prepared 9 natural C. Bovis, 2 artificial C. Bovis, and 2 in vitro cultured C. Bovis preparations for evaluation. Differences were noted between the three preparations relative to their effect on viability of cardiac fibroblasts from 1-day-old Wistar rats. Although natural C. Bovis had no effect on cell viability, 1-h treatment of the cells with 0.25 mg/ml of the substitutes significantly reduced cell viability, as detected by the MTS assay. Based on liquid chromatography and inductively coupled plasma mass spectrometry, the preparations also differed in composition. Indeed, the substitutes contained more taurine, cholic acid, iron, magnesium, and calcium than the natural preparations. They also differed spectroscopically.The present results reveal significant biological differences between natural C. Bovis and two of its substitutes. Since the substitutes appear to contain more taurine, cholic acid, and elements, these constituents may serve as markers to distinguish between natural C. Bovis and its substitutes.

  20. Taurine as a marker for the identification of natural Calculus Bovis and its substitutes.

    PubMed

    Shimada, Kayoko; Azuma, Yuko; Kawase, Masaya; Takahashi, Toshiharu; Schaffer, Stephen W; Takahashi, Kyoko

    2013-01-01

    Calculus Bovis (C. Bovis) is a commonly used animal-derived therapeutic preparation. To meet the increasing clinical demand for the preparation, two artificial substitutes for Bos Taurus have been introduced in China: artificial C. Bovis and in vitro cultured C. Bovis. However, information on their efficacy and safety is inadequate. Therefore, we investigated the biological differences between the commonly used natural preparation and its two substitutes, with the aim of not only identifying the differences but also providing a procedure to distinguish between the different preparations.In the study, we prepared 9 natural C. Bovis, 2 artificial C. Bovis, and 2 in vitro cultured C. Bovis preparations for evaluation. Differences were noted between the three preparations relative to their effect on viability of cardiac fibroblasts from 1-day-old Wistar rats. Although natural C. Bovis had no effect on cell viability, 1-h treatment of the cells with 0.25 mg/ml of the substitutes significantly reduced cell viability, as detected by the MTS assay. Based on liquid chromatography and inductively coupled plasma mass spectrometry, the preparations also differed in composition. Indeed, the substitutes contained more taurine, cholic acid, iron, magnesium, and calcium than the natural preparations. They also differed spectroscopically.The present results reveal significant biological differences between natural C. Bovis and two of its substitutes. Since the substitutes appear to contain more taurine, cholic acid, and elements, these constituents may serve as markers to distinguish between natural C. Bovis and its substitutes. PMID:23392879

  1. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia.

    PubMed

    Vargas-Hernández, G; André, M R; Faria, J L M; Munhoz, T D; Hernandez-Rodriguez, M; Machado, R Z; Tinucci-Costa, M

    2012-05-25

    Ehrlichiosis and babesiosis are tick-borne diseases, caused mainly by Ehrlichia canis and Babesia canis, respectively, with a worldwide occurrence in dogs, whose main vector is the brown-dog tick, Rhipicephalus sanguineus. The present work aimed to detect the presence of E. canis and Babesia sp. in 91 dog blood samples in Colombia, by molecular and serological techniques. We also performed sequence alignment to indicate the identity of the parasite species infecting these animals. The present work shows the first molecular detection of E. canis and B. vogeli in dogs from Colombia. Immunoglobulin-G (IgG) antibodies to E. canis and Babesia vogeli were found in 75 (82.4%) and 47 (51.6%) sampled dogs, respectively. Thirty-seven (40.6%) and 5 (5.5%) dogs were positive in PCR for E. canis and Babesia sp., respectively. After sequencing, amplicons showed 99% of identity with isolates of E. canis and B. vogeli. The phylogenetic trees based on 16S rRNA-Anaplasmataceae sequences and 18S rRNA-piroplasmid sequences supported the identity of the found E. canis and B. vogeli DNAs, respectively. The present work shows the first molecular detection of E. canis and B. vogeli in dogs in Colombia.

  2. Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia

    PubMed Central

    Vanstreels, Ralph Eric Thijl; Woehler, Eric J.; Ruoppolo, Valeria; Vertigan, Peter; Carlile, Nicholas; Priddel, David; Finger, Annett; Dann, Peter; Herrin, Kimberly Vinette; Thompson, Paul; Ferreira Junior, Francisco C.; Braga, Érika M.; Hurtado, Renata; Epiphanio, Sabrina; Catão-Dias, José Luiz

    2015-01-01

    Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected. PMID:25853053

  3. Frequency of piroplasms Babesia microti and Cytauxzoon felis in stray cats from northern Italy.

    PubMed

    Spada, Eva; Proverbio, Daniela; Galluzzo, Paola; Perego, Roberta; Bagnagatti De Giorgi, Giada; Roggero, Nora; Caracappa, Santo

    2014-01-01

    Emerging diseases caused by piroplasms pose a health risk for man and other animals, and domestic cats have been proposed as potential reservoirs for some piroplasm infections. The aim of this study was to identify the frequency of the piroplasms Babesia microti and Cytauxzoon felis in stray cats from northern Italy and to identify possible risk factors associated with these infections. Blood samples from 260 stray cats enrolled in a trap-neuter-release (TNR) program in northern Italy were examined with conventional PCR for the presence of Babesia microti and Cytauxzoon felis DNA. No sample (0.0%) tested positive for C. felis, whilst B. microti DNA was detected in two samples (0.8%). Both infected cats were in good clinical condition and recovered well from the neutering surgery. One of these two cats had a triple coinfection with Babesia microti, Candidatus Mycoplasma haemominutum, and Anaplasma phagocytophilum. Evidence presented in this study indicates that the blood borne protozoans Babesia microti and Cytauxzoon felis are not widely distributed in stray cat populations in Milan, northern Italy, and that the significance of cats as a reservoir host for B. microti in this area is limited.

  4. Frequency of Piroplasms Babesia microti and Cytauxzoon felis in Stray Cats from Northern Italy

    PubMed Central

    Spada, Eva; Proverbio, Daniela; Galluzzo, Paola; Perego, Roberta; Roggero, Nora; Caracappa, Santo

    2014-01-01

    Emerging diseases caused by piroplasms pose a health risk for man and other animals, and domestic cats have been proposed as potential reservoirs for some piroplasm infections. The aim of this study was to identify the frequency of the piroplasms Babesia microti and Cytauxzoon felis in stray cats from northern Italy and to identify possible risk factors associated with these infections. Blood samples from 260 stray cats enrolled in a trap-neuter-release (TNR) program in northern Italy were examined with conventional PCR for the presence of Babesia microti and Cytauxzoon felis DNA. No sample (0.0%) tested positive for C. felis, whilst B. microti DNA was detected in two samples (0.8%). Both infected cats were in good clinical condition and recovered well from the neutering surgery. One of these two cats had a triple coinfection with Babesia microti, Candidatus Mycoplasma haemominutum, and Anaplasma phagocytophilum. Evidence presented in this study indicates that the blood borne protozoans Babesia microti and Cytauxzoon felis are not widely distributed in stray cat populations in Milan, northern Italy, and that the significance of cats as a reservoir host for B. microti in this area is limited. PMID:24895629

  5. Epidemiology and molecular phylogeny of Babesia sp. in Little Penguins Eudyptula minor in Australia.

    PubMed

    Vanstreels, Ralph Eric Thijl; Woehler, Eric J; Ruoppolo, Valeria; Vertigan, Peter; Carlile, Nicholas; Priddel, David; Finger, Annett; Dann, Peter; Herrin, Kimberly Vinette; Thompson, Paul; Ferreira Junior, Francisco C; Braga, Érika M; Hurtado, Renata; Epiphanio, Sabrina; Catão-Dias, José Luiz

    2015-08-01

    Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected.

  6. Fecal volatile organic compound profiles from white-tailed deer (Odocoileus virginianus) as indicators of Mycobacterium bovis exposure or Mycobacterium bovis bacille Calmette-Guerin (BCG) vaccination

    Technology Transfer Automated Retrieval System (TEKTRAN)

    White-tailed deer (Odocoileus virginianus) serve as a reservoir for bovine tuberculosis, caused by Mycobacterium bovis, and can be a source of infection in cattle. Vaccination with M. bovis bacille Calmette-Guerin (BCG) is being considered for management of bovine tuberculosis in deer. Presently, no...

  7. The use of different diagnostic tools for Babesia and Theileria parasites in cattle in Menofia, Egypt.

    PubMed

    Nayel, Mohamed; El-Dakhly, Khaled Mohamed; Aboulaila, Mahmoud; Elsify, Ahmed; Hassan, Hany; Ibrahim, Elsayed; Salama, Akram; Yanai, Tokuma

    2012-09-01

    Bovine piroplasmosis is caused by tick-borne hemoprotozoans of the genera Babesia and Theileria and is the most prevalent in tropical and subtropical countries, causing a major economic impact worldwide. In the current study, a total of 405 cattle of different ages, sexes, and breeds were randomly sampled for surveying and diagnosis of babesiosis and theileriosis using three methods: direct microscopy (blood smears), indirect fluorescent antibody test (IFAT) and polymerase chain reaction (PCR). Giemsa-stained blood smears revealed that, out of 405 examined cattle, 33 (8.15 %) were infected with Babesia sp. and 65 (16.05 %) with Theileria sp. (total number of infected cattle was 98). Mixed infection was seen in 11 (2.72 %) animals. Moreover, application of the three diagnostic assays on 158 randomly sampled cattle indicated that 17 (10.76 %) and 33 (20.89 %) were positive for Babesia and Theileria spp. by the direct smear technique, 25 (15.82 %) and 33 (20.89 %) by IFAT (fluorescence was greenish yellow for Babesia and yellowish for Theileria), and 20 (12.66 %) and 38 (24.05 %) by PCR. Using primers specific for Babesia and Theileria spp., we found that diagnostic bands appeared at ~350 and ~370 bp, respectively indicating the presence of these piroplasms. Statistically, there was a non-significant difference of the positivity in response to the three techniques; thus, any of these methods can be described as useful for diagnosing blood parasites in both domesticated animals and birds. On the basis of the obtained results, it could be concluded that direct microscopy can be used in acute infections, whereas IFAT and PCR are useful in chronicity.

  8. Identification and Characterization of the Rhoptry Neck Protein 2 in Babesia divergens and B. microti

    PubMed Central

    Ord, Rosalynn L.; Rodriguez, Marilis; Cursino-Santos, Jeny R.; Hong, Hyunryung; Singh, Manpreet; Gray, Jeremy

    2016-01-01

    Apicomplexan parasites include those of the genera Plasmodium, Cryptosporidium, and Toxoplasma and those of the relatively understudied zoonotic genus Babesia. In humans, babesiosis, particularly transfusion-transmitted babesiosis, has been emerging as a major threat to public health. Like malaria, the disease pathology is a consequence of the parasitemia which develops through cyclical replication of Babesia parasites in host erythrocytes. However, there are no exoerythrocytic stages in Babesia, so targeting of the blood stage and associated proteins to directly prevent parasite invasion is the most desirable option for effective disease control. Especially promising among such molecules are the rhoptry neck proteins (RONs), whose homologs have been identified in many apicomplexan parasites. RONs are involved in the formation of the moving junction, along with AMA1, but no RON has been identified and characterized in any Babesia spp. Here we identify the RON2 proteins of Babesia divergens (BdRON2) and B. microti (BmRON2) and show that they are localized apically and that anti-BdRON2 antibodies are significant inhibitors of parasite invasion in vitro. Neither protein is immunodominant, as both proteins react only marginally with sera from infected animals. Further characterization of the direct role of both BdRON2 and BmRON2 in parasite invasion is required, but knowledge of the level of conformity of RON2 proteins within the apicomplexan phylum, particularly that of the AMA1-RON2 complex at the moving junction, along with the availability of an animal model for B. microti studies, provides a key to target this complex with a goal of preventing the erythrocytic invasion of these parasites and to further our understanding of the role of these conserved ligands in invasion. PMID:26953328

  9. Polymorphisms of 20 regulatory proteins between Mycobacterium tuberculosis and Mycobacterium bovis.

    PubMed

    Bigi, María M; Blanco, Federico Carlos; Araújo, Flabio R; Thacker, Tyler C; Zumárraga, Martín J; Cataldi, Angel A; Soria, Marcelo A; Bigi, Fabiana

    2016-08-01

    Mycobacterium tuberculosis and Mycobacterium bovis are responsible for tuberculosis in humans and animals, respectively. Both species are closely related and belong to the Mycobacterium tuberculosis complex (MTC). M. tuberculosis is the most ancient species from which M. bovis and other members of the MTC evolved. The genome of M. bovis is over >99.95% identical to that of M. tuberculosis but with seven deletions ranging in size from 1 to 12.7 kb. In addition, 1200 single nucleotide mutations in coding regions distinguish M. bovis from M. tuberculosis. In the present study, we assessed 75 M. tuberculosis genomes and 23 M. bovis genomes to identify non-synonymous mutations in 202 coding sequences of regulatory genes between both species. We identified species-specific variants in 20 regulatory proteins and confirmed differential expression of hypoxia-related genes between M. bovis and M. tuberculosis.

  10. Characterization of exochelins of the Mycobacterium bovis type strain and BCG substrains.

    PubMed

    Gobin, J; Wong, D K; Gibson, B W; Horwitz, M A

    1999-04-01

    Pathogenic mycobacteria must acquire iron in the host in order to multiply and cause disease. To do so, they release abundant quantities of siderophores called exochelins, which have the capacity to scavenge iron from host iron-binding proteins and deliver it to the mycobacteria. In this study, we have characterized the exochelins of Mycobacterium bovis, the causative agent of bovine and occasionally of human tuberculosis, and the highly attenuated descendant of M. bovis, bacillus Calmette-Guérin (BCG), widely used as a vaccine against human tuberculosis. The M. bovis type strain, five substrains of M. bovis BCG (Copenhagen, Glaxo, Japanese, Pasteur, and Tice), and two strains of virulent Mycobacterium tuberculosis all produce the same set of exochelins, although the relative amounts of individual exochelins may differ. Among these mycobacteria, the total amount of exochelins produced is greatest in M. tuberculosis, intermediate in M. bovis, and smallest in M. bovis BCG.

  11. Anatomical distribution of Mycobacterium bovis genotypes in experimentally infected white-tailed deer.

    PubMed

    Thacker, Tyler C; Palmer, Mitchell V; Robbe-Austerman, Suelee; Stuber, Tod P; Waters, W Ray

    2015-10-22

    Mycobacterium bovis (M. bovis) causes tuberculosis in white-tailed deer (WTD). Natural infection of WTD with M. bovis is most closely mimicked by instilling inoculum into palatine tonsillar crypts. One hundred fifty days after intratonsillar inoculation, M. bovis was cultured from 30 tissues originating from 14 deer. Whole-genome sequencing (WGS) was performed on the original inoculum, single colonies subcultured from the original inoculum, and M. bovis isolated from each culture positive tissue. Single nucleotide polymorphisms (SNP) were identified by comparing the derived sequences to the reference strain AF2122/97. Results indicate that the majority of the SNPs that were identified were homogeneous between the inoculum and the isolates from the tissues. The majority of individual tissues had different WGS genotypes from each other, suggesting that dissemination of M. bovis beyond the initial site of infection may require few mycobacteria representing a bottleneck. PMID:26243696

  12. Piroplasmosis in wildlife: Babesia and Theileria affecting free-ranging ungulates and carnivores in the Italian Alps

    PubMed Central

    2014-01-01

    Background Piroplasmosis are among the most relevant diseases of domestic animals. Babesia is emerging as cause of tick-borne zoonosis worldwide and free-living animals are reservoir hosts of several zoonotic Babesia species. We investigated the epidemiology of Babesia spp. and Theileria spp. in wild ungulates and carnivores from Northern Italy to determine which of these apicomplexan species circulate in wildlife and their prevalence of infection. Methods PCR amplification of the V4 hyper-variable region of the 18S rDNA of Babesia sp./Theileria sp was carried out on spleen samples of 1036 wild animals: Roe deer Capreolus capreolus (n = 462), Red deer Cervus elaphus (n = 52), Alpine Chamois Rupicapra rupicapra (n = 36), Fallow deer Dama dama (n = 17), Wild boar Sus scrofa (n = 257), Red fox Vulpes vulpes (n = 205) and Wolf Canis lupus (n = 7). Selected positive samples were sequenced to determine the species of amplified Babesia/Theileria DNA. Results Babesia/Theileria DNA was found with a mean prevalence of 9.94% (IC95% 8.27-11.91). The only piroplasms found in carnivores was Theileria annae, which was detected in two foxes (0.98%; IC95% 0.27-3.49). Red deer showed the highest prevalence of infection (44.23%; IC95% 31.6-57.66), followed by Alpine chamois (22.22%; IC95% 11.71-38.08), Roe deer (12.55%; IC95% 9.84-15.89), and Wild boar (4.67%; IC95% 2.69-7.98). Genetic analysis identified Babesia capreoli as the most prevalent piroplasmid found in Alpine chamois, Roe deer and Red deer, followed by Babesia bigemina (found in Roe deer, Red deer and Wild boar), and the zoonotic Babesia venatorum (formerly Babesia sp. EU1) isolated from 2 Roe deer. Piroplasmids of the genus Theileria were identified in Wild boar and Red deer. Conclusions The present study offers novel insights into the role of wildlife in Babesia/Theileria epidemiology, as well as relevant information on genetic variability of piroplasmids infecting wild ungulates and

  13. Multiplex-PCR for differentiation of Mycobacterium bovis from Mycobacterium tuberculosis complex.

    PubMed

    Spositto, F L E; Campanerut, P A Z; Ghiraldi, L D; Leite, C Q F; Hirata, M H; Hirata, R D C; Siqueira, V L D; Cardoso, R Fressatti

    2014-01-01

    We evaluated a multiplex-PCR to differentiate Mycobacterium bovis from M. tuberculosis Complex (MTC) by one step amplification based on simultaneous detection of pncA 169 C > G change in M. bovis and the IS6110 present in MTC species. Our findings showed the proposed multiplex-PCR is a very useful tool for complementation in differentiating M. bovis from other cultured MTC species.

  14. The transmission of Babesia canis to the wild dog Lycaon pictus (Temminck) and black-backed jackal Canis mesomelas Schreber.

    PubMed

    Van Heerden, J

    1980-06-01

    Babesia canis was successfully transmitted from the domestic dog to 3 wild dogs Lycaon pictus and 4 black-backed jackals Canis mesomelas. Both wild dogs and black-backed jackals showed no clinical signs or clinical pathological evidence of disease. Trophozoites of Babesia canis were found in peripheral blood smears from all experimental animals. The disease was also successfully transmitted from both black-backed jackals and wild dogs to the domestic dog.

  15. The transmission of Babesia canis to the wild dog Lycaon pictus (Temminck) and black-backed jackal Canis mesomelas Schreber.

    PubMed

    Van Heerden, J

    1980-06-01

    Babesia canis was successfully transmitted from the domestic dog to 3 wild dogs Lycaon pictus and 4 black-backed jackals Canis mesomelas. Both wild dogs and black-backed jackals showed no clinical signs or clinical pathological evidence of disease. Trophozoites of Babesia canis were found in peripheral blood smears from all experimental animals. The disease was also successfully transmitted from both black-backed jackals and wild dogs to the domestic dog. PMID:7252967

  16. Genetic variability of Babesia parasites in Haemaphysalis spp. and Ixodes persulcatus ticks in the Baikal region and Far East of Russia.

    PubMed

    Rar, V A; Epikhina, T I; Suntsova, O V; Kozlova, I V; Lisak, O V; Pukhovskaya, N M; Vysochina, N P; Ivanov, L I; Tikunova, N V

    2014-12-01

    To study Babesia diversity in Ixodid ticks in Russia, Ixodes persulcatus, Haemaphysalis japonica, Haemaphysalisconcinna, Dermacentor silvarum, and Dermacentor nuttalli ticks collected in the Far East and Baikal region were assayed for the presence of Babesia spp. using nested PCR. In total, Babesia DNA was detected in 30 of the 1125 (2.7%) I. persulcatus, 17 of the 573 (3.0%) H. concinna, and 12 of the 543 (2.2%) H. japonica but was undetectable in any of the 294 analyzed Dermacentor spp. Partial 18S rRNA gene sequences were determined for all of the positive samples. Among the positive ticks, nine I. persulcatus were infected by Babesia microti 'US'-type, five I. persulcatus were infected by Babesia divergens-like parasites, and 11 I. persulcatus were infected by Babesia venatorum. For all three of these species, the determined 18S rRNA gene sequences were identical to those of the Babesia genetic variants found previously in I. persulcatus in Russia. In addition, five I. persulcatus from the Baikal region and all of the positive Haemaphysalis spp. ticks carried 13 different sequence variants of Babesia sensu stricto belonging to distinct phylogenetic clusters. Babesia spp. from 29 ticks of different species collected in distinct locations belonged to the cluster of cattle and ovine parasites (Babesia crassa, Babesiamajor, Babesiamotasi, Babesiabigemina, etc.). Babesia spp. from four H. japonica ticks in the Far East belonged to the cluster formed by parasites of carnivores. One more Babesia sequence variant detected in an I. persulcatus tick from the Baikal region belonged to the cluster formed by parasites of cattle and wild cervids (B. divergens, Babesiacapreoli, B. venatorum, Babesiaodocoilei, etc.).

  17. Mycobacterium bovis infection in a wild sow (Sus scrofa): the first case in Korea

    PubMed Central

    Kim, Jae Myung; Jang, Young-Boo; Jang, Yunho; Yu, So Yoon; Kim, Jiro; Moon, Oun Kyung; Jung, Suk Chan; Lee, Min Kwon; Jeong, Tae Nam

    2016-01-01

    Mycobacterium (M.) bovis causes tuberculosis and has a broad host range, including humans, livestock, and wild animals. M. bovis infection of wild boar has been reported in several European countries. We report here the first case of M. bovis infection in a domesticated wild sow in Korea. Granulomatous and necrotizing lesions with small numbers of acid-fast bacilli were observed in nodules of the lung of wild sow. Furthermore, the M. bovis isolate from the wild sow had spoligotype SB0140 and a novel MIRU-VNTR allelic profile, which is not found in cattle and deer in Korea. PMID:26726026

  18. Mycobacterium bovis infection in a wild sow (Sus scrofa): first case in Korea.

    PubMed

    Ku, Bok Kyung; Jeon, Bo-Young; Kim, Jae Myung; Jang, Young-Boo; Jang, Yunho; Yu, So Yoon; Kim, Jiro; Moon, Oun Kyung; Jung, Suk Chan; Lee, Min Kwon; Jeong, Tae Nam

    2016-09-30

    Mycobacterium (M.) bovis causes tuberculosis and has a broad host range, including humans, livestock, and wild animals. M. bovis infection of wild boar has been reported in several European countries. We report here the first case of M. bovis infection in a domesticated wild sow in Korea. Granulomatous and necrotizing lesions with small numbers of acid-fast bacilli were observed in nodules of the lung of wild sow. Furthermore, the M. bovis isolate from the wild sow had spoligotype SB0140 and a novel MIRU-VNTR allelic profile, which is not found in cattle and deer in Korea.

  19. First molecular survey of Anaplasma bovis in small ruminants from Tunisia.

    PubMed

    Ben Said, Mourad; Belkahia, Hanène; Karaoud, Maroua; Bousrih, Maha; Yahiaoui, Mouna; Daaloul-Jedidi, Monia; Messadi, Lilia

    2015-09-30

    To date, no information is available regarding the presence of Anaplasma bovis in the South Mediterranean area. In this study, prevalence, risk factors, and genetic diversity of A. bovis were assessed in small ruminants. A total of 563 healthy small ruminants (260 sheep and 303 goats), from 25 randomly selected flocks located in 5 localities from two bioclimatic areas in Tunisia, were investigated for the detection of A. bovis in blood by nested polymerase chain reaction (nPCR) assay. The overall infection rates of A. bovis were 42.7 and 23.8% in sheep and goats, respectively. Goats located in a sub-humid area were statistically more infected than those located in a humid area. A. bovis prevalence rate varied significantly according to sheep and goat flocks, and to the sheep breed. Infection with A. bovis was validated by sequencing. Sequence analysis based on the 16S rRNA gene showed that A. bovis from Tunisian goats and sheep clustered with other strain sequences detected from wild and domestic animals and published in GenBank. This study gives the first insight of presence of A. bovis DNA in small ruminants in Tunisia and suggests that these animal species may be playing an important role in the bovine anaplasmosis natural cycle caused by A. bovis in the South Mediterranean ecosystem. PMID:26088935

  20. First molecular survey of Anaplasma bovis in small ruminants from Tunisia.

    PubMed

    Ben Said, Mourad; Belkahia, Hanène; Karaoud, Maroua; Bousrih, Maha; Yahiaoui, Mouna; Daaloul-Jedidi, Monia; Messadi, Lilia

    2015-09-30

    To date, no information is available regarding the presence of Anaplasma bovis in the South Mediterranean area. In this study, prevalence, risk factors, and genetic diversity of A. bovis were assessed in small ruminants. A total of 563 healthy small ruminants (260 sheep and 303 goats), from 25 randomly selected flocks located in 5 localities from two bioclimatic areas in Tunisia, were investigated for the detection of A. bovis in blood by nested polymerase chain reaction (nPCR) assay. The overall infection rates of A. bovis were 42.7 and 23.8% in sheep and goats, respectively. Goats located in a sub-humid area were statistically more infected than those located in a humid area. A. bovis prevalence rate varied significantly according to sheep and goat flocks, and to the sheep breed. Infection with A. bovis was validated by sequencing. Sequence analysis based on the 16S rRNA gene showed that A. bovis from Tunisian goats and sheep clustered with other strain sequences detected from wild and domestic animals and published in GenBank. This study gives the first insight of presence of A. bovis DNA in small ruminants in Tunisia and suggests that these animal species may be playing an important role in the bovine anaplasmosis natural cycle caused by A. bovis in the South Mediterranean ecosystem.

  1. Molecular evidence for the presence of new Babesia species in feral raccoons (Procyon lotor) in Hokkaido, Japan.

    PubMed

    Jinnai, Michio; Kawabuchi-Kurata, Takako; Tsuji, Masayoshi; Nakajima, Rui; Fujisawa, Kohei; Nagata, Shogo; Koide, Hikaru; Matoba, Yohei; Asakawa, Mitsuhiko; Takahashi, Kazuo; Ishihara, Chiaki

    2009-06-10

    We recently reported that feral raccoons (Procyon lotor) with splenomegaly native to Japan were carriers of a Babesia microti-like parasite identical to that found in the United States, which was likely introduced to Japan from North America via raccoons imported as pets. Thus, we attempted extensive molecular survey for piroplasma infections of feral raccoon with normal spleen in Hokkaido, Japan using nested PCR that target broadly to 18S ribosomal RNA gene (SSU-rDNA) of all the parasites in the genus Babesia, Theileria, Cytauxzoon and B. microti group. Of the 348 raccoon samples analyzed, 9 gave positive signals. Cloning and phylogenetic analysis on SSU-rDNA sequences revealed that six of nine positives were found to be infected with Babesia and the remaining three with previously unreported Sarcocystis. Babesia sequences were further separated into two distantly related groups, those that reside in a novel phylogenetic group were consisted solely of four parasites found in this study, while those which included one identical sequence found in the three of our specimens were assembled together with both Babesia parasites of tick's in Japan and of raccoon's in U.S. These results may indicate that not only a B. microti-like parasite but also at least two yet undescribed Babesia species are being established in their new life cycles in the feral raccoon populations in Japan.

  2. Mycobacterium bovis in Swine: Spoligotyping of Isolates from Argentina

    PubMed Central

    Barandiaran, Soledad; Martínez Vivot, Marcela; Moras, Eduardo Vicente; Cataldi, Angel Adrián; Zumárraga, Martín José

    2011-01-01

    A total of 143 Mycobacterium bovis isolates of pigs, from the most productive swine area in Argentina, were typed by spoligotyping. Twenty-two different spoligotypes were identified, and 133 (93%) isolates were grouped into 12 clusters. One of them, designed SB0140, was the most frequent because it held 83 (58%) isolates. This spoligotype also grouped 362 (43%) out of 841 isolates from previously typed cattle and, thus, constitutes the most frequent in our country. In addition, 135 (94%) isolates revealed spoligotypes identical to those of cattle, showing an epidemiological link. On the other hand, there were seven novel spoligotypes, six of which were also unique since they had only one isolate each. This study aimed to identify the spoligotypes of M. bovis isolated from pigs to contribute to a better understanding of the distribution of bovine tuberculosis in the main productive area of Argentina. PMID:21547236

  3. Structural definition of arabinomannans from Mycobacterium bovis BCG.

    PubMed

    Nigou, J; Gilleron, M; Brando, T; Vercellone, A; Puzo, G

    1999-06-01

    The structures of the hydrophilic parietal and cellular arabinomannans isolated from Mycobacterium bovis BCG cell wall [Nigou et al. (1997) J Biol Chem 272: 23094-103] were investigated. Their molecular mass as determined by MALDI-TOF mass spectrometry was around 16 kDa. Concerning cap structure, capillary electrophoresis analysis demonstrated that dimannoside (Manpalpha1-->2Manp) was the most abundant motif (65-75%). Using two-dimensional 1H-13C NMR spectroscopy, the mannan core was unambiguously demonstrated to be composed of -->6Manpalpha1--> backbone substituted at some O-2 by a single Manp unit. The branching degree was determined as 84%. Finally, arabinomannans were found to be devoid of the phosphatidyl-myo-inositol anchor and, by aminonaphthalene disulfonate tagging, the mannan core was shown to contain a reducing end. This constitutes the main difference between arabinomannans and lipoarabinomannans from Mycobacterium bovis BCG.

  4. Membrane proteins of Mycoplasma bovis and their role in pathogenesis.

    PubMed

    Adamu, James Y; Wawegama, Nadeeka K; Browning, Glenn F; Markham, Philip F

    2013-10-01

    Mycoplasma membrane proteins influence cell shape, cell division, motility and adhesion to host cells, and are thought to be integrally involved in the pathogenesis of mycoplasmoses. Many of the membrane proteins predicted from mycoplasma genome sequences remain hypothetical, as their presence in cellular protein preparations is yet to be established experimentally. Recent genome sequences of several strains of Mycoplasma bovis have provided further insight into the potential role of the membrane proteins of this pathogen in colonisation and infection. This review highlights recent advances in knowledge about the influence of M. bovis membrane proteins on the pathogenesis of infection with this species and identifies future research directions for enhancing our understanding of the role of these proteins. PMID:23810376

  5. Mycoplasma bovis mastitis and arthritis in a dairy heifer.

    PubMed

    2015-12-19

    Mycoplasma bovis causing mastitis and arthritis in a dairy heifer. Nutritional myopathy in a three-month-old suckler calf. Acute fasciolosis in ewes in Ayrshire. Cardiomyopathy of unknown aetiology causing death of a three-year-old Suffolk ram. Spinal aspergillosis in a seven-week-old pheasant poult These are among matters discussed in the disease surveillance report for August from SAC Consulting: Veterinary Services (SAC C VS).

  6. Mycoplasma bovis mastitis and arthritis in a dairy heifer.

    PubMed

    2015-12-19

    Mycoplasma bovis causing mastitis and arthritis in a dairy heifer. Nutritional myopathy in a three-month-old suckler calf. Acute fasciolosis in ewes in Ayrshire. Cardiomyopathy of unknown aetiology causing death of a three-year-old Suffolk ram. Spinal aspergillosis in a seven-week-old pheasant poult These are among matters discussed in the disease surveillance report for August from SAC Consulting: Veterinary Services (SAC C VS). PMID:26679914

  7. [Investigation of Mycobacterium bovis subsp. bovis among the strains of Mycobacterium tuberculosis complex isolated in Düzce Province, Turkey].

    PubMed

    Öztürk, Cihadiye Elif; Şahin, İdris; Öksüz, Şükrü; Kılıç, Nida; Kılınçel, Özge; Aydın, Leyla; Atik, Dursun; Afşin, Emine

    2016-07-01

    Throughout the history of mankind, tuberculosis (TB) has caused serious illness and still continues to do so. Archaeobiological studies indicated that TB in humans dates back to 4000-8000 BC, and cases were shown to be due to Mycobacterium bovis subsp.bovis rather than Mycobacterium tuberculosis. Moreover, this situation was thought to begin with domestication of animals, consumption of their milk, and living together in the same environment with them. Over time, with the consumption of boiled milk and with the establishment of separate animal shelters, M.bovis subsp. bovis infection began to be seen rarely. Today, M.bovis infection is mostly transmitted from animals to humans and very rarely from humans to other humans. The most significant means of transmission of the infection are to the gastrointestinal tract via consumption of raw milk and to the respiratory system via droplet infection from the animals with disease. In this study, it was planned to investigate the cause of occurrence of TB in cattles in Düzce in the past few years along with the presence of bovine type TB in cases of human tuberculosis. We aimed to carry out subtype determination of the M.tuberculosis complex (MTBC) strains isolated in our mycobacteriology laboratory between the years 2004-2014, and evaluate the clinical and sociodemographic data of patients in whom M.bovis subsp. bovis was detected. The strains that were selected for the study have been isolated from radiometric BACTEC™ 12B broth and/or Löwenstein-Jensen (LJ) media between 2004-2009, and BACTEC™ MGIT™ (Mycobacteria Growth Indicator Tube) and/or LJ media between 2009-2014 periods. The GenoType MTBC Kit (Hain-Lifescience GmbH, Germany) was used in the study for determination of the subspecies. Extraction and amplification of DNA and hybridizations were performed according to test procedure in order to investigate the presence of subtypes of the MTBC species in skimmed milk from collections stored at -20°C. In the

  8. [Investigation of Mycobacterium bovis subsp. bovis among the strains of Mycobacterium tuberculosis complex isolated in Düzce Province, Turkey].

    PubMed

    Öztürk, Cihadiye Elif; Şahin, İdris; Öksüz, Şükrü; Kılıç, Nida; Kılınçel, Özge; Aydın, Leyla; Atik, Dursun; Afşin, Emine

    2016-07-01

    Throughout the history of mankind, tuberculosis (TB) has caused serious illness and still continues to do so. Archaeobiological studies indicated that TB in humans dates back to 4000-8000 BC, and cases were shown to be due to Mycobacterium bovis subsp.bovis rather than Mycobacterium tuberculosis. Moreover, this situation was thought to begin with domestication of animals, consumption of their milk, and living together in the same environment with them. Over time, with the consumption of boiled milk and with the establishment of separate animal shelters, M.bovis subsp. bovis infection began to be seen rarely. Today, M.bovis infection is mostly transmitted from animals to humans and very rarely from humans to other humans. The most significant means of transmission of the infection are to the gastrointestinal tract via consumption of raw milk and to the respiratory system via droplet infection from the animals with disease. In this study, it was planned to investigate the cause of occurrence of TB in cattles in Düzce in the past few years along with the presence of bovine type TB in cases of human tuberculosis. We aimed to carry out subtype determination of the M.tuberculosis complex (MTBC) strains isolated in our mycobacteriology laboratory between the years 2004-2014, and evaluate the clinical and sociodemographic data of patients in whom M.bovis subsp. bovis was detected. The strains that were selected for the study have been isolated from radiometric BACTEC™ 12B broth and/or Löwenstein-Jensen (LJ) media between 2004-2009, and BACTEC™ MGIT™ (Mycobacteria Growth Indicator Tube) and/or LJ media between 2009-2014 periods. The GenoType MTBC Kit (Hain-Lifescience GmbH, Germany) was used in the study for determination of the subspecies. Extraction and amplification of DNA and hybridizations were performed according to test procedure in order to investigate the presence of subtypes of the MTBC species in skimmed milk from collections stored at -20°C. In the

  9. Guillain-Barré syndrome associated with Mycobacterium bovis lymphadenitis.

    PubMed

    Vergnon-Miszczycha, Delphine; Suy, Florence; Robert, Florence; Carricajo, Anne; Fresard, Anne; Cazorla, Céline; Guglielminotti, Claire; Lucht, Frédéric; Botelho-Nevers, Elisabeth

    2015-10-01

    Guillain-Barré syndrome (GBS) is an autoimmune disease that can be triggered by different infectious agents. Here we report the case of a 26-year-old Algerian woman who developed GBS associated with a Mycobacterium bovis cervical lymphadenitis. Following intravenous immunoglobulin therapy, the patient's neurologic state returned to normal after 3 months. The lymphadenitis responded more slowly to the antituberculous treatment and an excision of necrotic cervical lymph nodes had to be performed four times. Antibiotics were administered for 16 months: ethambutol was stopped after 2 months, and rifampicin and isoniazid pursued for 14 months. An extensive etiological investigation showed that, in this case, the only likely infectious trigger GBS was the concomitant M. bovis infection. To our knowledge, this is the first report of GBS triggered by M. bovis. We performed a literature review revealing that the association between tuberculosis and Guillain-Barré syndrome is very rare (only seven cases previously reported) but is not coincidental. Physicians should be aware that tuberculosis can be a cause of GBS.

  10. [Infection due to Mycobacterium bovis in common variable immunodeficiency].

    PubMed

    Herrera-Sánchez, Diana Andrea; Castilla-Rodríguez, Jaisel Luz; Castrejón-Vázquez, María Isabel; Vargas-Camaño, María Eugenia; Medina-Torres, Edgar Alejandro; Blancas-Galicia, Lizbeth; Espinosa-Padilla, Sara Elva

    2015-01-01

    Common variable immunodeficiency (CVID) is an heterogeneous group of disorders characterized by impaired antibody production. It shows a wide spectrum of manifestations including severe and recurrent respiratory infections (Streptococcus pneumoniae, Haemophilus) and gastrointestinal (Campylobacter jejuni, rotavirus and Giardia lamblia). Viral infections caused by herpes zoster, cytomegalovirus (CMV) and hepatitis C are rare. The opportunistic agents such as CMV, Pneumocystis jirovecii, cryptococcus and atypical mycobacteria have been reported as isolated cases. This paper reports the case of a 38-year-old female patient, who began six years before with weight loss of 7 kg in six months, fatigue, weakness, sweating, fever and abdominal pain. Furthermore, patient had intestinal obstruction and abdominal CT showed mesenteric lymph growth. The mesenteric lymph node biopsy revealed positives Mycobacterium PCR, Ziehl-Neelsen staining and culture for M. bovis. In the laparotomy postoperative period was complicated with nosocomial pneumonia, requiring mechanical ventilation and tracheostomy. Two years later, she developed right renal abscess that required surgical drainage, once again with a positive culture for Mycobacterium bovis. She was referred to highly specialized hospital and we documented panhypogammaglobulinemia and lymphopenia. Secondary causes of hypogammaglobulinemia were ruled out and common variable immunodeficiency (CVID) was confirmed, we started IVIG replacement. Four years later she developed mixed cellularity Hodgkin's lymphoma. Until today she continues with IVIG and chemotherapy. This report of a patient with CVID and Mycobacterium bovis infection, a unusual association, shows the cellular immunity susceptibility in this immunodeficiency, additional to the humoral defect.

  11. [Infection due to Mycobacterium bovis in common variable immunodeficiency].

    PubMed

    Herrera-Sánchez, Diana Andrea; Castilla-Rodríguez, Jaisel Luz; Castrejón-Vázquez, María Isabel; Vargas-Camaño, María Eugenia; Medina-Torres, Edgar Alejandro; Blancas-Galicia, Lizbeth; Espinosa-Padilla, Sara Elva

    2015-01-01

    Common variable immunodeficiency (CVID) is an heterogeneous group of disorders characterized by impaired antibody production. It shows a wide spectrum of manifestations including severe and recurrent respiratory infections (Streptococcus pneumoniae, Haemophilus) and gastrointestinal (Campylobacter jejuni, rotavirus and Giardia lamblia). Viral infections caused by herpes zoster, cytomegalovirus (CMV) and hepatitis C are rare. The opportunistic agents such as CMV, Pneumocystis jirovecii, cryptococcus and atypical mycobacteria have been reported as isolated cases. This paper reports the case of a 38-year-old female patient, who began six years before with weight loss of 7 kg in six months, fatigue, weakness, sweating, fever and abdominal pain. Furthermore, patient had intestinal obstruction and abdominal CT showed mesenteric lymph growth. The mesenteric lymph node biopsy revealed positives Mycobacterium PCR, Ziehl-Neelsen staining and culture for M. bovis. In the laparotomy postoperative period was complicated with nosocomial pneumonia, requiring mechanical ventilation and tracheostomy. Two years later, she developed right renal abscess that required surgical drainage, once again with a positive culture for Mycobacterium bovis. She was referred to highly specialized hospital and we documented panhypogammaglobulinemia and lymphopenia. Secondary causes of hypogammaglobulinemia were ruled out and common variable immunodeficiency (CVID) was confirmed, we started IVIG replacement. Four years later she developed mixed cellularity Hodgkin's lymphoma. Until today she continues with IVIG and chemotherapy. This report of a patient with CVID and Mycobacterium bovis infection, a unusual association, shows the cellular immunity susceptibility in this immunodeficiency, additional to the humoral defect. PMID:25758115

  12. Differentiation of Mycobacterium bovis isolates from animals by DNA typing.

    PubMed Central

    Skuce, R A; Brittain, D; Hughes, M S; Neill, S D

    1996-01-01

    The insertion sequence IS6110 and the direct repeat (DR) specific to tuberculosis complex mycobacteria and the highly repeated DNA sequence, the polymorphic GC-rich repeat sequence (PGRS), were systematically used to identify restriction fragment length polymorphisms (RFLPs) within 210 isolates of Mycobacterium bovis. The isolates were primarily of bovine origin, but isolates from badgers, feral deer, sheep, humans, and a pig were included. The RFLP probes IS6110, DR, and PGRS individually identified 17, 18, and 18 different RFLP types, respectively, but in combination these probes identified a total of 39 different M. bovis RFLP types. The recommendations (J. D. A. van Embden, M. D. Cave, J. T. Crawford, J. W. Dale, K. D. Eisenach, B. Gicquel, P. W. M. Hermans, C. Martin, R. McAdam, T. M. Shinnick, and P. M. Small, J. Clin. Microbiol. 31:406-409, 1993) for a standardized RFLP analysis for M. tuberculosis were adapted to facilitate gel documentation, image analysis, and construction of a database of RFLP types. In the present study the same M. bovis RFLP types were evident in the various animal species included, indicating that the strains were not host restricted. Application of these techniques to defined field studies should help elucidate more accurately aspects of the epidemiology of bovine tuberculosis in different countries. PMID:8880502

  13. The transmission of Mycobacterium bovis infection to cattle.

    PubMed

    Phillips, C J C; Foster, C R W; Morris, P A; Teverson, R

    2003-02-01

    The prevalence of Mycobacterium bovis infection in cattle is increasing rapidly in some countries, including the UK and Ireland. The organism infects a wide range of mammalian hosts, and eradication of the disease is difficult if there is an extensive reservoir in the wildlife population. Existing evidence suggests that wildlife vectors include the European badger in the UK and Ireland, the brush-tailed possum and ferret in New Zealand and ungulates in some other countries. Cattle grazing field boundaries or short swards are at particularly high risk, since the chance of contact with the intermediate host or their excreta is increased. There is evidence that the transmission of the disease between cattle following movement accounts for 10-15% of outbreaks in the British Isles and that transmission can occur across farm boundaries. The prevalence the prevalence of single reactors in herds suggested that within-herd transmission was not common. In herds with infected cattle, spreading slurry is a risk factor, which can be minimised by prolonged storage of the slurry, by spreading it on fields not used for grazing or by soil injection. M. bovis also survives in water and may enter the respiratory tract during drinking. It is concluded that M. bovis infection in cattle can be transmitted by a number of routes, some of which can be controlled by appropriate husbandry, but that circumstantial evidence suggests that the existence of a widespread intermediate host is the greatest contributor to infection in cattle.

  14. Bovine Tuberculosis (Mycobacterium bovis) in Wildlife in Spain

    PubMed Central

    Aranaz, Alicia; de Juan, Lucía; Montero, Natalia; Sánchez, Celia; Galka, Margarita; Delso, Consuelo; Álvarez, Julio; Romero, Beatriz; Bezos, Javier; Vela, Ana I.; Briones, Victor; Mateos, Ana; Domínguez, Lucas

    2004-01-01

    Mycobacterium bovis infection in wildlife and feral species is a potential source of infection for livestock and a threat to protected and endangered species. The aim of this study was to identify Spanish wild animal species infected with M. bovis through bacteriological culture and spacer oligonucleotide typing (spoligotyping) of isolates for epidemiological purposes. This study included samples from red deer (Cervus elaphus), fallow deer (Dama dama), wild boar (Sus scrofa), Iberian lynx (Lynx pardina), hare (Lepus europaeus), and cattle (Bos taurus). They were collected in several geographical areas that were selected for their unique ecological value and/or known relationships between wildlife and livestock. In the areas included in this survey, M. bovis strains with the same spoligotyping pattern were found infecting several wild species and livestock, which indicates an epidemiological link. A locally predominant spoligotype was found in these areas. Better understanding of the transmission and distribution of disease in these populations will permit more precise targeting of control measures. PMID:15184440

  15. Molecular phylogeny of Babesia poelea from brown boobies (Sula leucogaster) from Johnston Atoll, central Pacific

    USGS Publications Warehouse

    Yabsley, M.J.; Work, T.M.; Rameyer, R.A.

    2006-01-01

    The phylogenetic relationship of avian Babesia with other piroplasms remains unclear, mainly because of a lack of objective criteria such as molecular phylogenetics. In this study, our objective was to sequence the entire 18S, ITS-1, 5.8S, and ITS-2 regions of the rRNA gene and partial ??-tubulin gene of B. poelea, first described from brown boobies (Sula leucogaster) from the central Pacific, and compare them to those of other piroplasms. Phylogenetic analyses of the entire 18S rRNA gene sequence revealed that B. poelea belonged to the clade of piroplasms previously detected in humans, domestic dogs, and wild ungulates in the western United States. The entire ITS-1, 5.8S, ITS-2, and partial ??-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the ??-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm. ?? American Society of Parasitologists 2006.

  16. Molecular phylogeny of Babesia poelea from brown boobies (Sula leucogaster) from Johnston Atoll, Central Pacific

    USGS Publications Warehouse

    Yabsley, Michael J.; Work, Thierry M.; Rameyer, Robert A.

    2006-01-01

    The phylogenetic relationship of avian Babesia with other piroplasms remains unclear, mainly because of a lack of objective criteria such as molecular phylogenetics. In this study, our objective was to sequence the entire 18S, ITS-1, 5.8S, and ITS-2 regions of the rRNA gene and partial β-tubulin gene of B. poelea, first described from brown boobies (Sula leucogaster) from the central Pacific, and compare them to those of other piroplasms. Phylogenetic analyses of the entire 18S rRNA gene sequence revealed that B. poelea belonged to the clade of piroplasms previously detected in humans, domestic dogs, and wild ungulates in the western United States. The entire ITS-1, 5.8S, ITS-2, and partial β-tubulin gene sequence shared conserved regions with previously described Babesia and Theileria species. The intron of the β-tubulin gene was 45 bp. This is the first molecular characterization of an avian piroplasm.

  17. Babesia odocoilei infection in a Saskatchewan elk (Cervus elaphus canadensis) herd.

    PubMed

    Pattullo, Kimberly M; Wobeser, Gary; Lockerbie, Betty P; Burgess, Hilary J

    2013-07-01

    An 8-year-old female elk (Cervus elaphus canadensis) cow, presented for chronic severe weight loss and unthriftiness, was diagnosed with Babesia odocoilei infection based on blood smear evaluation, polymerase chain reaction (PCR), and DNA sequence analysis. Subsequently, velvet antler from a male that died acutely on the same farm was also PCR positive for Babesia spp. Both animals originated from a game ranch of Saskatchewan-bred and -raised animals with no known history of tick exposure, but with a history of numerous sudden deaths of unknown etiology. The presence of B. odocoilei in Canada might be a result of a recent introduction that could have deleterious effects on local wild ungulates or may represent discovery of a previously unrecognized endemic disease in local wildlife.

  18. First molecular identification of Babesia gibsoni in dogs from Slovakia, central Europe.

    PubMed

    Víchová, Bronislava; Horská, Mária; Blaňarová, Lucia; Švihran, Milan; Andersson, Martin; Peťko, Branislav

    2016-02-01

    Canine babesiosis is a severe and potentially life threatening infection. In Europe, Babesia canis is considered to be the most common species responsible for the disease. We report two cases of babesiosis caused by Babesia gibsoni. The polymerase chain reaction, restriction fragment length polymorphism analysis and further sequencing of 18S rRNA gene fragments from blood samples of both dogs revealed the identity of isolates with B. gibsoni genotypes from other dogs worldwide. This species was previously not known to infect dogs in Slovakia. It is resistant to traditional anti-babesial therapy. Therefore, correct diagnosis is crucial for the successful treatment, especially in dogs with hemolytic anemia and febrile conditions. PMID:26304014

  19. Molecular Characterization of a Non–Babesia divergens Organism Causing Zoonotic Babesiosis in Europe

    PubMed Central

    Cacciò, Simone; Gherlinzoni, Filippo; Aspöck, Horst; Slemenda, Susan B.; Piccaluga, PierPaolo; Martinelli, Giovanni; Edelhofer, Renate; Hollenstein, Ursula; Poletti, Giovanni; Pampiglione, Silvio; Löschenberger, Karin; Tura, Sante; Pieniazek, Norman J.

    2003-01-01

    In Europe, most reported human cases of babesiosis have been attributed, without strong molecular evidence, to infection with the bovine parasite Babesia divergens. We investigated the first known human cases of babesiosis in Italy and Austria, which occurred in two asplenic men. The complete 18S ribosomal RNA (18S rRNA) gene was amplified from specimens of their whole blood by polymerase chain reaction (PCR). With phylogenetic analysis, we compared the DNA sequences of the PCR products with those for other Babesia spp. The DNA sequences were identical for the organism from the two patients. In phylogenetic analysis, the organism clusters with B. odocoilei, a parasite of white-tailed deer; these two organisms form a sister group with B. divergens. This evidence indicates the patients were not infected with B. divergens but with an organism with previously unreported molecular characteristics for the 18S rRNA gene. PMID:12967491

  20. Vertical Transmission of Babesia microti in BALB/c Mice: Preliminary Report

    PubMed Central

    Bednarska, Malgorzata; Bajer, Anna; Drozdowska, Anna; Mierzejewska, Ewa J.; Tolkacz, Katarzyna; Welc-Falęciak, Renata

    2015-01-01

    Babesia spp. (Apicomplexa, Piroplasmida) are obligate parasites of many species of mammals, causing a malaria-like infection- babesiosis. Three routes of Babesia infection have been recognized to date. The main route is by a tick bite, the second is via blood transfusion. The third, vertical route of infection is poorly recognized and understood. Our study focused on vertical transmission of B. microti in a well-established mouse model. We assessed the success of this route of infection in BALB/c mice with acute and chronic infections of B. microti. In experimental groups, females were mated on the 1st day of Babesia infection (Group G0); on the 28th day post infection (dpi) in the post- acute phase of the parasite infection (G28); and on the 90th and 150th dpi (G90 and G150 group, respectively), in the chronic phase of the parasite infection. Pups were obtained from 58% of females mated in the post-acute phase (G28) and from 33% of females in groups G90 and G150. Mice mated in the pre-acute phase of infection (G0) did not deliver pups. Congenital B. microti infections were detected by PCR amplification of Babesia 18S rDNA in almost all pups (96%) from the experimental groups G28, G90 and G150. Parasitaemia in the F1 generation was low and varied between 0.01–0.001%. Vertical transmission of B. microti was demonstrated for the first time in BALB/c mice. PMID:26372043

  1. In situ cytokine expression in pulmonary granulomas of cattle experimentally infected by aerosolized Mycobacterium bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium bovis is the cause of tuberculosis in most animal species, including cattle and is a serious zoonotic pathogen. In humans, M. bovis infection can result in disease clinically indistinguishable from that caused by Mycobacterium tuberculosis, the cause of most tuberculosis in humans. Reg...

  2. Effect of skin test on serum antibody responses to Mycobacterium bovis infection in cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recently, several serologic tests designed to detect immunodominant antibodies to M. bovis antigens (e.g., MPB83, MPB70, ESAT-6, and CFP10) have emerged for potential use with samples from cattle. Of these, a commercial ELISA to MPB83/MPB70 (M. bovis antibody ELISA) has gained approval for use in ca...

  3. Polymorphisms of twenty regulatory proteins between Mycobacterium tuberculosis and Mycobacterium bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycobacterium tuberculosis and Mycobacterium bovis are responsible for tuberculosis in humans or animals, respectively. Both species are closely related and belong to the Mycobacterium tuberculosis complex (MTC). M. tuberculosis is the most ancient species from which M. bovis and the other members o...

  4. A multilocus sequence typing method and curated database for Mycoplasma bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant problem in bison, causing necrotic pha...

  5. Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant health problem in bison, causing necro...

  6. Complete Genome Sequences of Field Isolates of Mycobacterium bovis and Mycobacterium caprae.

    PubMed

    de la Fuente, José; Díez-Delgado, Iratxe; Contreras, Marinela; Vicente, Joaquín; Cabezas-Cruz, Alejandro; Manrique, Marina; Tobes, Raquel; López, Vladimir; Romero, Beatriz; Domínguez, Lucas; Garrido, Joseba M; Juste, Ramón; Gortazar, Christian

    2015-06-25

    Here we report the complete genome sequences of field isolates of Mycobacterium bovis and the related mycobacterial species, Mycobacterium caprae. The genomes of three M. bovis (MB1, MB3, MB4) and one M. caprae (MB2) field isolates with different virulence, prevalence, and host distribution phenotypes were sequenced.

  7. Mycobacterium bovis Infection in Humans and Cats in Same Household, Texas, USA, 2012

    PubMed Central

    Lyashchenko, Konstantin P.; Greenwald, Rena; Robbe-Austerman, Suelee; McManis, Cynthia; Waters, W. Ray

    2015-01-01

    Mycobacterium bovis infection of cats is exceedingly rare in regions where bovine tuberculosis is not endemic. We describe the diagnosis and clinical management of pulmonary M. bovis infection in 2 indoor-housed cats and their association with at least 1 M. bovis–infected human in Texas, USA, in September 2012. PMID:25695666

  8. Rapid dissemination of Mycobacterium bovis from cattle dung to soil by the earthworm Lumbricus terrestris.

    PubMed

    Barbier, Elodie; Chantemesse, Benoit; Rochelet, Murielle; Fayolle, Léon; Bollache, Loïc; Boschiroli, Maria Laura; Hartmann, Alain

    2016-04-15

    Indirect transmission of Mycobacterium bovis, the causative agent of bovine tuberculosis (bTB), between wildlife and livestock is thought to occur by inhalation or ingestion of environmental substrates contaminated through animal shedding. The role of the soil fauna, such as earthworms, in the circulation of M. bovis from contaminated animal feces is of interest in the epidemiology of bTB. The objective of this study was to assess the impact of earthworm activity on M. bovis transfer from animal dung to castings and the surrounding soil. For this purpose, microcosms of soil containing the anecic earthworms Lumbricus terrestris were prepared and covered with cattle feces spiked with the M. bovis BCG strain Pasteur to carry out two separate experiments. The dissemination, the gut carriage and the excretion of M. bovis were all monitored using a specific qPCR-based assay. Our results showed that the earthworm L. terrestris was able to rapidly disseminate M. bovis from the contaminated cattle feces to the surrounding soil through casting egestion. Moreover, contaminated earthworms were shown to shed the bacteria for 4 days when transferred to a M. bovis-free soil. This study highlights for the first time the possible role of earthworms in the dissemination and the persistence of M. bovis in soils within bTB endemic areas. PMID:27016750

  9. A study on ovine tick-borne hemoprotozoan parasites (Theileria and Babesia) in the East Black Sea Region of Turkey.

    PubMed

    Altay, Kursat; Dumanli, Nazir; Aktas, Munir

    2012-07-01

    In this study, the frequency of Theileria and Babesia species was assessed via reverse line blotting and blood smear-based diagnostic methods in small ruminants. A total of 201 apparently healthy animals from 26 randomly selected herds located in 4 locations (Artvin, Giresun, Gumushane, and Tokat) of East Black Sea Region of Turkey were investigated for the blood protozoans. In a polymerase chain reaction (PCR), the hypervariable V4 region of the 18S ribosomal RNA gene was amplified with a set of general primers specific for all Theileria and Babesia species. The PCR products were hybridized against catchall and species-specific (Theileria spp., Theileria lestoquardi, Theileria ovis, Theileria sp. OT1, Theileria sp., OT3, Theileria sp., MK, Theileria luwenshuni, Theileria uilenbergi, Babesia spp., Babesia ovis, Babesia motasi, and Babesia crassa) probes. Theileria piroplasms were identified in nine (4.47%) samples by microscopic examination. Reverse line blotting (RLB) detected the infection in 19.90% of the samples. The infection rate of sheep (28.90%) was higher than goats (4.10%). T. ovis, Theileria sp., MK, and Theileria sp. OT3 were detected by RLB. The most prevalent Theileria species was T. ovis (18.90%) followed by Theileria sp. MK (0.99%). Theileria sp. OT3 was detected in one sample (0.43%). A single animal was infected as mix with T. ovis and Theileria sp. MK. The other Theileria (T. lestoquardi, Theileria sp. OT1, T. luwenshuni, and T. uilenbergi) and Babesia (B. ovis, B. motasi, and B. crassa) species were not detected. This study is the first molecular survey on ovine tick-borne protozoans in East Black Sea Region of Turkey.

  10. Genetic diversity and molecular characterization of Babesia motasi-like in small ruminants and ixodid ticks from China.

    PubMed

    Niu, Qingli; Liu, Zhijie; Yang, Jifei; Yu, Peifa; Pan, Yuping; Zhai, Bintao; Luo, Jianxun; Yin, Hong

    2016-07-01

    Ovine babesioses, an important tick-borne disease of sheep and goats in China, is caused by the reproduction of intraerythrocytic protozoa of the Babesia genus. Babesia motasi-like is a Babesia parasite that infects small ruminant in China, and two sub-groups of B. motasi-like can be subdivided based on differences in the rhoptry-associated-protein-1 gene. This study aimed to characterize the distribution, epidemiology and genetics of B. motasi-like in animals and ticks. A molecular investigation was carried out from 2009 to 2015 in 16 provinces in China. In total, 1081 blood samples were collected from sheep and goats originating from 27 different regions, and 778 ixodid tick samples were collected from 8 regions; the samples were tested for the presence of B. motasi-like using a specific nested PCR assay based on the rap-1b gene. The results indicated that 139 (12.9%), 91 (8.4%), 48 (4.4%) and 6 (0.7%) of the blood samples were positive for general B. motasi-like, Babesia sp. BQ1 (Lintan and Ningxian), Babesia sp. Tianzhu and Babesia sp. Hebei sub-groups, mixed infections, respectively. Among the collected 778 ixodid ticks (including Haemaphysalis longicornis, Haemaphysalis qinghaiensis, Dermacentor silvarum, Ixodes persulcatus, Rhipicephalus sanguineus and Rhipicephalus (Boophilus) microplus), the most frequently infected with Babesia were D. silvarum and I. persulcatus (35.7%), followed by H. longicornis (26.8%), H. qinghaiensis (24.8%) and R. sanguineus (9.3%). The PCR results were confirmed by DNA sequencing. The positive rates of B. motasi-like infection in ticks were found to be higher in China, compared with previous studies in other countries. B. motasi-like infections have not previously been reported in D. silvarum, I. persulcatus or R. sanguineus. The findings obtained in this study could be used for planning effective control strategies against babesiosis in China. PMID:26976477

  11. Molecular characterization of Babesia vogeli in dogs from Belém, northern Brazil.

    PubMed

    Moraes, P H G; Rufino, C P; Baraúna, A R F; Reis, T; Agnol, L T D; Meneses, A M C; Aguiar, D C F; Nunes, M R T; Gonçalves, E C

    2015-01-01

    Babesiosis is an infectious hemolytic disease that occurs worldwide, and is caused by a protozoan of the Babesia genus (Apicomplexa). Little is known about this genus; therefore, this study conducted a molecular characterization of Babesia spp in naturally infected dogs in northern Brazil. Blood samples were collected from 172 dogs from metropolitan Belém, and screened for Babesia spp using semi-nested polymerase chain reactions. Subsequently, 18S rDNA gene fragments were amplified and sequenced. Alignments of the 27 nucleotide sequences returned fragments measuring 1023 bp in size, which did not include any polymorphic sites (100% identity). This genotype was very similar to the 18S rDNA gene in B. vogeli. This study provides an important molecular characterization of the parasite responsible for canine babesiosis in naturally infected dogs in metropolitan Belém. In addition, the 18S rDNA gene in B. vogeli may represent the most common genotype occurring in South America. PMID:26662431

  12. Identification and Characterization of the RouenBd1987 Babesia divergens Rhopty-Associated Protein 1

    PubMed Central

    Rodriguez, Marilis; Alhassan, Andy; Ord, Rosalynn L.; Cursino-Santos, Jeny R.; Singh, Manpreet; Gray, Jeremy; Lobo, Cheryl A.

    2014-01-01

    Human babesiosis is caused by one of several babesial species transmitted by ixodid ticks that have distinct geographical distributions based on the presence of competent animal hosts. The pathology of babesiosis, like malaria, is a consequence of the parasitaemia which develops through the cyclical replication of Babesia parasites in a patient's red blood cells, though symptoms typically are nonspecific. We have identified the gene encoding Rhoptry-Associated Protein −1 (RAP-1) from a human isolate of B. divergens, Rouen1987 and characterized its protein product at the molecular and cellular level. Consistent with other Babesia RAP-1 homologues, BdRAP-1 is expressed as a 46 kDa protein in the parasite rhoptries, suggesting a possible role in red cell invasion. Native BdRAP-1 binds to an unidentified red cell receptor(s) that appears to be non-sialylated and non-proteinacious in nature, but we do not find significant reduction in growth with anti-rRAP1 antibodies in vitro, highlighting the possibility the B. divergens is able to use alternative pathways for invasion, or there is an alternative, complementary, role for BdRAP-1 during the invasion process. As it is the parasite's ability to recognize and then invade host cells which is central to clinical disease, characterising and understanding the role of Babesia-derived proteins involved in these steps are of great interest for the development of an effective prophylaxis. PMID:25226276

  13. Molecular and serological detection of Babesia spp. in neotropical and exotic carnivores in Brazilian zoos.

    PubMed

    André, Marcos Rogério; Adania, Cristina Harumi; Teixeira, Rodrigo Hidalgo Friciello; Allegretti, Silmara Marques; Machado, Rosangela Zacarias

    2011-03-01

    Large and small piroplasms have been observed in the blood smears of various wild carnivores, but few studies utilizing molecular characterization have been done. The goal of this present study was to investigate the presence of Babesia sp. by molecular and serologic techniques in exotic and neotropical carnivores maintained in captivity at Brazilian zoos. Blood and sera samples were collected from 146 Brazilian wild felids, 21 exotic felids, 1 genet (Genetta tigrina), 3 European wolves (Canis lupus), and 94 Brazilian wild canids in Brazilian zoos in the São Paulo and Mato Grosso states and in the Federal District. A total of 53 wild felids (31.74%) and 10 wild canids (10.31%) were seropositive for Babesia canis by Indirect Fluorescent Antibody Test (IFAT). Antibodies were detected in ocelots, little-spotted cats, margays, pampas cats, jaguars, pumas, jaguarundis, crab-eating foxes, and bush dogs. Babesia sp. DNA, with high similarity to B. leo, was detected in one pampas cat and one genet.

  14. A Cysteine Protease Is Critical for Babesia spp. Transmission in Haemaphysalis Ticks

    PubMed Central

    Tsuji, Naotoshi; Miyoshi, Takeharu; Battsetseg, Badger; Matsuo, Tomohide; Xuan, Xuenan; Fujisaki, Kozo

    2008-01-01

    Vector ticks possess a unique system that enables them to digest large amounts of host blood and to transmit various animal and human pathogens, suggesting the existence of evolutionally acquired proteolytic mechanisms. We report here the molecular and reverse genetic characterization of a multifunctional cysteine protease, longipain, from the babesial parasite vector tick Haemaphysalis longicornis. Longipain shares structural similarity with papain-family cysteine proteases obtained from invertebrates and vertebrates. Endogenous longipain was mainly expressed in the midgut epithelium and was specifically localized at lysosomal vacuoles and possibly released into the lumen. Its expression was up-regulated by host blood feeding. Enzymatic functional assays using in vitro and in vivo substrates revealed that longipain hydrolysis occurs over a broad range of pH and temperature. Haemoparasiticidal assays showed that longipain dose-dependently killed tick-borne Babesia parasites, and its babesiacidal effect occurred via specific adherence to the parasite membranes. Disruption of endogenous longipain by RNA interference revealed that longipain is involved in the digestion of the host blood meal. In addition, the knockdown ticks contained an increased number of parasites, suggesting that longipain exerts a killing effect against the midgut-stage Babesia parasites in ticks. Our results suggest that longipain is essential for tick survival, and may have a role in controlling the transmission of tick-transmittable Babesia parasites. PMID:18483546

  15. Cytokine expression in lungs of calves spontaneously infected with Mycoplasma bovis.

    PubMed

    Rodríguez, Francisco; González, Jorge F; Arbelo, Manuel; Zucca, Daniele; Fernández, Antonio

    2015-03-01

    Cytokine expression in the lung can play an important role during Mycoplasma bovis infection through leukocyte recruitment and activation, and the induction of a broad array of inflammatory mediators. To gain further insight into the pathogenesis of M. bovis-associated pneumonia, cytokine expression was examined, by immunohistochemical methods in formalin-fixed, paraffin wax-embedded tissues, in the lung of 20 calves spontaneously infected. Immunolabelling for tumor necrosis factor alpha (TNF)-α, interleukin-4 (IL-4), IL-10 and interferon-gamma (IFN-γ), was usually associated with pneumonia, particularly in macrophages and lymphocytes, and with the presence of M. bovis antigen. The expression was minimal in lungs from negative controls. The results demonstrated consistent upregulation of TNF-α, IL-4, IL-10 and IFN-γ expression during M. bovis-associated pneumonic lesions. These cytokines can participate in the immune and inflammatory responses during the pulmonary defense mechanisms against M. bovis infection. PMID:25331253

  16. Absence of Mycobacterium bovis in feral swine (Sus scrofa) from the southern Texas border region.

    PubMed

    Campbell, Tyler A; Long, David B; Bazan, Luis R; Thomsen, Bruce V; Robbe-Austerman, Suelee; Davey, Ronald B; Soliz, Liza A; Swafford, Seth R; VerCauteren, Kurt C

    2011-10-01

    Free-ranging wildlife, such as feral swine (Sus scrofa), harbor a variety of diseases that are transmissible to livestock and could negatively impact agricultural production. Information is needed regarding the exposure and infection rates of Mycobacterium bovis and many other diseases and parasites in feral swine occurring in the Texas border region. Our main objective was to determine exposure rates and possible infection rates of M. bovis in feral swine by opportunistically sampling animals from the Texas border region. From June to September 2010, we obtained samples from 396 feral swine and tested 98 samples for M. bovis by histopathology and mycobacteriologic culture. We found no evidence of M. bovis infection. We believe that it is important to periodically and strategically sample feral swine for M. bovis in high-risk areas of the United States because they are capable of becoming reservoirs of the disease.

  17. Development of a specific DNA probe and PCR for the detection of Mycoplasma bovis.

    PubMed

    Ghadersohi, A; Coelen, R J; Hirst, R G

    1997-05-01

    Mycoplasma bovis is responsible for several production diseases in cattle, including mastitis, arthritis, pneumonia, abortion and infertility. Current methodologies for detecting and identifying M. bovis are time consuming and difficult. Tests which rely on antigen or antibody detection have poor sensitivity and specificity. In this paper associated protocols for the development of a hybridization probe and PCR are described. A genomic library (SauIIIA digested) was prepared from M. bovis DNA (Colindale Reference Strain: NC10131:02) and cloned into pUC19. Colony hybridization, using a probe preparation made from purified M. bovis DNA, was used to identify colonies of interest. M. bovis DNA fragments were retrieved from recombinant plasmids by digestion with EcoRI and HindIII. This DNA was used to prepare randomly primed probes for dot blot hybridization analysis with immobilized DNA from M. bovis (two strains), M. dispar, M. agalactiae, M. bovigenitalium (two strains), M. ovipneumoniae, a Group 7 strain, M. arginini and bacteria belonging to different genera. Four probes were found to hybridize only with M. bovis and M. ovipneumoniae DNA, whereas one probe reacted with genomic DNA from only one of the two M. bovis strains. The level of sensitivity of the dot blot hybridization assay was 200 CFU (colony forming units)/mL. To enhance the sensitivity further, an M. bovis-specific PCR assay was developed. The primers were designed using sequences obtained from the probe DNA which discriminated M. bovis from all other Mycoplasma DNA tested. The minimum amount of target DNA that could be detected by the PCR assay was that isolated from 10-20 CFU/mL. The PCR assay was therefore 10 times more sensitive than dot blot hybridization.

  18. Oral vaccination of guinea pigs with a Mycobacterium bovis bacillus Calmette-Guerin vaccine in a lipid matrix protects against aerosol infection with virulent M. bovis.

    PubMed

    Clark, Simon; Cross, Martin L; Nadian, Allan; Vipond, Julia; Court, Pinar; Williams, Ann; Hewinson, R Glyn; Aldwell, Frank E; Chambers, Mark A

    2008-08-01

    Increased incidence of bovine tuberculosis (TB) in the United Kingdom caused by infection with Mycobacterium bovis is a cause of considerable economic loss to farmers and the government. The Eurasian badger (Meles meles) represents a wildlife source of recurrent M. bovis infections of cattle in the United Kingdom, and its vaccination against TB with M. bovis bacillus Calmette-Guérin (BCG) is an attractive disease control option. Delivery of BCG in oral bait holds the best prospect for vaccinating badgers over a wide geographical area. Using a guinea pig pulmonary challenge model, we evaluated the protective efficacy of candidate badger oral vaccines, based on broth-grown or ball-milled BCG, delivered either as aqueous suspensions or formulated in two lipids with differing fatty acid profiles (one being animal derived and the other being vegetable derived). Protection was determined in terms of increasing body weight after aerosol challenge with virulent M. bovis, reduced dissemination of M. bovis to the spleen, and, in the case of one oral formulation, restricted growth of M. bovis in the lungs. Only oral BCG formulated in lipid gave significant protection. These data point to the potential of the BCG-lipid formulation for further development as a tool for controlling tuberculosis in badgers.

  19. Epidemiological studies on Schistosoma bovis in Iringa Region, Tanzania.

    PubMed

    Kassuku, A; Christensen, N O; Monrad, J; Nansen, P; Knudsen, J

    1986-06-01

    Various aspects of the epidemiology of Schistosoma bovis were studied over a one-year period in Iringa Region, Tanzania. An abattoir survey revealed an overall prevalence rate of 30.8% in cattle and 3.8% in goats in the area, and field studies on two dairy farms both providing good opportunities for schistosome transmission provided information concerning the transmission ecology of S. bovis in relation to different types of grazing and water supply. The traditional management system on one farm with a large number of cattle utilizing a limited water resource highly suitable for sustaining populations of the snail host Bulinus africanus resulted in intensive transmission as evidenced by uptake of massive infections in calves and development of resistance to S. bovis challenge in dairy cows. On another farm, appropriate management comprising watering of cattle at a B. africanus-free pond provided the background for less intensive transmission in that transmission risk was confined to occasional contact with water contact sites of secondary importance. Besides, the transmission pattern as regards intensity and seasonality was affected markedly by the geographical and seasonal distribution of the host snail B. africanus. Thus, transmission in canals and temporary ponds was limited mainly to the dry season and the end of the rainy season, respectively, while transmission in permanent ponds occurred intermittently throughout at least most of the year. It is concluded that prevention of severe loss of productivity in domestic ruminants due to schistosome infections should be possible using strategic management procedures provided that essential information is available concerning the pattern of transmission in the particular area. PMID:2874712

  20. Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil.

    PubMed

    Gottlieb, Juliana; André, Marcos Rogério; Soares, João Fábio; Gonçalves, Luiz Ricardo; Tonial de Oliveira, Mateus; Costa, Marcio Machado; Labruna, Marcelo Bahia; Bortolini, Carlos Eduardo; Machado, Rosangela Zacarias; Vieira, Maria Isabel Botelho

    2016-06-14

    Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed. PMID:27334817

  1. Rangelia vitalii, Babesia spp. and Ehrlichia spp. in dogs in Passo Fundo, state of Rio Grande do Sul, Brazil.

    PubMed

    Gottlieb, Juliana; André, Marcos Rogério; Soares, João Fábio; Gonçalves, Luiz Ricardo; Tonial de Oliveira, Mateus; Costa, Marcio Machado; Labruna, Marcelo Bahia; Bortolini, Carlos Eduardo; Machado, Rosangela Zacarias; Vieira, Maria Isabel Botelho

    2016-06-14

    Pathogens transmitted by ticks are an emerging problem worldwide, this study aimed to diagnose the causal agents of infection in dogs presenting suspected hemoparasitoses. Fifty-eight dogs with clinical signs such as depression, hemorrhagic diathesis and fever were evaluated regarding clinical presentation, hemogram, blood smears and serological tests, using the indirect immunofluorescence method for the agents Babesia vogeli and Ehrlichia canis and conventional PCR for Babesia spp. (gene 18S rRNA), Rangelia vitalii (gene 18S rRNA) and Ehrlichia spp. (gene dsb). Five (8.6%) of the 58 dogs were serologically positive for Babesia spp. and three (5.1%) for E. canis. Four dogs (6.8%) were positive for R. vitalii through the molecular diagnosis. The PCR products were sequenced and the DNA from R. vitalii was found to be 99% genetically identical to samples of R. vitalii that had been isolated in Brazil. No presence of Babesia spp. or E. canis was observed through PCR on the dogs evaluated here. The results indicate the presence of R. vitalii and exposure to Babesia spp. and Ehrlichia spp. among the dogs analyzed. PMID:27304518

  2. Sympatric occurrence of Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks and Rickettsia and Babesia species in Slovakia.

    PubMed

    Svehlová, Andrea; Berthová, Lenka; Sallay, Balázs; Boldiš, Vojtech; Sparagano, Olivier A E; Spitalská, Eva

    2014-09-01

    Vojka nad Dunajom in the south-west of the Slovak Republic is a locality with sympatric occurrence of 3 species of ticks. This study investigated the spatial distribution of Dermacentor reticulatus, Ixodes ricinus, and Haemaphysalis concinna ticks in this area and determined the prevalence of Babesia and Rickettsia species in questing adults of these tick species considered as potential risk for humans and animals. Ticks were collected by blanket dragging over the vegetation from September 2011 to October 2012. All ticks were subjected to DNA extraction and individually assayed with PCR-based methods targeting the gltA, sca4, 23S rRNA genes of Rickettsia spp. and the 18S rRNA gene of Babesia spp. D. reticulatus was the dominant species occurring in this area (67.7%, n=600), followed by I. ricinus (31.8%, n=282) and H. concinna (0.5%, n=4) ticks. Rickettsial infection was determined in 10.8% (n=65) and 11.7% (n=33) of D. reticulatus and I. ricinus ticks, respectively. Babesia spp. infection was confirmed in 1.8% (n=11) of D. reticulatus and 0.4% (n=1) of I. ricinus ticks. DNA of 6 different pathogenic tick-borne species, Rickettsia helvetica, Rickettsia monacensis, Rickettsia slovaca, Rickettsia raoultii, Babesia canis, and Babesia venatorum were identified in this locality with sympatric occurrence of I. ricinus, D. reticulatus, and H. concinna ticks.

  3. Mycobacterium bovis infection in a horse with granulomatous enterocolitis.

    PubMed

    Sarradell, Javier E; Alvarez, Julio; Biscia, Mariana; Zumarraga, Martin; Wunschmann, Arno; Armien, Anibal G; Perez, Andres M

    2015-03-01

    A 2-year-old dappled Percheron horse had a wasting condition that did not respond to antibiotic treatments and ultimately resulted in death. Thickening of the wall of the large colon and enlargement of the mesenteric lymph nodes were observed at postmortem examination, along with the presence of pinpoint whitish foci in the liver. Microscopic examination of affected tissues revealed diffuse chronic granulomatous enterocolitis, granulomatous mesenteric lymphadenitis, and multifocal granulomatous hepatitis. The DNA extracted from paraffin-embedded intestinal and lymph node samples was analyzed using both a polymerase chain reaction (PCR) assay and PCR-restriction endonuclease analysis and demonstrated the presence of Mycobacterium bovis.

  4. Intracranial myiasis by Hypoderma bovis (Linnaeus) in a horse.

    PubMed

    Hadlow, W J; Ward, J K; Krinsky, W L

    1977-04-01

    Acute neurologic disease associated with intracranial migration of a first instar larva of a warble fly, Hypoderma bovis (Linnaeus), was observed in a 14-year-old Quarter Horse gelding in western Montana. The disease was characterized by incoordination of gait, circling to the left, head tilt to the right, partial paralysis of the right side of the face, and impaired vision in the right eye. Two and one-half hours after it was first noticed sick, the horse collapsed and was euthanized. Massive hemorrhage unaccompanied by necrosis or significant cellular response was present in the right side of the midbrain and pons.

  5. Zoonotic tuberculosis due to Mycobacterium bovis in developing countries.

    PubMed Central

    Cosivi, O.; Grange, J. M.; Daborn, C. J.; Raviglione, M. C.; Fujikura, T.; Cousins, D.; Robinson, R. A.; Huchzermeyer, H. F.; de Kantor, I.; Meslin, F. X.

    1998-01-01

    The World Health Organization (WHO) estimates that human tuberculosis (TB) incidence and deaths for 1990 to 1999 will be 88 million and 30 million, respectively, with most cases in developing countries. Zoonotic TB (caused by Mycobacterium bovis) is present in animals in most developing countries where surveillance and control activities are often inadequate or unavailable; therefore, many epidemiologic and public health aspects of infection remain largely unknown. We review available information on zoonotic TB in developing countries, analyze risk factors that may play a role in the disease, review recent WHO activities, and recommend actions to assess the magnitude of the problem and control the disease in humans and animals. PMID:9452399

  6. Social group size affects Mycobacterium bovis infection in European badgers (Meles meles).

    PubMed

    Woodroffe, Rosie; Donnelly, Christl A; Wei, Gao; Cox, D R; Bourne, F John; Burke, Terry; Butlin, Roger K; Cheeseman, C L; Gettinby, George; Gilks, Peter; Hedges, Simon; Jenkins, Helen E; Johnston, W Thomas; McInerney, John P; Morrison, W Ivan; Pope, Lisa C

    2009-07-01

    1. In most social animals, the prevalence of directly transmitted pathogens increases in larger groups and at higher population densities. Such patterns are predicted by models of Mycobacterium bovis infection in European badgers (Meles meles). 2. We investigated the relationship between badger abundance and M. bovis prevalence, using data on 2696 adult badgers in 10 populations sampled at the start of the Randomized Badger Culling Trial. 3. M. bovis prevalence was consistently higher at low badger densities and in small social groups. M. bovis prevalence was also higher among badgers whose genetic profiles suggested that they had immigrated into their assigned social groups. 4. The association between high M. bovis prevalence and small badger group size appeared not to have been caused by previous small-scale culling in study areas, which had been suspended, on average, 5 years before the start of the current study. 5. The observed pattern of prevalence might occur through badgers in smaller groups interacting more frequently with members of neighbouring groups; detailed behavioural data are needed to test this hypothesis. Likewise, longitudinal data are needed to determine whether the size of infected groups might be suppressed by disease-related mortality. 6. Although M. bovis prevalence was lower at high population densities, the absolute number of infected badgers was higher. However, this does not necessarily mean that the risk of M. bovis transmission to cattle is highest at high badger densities, since transmission risk depends on badger behaviour as well as on badger density. PMID:19486382

  7. Histopathological and immunohistochemical study of lambs experimentally infected with Fasciola hepatica and Schistosoma bovis.

    PubMed

    Ferreras, M C; García-Iglesias, M J; Manga-González, M Y; Pérez-Martínez, C; Mizinska, Y; Ramajo, V; González-Lanza, M C; Escudero, A; García-Marín, J F

    2000-12-01

    The aim of this study was to investigate the cross-resistance between Fasciola hepatica and Schistosoma bovis in lambs assessing parasitologic, gross pathologic, histopathologic and immunohistochemical changes in liver and small intestine. Thirty Castellana breed lambs were divided into five comparable groups and exposed to F. hepatical S. bovis (group F/S), S. bovis/F. hepatica (group S/F), S. bovis (group S) or F. hepatica (group F) and six unexposed lambs were used as non-infected controls (group C). Primary patent infection with F. hepatica induced a lower number of schistosome eggs and a higher number of lymphocytes in intestinal and liver schistosome egg-induced granulomas in group F/S than in the groups S/F and S, liver damage being mainly attributed to F. hepatica. S. bovis infection followed by challenge with F. hepatica particularly increased the severity of the most significant liver alterations (cholangiohepatitis by F. hepatica and mesoendophlebitis by S. bovis) and F. hepatica seemed not to have an influence on established S. bovis infection. In addition, immunohistochemical results suggested that the predominant local immune response in both double-infected groups was different, being mainly a cell-mediated immune response in group F/S and a mucosal response in group S/F.

  8. Herd-specific strains of Mycoplasma bovis in outbreaks of mycoplasmal mastitis and pneumonia.

    PubMed

    Aebi, Marlis; Bodmer, Michèle; Frey, Joachim; Pilo, Paola

    2012-06-15

    Mycoplasma bovis causes severe economic losses in livestock production, particularly on the Northern American continent and more recently also in continental Europe. The aim of the current study was to evaluate whether the recently emerging outbreaks were due to a particular clone or strain of M. bovis or whether these outbreaks are due to multiple infectious strains of M. bovis. The study is based on the analysis M. bovis isolated from cattle of herds with outbreaks of mycoplasmal mastitis or pneumonia from geographically non related parts of Switzerland. M. bovis isolates were typed by insertion sequence (IS) element analysis based upon ISMbov1 and ISMbov2 southern-blot hybridization. We observed a strong divergence of M. bovis strains among affected herds which mostly were herd specific. This argues against the assumption that a recent infiltration of a particular clone of M. bovis is the cause of the perilous emerging outbreaks. The study suggests that transmission occurs from animal to animal most probably via milk. PMID:22306036

  9. Spatial relationship between Mycobacterium bovis strains in cattle and badgers in four areas in Ireland.

    PubMed

    Olea-Popelka, F J; Flynn, O; Costello, E; McGrath, G; Collins, J D; O'keeffe, J; Kelton, D F; Berke, O; Martin, S W

    2005-09-30

    We investigated whether strains (restriction fragment length polymorphism, RFLP-types) of Mycobacterium bovis isolated from badgers and from cattle clustered among and within four areas in Ireland. The spatial scan test and nearest-neighbor analysis were used as the spatial cluster-detection techniques. In addition, for each of the major strains, associations between the distance to badger setts and the "centroid" of the cattle farm were assessed in a logistic model. Overall, between September 1997 and May 2000, 316 and 287 M. bovis samples, from badgers and cattle, respectively, were strain-typed. The distribution of strains in badgers, and separately in cattle, differed among areas. Within each of the four large areas, badgers and cattle tended to have similar strains; this is consistent with the sharing of M. bovis strains within an area. In more detailed within-area analyses, some spatial clusters of M. bovis strains were detected, separately, in both cattle and badgers. Almost half of the infected badger setts with a specific strain were located outside of the "detected" clusters. There was no association between the number of infected badgers with a specific M. bovis strain within 2 or 5 km distances to cattle herds, and the risk of the same strain in cattle. We speculate about the dynamic nature of badger movements, as an explanation for the absence of more clusters of most of the strains of M. bovis isolated from badgers, and its impact on trying to study transmission of M. bovis between cattle and badger.

  10. Social group size affects Mycobacterium bovis infection in European badgers (Meles meles).

    PubMed

    Woodroffe, Rosie; Donnelly, Christl A; Wei, Gao; Cox, D R; Bourne, F John; Burke, Terry; Butlin, Roger K; Cheeseman, C L; Gettinby, George; Gilks, Peter; Hedges, Simon; Jenkins, Helen E; Johnston, W Thomas; McInerney, John P; Morrison, W Ivan; Pope, Lisa C

    2009-07-01

    1. In most social animals, the prevalence of directly transmitted pathogens increases in larger groups and at higher population densities. Such patterns are predicted by models of Mycobacterium bovis infection in European badgers (Meles meles). 2. We investigated the relationship between badger abundance and M. bovis prevalence, using data on 2696 adult badgers in 10 populations sampled at the start of the Randomized Badger Culling Trial. 3. M. bovis prevalence was consistently higher at low badger densities and in small social groups. M. bovis prevalence was also higher among badgers whose genetic profiles suggested that they had immigrated into their assigned social groups. 4. The association between high M. bovis prevalence and small badger group size appeared not to have been caused by previous small-scale culling in study areas, which had been suspended, on average, 5 years before the start of the current study. 5. The observed pattern of prevalence might occur through badgers in smaller groups interacting more frequently with members of neighbouring groups; detailed behavioural data are needed to test this hypothesis. Likewise, longitudinal data are needed to determine whether the size of infected groups might be suppressed by disease-related mortality. 6. Although M. bovis prevalence was lower at high population densities, the absolute number of infected badgers was higher. However, this does not necessarily mean that the risk of M. bovis transmission to cattle is highest at high badger densities, since transmission risk depends on badger behaviour as well as on badger density.

  11. Bartonella bovis isolated from a cow with endocarditis.

    PubMed

    Erol, Erdal; Jackson, Carney; Bai, Ying; Sells, Stephen; Locke, Steve; Kosoy, Michael

    2013-03-01

    A 7-year-old pregnant Angus cow was found dead in the field. At necropsy, the aortic valve was expanded by moderate fibrous connective tissue and acidophilic coagulum containing multifocal marked bacteria, mineral, neutrophils, and red blood cells. Numerous tiny grayish, opaque bacterial colonies were detected on blood agar plates at 7 days after inoculation with a swab of the heart valve of the cow. The bacterium was a Gram-negative, very small coccobacillus that was catalase, oxidase, and urease negative, and did not change litmus milk, triple sugar iron agar, and sulfide-indole-motility medium. The bacterium was negative for esculin hydrolysis, phenylalanine deaminase, nitrate reduction, and gelatin hydrolysis. The isolate did not produce acid from glycerol, inulin, lactose, maltose, mannose, raffinose, salicin, sorbitol, sucrose, trehalose, glycogen, ribose, or starch. Polymerase chain reaction tests for the gltA, ssrA, ftsZ, ribC, rpoB, and 16S ribosomal RNA genes of Bartonella species were positive for the isolate. Amplicons were sequenced, and the gltA, ribC, ssrA, and 16S ribosomal RNA gene sequences were found to have 100% homology to the type strain of Bartonella bovis, whereas the fts and rpoB sequences showed 99.9% and 99.6% homology, respectively, to the type strain of Bartonella bovis. Diagnosticians should be aware of slow-growing microorganisms, and culture media should be incubated beyond the standard period to enhance the recovery of Bartonella species.

  12. Alternative activation modifies macrophage resistance to Mycobacterium bovis.

    PubMed

    Castillo-Velázquez, Uziel; Aranday-Cortés, Elihú; Gutiérrez-Pabello, José A

    2011-07-01

    The aim of this study was to evaluate the influence of macrophage alternative activation in the intracellular pathogen natural disease resistance phenotype of the host. Macrophage monolayers from resistant (R) (3) or susceptible (S) (3) cattle donors were treated with 10 ng/ml of bovine recombinant IL-4 (rbIL-4), and infected with virulent and avirulent Mycobacterium bovis (MOI 10:1). Bactericidal assays were performed to assess the bacterial phagocytic index and intracellular survival. Total RNA was reverse transcribed and used to analyze the relative changes in gene expression of IL-10, IL-12, IL-18 IL-1β, TNF-α, MCP-1, MCP-2, IL-6, MIP-1, MIP-3, iNOS, ARGII and SLAM by real time PCR. Cell supernatants were collected and nitric oxide and arginase production was assessed. Apoptosis induction was measured by TUNEL. IL-4 treatment increased the phagocytic index in both R and S macrophages; however intracellular survival was augmented mainly in S macrophages. Alternative activation decreased gene expression of pro-inflammatory cytokines, nitric oxide production and DNA fragmentation mainly in R macrophages. On the other hand, arginase production was not different between R and S macrophages. Alternative activation modifies the macrophage response against M. bovis. IL-4 treatment minimized the functional differences that exist between R and S macrophages.

  13. Surveillance of a Ventilated Rack System for Corynebacterium bovis by Sampling Exhaust-Air Manifolds

    PubMed Central

    Manuel, Christopher A; Pugazhenthi, Umarani; Leszczynski, Jori K

    2016-01-01

    Corynebacterium bovis causes an opportunistic infection of nude (Foxn1, nu/nu) mice, leading to nude mouse hyperkeratotic dermatitis (scaly skin disease). Enzootic in many nude mouse colonies, C. bovis spreads rapidly to naive nude mice, despite modern husbandry practices, and is very difficult to eradicate. To facilitate rapid detection in support of eradication efforts, we investigated a surveillance method based on quantitative real-time PCR (qPCR) evaluation of swabs collected from the horizontal exhaust manifold (HEM) of an IVC rack system. We first evaluated the efficacy of rack sanitation methods for removing C. bovis DNA from the HEM of racks housing endemic colonies of infected nude mice. Pressurized water used to flush the racks’ air exhaust system followed by a standard rack-washer cycle was ineffective in eliminating C. bovis DNA. Only after autoclaving did all sanitized racks test negative for C. bovis DNA. We then measured the effects of stage of infection (early or established), cage density, and cage location on the rack on time-to-detection at the HEM. Stage of infection significantly affected time-to-detection, independent of cage location. Early infections required 7.3 ± 1.2 d whereas established infections required 1 ± 0 d for detection of C. bovis at the HEM. Cage density influenced the quantity of C. bovis DNA detected but not time-to-detection. The location of the cage on the rack affected the time-to-detection only during early C. bovis infections. We suggest that qPCR swabs of HEM are useful during the routine surveillance of nude mouse colonies for C. bovis infection. PMID:26817981

  14. Mycobacterium bovis in Burkina Faso: Epidemiologic and Genetic Links between Human and Cattle Isolates

    PubMed Central

    Sanou, Adama; Tarnagda, Zekiba; Kanyala, Estelle; Zingué, Dezemon; Nouctara, Moumini; Ganamé, Zakaria; Combary, Adjima; Hien, Hervé; Dembele, Mathurin; Kabore, Antoinette; Meda, Nicolas; Van de Perre, Philippe; Neveu, Dorine

    2014-01-01

    Background In sub-Saharan Africa, bovine tuberculosis (bTB) is a potential hazard for animals and humans health. The goal of this study was to improve our understanding of bTB epidemiology in Burkina Faso and especially Mycobacterium bovis transmission within and between the bovine and human populations. Methodology/principal findings Twenty six M. bovis strains were isolated from 101 cattle carcasses with suspected bTB lesions during routine meat inspections at the Bobo Dioulasso and Ouagadougou slaughterhouses. In addition, 7 M. bovis strains were isolated from 576 patients with pulmonary tuberculosis. Spoligotyping, RDAf1 deletion and MIRU-VNTR typing were used for strains genotyping. The isolation of M. bovis strains was confirmed by spoligotyping and 12 spoligotype signatures were detected. Together, the spoligotyping and MIRU-VNTR data allowed grouping the 33 M. bovis isolates in seven clusters including isolates exclusively from cattle (5) or humans (1) or from both (1). Moreover, these data (genetic analyses and phenetic tree) showed that the M. bovis isolates belonged to the African 1 (Af1) clonal complex (81.8%) and the putative African 5 (Af5) clonal complex (18.2%), in agreement with the results of RDAf1 deletion typing. Conclusions/Significance This is the first detailed molecular characterization of M. bovis strains from humans and cattle in Burkina Faso. The distribution of the two Af1 and putative Af5 clonal complexes is comparable to what has been reported in neighbouring countries. Furthermore, the strain genetic profiles suggest that M. bovis circulates across the borders and that the Burkina Faso strains originate from different countries, but have a country-specific evolution. The genetic characterization suggests that, currently, M. bovis transmission occurs mainly between cattle, occasionally between cattle and humans and potentially between humans. This study emphasizes the bTB risk in cattle but also in humans and the difficulty to set up

  15. Classification of Babesia canis strains in Europe based on polymorphism of the Bc28.1-gene from the Babesia canis Bc28 multigene family.

    PubMed

    Carcy, B; Randazzo, S; Depoix, D; Adaszek, L; Cardoso, L; Baneth, G; Gorenflot, A; Schetters, T P

    2015-07-30

    The vast majority of clinical babesiosis cases in dogs in Europe is caused by Babesia canis. Although dogs can be vaccinated, the level of protection is highly variable, which might be due to genetic diversity of B. canis strains. One of the major merozoite surface antigens of B. canis is a protein with a Mr of 28 kDa that belongs to the Bc28 multigene family, that comprises at least two genes, Bc28.1 and a homologous Bc28.2 gene. The two genes are relatively conserved but they are very distinct in their 3' ends, enabling the design of specific primers. Sequencing of the Bc28.1 genes from 4 genetically distinct B. canis laboratory strains (A8, B, 34.01 and G) revealed 20 mutations at conserved positions of which three allowed the classification of B. canis strains into three main groups (A, B and 34.01/G) by RFLP. This assay was subsequently used to analyze blood samples of 394 dogs suspected of clinical babesiosis from nine countries in Europe. All blood samples were first analyzed with a previously described assay that allowed detection of the different Babesia species that infect dogs. Sixty one percent of the samples contained detectable levels of Babesia DNA. Of these, 98.3% were positive for B. canis, the remaining cases were positive for B. vogeli. Analysis of the Bc28.1 gene, performed on 178 of the B. canis samples, revealed an overall dominance of genotype B (62.4%), followed by genotypes A (37.1%) and 34 (11.8%). Interestingly, a great variation in the geographical distribution and prevalence of the three B. canis genotypes was observed; in the North-East genotype A predominated (72.1% A against 27.9% B), in contrast to the South-West where genotype B predominated (10.3% A against 89.7% B). In the central part of Europe intermediate levels were found (26.0-42.9% A against 74.0-57.1% B, from West to East). Genotype 34 was only identified in France (26.9% among 78 samples) and mostly as co-infection with genotypes A or B (61.9%). A comparative analysis of

  16. Identification of novel Babesia and Theileria genotypes in the endangered marsupials, the woylie (Bettongia penicillata ogilbyi) and boodie (Bettongia lesueur).

    PubMed

    Paparini, Andrea; Ryan, Una M; Warren, Kris; McInnes, Linda M; de Tores, Paul; Irwin, Peter J

    2012-05-01

    Piroplasms, which include the genera Theileria and Babesia, are blood-borne parasites transmitted mainly by tick vectors. Relatively little is known about their prevalence and clinical impact in Australian marsupials. In the present study the occurrence and molecular phylogeny of these parasites were studied in both wild and captive marsupials from Western Australia (WA) and Queensland (QLD). Blood samples were screened by microscopy and molecular methods, using PCR and DNA sequencing of the 18S ribosomal RNA gene (18S rDNA). Overall, 7.1% of the blood samples (8/113) were positive for piroplasm 18S rDNA. Theileria and Babesia rDNA was detected in 0.9% (1/113) and 6.2% (7/113) of the animals, respectively. The single Theileria positive was identified in one of three boodies (Bettongia lesueur) screened from a wildlife rehabilitation centre in WA, while all seven Babesia positives were detected in WA in wild captured woylies (Bettongia penicillata ogilbyi). Small intraerythrocytic inclusions were observed in blood films made from six of these individuals. This is the first report of a Babesia sp. in woylies, and Theileria sp. in boodies. Phylogenetic analysis indicated that the woylie-derived Babesia was genetically distinct and most closely related to Babesia occultans, the causative agent of a benign form of cattle babesiosis (genetic similarity 98.4%). The Theileria identified was most closely related to the marsupial-derived species Theileria penicillata from the woylie, Theileria brachyuri from the quokka (Setonix brachyurus), and Theileria sp. from the long-nosed potoroo (Potorous tridactylus).

  17. Detection of Corynebacterium bovis infection in athymic nude mice from a research animal facility in Korea.

    PubMed

    Kim, Tae-Hyoun; Kim, Dong-Su; Han, Ju-Hee; Chang, Seo-Na; Kim, Kyung-Sul; Seok, Seung-Hyeok; Kim, Dong-Jae; Park, Jong-Hwan; Park, Jae-Hak

    2014-12-01

    Corynebacterium (C.) bovis infection in nude mice causes hyperkeratosis and weight loss and has been reported worldwide but not in Korea. In 2011, nude mice from an animal facility in Korea were found to have white flakes on their dorsal skin. Histopathological testing revealed that the mice had hyperkeratosis and Gram-positive bacteria were found in the skin. We identified isolated bacteria from the skin lesions as C. bovis using PCR and 16S rRNA sequencing. To the best of our knowledge, this is the first report of C. bovis infection in nude mice from Korea.

  18. Molecular detection of Babesia rossi and Hepatozoon sp. in African wild dogs (Lycaon pictus) in South Africa.

    PubMed

    Matjila, Paul Tshepo; Leisewitz, Andrew L; Jongejan, Frans; Bertschinger, Henk J; Penzhorn, Barend L

    2008-10-20

    Blood specimens from wild dogs (n=301) were obtained from De Wildt Cheetah and Wildlife Centre (Pretoria) and five game reserves (4 in the North-West Province and 1 in Limpopo Province), South Africa. Specimens were screened for Babesia, Theileria, Hepatozoon and Ehrlichia/Anaplasma species using PCR and Reverse Line Blot (RLB) assays. Positive results were obtained in 18 (6%) wild dogs. Sixteen specimens were found positive for Babesia rossi and two dogs were Hepatozoon sp. positive. It appears that these tick-borne pathogens are not widely distributed in wild dog populations.

  19. Optimization of modified Middlebrook 7H11 agar for isolation of Mycobacterium bovis from raw milk cheese.

    PubMed

    Forgrave, R; Donaghy, J A; Fisher, A; Rowe, M T

    2014-10-01

    Reports have highlighted the absence of contemporary peer reviewed publications pertaining to Mycobacterium bovis culture from raw milk and cheese. By replicating traditional methods, cheese-making methodology and equipment were devised to produce Cheddar (n = 6) and Caerphilly (n = 3) artificially contaminated with M. bovis (three genotypes) under stringent laboratory-containment guidelines for handling hazardous microbiological material. Middlebrook 7H11, modified for M. bovis isolation, was assessed for capacity to enumerate M. bovis despite changing cheese microflora and prolonged M. bovis exposure to the cheese matrix using maturing cheese test portions (n = 63; up to 16 weeks). Malachite green (MG) containing media isolated M. bovis at significantly (P < 0·05) lower levels than unmodified Middlebrook 7H11 agar despite MG being a common adjunct of Middlebrook 7H11 agar modified for M. bovis growth. Subsequently, a selective MG-free Middlebrook 7H11 agar modified using haemolysed red cells and calf serum was demonstrated as the best performing (P < 0·05) medium for recovery of M. bovis from typical UK cheese types, Cheddar and Caerphilly. Significance and impact of the study: Following increased M. bovis infection of UK cattle, the risk posed to consumers from consumption of unpasteurized milk and dairy products has changed. Furthermore, published methods for the culture and molecular detection of M. bovis in raw milk products are limited. Cheese-making protocols and M. bovis culture media reported here provide tools for further investigation of M. bovis survival during all stages of cheese manufacture and could inform future assessment of the risk to consumers from M. bovis contamination of unpasteurized dairy products.

  20. Optimization of modified Middlebrook 7H11 agar for isolation of Mycobacterium bovis from raw milk cheese.

    PubMed

    Forgrave, R; Donaghy, J A; Fisher, A; Rowe, M T

    2014-10-01

    Reports have highlighted the absence of contemporary peer reviewed publications pertaining to Mycobacterium bovis culture from raw milk and cheese. By replicating traditional methods, cheese-making methodology and equipment were devised to produce Cheddar (n = 6) and Caerphilly (n = 3) artificially contaminated with M. bovis (three genotypes) under stringent laboratory-containment guidelines for handling hazardous microbiological material. Middlebrook 7H11, modified for M. bovis isolation, was assessed for capacity to enumerate M. bovis despite changing cheese microflora and prolonged M. bovis exposure to the cheese matrix using maturing cheese test portions (n = 63; up to 16 weeks). Malachite green (MG) containing media isolated M. bovis at significantly (P < 0·05) lower levels than unmodified Middlebrook 7H11 agar despite MG being a common adjunct of Middlebrook 7H11 agar modified for M. bovis growth. Subsequently, a selective MG-free Middlebrook 7H11 agar modified using haemolysed red cells and calf serum was demonstrated as the best performing (P < 0·05) medium for recovery of M. bovis from typical UK cheese types, Cheddar and Caerphilly. Significance and impact of the study: Following increased M. bovis infection of UK cattle, the risk posed to consumers from consumption of unpasteurized milk and dairy products has changed. Furthermore, published methods for the culture and molecular detection of M. bovis in raw milk products are limited. Cheese-making protocols and M. bovis culture media reported here provide tools for further investigation of M. bovis survival during all stages of cheese manufacture and could inform future assessment of the risk to consumers from M. bovis contamination of unpasteurized dairy products. PMID:24888395

  1. Distribution patterns of Babesia gibsoni infection in hunting dogs from nine Japanese islands.

    PubMed

    El-Dakhly, Khaled Mohamed; Goto, Minami; Noishiki, Kaori; El-Nahass, El-Shaymaa; Sakai, Hiroki; Yanai, Tokuma; Takashima, Yasuhiro

    2015-04-01

    Canine babesiosis constitutes a major global veterinary medical problem caused by tick-borne hemoparasites Babesia gibsoni and Babesia canis. Babesia gibsoni induces more severe clinical signs and is mainly transmitted by the ixodid Haemaphysalis longicornis. In Japan, B. gibsoni is primarily found in the western districts, with few records in the eastern parts. The aim of the current investigation was to evaluate distribution patterns of B. gibsoni infection in 9 Japanese islands and peninsulas using direct microscopy and PCR. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011. Ages and sexes of dogs were identified. Direct microscopy of Giemsa-stained blood smear revealed pear-shaped piroplasms of B. gibsoni in 3 (1.6%) dogs. PCR was done initially with the universal primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, followed by the specific primer set (Bg18F1 and Bg18R2) amplifying 2,363-bp fragments of the same gene. Accordingly, 84 (42.9%) and 8 (4.1%) dogs were positive, respectively. The current investigation shows that canine babesiosis was recorded in all islands except for Sado Island, Atsumi Peninsula, and Tanegashima Island. The highest infection rate was detected in the main island of Okinawa, while the lowest was on Ishigaki Island. Both sexes were non-significantly infected. However, the diversity of infection in islands was significantly different (P < 0.05). Although B. gibsoni has been previously found in western and eastern Japan, the present work highlights the prevalence of infection in many Japanese districts, including islands and peninsulas, giving realistic data that can facilitate treatment and control.

  2. Molecular detection of Theileria, Babesia, and Hepatozoon spp. in ixodid ticks from Palestine.

    PubMed

    Azmi, Kifaya; Ereqat, Suheir; Nasereddin, Abedelmajeed; Al-Jawabreh, Amer; Baneth, Gad; Abdeen, Ziad

    2016-07-01

    Ixodid ticks transmit various infectious agents that cause disease in humans and livestock worldwide. A cross-sectional survey on the presence of protozoan pathogens in ticks was carried out to assess the impact of tick-borne protozoa on domestic animals in Palestine. Ticks were collected from herds with sheep, goats and dogs in different geographic districts and their species were determined using morphological keys. The presence of piroplasms and Hepatozoon spp. was determined by PCR amplification of a 460-540bp fragment of the 18S rRNA gene followed by RFLP or DNA sequencing. A PCR-RFLP method based on the 18S rRNA was used in order to detect and to identify Hepatozoon, Babesia and Theileria spp. A total of 516 ticks were collected from animals in six Palestinian localities. Five tick species were found: Rhipicephalus sanguineus sensu lato, Rhipicephalus turanicus, Rhipicephalus bursa, Haemaphysalis parva and Haemaphysalis adleri. PCR-based analyses of the ticks revealed Theileria ovis (5.4%), Hepatozoon canis (4.3%), Babesia ovis (0.6%), and Babesia vogeli (0.4%). Theileria ovis was significantly associated with ticks from sheep and with R. turanicus ticks (p<0.01). H. canis was detected only in R. sanguineus s.l. and was significantly associated with ticks from dogs (p<0.01). To our knowledge, this is the first report describing the presence of these pathogens in ticks collected from Palestine. Communicating these findings with health and veterinary professionals will increase their awareness, and contribute to improved diagnosis and treatment of tick-borne diseases. PMID:26969489

  3. Distribution patterns of Babesia gibsoni infection in hunting dogs from nine Japanese islands.

    PubMed

    El-Dakhly, Khaled Mohamed; Goto, Minami; Noishiki, Kaori; El-Nahass, El-Shaymaa; Sakai, Hiroki; Yanai, Tokuma; Takashima, Yasuhiro

    2015-04-01

    Canine babesiosis constitutes a major global veterinary medical problem caused by tick-borne hemoparasites Babesia gibsoni and Babesia canis. Babesia gibsoni induces more severe clinical signs and is mainly transmitted by the ixodid Haemaphysalis longicornis. In Japan, B. gibsoni is primarily found in the western districts, with few records in the eastern parts. The aim of the current investigation was to evaluate distribution patterns of B. gibsoni infection in 9 Japanese islands and peninsulas using direct microscopy and PCR. Therefore, 196 hunting dogs were randomly sampled during the period from March to September 2011. Ages and sexes of dogs were identified. Direct microscopy of Giemsa-stained blood smear revealed pear-shaped piroplasms of B. gibsoni in 3 (1.6%) dogs. PCR was done initially with the universal primer set (B18S-F and B18S-R) amplifying the 1,665-bp portion of the 18S rRNA gene, followed by the specific primer set (Bg18F1 and Bg18R2) amplifying 2,363-bp fragments of the same gene. Accordingly, 84 (42.9%) and 8 (4.1%) dogs were positive, respectively. The current investigation shows that canine babesiosis was recorded in all islands except for Sado Island, Atsumi Peninsula, and Tanegashima Island. The highest infection rate was detected in the main island of Okinawa, while the lowest was on Ishigaki Island. Both sexes were non-significantly infected. However, the diversity of infection in islands was significantly different (P < 0.05). Although B. gibsoni has been previously found in western and eastern Japan, the present work highlights the prevalence of infection in many Japanese districts, including islands and peninsulas, giving realistic data that can facilitate treatment and control. PMID:25419881

  4. Primary Babesia rodhaini infection followed by recovery confers protective immunity against B. rodhaini reinfection and Babesia microti challenge infection in mice.

    PubMed

    Wang, Guanbo; Efstratiou, Artemis; Adjou Moumouni, Paul Franck; Liu, Mingming; Jirapattharasate, Charoonluk; Guo, Huanping; Gao, Yang; Cao, Shinuo; Zhou, Mo; Suzuki, Hiroshi; Igarashi, Ikuo; Xuan, Xuenan

    2016-10-01

    In the present study, we investigated the protective immunity against challenge infections with Babesia rodhaini and Babesia microti in the mice recovered from B. rodhaini infection. Six groups with 5 test mice in each group were used in this study, and were intraperitoneally immunized with alive and dead B. rodhaini. The challenge infections with B. rodhaini or B. microti were performed using different time courses. Our results showed that the mice recovered from primary B. rodhaini infection exhibited low parasitemia and no mortalities after the challenge infections, whereas mock mice which had received no primary infection showed a rapid increase of parasitemia and died within 7 days after the challenge with B. rodhaini. Mice immunized with dead B. rodhaini were not protected against either B. rodhaini or B. microti challenge infections, although high titers of antibody response were induced. These results indicate that only mice immunized with alive B. rodhaini could acquire protective immunity against B. rodhaini or B. microti challenge infection. Moreover, the test mice produced high levels of antibody response and low levels of cytokines (INF-γ, IL-4, IL-12, IL-10) against B. rodhaini or B. microti after challenge infection. Mock mice, however, showed rapid increases of these cytokines, which means disordered cytokines secretion occurred during the acute stage of challenge infection. The above results proved that mice immunized with alive B. rodhaini could acquire protective immunity against B. rodhaini and B. microti infections. PMID:27423972

  5. Vaccination with recombinant Mycoplasma bovis GAPDH results in a strong humoral immune response but does not protect feedlot cattle from an experimental challenge with M. bovis.

    PubMed

    Prysliak, Tracy; van der Merwe, Jacques; Perez-Casal, Jose

    2013-02-01

    Mycoplasma bovis continues to cause significant disease in feedlots and dairy farms. The ability of the micro-organism to evade the immune system of the host combined with the lack of effective vaccines makes this disease difficult to control. Bacterin-based vaccines have not been successful in field trials and in some cases enhance the disease. In an attempt to develop a sub-unit vaccine, we used the conserved M. bovis glyceraldehyde-3-phosphate (GAPDH) protein in combination with a protein extract prepared from three M. bovis isolates to immunize feedlot animals. After challenge with a combination of three M. bovis isolates, there were differences in the proportion of weight loss between the control and vaccinated groups but no differences in rectal temperature and survival rate in all the groups. In addition, there were no significant differences between the proportions of lungs lesions in all the groups despite the percentages of lesions being higher in the vaccinated groups. These findings indicate that the M. bovis GAPDH protein is not a suitable antigen for a vaccine against this pathogen.

  6. New Molecules in Babesia gibsoni and Their Application for Diagnosis, Vaccine Development, and Drug Discovery

    PubMed Central

    Goo, Youn-Kyoung

    2014-01-01

    Babesia gibsoni is an intraerythrocytic apicomplexan parasite that causes piroplasmosis in dogs. B. gibsoni infection is characterized clinically by fever, regenerative anemia, splenomegaly, and sometimes death. Since no vaccine is available, rapid and accurate diagnosis and prompt treatment of infected animals are required to control this disease. Over the past decade, several candidate molecules have been identified using biomolecular techniques in the authors' laboratory for the development of a serodiagnostic method, vaccine, and drug for B. gibsoni. This review article describes newly identified candidate molecules and their applications for diagnosis, vaccine production, and drug development of B. gibsoni. PMID:25246713

  7. Serological investigations on Babesia orientalis infection. Status of water buffaloes in Hubei Province.

    PubMed

    Yao, Baoan; Zhao, Junlong; Liu, Enyong; Ding, Sutian; Shi, Junhua; Liu, Zhongling

    2002-05-01

    A total of 230 water buffaloes were sampled for the determination of the epidemiological situation of Babesia orientalis infection in Hubei Province, using the latex agglutination test. This serological test revealed that 74.7% of the collected sera were positive for antibodies against B. orientalis in endemic areas. In contrast, only 1.79% of water buffaloes were positive in non-epidemic areas. The incidence of the infection was substantially higher in older than in younger animals. The results were analysed using mathematical model and indicated that these areas were in a stable situation.

  8. Genetic characterization and phylogenetic relationships based on 18S rRNA and ITS1 region of small form of canine Babesia spp. from India.

    PubMed

    Mandal, M; Banerjee, P S; Garg, Rajat; Ram, Hira; Kundu, K; Kumar, Saroj; Kumar, G V P P S Ravi

    2014-10-01

    Canine babesiosis is a vector borne disease caused by intra-erythrocytic apicomplexan parasites Babesia canis (large form) and Babesia gibsoni (small form), throughout the globe. Apart from few sporadic reports on the occurrence of B. gibsoni infection in dogs, no attempt has been made to characterize Babesia spp. of dogs in India. Fifteen canine blood samples, positive for small form of Babesia, collected from northern to eastern parts of India, were used for amplification of 18S rRNA gene (∼1665bp) of Babesia sp. and partial ITS1 region (∼254bp) of B. gibsoni Asian genotype. Cloning and sequencing of the amplified products of each sample was performed separately. Based on sequences and phylogenetic analysis of 18S rRNA and ITS1 sequences, 13 were considered to be B. gibsoni. These thirteen isolates shared high sequence identity with each other and with B. gibsoni Asian genotype. The other two isolates could not be assigned to any particular species because of the difference(s) in 18S rRNA sequence with B. gibsoni and closer identity with Babesiaoccultans and Babesiaorientalis. In the phylogenetic tree, all the isolates of B. gibsoni Asian genotype formed a separate major clade named as Babesia spp. sensu stricto clade with high bootstrap support. The two unnamed Babesia sp. (Malbazar and Ludhiana isolates) clustered close together with B. orientalis, Babesia sp. (Kashi 1 isolate) and B. occultans of bovines. It can be inferred from this study that 18S rRNA gene and ITS1 region are highly conserved among 13 B. gibsoni isolates from India. It is the maiden attempt of genetic characterization by sequencing of 18S rRNA gene and ITS1 region of B. gibsoni from India and is also the first record on the occurrence of an unknown Babesia sp. of dogs from south and south-east Asia.

  9. Multinucleated giant cell cytokine expression in pulmonary granulomas of cattle experimentally infected with Mycobacterium bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Pathogenic mycobacteria of the Mycobacterium tuberculosis complex such as Mycobacterium bovis, induce a characteristic lesion known as a granulomas. Granulomas represent a specific host response to chronic antigenic stimuli, such as foreign bodies, certain bacterial components, or persistent pathoge...

  10. Circulating Mycobacterium bovis peptides and host response proteins as biomarkers for unambiguous detection of subclinical infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Bovine tuberculosis remains one of the most damaging zoonotic diseases. A critical need exists for rapid and inexpensive diagnostics capable of detecting and differentiating M. bovis infection from other pathogenic and environmental mycobacteria at multiple surveillance levels. Method...

  11. Phenolic glycolipids of Mycobacterium bovis: new structures and synthesis of a corresponding seroreactive neoglycoprotein.

    PubMed Central

    Chatterjee, D; Bozic, C M; Knisley, C; Cho, S N; Brennan, P J

    1989-01-01

    The glycolipid that characterizes the majority of isolates of Mycobacterium bovis and that has come to be known as M. bovis-identifying lipid is the phenolic glycolipid mycoside B described in the literature by others. However, when mycoside B obtained from M. bovis BCG, field isolates, and infected tissues was examined in detail, it was shown to be different from that described in the literature in some important respects. In particular, the glycosyl substituent is 2-O-methyl-alpha-L-rhamnopyranose rather than 2-O-methyl-beta-D-rhamnopyranose. With this information, a seroreactive neoglycoprotein (neoantigen) containing the 2-O-methyl-alpha-L-rhamnopyranosyl substituent suitable for the serodiagnosis of bovine tuberculosis was synthesized. M. bovis also contains other minor seroreactive phenolic glycolipids, one of which is a deacylated form of mycoside B and another of which contains an alpha-L-rhamnopyranosyl unit rather than 2-O-methyl-alpha-L-rhamnopyranose. Images PMID:2643563

  12. Amplification of a 500-Base-Pair Fragment from Cultured Isolates of Mycobacterium bovis

    PubMed Central

    Rodríguez, Juan Germán; Fissanoti, Juan Carlos; Del Portillo, Patricia; Patarroyo, Manuel Elkin; Romano, María Isabel; Cataldi, Angel

    1999-01-01

    The presence of a 500-bp fragment which amplifies a region from the genome of Mycobacterium bovis (J. G. Rodriguez, G. A. Meija, P. Del Portillo, M. E. Patarroyo, and L. A. Murillo, Microbiology 141:2131–2138, 1995) was evaluated by carrying out PCR on 121 M. bovis isolates. The M. bovis strains, previously characterized by culture and biochemical tests, were isolated from cattle in different regions of Argentina, Mexico, and Colombia. Four additional strains isolated from sea lions that belong to the M. tuberculosis complex were also included in the study. All of the isolates tested were PCR positive, rendering the expected 500-bp band and giving a correlation of 100% with previous microbiological characterization. Southern blot analysis revealed a common band of 1,800 bp and a polymorphic high-molecular-mass hybridization pattern. The results show that this assay may be useful for diagnosis and identification of M. bovis in cattle. PMID:10364607

  13. Pulmonary infection due to Mycobacterium bovis in a black rhinoceros (Diceros bicornis minor) in South Africa.

    PubMed

    Espie, Ian W; Hlokwe, Tiny M; Gey van Pittius, Nicolaas C; Lane, Emily; Tordiffe, Adrian S W; Michel, Anita L; Müller, Annélle; Kotze, Antoinette; van Helden, Paul D

    2009-10-01

    We report a case of tuberculosis due to infection with Mycobacterium bovis in an elderly male black rhinoceros (Diceros bicornis minor) from the Limpopo Province in South Africa. The animal was euthanized due to very poor condition, old age, and dental attrition. Necropsy examination revealed two small nonencapsulated granulomas (approximately 40-mm diameter) in the dorsocaudal lobe of the left lung. Sequencing of isolated crude lung tissue PCR product and boiled lung culture samples confirmed that the causative organism was M. bovis. Genotyping revealed limited similarities with M. bovis strains isolated thus far from South African cattle or wildlife. The source of the infection could not be determined. This case illustrates that M. bovis could impact conservation of free-ranging rare and endangered species. Effective diagnostics are urgently needed for different animal species, such as white or black rhinoceroses, to certify with a reasonable degree of certainty that these animals are free of tuberculosis in natural habitats. PMID:19901395

  14. An improved loop-mediated isothermal amplification assay for the detection of Mycoplasma bovis

    PubMed Central

    HIGA, Yumiko; UEMURA, Ryoko; YAMAZAKI, Wataru; GOTO, Shinya; GOTO, Yoshitaka; SUEYOSHI, Masuo

    2016-01-01

    We improved a loop-mediated isothermal amplification (LAMP) assay permitting sensitive and rapid Mycoplasma bovis detection. A total of 55 bacterial strains were examined in this study, including 33 M. bovis strains, 14 non-M. bovis mycoplasmas and eight non-mycoplasma bacterial strains. M. bovis was successfully detected by the LAMP assay within 60 min without cross-reaction to any other bacteria. Furthermore, a total of 135 nasal swab samples were tested directly using our LAMP assays, the previously reported LAMP assay, conventional PCR assay without pre-culture and comparing standard culture methods. The improved LAMP assay showed sensitivity and specificity of 97.2% and 90.9%, respectively (with a kappa coefficient of 0.8231), and the sensitivity of our revised LAMP assay was increased compared to existing methods. PMID:27109067

  15. [Interlaboratory test: Isolation of Mycobacterium bovis from granulomatous lesions in bovine].

    PubMed

    Garbaccio, Sergio; Barandiaran, Soledad; Fernandez, Analía; Macias, Analía; Magnano, Gabriel; Martinez Vivot, Marcela; Peyrú, Maite; Cataldi, Angel

    2016-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis. The diagnostic laboratory confirmation is made through bacterial isolation. The aim of interlaboratory tests is to assess the performance of each participant in comparison with other of similar capacities. The test objective was to determine the efficiency of isolation of M. bovis. Four laboratories were part of the test and processed 25 blind tissue samples from granulomatous lesions and with previous M. bovis isolation. The laboratory that had the highest proportion of isolates was A (68%), followed by C (60%) and then B and D (both with 52%). The greatest concordance was observed between B-D and B-C laboratories (68%). The differences could be due to specific factors in each laboratory procedures. This type of interlaboratory tests highlights errors in the bacteriology and identifies critical points in the process to detect M. bovis accurately. PMID:27237425

  16. [Interlaboratory test: Isolation of Mycobacterium bovis from granulomatous lesions in bovine].

    PubMed

    Garbaccio, Sergio; Barandiaran, Soledad; Fernandez, Analía; Macias, Analía; Magnano, Gabriel; Martinez Vivot, Marcela; Peyrú, Maite; Cataldi, Angel

    2016-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis. The diagnostic laboratory confirmation is made through bacterial isolation. The aim of interlaboratory tests is to assess the performance of each participant in comparison with other of similar capacities. The test objective was to determine the efficiency of isolation of M. bovis. Four laboratories were part of the test and processed 25 blind tissue samples from granulomatous lesions and with previous M. bovis isolation. The laboratory that had the highest proportion of isolates was A (68%), followed by C (60%) and then B and D (both with 52%). The greatest concordance was observed between B-D and B-C laboratories (68%). The differences could be due to specific factors in each laboratory procedures. This type of interlaboratory tests highlights errors in the bacteriology and identifies critical points in the process to detect M. bovis accurately.

  17. Molecular characterization of Mycobacterium bovis isolates from patients with tuberculosis in Baja California, Mexico.

    PubMed

    Laniado-Laborín, Rafael; Muñiz-Salazar, Raquel; García-Ortiz, Rosa Alejandra; Vargas-Ojeda, Adriana Carolina; Villa-Rosas, Cecilia; Oceguera-Palao, Lorena

    2014-10-01

    The incidence of tuberculosis (TB) from Mycobacterium bovis in humans is likely to be underestimated and in some cases even ignored in most developing countries. This may be due to the difficulty of differentiating TB caused by either Mycobacteriumtuberculosis or M. bovis. Our objectives were to determine the prevalence of M. bovis human disease among the patients referred for study to the Tuberculosis Laboratory of the Tijuana General Hospital in Baja California, Mexico and to characterize molecularly the clinical isolates using 8 loci of MIRU-VNTR. A cross-sectional analysis of all culture-proven cases of tuberculosis was conducted during the period from January 1, 2011 through June 30, 2013. Clinical isolates that exhibited resistance to pyrazinamide (Z) were submitted for molecular analysis. A total of 2699 clinical samples were cultured during the study period and 600 (22%) that tested positive were processed for drug susceptibility for first line drugs. Sixty-four (10.7%) of the tested isolates tested were resistant to Z, and 27 (4.5%) of those were subsequently identified molecularly as M. bovis. Three of the M. bovis isolates were polyresistant to Z, isoniazid (H), ethambutol (E) and rifampicin (R) (Z+H+E, Z+E and Z+R); the rest were only resistant only to Z. VNTR typing, based on the 8 VNTR loci commonly tested for M.bovis, detected 12 allelic profiles (genotypes). The real burden of M. bovis cases among the total reported human tuberculosis cases can only be known from especially designed studies in which, during a specific period, all specimens submitted to tuberculosis diagnosis in one or more laboratories are cultured on the appropriate media and the isolated mycobacteria are analyzed to differentiate M. bovis from M. tuberculosis and other Mycobacterium species.

  18. Molecular Typing of Mycobacterium bovis from Cattle Reared in Midwest Brazil

    PubMed Central

    Carvalho, Ricardo César Tavares; Vasconcellos, Sidra Ezidio Gonçalves; Issa, Marina de Azevedo; Soares Filho, Paulo Martins; Mota, Pedro Moacyr Pinto Coelho; de Araújo, Flábio Ribeiro; Carvalho, Ana Carolina da Silva; Gomes, Harrison Magdinier; Suffys, Philip Noel; Paschoalin, Vânia Margaret Flosi

    2016-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), the pathogen responsible for serious economic impact on the livestock sector. In order to obtain data on isolated M. bovis strains and assist in the control and eradication program for BTB, a cross sectional descriptive molecular epidemiology study in the Brazilian Midwest was conducted. Through spoligotyping and 24-loci MIRU-VNTR methods, 37 clinical isolates of M. bovis circulating in the region were analyzed, 10 isolated from the state of Mato Grosso, 12 from the state of Mato Grosso do Sul and 15 from the state of Goiás. The spoligotyping analysis identified 10 distinct M. bovis profiles (SB0121 n = 14, SB0295 n = 6, SB0140 n = 6, SB0881 n = 3, SB1144 n = 2, SB1145 n = 2, SB0134 n = 1, SB1050 n = 1, SB1055 n = 1, SB1136 n = 1) grouped in six clusters and four orphan patterns. The MIRU-VNTR 24-loci grouped the same isolates in six clusters and 22 unique orphan patterns, showing higher discriminatory power than spoligotyping. When associating the results of both techniques, the isolates were grouped in five clusters and 24 unique M. bovis profiles. Among the 24-loci MIRU-VNTR evaluated, two, ETR-A and QUB 11b loci, showed high discriminatory ability (h = ≥ 0.50), while MIRU 16, MIRU 27, ETR-B, ETR-C, Mtub21 and QUB 26 loci showed moderate ability (h = 0.33 or h = 0.49) and were the most effective in evaluating the genotypic similarities among the clinical M. bovis isolate samples. Herein, the 29 patterns found amongst the 37 isolates of M. bovis circulating in the Brazilian Midwest can be due to the animal movement between regions, municipalities and farms, thus causing the spread of various M. bovis strains in herds from Midwest Brazil. PMID:27631383

  19. Molecular Typing of Mycobacterium bovis from Cattle Reared in Midwest Brazil.

    PubMed

    Carvalho, Ricardo César Tavares; Vasconcellos, Sidra Ezidio Gonçalves; Issa, Marina de Azevedo; Soares Filho, Paulo Martins; Mota, Pedro Moacyr Pinto Coelho; Araújo, Flábio Ribeiro de; Carvalho, Ana Carolina da Silva; Gomes, Harrison Magdinier; Suffys, Philip Noel; Figueiredo, Eduardo Eustáquio de Souza; Paschoalin, Vânia Margaret Flosi

    2016-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis (BTB), the pathogen responsible for serious economic impact on the livestock sector. In order to obtain data on isolated M. bovis strains and assist in the control and eradication program for BTB, a cross sectional descriptive molecular epidemiology study in the Brazilian Midwest was conducted. Through spoligotyping and 24-loci MIRU-VNTR methods, 37 clinical isolates of M. bovis circulating in the region were analyzed, 10 isolated from the state of Mato Grosso, 12 from the state of Mato Grosso do Sul and 15 from the state of Goiás. The spoligotyping analysis identified 10 distinct M. bovis profiles (SB0121 n = 14, SB0295 n = 6, SB0140 n = 6, SB0881 n = 3, SB1144 n = 2, SB1145 n = 2, SB0134 n = 1, SB1050 n = 1, SB1055 n = 1, SB1136 n = 1) grouped in six clusters and four orphan patterns. The MIRU-VNTR 24-loci grouped the same isolates in six clusters and 22 unique orphan patterns, showing higher discriminatory power than spoligotyping. When associating the results of both techniques, the isolates were grouped in five clusters and 24 unique M. bovis profiles. Among the 24-loci MIRU-VNTR evaluated, two, ETR-A and QUB 11b loci, showed high discriminatory ability (h = ≥ 0.50), while MIRU 16, MIRU 27, ETR-B, ETR-C, Mtub21 and QUB 26 loci showed moderate ability (h = 0.33 or h = 0.49) and were the most effective in evaluating the genotypic similarities among the clinical M. bovis isolate samples. Herein, the 29 patterns found amongst the 37 isolates of M. bovis circulating in the Brazilian Midwest can be due to the animal movement between regions, municipalities and farms, thus causing the spread of various M. bovis strains in herds from Midwest Brazil. PMID:27631383

  20. Protection against tuberculosis in Eurasian wild boar vaccinated with heat-inactivated Mycobacterium bovis.

    PubMed

    Garrido, Joseba M; Sevilla, Iker A; Beltrán-Beck, Beatriz; Minguijón, Esmeralda; Ballesteros, Cristina; Galindo, Ruth C; Boadella, Mariana; Lyashchenko, Konstantin P; Romero, Beatriz; Geijo, Maria Victoria; Ruiz-Fons, Francisco; Aranaz, Alicia; Juste, Ramón A; Vicente, Joaquín; de la Fuente, José; Gortázar, Christian

    2011-01-01

    Tuberculosis (TB) caused by Mycobacterium bovis and closely related members of the Mycobacterium tuberculosis complex continues to affect humans and animals worldwide and its control requires vaccination of wildlife reservoir species such as Eurasian wild boar (Sus scrofa). Vaccination efforts for TB control in wildlife have been based primarily on oral live BCG formulations. However, this is the first report of the use of oral inactivated vaccines for controlling TB in wildlife. In this study, four groups of 5 wild boar each were vaccinated with inactivated M. bovis by the oral and intramuscular routes, vaccinated with oral BCG or left unvaccinated as controls. All groups were later challenged with a field strain of M. bovis. The results of the IFN-gamma response, serum antibody levels, M. bovis culture, TB lesion scores, and the expression of C3 and MUT genes were compared between these four groups. The results suggested that vaccination with heat-inactivated M. bovis or BCG protect wild boar from TB. These results also encouraged testing combinations of BCG and inactivated M. bovis to vaccinate wild boar against TB. Vaccine formulations using heat-inactivated M. bovis for TB control in wildlife would have the advantage of being environmentally safe and more stable under field conditions when compared to live BCG vaccines. The antibody response and MUT expression levels can help differentiating between vaccinated and infected wild boar and as correlates of protective response in vaccinated animals. These results suggest that vaccine studies in free-living wild boar are now possible to reveal the full potential of protecting against TB using oral M. bovis inactivated and BCG vaccines.

  1. Pathogenicity and genetic variation of 3 strains of Corynebacterium bovis in immunodeficient mice.

    PubMed

    Dole, Vandana S; Henderson, Kenneth S; Fister, Richard D; Pietrowski, Michael T; Maldonado, Geomaris; Clifford, Charles B

    2013-07-01

    Corynebacterium bovis has been associated with hyperkeratotic dermatitis and acanthosis in mice. We studied 3 different strains of C. bovis: one previously described to cause hyperkeratotic dermatitis (HAC), one that infected athymic nude mice without leading to the classic clinical signs, and one of bovine origin (ATCC 7715). The 3 strains showed a few biochemical and genetic differences. Immunodeficient nude mice were housed in 3 independent isolators and inoculated with pure cultures of the 3 strains. We studied the transmission of these C. bovis studies to isolator-bedding and contact sentinels housed for 5 to 12 wk in filter-top or wire-top cages in the respective isolators. Using a 16S rRNA-based qPCR assay, we did not find consistent differences in growth and transmission among the 3 C. bovis strains, and neither the incidence nor severity of hyperkeratosis or acanthosis differed between strains. Housing in filter-top compared with wire-top cages did not alter the morbidity associated with any of the strains. Our findings confirmed the variability in the gross and histologic changes associated with C. bovis infection of mice. Although bacteriology was a sensitive method for the detection of Corynebacterium spp., standard algorithms occasionally misidentified C. bovis and several related species. Our study demonstrates that PCR of skin swabs or feces is a sensitive and specific method for the detection of C. bovis infection in mice. An rpoB-based screen of samples from North American vivaria revealed that HAC is the predominant C. bovis strain in laboratory mice. PMID:23849444

  2. A Study of the Persistence of Mycobacterium bovis in the Environment under Natural Weather Conditions in Michigan, USA

    PubMed Central

    Fine, Amanda E.; Bolin, Carole A.; Gardiner, Joseph C.; Kaneene, John B.

    2011-01-01

    Reisolation of Mycobacterium bovis from inoculated substrates was used to follow the persistence of viable M. bovis bacteria exposed to natural weather conditions over a 12-month period. Environmental factors were recorded continuously, and factors affecting M. bovis persistence (i.e., temperature, season, and substrate) were studied using survival analysis and Cox's proportional hazards regression. Persistence of M. bovis in the environment was significantly shorter in the spring/summer season, characterized by the highest average daily temperatures over the 12-month period. M. bovis persisted up to 88 days in soil, 58 days in water and hay, and 43 days on corn. These studies demonstrate that M. bovis bacteria persist long enough to represent a risk of exposure for cattle and/or wildlife and strengthen evidence that suggests cattle farm biosecurity and efforts to eliminate supplemental feeding of white-tailed deer will decrease the risk of bovine TB transmission among and between cattle and deer populations. PMID:21547222

  3. Use of released pigs as sentinels for Mycobacterium bovis.

    PubMed

    Nugent, Graham; Whitford, Jackie; Young, Nigel

    2002-10-01

    Identifying the presence of bovine tuberculosis (TB; Mycobacterium bovis) in wildlife is crucial in guiding management aimed at eradicating the disease from New Zealand. Unfortunately, surveys of the principal wildlife host, the introduced brushtail possum (Trichosurus vulpecula), require large samples (> 95% of the population) before they can provide reasonable confidence that the disease is absent. In this study, we tested the feasibility of using a more wide-ranging species, feral pig (Sus scrofa), as an alternative sentinel capable of indicating TB presence. In January 2000, 17 pigs in four groups were released into a forested area with a low density of possums in which TB was known to be present. The pigs were radiotracked at 2 wk intervals from February to October 2000, and some of them were killed and necropsied at various intervals after release. Of the 15 pigs successfully recovered and necropsied, one killed 2 mo after release had no gross lesions typical of TB, and the only other pig killed at that time had greatly enlarged mandibular lymph nodes. The remainder were killed at longer intervals after release and all had gross lesions typical of TB. Mycobacterium bovis was isolated from all 15 pigs by mycobacterial culture. Home range sizes of pigs varied widely and increased with the length of time the pigs were in the forest, with minimum convex polygon range-size estimates averaging 10.7 km2 (range 4.7-20.3 km2) for the pigs killed after 6 mo. A 6 km radius around the kill site of each pig would have encompassed 95% of all of their previous locations at which they could have become infected. However, one pig shifted 35 km, highlighting the main limitation of using unmarked feral pigs as sentinels. This trial indicates use of resident and/or released free-ranging pigs is a feasible alternative to direct prevalence surveys of possums for detecting TB presence.

  4. The distribution of Mycobacterium bovis infection in naturally infected badgers.

    PubMed

    Corner, Leigh A L; O'Meara, D; Costello, E; Lesellier, S; Gormley, E

    2012-11-01

    Populations of Eurasian badgers (Meles meles) with tuberculosis (Mycobacterium bovis infection) are a significant reservoir of infection for cattle in Ireland and the United Kingdom. In this study the distribution of infection, histological lesions and gross lesions was determined in a sample of 132 culled badgers from naturally-infected wild populations. Badgers were culled when an epidemiological investigation following a tuberculosis breakdown in a cattle herd implicated badgers as the probable source of infection. The definition of tuberculosis infection was based on the isolation of M. bovis from tissues or clinical samples. An accurate diagnosis of infection was achieved by culturing a wide range of lymph nodes (LN) and organ tissues (mean 32.1) and clinical samples (faeces and urine) from each badger. Infection was detected in 57/132 badgers (43.2%). Histological lesions consistent with tuberculosis were seen in 39/57 (68.4%) culture-positive and 7/75 (9.3%) culture-negative animals. Gross lesions were seen in only 30/57 (52.6%) infected badgers, leaving a high proportion (47.4%) of infected animals with latent infection (no grossly visible lesions). The most frequently infected tissues were the lungs and axillary LN, followed by the deep cervical LN, parotid LN and tracheobronchial LN. The data support the hypotheses that in badgers there are only two significant routes of infection, namely, the lower respiratory tract and bite wounds, and that badgers are very susceptible to infection but resistant to the development and progression of the disease. At all levels of disease severity, infection was found in widely dispersed anatomical locations suggesting that there is early dissemination of infection in the period preceding the development of active immunity.

  5. Study on bioactive compounds of in vitro cultured Calculus Suis and natural Calculus Bovis.

    PubMed

    Wan, Tien-Chun; Cheng, Fu-Yuan; Liu, Yu-Tse; Lin, Liang-Chuan; Sakata, Ryoichi

    2009-12-01

    The purpose of the study was to investigate bioactive compounds of in vitro cultured Calculus Suis and natural Calculus Bovis obtained as valuable by-products from animals used for meat production. The results showed that the components of natural Calculus Bovis were rich in bilirubin and biliverdin and had higher content of essential amino acids. The major amino acids of in vitro cultured Calculus Suis were identified as glycine, alanine, glutamic acid and aspartic acid, and those for natural Calculus Bovis were found to be glutamic acid, aspartic acid, proline, and arginine. The methionine and cysteine contents of precursors for glutathione in natural Calculus Bovis were significantly higher than those of in vitro cultured Calculus Suis. The mineral contents of zinc, iron and manganese of natural Calculus Bovis were significantly higher than those of in vitro cultured Calculus Suis. The major bile acids in both products were cholic acid and dehydrocholic acid, respectively. The chenodeoxycholic and ursodeoxycholic acid content of in vitro cultured Calculus Suis was significantly higher than that of natural Calculus Bovis.

  6. Mycobacterium bovis in free-living and captive wildlife, including farmed deer.

    PubMed

    de Lisle, G W; Mackintosh, C G; Bengis, R G

    2001-04-01

    Mycobacterium bovis has been isolated from a wide range of wildlife species, in addition to domestic animals. This review examines the role played by various species in the maintenance of M. bovis in wildlife communities and the spread to domestic animals. Badgers (Meles meles), brushtail possums (Trichosurus vulpecula), deer (Odocoileus virginianus), bison (Bison bison) and African buffalo (Syncerus caffer) are examples of wildlife that are maintenance hosts of M. bovis. The importance of these hosts has been highlighted by the growing realisation that these animals can represent the principal source of infection for both domestic animals and protected wildlife species. The range of methods for controlling M. bovis in wildlife is limited. While population control has been used in some countries, this approach is not applicable in many situations where protected wildlife species are concerned. Vaccination is a potential alternative control method, although as yet, no practical, effective system has been developed for vaccinating wildlife against bovine tuberculosis. Tuberculosis caused by M. bovis has also been a problem in captive wildlife and in recently domesticated animals such as farmed deer. Control of M. bovis in this group of animals is dependent on the judicious use of diagnostic tests and the application of sound disease control principles. The advances in the development of bovine tuberculosis vaccines for cattle and farmed deer may offer valuable insights into the use of vaccination for the control of tuberculosis in a range of captive wildlife species. PMID:11288522

  7. Antimicrobial susceptibility of Mycoplasma bovis isolates from veal calves and dairy cattle in the Netherlands.

    PubMed

    Heuvelink, Annet; Reugebrink, Constance; Mars, Jet

    2016-06-30

    Control of Mycoplasma bovis infections depends on good husbandry practices and antibiotic treatment. To allow more prudent use of antimicrobial drugs, there is a need for information on the susceptibility profile of this pathogen. The objective of the present study was to analyse the in vitro antimicrobial susceptibility of clinical M. bovis isolates in the Netherlands. The collection comprised 95 bovine isolates, originating from lungs (n=56), mastitis milk (n=27), and synovial fluid (n=12), collected between 2008 and 2014. Minimal inhibitory concentrations (MICs) were assessed by broth microdilution, both by using in-house prepared MIC plates and by using commercially available MIC plates. For each antimicrobial agent, the range of MIC results, the MIC50, and MIC90 values were calculated. M. bovis strains recently isolated in the Netherlands appeared to be characterized by relatively high MIC values for antimicrobial agents that, until now, have been recommended by the Dutch Association of Veterinarians for treating pneumonia caused by Mycoplasma species. Fluoroquinolones appeared to be the most efficacious in inhibiting M. bovis growth, followed by tulathromycin and oxytetracycline. The highest MIC values were obtained for erythromycin, tilmicosin, and tylosin. Future studies should be done on determining M. bovis specific clinical breakpoints, standardization of methods to determine MIC values as well as molecular studies on detection of antimicrobial resistance mechanisms of M. bovis isolates to develop PCR assays for determining resistance.

  8. Antimicrobial susceptibility of Mycoplasma bovis isolates from veal calves and dairy cattle in the Netherlands.

    PubMed

    Heuvelink, Annet; Reugebrink, Constance; Mars, Jet

    2016-06-30

    Control of Mycoplasma bovis infections depends on good husbandry practices and antibiotic treatment. To allow more prudent use of antimicrobial drugs, there is a need for information on the susceptibility profile of this pathogen. The objective of the present study was to analyse the in vitro antimicrobial susceptibility of clinical M. bovis isolates in the Netherlands. The collection comprised 95 bovine isolates, originating from lungs (n=56), mastitis milk (n=27), and synovial fluid (n=12), collected between 2008 and 2014. Minimal inhibitory concentrations (MICs) were assessed by broth microdilution, both by using in-house prepared MIC plates and by using commercially available MIC plates. For each antimicrobial agent, the range of MIC results, the MIC50, and MIC90 values were calculated. M. bovis strains recently isolated in the Netherlands appeared to be characterized by relatively high MIC values for antimicrobial agents that, until now, have been recommended by the Dutch Association of Veterinarians for treating pneumonia caused by Mycoplasma species. Fluoroquinolones appeared to be the most efficacious in inhibiting M. bovis growth, followed by tulathromycin and oxytetracycline. The highest MIC values were obtained for erythromycin, tilmicosin, and tylosin. Future studies should be done on determining M. bovis specific clinical breakpoints, standardization of methods to determine MIC values as well as molecular studies on detection of antimicrobial resistance mechanisms of M. bovis isolates to develop PCR assays for determining resistance. PMID:27259820

  9. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites

    PubMed Central

    Terkawi, Mohamed Alaa; Youssef, Mohamed Ahmed; El Said, El Said El Shirbini; Elsayed, Gehad; El-Khodery, Sabry; El-Ashker, Maged; Elsify, Ahmed; Omar, Mosaab; Salama, Akram; Yokoyama, Naoaki; Igarashi, Ikuo

    2015-01-01

    A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses. PMID:25915529

  10. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.

    PubMed

    Rizk, Mohamed Abdo; El-Sayed, Shimaa Abd El-Salam; Terkawi, Mohamed Alaa; Youssef, Mohamed Ahmed; El Said, El Said El Shirbini; Elsayed, Gehad; El-Khodery, Sabry; El-Ashker, Maged; Elsify, Ahmed; Omar, Mosaab; Salama, Akram; Yokoyama, Naoaki; Igarashi, Ikuo

    2015-01-01

    A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.

  11. Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.

    PubMed

    Rizk, Mohamed Abdo; El-Sayed, Shimaa Abd El-Salam; Terkawi, Mohamed Alaa; Youssef, Mohamed Ahmed; El Said, El Said El Shirbini; Elsayed, Gehad; El-Khodery, Sabry; El-Ashker, Maged; Elsify, Ahmed; Omar, Mosaab; Salama, Akram; Yokoyama, Naoaki; Igarashi, Ikuo

    2015-01-01

    A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses. PMID:25915529

  12. Assessment of theileria equi and babesia caballi infections in equine populations in Egypt by molecular, serological and hematological approaches

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Equine piroplasmosis caused by Theileria equi, Babesia caballi, or both, cause significant economic losses in the equine industry and remains uncontrolled in Egypt. Methods: T. equi and B. caballi infections were assessed in blood from 88 horses and 51 donkeys from different localities ...

  13. Fecal Volatile Organic Ccompound Profiles from White-Tailed Deer (Odocoileus virginianus) as Indicators of Mycobacterium bovis Exposure or Mycobacterium bovis Bacille Calmette-Guerin (BCG) Vaccination.

    PubMed

    Stahl, Randal S; Ellis, Christine K; Nol, Pauline; Waters, W Ray; Palmer, Mitchell; VerCauteren, Kurt C

    2015-01-01

    White-tailed deer (Odocoileus virginianus) serve as a reservoir for bovine tuberculosis, caused by Mycobacterium bovis, and can be a source of infection in cattle. Vaccination with M. bovis Bacille Calmette Guerin (BCG) is being considered for management of bovine tuberculosis in deer. Presently, no method exists to non-invasively monitor the presence of bovine tuberculosis in deer. In this study, volatile organic compound profiles of BCG-vaccinated and non-vaccinated deer, before and after experimental challenge with M. bovis strain 95-1315, were generated using solid phase microextraction fiber head-space sampling over suspended fecal pellets with analysis by gas chromatography/mass spectrometry. Chromatograms were processed using XCMS Online to characterize ion variation among treatment groups. The principal component scores resulting from significant (α = 0.05) ion responses were used to build linear discriminant analysis models. The sensitivity and specificity of these models were used to evaluate the feasibility of using this analytical approach to distinguish within group comparisons between pre- and post-M. bovis challenge: non-vaccinated male or female deer, BCG-vaccinated male deer, and the mixed gender non-vaccinated deer data. Seventeen compounds were identified in this analysis. The peak areas for these compounds were used to build a linear discriminant classification model based on principal component analysis scores to evaluate the feasibility of discriminating between fecal samples from M. bovis challenged deer, irrespective of vaccination status. The model best representing the data had a sensitivity of 78.6% and a specificity of 91.4%. The fecal head-space sampling approach presented in this pilot study provides a non-invasive method to discriminate between M. bovis challenged deer and BCG-vaccinated deer. Additionally, the technique may prove invaluable for BCG efficacy studies with free-ranging deer as well as for use as a non

  14. Fecal Volatile Organic Ccompound Profiles from White-Tailed Deer (Odocoileus virginianus) as Indicators of Mycobacterium bovis Exposure or Mycobacterium bovis Bacille Calmette-Guerin (BCG) Vaccination

    PubMed Central

    Stahl, Randal S.; Ellis, Christine K.; Nol, Pauline; Waters, W. Ray; Palmer, Mitchell; VerCauteren, Kurt C.

    2015-01-01

    White-tailed deer (Odocoileus virginianus) serve as a reservoir for bovine tuberculosis, caused by Mycobacterium bovis, and can be a source of infection in cattle. Vaccination with M. bovis Bacille Calmette Guerin (BCG) is being considered for management of bovine tuberculosis in deer. Presently, no method exists to non-invasively monitor the presence of bovine tuberculosis in deer. In this study, volatile organic compound profiles of BCG-vaccinated and non-vaccinated deer, before and after experimental challenge with M. bovis strain 95–1315, were generated using solid phase microextraction fiber head-space sampling over suspended fecal pellets with analysis by gas chromatography/mass spectrometry. Chromatograms were processed using XCMS Online to characterize ion variation among treatment groups. The principal component scores resulting from significant (α = 0.05) ion responses were used to build linear discriminant analysis models. The sensitivity and specificity of these models were used to evaluate the feasibility of using this analytical approach to distinguish within group comparisons between pre- and post-M. bovis challenge: non-vaccinated male or female deer, BCG-vaccinated male deer, and the mixed gender non-vaccinated deer data. Seventeen compounds were identified in this analysis. The peak areas for these compounds were used to build a linear discriminant classification model based on principal component analysis scores to evaluate the feasibility of discriminating between fecal samples from M. bovis challenged deer, irrespective of vaccination status. The model best representing the data had a sensitivity of 78.6% and a specificity of 91.4%. The fecal head-space sampling approach presented in this pilot study provides a non-invasive method to discriminate between M. bovis challenged deer and BCG-vaccinated deer. Additionally, the technique may prove invaluable for BCG efficacy studies with free-ranging deer as well as for use as a non

  15. Fecal Volatile Organic Ccompound Profiles from White-Tailed Deer (Odocoileus virginianus) as Indicators of Mycobacterium bovis Exposure or Mycobacterium bovis Bacille Calmette-Guerin (BCG) Vaccination.

    PubMed

    Stahl, Randal S; Ellis, Christine K; Nol, Pauline; Waters, W Ray; Palmer, Mitchell; VerCauteren, Kurt C

    2015-01-01

    White-tailed deer (Odocoileus virginianus) serve as a reservoir for bovine tuberculosis, caused by Mycobacterium bovis, and can be a source of infection in cattle. Vaccination with M. bovis Bacille Calmette Guerin (BCG) is being considered for management of bovine tuberculosis in deer. Presently, no method exists to non-invasively monitor the presence of bovine tuberculosis in deer. In this study, volatile organic compound profiles of BCG-vaccinated and non-vaccinated deer, before and after experimental challenge with M. bovis strain 95-1315, were generated using solid phase microextraction fiber head-space sampling over suspended fecal pellets with analysis by gas chromatography/mass spectrometry. Chromatograms were processed using XCMS Online to characterize ion variation among treatment groups. The principal component scores resulting from significant (α = 0.05) ion responses were used to build linear discriminant analysis models. The sensitivity and specificity of these models were used to evaluate the feasibility of using this analytical approach to distinguish within group comparisons between pre- and post-M. bovis challenge: non-vaccinated male or female deer, BCG-vaccinated male deer, and the mixed gender non-vaccinated deer data. Seventeen compounds were identified in this analysis. The peak areas for these compounds were used to build a linear discriminant classification model based on principal component analysis scores to evaluate the feasibility of discriminating between fecal samples from M. bovis challenged deer, irrespective of vaccination status. The model best representing the data had a sensitivity of 78.6% and a specificity of 91.4%. The fecal head-space sampling approach presented in this pilot study provides a non-invasive method to discriminate between M. bovis challenged deer and BCG-vaccinated deer. Additionally, the technique may prove invaluable for BCG efficacy studies with free-ranging deer as well as for use as a non

  16. Occurrence of Babesia spp., Rickettsia spp. and Bartonella spp. in Ixodes ricinus in Bavarian public parks, Germany

    PubMed Central

    2011-01-01

    Background Only limited information is available about the occurrence of ticks and tick-borne pathogens in public parks, which are areas strongly influenced by human beings. For this reason, Ixodes ricinus were collected in public parks of different Bavarian cities in a 2-year survey (2009 and 2010) and screened for DNA of Babesia spp., Rickettsia spp. and Bartonella spp. by PCR. Species identification was performed by sequence analysis and alignment with existing sequences in GenBank. Additionally, coinfections with Anaplasma phagocytophilum were investigated. Results The following prevalences were detected: Babesia spp.: 0.4% (n = 17, including one pool of two larvae) in 2009 and 0.5 to 0.7% (n = 11, including one pool of five larvae) in 2010; Rickettsia spp.: 6.4 to 7.7% (n = 285, including 16 pools of 76 larvae) in 2009. DNA of Bartonella spp. in I. ricinus in Bavarian public parks could not be identified. Sequence analysis revealed the following species: Babesia sp. EU1 (n = 25), B. divergens (n = 1), B. divergens/capreoli (n = 1), B. gibsoni-like (n = 1), R. helvetica (n = 272), R. monacensis IrR/Munich (n = 12) and unspecified R. monacensis (n = 1). The majority of coinfections were R. helvetica with A. phagocytophilum (n = 27), but coinfections between Babesia spp. and A. phagocytophilum, or Babesia spp. and R. helvetica were also detected. Conclusions I. ricinus ticks in urban areas of Germany harbor several tick-borne pathogens and coinfections were also observed. Public parks are of particularly great interest regarding the epidemiology of tick-borne pathogens, because of differences in both the prevalence of pathogens in ticks as well as a varying species arrangement when compared to woodland areas. The record of DNA of a Babesia gibsoni-like pathogen detected in I. ricinus suggests that I. ricinus may harbor and transmit more Babesia spp. than previously known. Because of their high recreational value for human beings, urban green areas are likely to

  17. PCR-Based Detection of Babesia ovis in Rhipicephalus bursa and Small Ruminants.

    PubMed

    Esmaeilnejad, Bijan; Tavassoli, Mousa; Asri-Rezaei, Siamak; Dalir-Naghadeh, Bahram; Mardani, Karim; Jalilzadeh-Amin, Ghader; Golabi, Mostafa; Arjmand, Jafar

    2014-01-01

    This study aimed to assess the prevalence of Babesia ovis infection in adult Rhipicephalus bursa and small ruminants in West Azerbaijan province, Iran. Blood samples were collected from 280 sheep and 122 goats of forty randomly selected flocks. Specific B. ovis fragment was detected in 67 animals (16.7%), of which 52 animals (18.6%) were sheep and 15 animals (12.2%) goats (P < 0.05). Of the 848 R. bursa collected from naturally infested small ruminants and farm dogs, Babesia ovis was detected by PCR in salivary glands of 94 adult ticks. The frequency of B. ovis infection was higher in flocks with tick in comparison with animals without tick (P < 0.05). Positive amplification from blood of ruminants, ticks, oviposition ticks, eggs, and larvae was subjected to restriction digestion with HphI. One RFLP profile was produced. The PCR-RFLP results indicated that one strain of B. ovis exists in this area. The results showed that the PCR was useful method to investigate the epidemiology of small ruminants' babesiosis. Furthermore, R. Bursa, which can transovarially transmit B. ovis and as well as being widely distributed in West Azerbaijan province, Iran, might play an important role in the field as a natural vector of B. ovis.

  18. Development of loop-mediated isothermal amplification (LAMP) for detection of Babesia gibsoni infection in dogs.

    PubMed

    Mandal, Mrityunjay; Banerjee, Partha S; Kumar, Saroj; Ram, Hira; Garg, Rajat; Pawde, Abhijit M

    2015-04-15

    Diagnosis of canine babesiosis, caused by Babesia gibsoni is difficult, especially in chronically infected dogs. A loop mediated isothermal amplification (LAMP) assay was developed and standardized by using four oligonucleotide primers targeting the hypervariable region of 18S rRNA gene (GenBank Acc. no. KC461261). The primers specifically amplified B. gibsoni DNA, while no amplification was detected with DNA from non-infected dogs as well as from dogs infected with Babesia canis vogeli, Hepatozoon canis, Ehrlichia canis and Trypanosoma evansi. The assay could detect 1.35 × 10(-7) parasitaemia and 10(-4) dilution of recombinant plasmid, equivalent to 12 pg of target DNA. All the samples were tested by nested PCR as well as LAMP assay. LAMP was found to be 10 times more sensitive than nested PCR targeting the same gene. Out of 75 suspected field samples, collected from different parts of the country, LAMP could detect B. gibsoni in 43 samples, while nested PCR and microscopy could detect 37 and 23 samples, respectively. High sensitivity, specificity and rapidity of LAMP assay may be exploited for screening large number of samples in a field setting. PMID:25749021

  19. Microscopic and Molecular Detection of Theileria (Babesia) Equi Infection in Equids of Kurdistan Province, Iran

    PubMed Central

    HABIBI, Gholamreza; ESMAEILNIA, Kasra; HABLOLVARID, Mohammad Hasan; AFSHARI, Asghar; ZAMEN, Mohsen; BOZORGI, Soghra

    2016-01-01

    Background: Equine piroplasmosis (EP) is the cause of persistent tick-borne infection with no symptoms, but the most important problem of EP is due to the persistent carrier state. Carrier animals to Babesia (Theileria) equi (Laveran 1901) and B. caballi (Nuttall, 1910) infestation could be identified by extremely sensitive PCR-based method. The purpose of this study was to identify the causative agents of equine piroplasmosis based on molecular and microscopic assays in equids from Kurdistan Province, Iran. Methods: Thirty one horse and mule blood samples were used with history of living in Kurdistan Province of Iran. The blood specimens were utilized for T. equi and B. caballi DNA identification by PCR and Giemsa stained smears for microscopic observation. Results: The results clearly showed the presence of B. (Theileria) equi DNA in 30 of 31 blood samples (96.77%), but the microscopic examination revealed the 3 of 31 positive Babesia like organisms in the red blood cells (9.67%). Conclusion: The obtained results demonstrated the presence of hidden B. (Theileria) equi infection in horses with previous habitance in Kurdistan Province of Iran. The carrier animals became a main source of infection and can transmit the disease. Therefore, hidden infection might be considered as a health threatening and limiting factor in animals used in therapeutic antisera research and production centers. PMID:27095973

  20. Coinfection of sheep with Anaplasma, Theileria and Babesia species in the Kurdistan Region, Iraq.

    PubMed

    Renneker, S; Abdo, J; Bakheit, M A; Kullmann, B; Beyer, D; Ahmed, J; Seitzer, U

    2013-11-01

    Infections of small ruminants with Anaplasma, Theileria and Babesia species are widely distributed in the old world and are of great economic impact. In Iraq, data on disease occurrence in sheep caused by above-mentioned infectious agents are scarce. This study provides information on various haemoparasitic agents infecting sheep in the Kurdistan Region, Iraq, using molecular diagnostic tools. Altogether, 195 samples originating from three governorates in the Kurdistan Region, namely Duhok, Erbil and Sulaimaniya, were analysed. The following pathogens were identified: Anaplasma ovis (62.6%), Theileria ovis (14.35%), T. lestoquardi (7.7%), T. uilenbergi (5.6%) and Babesia ovis (1.5%). T. uilenbergi is detected for the first time in Iraq. Coinfection of sheep with different pathogens could be observed in this study, and it was found that 45 of 195 (23%) of the samples contained more than one pathogen. Even triple-positive samples were identified in 3% of the investigated animals. In conclusion, we confirm the coinfection of sheep with various haemoparasitic pathogen species in the Kurdistan Region of Iraq. Further investigations are needed to reveal the epidemiology of the diseases, the respective tick vectors, and, in the case of coinfection, pathogens' interaction and possible cross-protection.

  1. Detection of Leishmania infantum, Babesia canis, and rickettsiae in ticks removed from dogs living in Italy.

    PubMed

    Trotta, Michele; Nicetto, Martina; Fogliazza, Alessandro; Montarsi, Fabrizio; Caldin, Marco; Furlanello, Tommaso; Solano-Gallego, Laia

    2012-12-01

    The aims of this study were to determine natural infections by Anaplasma phagocytophilum/Anaplasma platys, Bartonella henselae, Ehrlichia canis, Leishmania infantum, Rickettsia spp., Babesia spp., and Hepatozoon spp. by molecular methods in ticks (n=91) removed from dogs with clinical signs and laboratory abnormalities compatible with tick-borne diseases (n=22) living in Italy and to assess the distribution and species of ticks encountered. Ticks from dogs living in southern Italy were all identified as Rhipicephalus sanguineus (n=25), ticks from central Italy included Rh. sanguineus (n=8) and Ixodes ricinus (n=9), ticks from northern Italy included Rh. sanguineus (n=45), Dermacentor marginatus (n=3), and one I. ricinus. Leishmania infantum, Rickettsia spp., and Babesia canis were the only pathogens detected in 7 (8%), 4 (4%), and 2 (2%) out of 91 ticks, respectively. L. infantum was detected in I. ricinus from central Italy and in Rh. sanguineus from northern and central Italy. Rickettsia conorii and Ri. massiliae were detected in Rh. sanguineus ticks from central and southern Italy (Sicily), respectively. Bab. canis was detected in D. marginatus ticks from northern Italy.

  2. Morphological and Molecular Descriptors of the Developmental Cycle of Babesia divergens Parasites in Human Erythrocytes

    PubMed Central

    Rossouw, Ingrid; Maritz-Olivier, Christine; Niemand, Jandeli; van Biljon, Riette; Smit, Annel; Olivier, Nicholas A.; Birkholtz, Lyn-Marie

    2015-01-01

    Human babesiosis, especially caused by the cattle derived Babesia divergens parasite, is on the increase, resulting in renewed attentiveness to this potentially life threatening emerging zoonotic disease. The molecular mechanisms underlying the pathophysiology and intra-erythrocytic development of these parasites are poorly understood. This impedes concerted efforts aimed at the discovery of novel anti-babesiacidal agents. By applying sensitive cell biological and molecular functional genomics tools, we describe the intra-erythrocytic development cycle of B. divergens parasites from immature, mono-nucleated ring forms to bi-nucleated paired piriforms and ultimately multi-nucleated tetrads that characterizes zoonotic Babesia spp. This is further correlated for the first time to nuclear content increases during intra-erythrocytic development progression, providing insight into the part of the life cycle that occurs during human infection. High-content temporal evaluation elucidated the contribution of the different stages to life cycle progression. Moreover, molecular descriptors indicate that B. divergens parasites employ physiological adaptation to in vitro cultivation. Additionally, differential expression is observed as the parasite equilibrates its developmental stages during its life cycle. Together, this information provides the first temporal evaluation of the functional transcriptome of B. divergens parasites, information that could be useful in identifying biological processes essential to parasite survival for future anti-babesiacidal discoveries. PMID:25955414

  3. Molecular detection and genetic diversity of Babesia gibsoni in dogs in Bangladesh.

    PubMed

    Terao, Masashi; Akter, Shirin; Yasin, Md Golam; Nakao, Ryo; Kato, Hirotomo; Alam, Mohammad Zahangir; Katakura, Ken

    2015-04-01

    Babesia gibsoni is a tick-borne hemoprotozoan parasite of dogs that often causes fever and hemolytic illness. Detection of B. gibsoni has been predominantly reported in Asian countries, including Japan, Korea, Taiwan, Malaysia, Bangladesh and India. The present study shows the first molecular characterization of B. gibsoni detected from dogs in Bangladesh. Blood samples were collected on FTA® Elute cards from 50 stray dogs in Mymensingh District in Bangladesh. DNA eluted from the cards was subjected to nested PCR for the 18S rRNA gene of Babesia species. Approximately 800bp PCR products were detected in 15 of 50 dogs (30%). Based on restriction fragment length polymorphism (RFLP) and direct sequencing of the PCR products, all parasite isolates were identified as B. gibsoni. Furthermore, the BgTRAP (B. gibsoni thrombospondin-related adhesive protein) gene fragments were detected in 13 of 15 18S rRNA gene PCR positive blood samples. Phylogenetic analysis of the BgTRAP gene revealed that B. gibsoni parasites in Bangladesh formed a cluster, which was genetically different from other Asian B. gibsoni isolates. In addition, tandem repeat analysis of the BgTRAP gene clearly showed considerable genetic variation among Bangladeshi isolates. These results suggested that B. gibsoni parasites in a different genetic clade are endemic in dogs in Bangladesh. Further studies are required to elucidate the origin, distribution, vector and pathogenesis of B. gibsoni parasites circulating in dogs in Bangladesh.

  4. Detecting DNAs of Anaplasma phagocytophilum and Babesia in the blood of patients suspected of Lyme disease.

    PubMed

    Hermanowska-Szpakowicz, Teresa; Skotarczak, Bogumiła; Kondrusik, Maciej; Rymaszewska, Anna; Sawczuk, Marek; Maciejewska, Agnieszka; Adamska, Małgorzata; Pancewicz, Sławomir; Zajkowska, Joanna

    2004-01-01

    Co-occurrence of granulocytic anaplasmosis, borreliosis and babesiosis in humans is a result of common vectors for the respective pathogens of these diseases, most commonly ticks from the genus Ixodes. Studies on ticks in Europe and also in Poland have shown that several pathogens may co-occur in individuals of I. ricnus. A total of 96 hospitalised patients infected or suspected of being infected with borreliosis were screened for A. phagocytophilum and Babesia sp. DNA. Positive results of PCRs for A. phagocytophilum DNA were obtained for 10 patients, 8 of whom were diagnosed with borreliosis earlier, and 4 of whom were diagnosed with tick-borne encephalitis (on the basis of serological studies of serum and cerebrospinal fluid). None of the 10 patients had clinical or biochemical markers of anaplasmosis, corroborating the existence of asymptomatic anaplasmosis or self-limiting course. in Europe. Similarly, Babesia DNA was not found in the blood of any of the patients. The results of the studies show that in diagnosing tick-borne diseases, clinical examinations should consider infection by two or even three tick-borne pathogens. PMID:15627349

  5. Survey of babesiosis in symptomatic dogs from Romania: occurrence of Babesia gibsoni associated with breed.

    PubMed

    Imre, Mirela; Farkas, Róbert; Ilie, Marius Stelian; Imre, Kálmán; Dărăbuş, Gheorghe

    2013-12-01

    Blood samples from 49 symptomatic dogs from 5 western and north-western counties of Romania were screened using microscopic examination, polymerase-chain-reaction-restriction-fragment-length-polymorphism procedure (PCR-RFLP), and sequence analysis. Results of the microscopic evaluation of stained blood smears revealed 45 positive samples with the presence of large and small intraerythrocytic piroplasms in 35 and 10 samples, respectively. Babesia canis (35/49, 71.4%) and Babesia gibsoni (14/49, 28.6%) were identified and differentiated by PCR-RFLP targeting the 18S rRNA gene. Results of the sequence analysis of all B. gibsoni and 17 randomly selected B. canis PCR products confirmed the PCR-RFLP-diagnosed species. The distribution of B. gibsoni infection was positively associated (p<0.001) with fighting dog breeds including infection in 12 American Pit Bull Terriers and one American Staffordshire Terrier. This report is the first to present molecular evidence of the occurrence of B. gibsoni in Romania confirmed by sequencing.

  6. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species☆

    PubMed Central

    Springer, Andrea; Fichtel, Claudia; Calvignac-Spencer, Sébastien; Leendertz, Fabian H.; Kappeler, Peter M.

    2015-01-01

    Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites. PMID:26767166

  7. Structural and Functional Characterization of Bc28.1, Major Erythrocyte-binding Protein from Babesia canis Merozoite Surface*

    PubMed Central

    Yang, Yin-Shan; Murciano, Brice; Moubri, Karina; Cibrelus, Prisca; Schetters, Theo; Gorenflot, André; Delbecq, Stéphane; Roumestand, Christian

    2012-01-01

    Babesiosis (formerly known as piroplasmosis) is a tick-borne disease caused by the intraerythrocytic development of protozoa parasites from the genus Babesia. Like Plasmodium falciparum, the agent of malaria, or Toxoplasma gondii, responsible for human toxoplasmosis, Babesia belongs to the Apicomplexa family. Babesia canis is the agent of the canine babesiosis in Europe. Clinical manifestations of this disease range from mild to severe and possibly lead to death by multiple organ failure. The identification and characterization of parasite surface proteins represent major goals, both for the understanding of the Apicomplexa invasion process and for the vaccine potential of such antigens. Indeed, we have already shown that Bd37, the major antigenic adhesion protein from Babesia divergens, the agent of bovine babesiosis, was able to induce complete protection against various parasite strains. The major merozoite surface antigens of Babesia canis have been described as a 28-kDa membrane protein family, anchored at the surface of the merozoite. Here, we demonstrate that Bc28.1, a major member of this multigenic family, is expressed at high levels at the surface of the merozoite. This protein is also found in the parasite in vitro culture supernatants, which are the basis of effective vaccines against canine babesiosis. We defined the erythrocyte binding function of Bc28.1 and determined its high resolution solution structure using NMR spectroscopy. Surprisingly, although these proteins are thought to play a similar role in the adhesion process, the structure of Bc28.1 from B. canis appears unrelated to the previously published structure of Bd37 from B. divergens. Site-directed mutagenesis experiments also suggest that the mechanism of the interaction with the erythrocyte membrane could be different for the two proteins. The resolution of the structure of Bc28 represents a milestone for the characterization of the parasite erythrocyte binding and its interaction with

  8. Molecular identification of Theileria and Babesia in sheep and goats in the Black Sea Region in Turkey.

    PubMed

    Aydin, Mehmet Fatih; Aktas, Munir; Dumanli, Nazir

    2013-08-01

    This study was carried out to investigate presence and distribution of Theileria and Babesia species via microscopic examination and reverse line blotting (RLB) techniques in sheep and goats in the Black Sea region of Turkey. For this purpose, 1,128 blood samples (869 sheep and 259 goats) were collected by active surveillance from sheep and goats in different provinces of various cities in the region in the years 2010 and 2011. Smears were prepared from the blood samples, stained with Giemsa, and examined under the light microscope for Theileria and Babesia piroplasms. The genomic DNAs were extracted from blood samples. The length of 360-430-bp fragment in the variable V4 region of 18S SSU rRNA gene of Theileria and Babesia species was amplified using the gDNAs. The polymerase chain reaction products were hybridized to the membrane-connected species-specific probes. A total of 38 animals (3.37%) including 34 sheep (3.91%) and 4 goats (1.54%) were found to be positive for Theileria spp. piroplasms in microscopic examination of smears while Babesia spp. piroplasm could not detected. Infection rates were 34.64% in sheep, 10.04% in goats, and totally 28.99% for Theileria ovis while 0.58% in sheep and totally 0.44% for Babesia ovis. However, Theileria sp. OT3 was detected in 2.65% of sheep and 2.04% of all animals; besides Theileria sp., MK had 0.58% prevalence in sheep and 0.77% in goats, with a total 0.62% with RLB. Although T. ovis and Theileria sp. MK were determined in both sheep and goats, B. ovis and Theileria sp. OT3 were observed only in the sheep. These results provide the first detailed molecular data for sheep and goat theileriosis and babesiosis in the region.

  9. Molecular identification of Theileria and Babesia in ticks collected from sheep and goats in the Black Sea region of Turkey.

    PubMed

    Aydin, Mehmet Fatih; Aktas, Munir; Dumanli, Nazir

    2015-01-01

    A molecular survey was undertaken in the Black Sea region of Turkey to determine the presence of Theileria and Babesia species of medical and veterinary importance. The ticks were removed from sheep and goats, pooled according to species and locations, and analyzed by PCR-based reverse line blot (RLB) and sequencing. A total of 2241 ixodid ticks belonging to 5 genus and 12 species were collected and divided into 310 pools. Infection rates were calculated as the maximum likelihood estimation (MLE) with 95% confidence intervals (CI). Of the 310 pools tested, 46 (14.83%) were found to be infected with Theileria or Babesia species, and the overall MLE of the infection rate was calculated as 2.27% (CI 1.67-2.99). The MLE of the infection rates were calculated as 0.691% (CI 0.171-1.78) in Haemaphysalis parva, 1.47% (CI 0.081-6.37) in Rhipicephalus sanguineus, 1.84% (CI 0.101-7.87) in Ixodes ricinus, 2.86% (CI 1.68-4.48) in Rhipicephalus turanicus, 5.57% (CI 0.941-16.3) in Hyalomma marginatum, and 6.2% (CI 4.02-9.02) in Rhipicephalus bursa. Pathogens identified in ticks included Theileria ovis, Babesia ovis, Babesia bigemina, and Babesia microti. Most tick pools were infected with a single pathogen. However, five pools displayed mixed infections with T. ovis and B. ovis. This study provides the first molecular evidence for the presence of B. microti in ticks in Turkey.

  10. Molecular detection of Theileria spp. and Babesia spp. in sheep and ixodid ticks from the northeast of Iran.

    PubMed

    Razmi, Gholamreza; Pourhosseini, Moslem; Yaghfouri, Saeed; Rashidi, Ahmad; Seidabadi, Mohsen

    2013-02-01

    Theilerioses and babesioses are important diseases in Iranian sheep. The present study was undertaken to identify and classify/specify Theileria spp. and Babesia spp. in sheep and vector ticks. Investigation was carried out from 2009 to 2011 in the Khorasan Razavi Province, Iran. In total, 302 sheep originating from 60 different flocks were clinically examined and their blood collected. In addition, from the same flocks, ixodid ticks were sampled. Stained blood smears were microscopically examined for the presence of Theileria and Babesia organisms, and a semi-nested PCR was used for subsequent molecular specification. From the ticks, salivary glands and uterus were isolated and subsequently analyzed by semi-nested PCR. Piroplasm organisms were observed in 29% of the blood smears with low parasitemia, whereas 65% of the blood samples yielded positive PCR findings. The presence of Theileria ovis (55.6%), Theileria lestoquardi, and mixed infection with Theileria spp. and Babesia ovis were detected by semi-nested PCR in 0.3%, 5.6%, and 0.99%, respectively. In total, 429 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 376; 87.6% of the total), followed by Hyalomma marginatum turanicum (n = 30; 7.0%), Dermacentor raskemensis (n = 12; 2.8%), Hyalomma anatolicum anatolicum (n = 7; 1.6%), Dermacentor marginatus (n = 2; 0.5%), Rhipicephalus bursa (n = 1; 0.2%), and Haemaphysalis sp. (n = 1; 0.2%). Of the positive R. turanicus samples, 5 (5.7%) were infected with T. ovis and 2 (2.9%) with T. lestoquardi. Neither Babesia ovis nor Babesia motasi infection was detected in salivary glands or uterine samples of the ticks. The results also suggest that R. turanicus could be the vector responsible for transmission of the 2 Theileria species.

  11. Hemoparasites in a wild primate: Infection patterns suggest interaction of Plasmodium and Babesia in a lemur species.

    PubMed

    Springer, Andrea; Fichtel, Claudia; Calvignac-Spencer, Sébastien; Leendertz, Fabian H; Kappeler, Peter M

    2015-12-01

    Hemoparasites can cause serious morbidity in humans and animals and often involve wildlife reservoirs. Understanding patterns of hemoparasite infections in natural populations can therefore inform about emerging disease risks, especially in the light of climate change and human disruption of natural ecosystems. We investigated the effects of host age, sex, host group size and season on infection patterns of Plasmodium sp., Babesia sp. and filarial nematodes in a population of wild Malagasy primates, Verreaux's sifakas (Propithecus verreauxi), as well as the effects of these infections on hematological variables. We tested 45 blood samples from 36 individuals and identified two species of Plasmodium, one species of Babesia and two species of filarial nematodes. Plasmodium spp. and Babesia sp. infections showed opposite patterns of age-dependency, with babesiosis being prevalent among young animals, while older animals were infected with Plasmodium sp. In addition, Babesia sp. infection was a statistically significant negative predictor of Plasmodium sp. infection. These results suggest that Plasmodium and Babesia parasites may interact within the host, either through cross-immunity or via resource competition, so that Plasmodium infections can only establish after babesiosis has resolved. We found no effects of host sex, host group size and season on hemoparasite infections. Infections showed high prevalences and did not influence hematological variables. This preliminary evidence supports the impression that the hosts and parasites considered in this study appear to be well-adapted to each other, resulting in persistent infections with low pathogenic and probably low zoonotic potential. Our results illustrate the crucial role of biodiversity in host-parasite relationships, specifically how within-host pathogen diversity may regulate the abundance of parasites. PMID:26767166

  12. Mycobacterium bovis DNA detection in colostrum as a potential indicator of vaccination effectiveness against bovine tuberculosis.

    PubMed

    Herrera-Rodríguez, Sara E; Gordiano-Hidalgo, María Alejandra; López-Rincón, Gonzálo; Bojorquez-Narváez, Luis; Padilla-Ramírez, Francisco Javier; Pereira-Suárez, Ana Laura; Flores-Valdez, Mario Alberto; Estrada-Chávez, Ciro

    2013-04-01

    Bovine tuberculosis (bTB) remains a problem on many dairy farms in Mexico, as well as a public health risk. We previously found a high frequency of Mycobacterium bovis DNA in colostrum from dairy cows using a nested PCR to detect mpb70. Since there are no reliable in vivo tests to determine the effectiveness of booster Mycobacterium bovis BCG vaccination against bTB, in this work we monitored M. bovis DNA in colostrum by using this nested PCR. In order to decrease the risk of adverse reactions in animals likely containing viable M. bovis, a single application of BCG and a subunit vaccine (EEP-1) formulated with M. bovis culture filtrate proteins (CFP) and a copolymer as the adjuvant was performed in tuberculin skin test-negative cattle (TST(-)), while TST reactor animals (TST(+)) received EEP-1 only. Booster immunization using EEP-1 was applied to both groups, 2 months after primary vaccination to whole herds and 12 months later to lactating cows. Colostrum samples were collected from 6 farms where the cows were vaccinated over a 12-month period postvaccination and, for comparison, from one control farm where the cows were not vaccinated with comparable bTB prevalence. We observed an inverse relationship between the frequency of M. bovis DNA detection and time postvaccination at the first (P < 0.001) and second (P < 0.0001) 6-month periods. Additionally, the concentration of gamma interferon (IFN-γ) was higher in mpb70 PCR-positive colostrum samples (P = 0.0003). These results suggest that M. bovis DNA frequency in colostrum could be a potentially useful biomarker for bTB vaccine efficacy on commercial dairy farms. PMID:23425597

  13. Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum strains to antimicrobial agents in vitro.

    PubMed Central

    ter Laak, E A; Noordergraaf, J H; Verschure, M H

    1993-01-01

    The purpose of this study was to determine the susceptibility of various strains of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum, which are prevalent causes of pneumonia in calves, to 16 antimicrobial agents in vitro. The MICs of the antimicrobial agents were determined by a serial broth dilution method for 16 field strains and the type strain of M. bovis, for 19 field strains and the type strain of M. dispar, and for 17 field strains of U. diversum. Final MICs for M. bovis and M. dispar were read after 7 days and final MICs for U. diversum after 1 to 2 days. All strains tested were susceptible to tylosin, kitasamycin, and tiamulin but were resistant to nifuroquine and streptomycin. Most strains of U. diversum were intermediately susceptible to oxytetracycline but fully susceptible to chlortetracycline; most strains of M. bovis and M. dispar, however, were resistant to both agents. Strains of M. dispar and U. diversum were susceptible to doxycycline and minocycline, but strains of M. bovis were only intermediately susceptible. Susceptibility or resistance to chloramphenicol, spiramycin, spectinomycin, lincomycin, or enrofloxacin depended on the species but was not equal for the three species. The type strains of M. bovis and M. dispar were more susceptible to various antimicrobial agents, including tetracyclines, than the field strains. This finding might indicate that M. bovis and M. dispar strains are becoming resistant to these agents. Antimicrobial agents that are effective in vitro against all three mycoplasma species can be considered for treating mycoplasma infections in pneumonic calves. Therefore, tylosin, kitasamycin, and tiamulin may be preferred over oxytetracycline and chlortetracycline. PMID:8452363

  14. Development and host compatibility of plasmids for two important ruminant pathogens, Mycoplasma bovis and Mycoplasma agalactiae.

    PubMed

    Sharma, Shukriti; Citti, Chistine; Sagné, Eveline; Marenda, Marc S; Markham, Philip F; Browning, Glenn F

    2015-01-01

    Mycoplasma bovis is a cause of pneumonia, mastitis, arthritis and otitis media in cattle throughout the world. However, despite its clinical significance, there is a paucity of tools to genetically manipulate it, impeding our capacity to further explore the molecular basis of its virulence. To address this limitation, we developed a series of homologous and heterologous replicable plasmids from M. bovis and M. agalactiae. The shortest replicable oriC plasmid based on the region downstream of dnaA in M. bovis was 247 bp and contained two DnaA boxes, while oriC plasmids based on the region downstream of dnaA in M. agalactiae strains 5632 and PG2 were 219 bp and 217 bp in length, respectively, and contained only a single DnaA box. The efficiency of transformation in M. bovis and M. agalactiae was inversely correlated with the size of the oriC region in the construct, and, in general, homologous oriC plasmids had a higher transformation efficiency than heterologous oriC plasmids. The larger pWholeoriC45 and pMM21-7 plasmids integrated into the genomic oriC region of M. bovis, while the smaller oriC plasmids remained extrachromosomal for up to 20 serial passages in selective media. Although specific gene disruptions were not be achieved in M. bovis in this study, the oriC plasmids developed here could still be useful as tools in complementation studies and for expression of exogenous genes in both M. bovis and M. agalactiae.

  15. Mycobacterium bovis DNA detection in colostrum as a potential indicator of vaccination effectiveness against bovine tuberculosis.

    PubMed

    Herrera-Rodríguez, Sara E; Gordiano-Hidalgo, María Alejandra; López-Rincón, Gonzálo; Bojorquez-Narváez, Luis; Padilla-Ramírez, Francisco Javier; Pereira-Suárez, Ana Laura; Flores-Valdez, Mario Alberto; Estrada-Chávez, Ciro

    2013-04-01

    Bovine tuberculosis (bTB) remains a problem on many dairy farms in Mexico, as well as a public health risk. We previously found a high frequency of Mycobacterium bovis DNA in colostrum from dairy cows using a nested PCR to detect mpb70. Since there are no reliable in vivo tests to determine the effectiveness of booster Mycobacterium bovis BCG vaccination against bTB, in this work we monitored M. bovis DNA in colostrum by using this nested PCR. In order to decrease the risk of adverse reactions in animals likely containing viable M. bovis, a single application of BCG and a subunit vaccine (EEP-1) formulated with M. bovis culture filtrate proteins (CFP) and a copolymer as the adjuvant was performed in tuberculin skin test-negative cattle (TST(-)), while TST reactor animals (TST(+)) received EEP-1 only. Booster immunization using EEP-1 was applied to both groups, 2 months after primary vaccination to whole herds and 12 months later to lactating cows. Colostrum samples were collected from 6 farms where the cows were vaccinated over a 12-month period postvaccination and, for comparison, from one control farm where the cows were not vaccinated with comparable bTB prevalence. We observed an inverse relationship between the frequency of M. bovis DNA detection and time postvaccination at the first (P < 0.001) and second (P < 0.0001) 6-month periods. Additionally, the concentration of gamma interferon (IFN-γ) was higher in mpb70 PCR-positive colostrum samples (P = 0.0003). These results suggest that M. bovis DNA frequency in colostrum could be a potentially useful biomarker for bTB vaccine efficacy on commercial dairy farms.

  16. Mycobacterium bovis DNA Detection in Colostrum as a Potential Indicator of Vaccination Effectiveness against Bovine Tuberculosis

    PubMed Central

    Herrera-Rodríguez, Sara E.; Gordiano-Hidalgo, María Alejandra; López-Rincón, Gonzálo; Bojorquez-Narváez, Luis; Padilla-Ramírez, Francisco Javier; Pereira-Suárez, Ana Laura; Flores-Valdez, Mario Alberto

    2013-01-01

    Bovine tuberculosis (bTB) remains a problem on many dairy farms in Mexico, as well as a public health risk. We previously found a high frequency of Mycobacterium bovis DNA in colostrum from dairy cows using a nested PCR to detect mpb70. Since there are no reliable in vivo tests to determine the effectiveness of booster Mycobacterium bovis BCG vaccination against bTB, in this work we monitored M. bovis DNA in colostrum by using this nested PCR. In order to decrease the risk of adverse reactions in animals likely containing viable M. bovis, a single application of BCG and a subunit vaccine (EEP-1) formulated with M. bovis culture filtrate proteins (CFP) and a copolymer as the adjuvant was performed in tuberculin skin test-negative cattle (TST−), while TST reactor animals (TST+) received EEP-1 only. Booster immunization using EEP-1 was applied to both groups, 2 months after primary vaccination to whole herds and 12 months later to lactating cows. Colostrum samples were collected from 6 farms where the cows were vaccinated over a 12-month period postvaccination and, for comparison, from one control farm where the cows were not vaccinated with comparable bTB prevalence. We observed an inverse relationship between the frequency of M. bovis DNA detection and time postvaccination at the first (P < 0.001) and second (P < 0.0001) 6-month periods. Additionally, the concentration of gamma interferon (IFN-γ) was higher in mpb70 PCR-positive colostrum samples (P = 0.0003). These results suggest that M. bovis DNA frequency in colostrum could be a potentially useful biomarker for bTB vaccine efficacy on commercial dairy farms. PMID:23425597

  17. Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum strains to antimicrobial agents in vitro.

    PubMed

    ter Laak, E A; Noordergraaf, J H; Verschure, M H

    1993-02-01

    The purpose of this study was to determine the susceptibility of various strains of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum, which are prevalent causes of pneumonia in calves, to 16 antimicrobial agents in vitro. The MICs of the antimicrobial agents were determined by a serial broth dilution method for 16 field strains and the type strain of M. bovis, for 19 field strains and the type strain of M. dispar, and for 17 field strains of U. diversum. Final MICs for M. bovis and M. dispar were read after 7 days and final MICs for U. diversum after 1 to 2 days. All strains tested were susceptible to tylosin, kitasamycin, and tiamulin but were resistant to nifuroquine and streptomycin. Most strains of U. diversum were intermediately susceptible to oxytetracycline but fully susceptible to chlortetracycline; most strains of M. bovis and M. dispar, however, were resistant to both agents. Strains of M. dispar and U. diversum were susceptible to doxycycline and minocycline, but strains of M. bovis were only intermediately susceptible. Susceptibility or resistance to chloramphenicol, spiramycin, spectinomycin, lincomycin, or enrofloxacin depended on the species but was not equal for the three species. The type strains of M. bovis and M. dispar were more susceptible to various antimicrobial agents, including tetracyclines, than the field strains. This finding might indicate that M. bovis and M. dispar strains are becoming resistant to these agents. Antimicrobial agents that are effective in vitro against all three mycoplasma species can be considered for treating mycoplasma infections in pneumonic calves. Therefore, tylosin, kitasamycin, and tiamulin may be preferred over oxytetracycline and chlortetracycline.

  18. Isolation and molecular characterization of Mycobacterium bovis from Kafue lechwe (Kobus leche kafuensis) from Zambia.

    PubMed

    Malama, Sydney; Johansen, Tone Bjordal; Muma, John Bwalya; Mwanza, Sydney; Djønne, Berit; Godfroid, Jacques

    2014-01-01

    Bovine tuberculosis (BTB) is a chronic bacterial disease caused by Mycobacterium bovis. Infections due to M. bovis, which serves as a stable reservoir, can pose serious challenge to control and eradicate in both wildlife and livestock at the interface. This study aimed at isolating and characterizing M. bovis from Kafue lechwe (Kobus leche kafuensis) and black lechwe (Kobus leche smithemani) at the animal/human interface in Zambia. The samples with lesions compatible with BTB collected during the hunting seasons of 2009 and 2010 were cultured for isolation of mycobacteria using Stonebrink with pyruvate (BD Diagnostics, MD, USA) and Middlebrook 7H10 (BD Diagnostics) slants. Isolated mycobacteria were identified using IS6110 polymerase chain reaction and deletion analysis. Molecular characterization of the isolates was performed using spoligotyping and mycobacteria interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) with nine loci. Data was analyzed using BioNumerics software 6.1. Out of the 39 samples, acid fast bacilli were detected in 27 (69.2 %) based on smear microscopy. Seven isolates were found to belong to Mycobacterium tuberculosis complex, and all were identified as M. bovis based on deletion analysis. All seven isolates were identical on spoligotyping as belonging to the SB0120 (SIT 482). MIRU-VNTR differentiated the isolates into five different patterns. This study has confirmed that M. bovis circulates in the Kafue lechwe, and non-tuberculous mycobacteria were detected in the black lechwe in Zambia which represents a wildlife reservoir, with a potential to spillover to cattle and humans. Isolates of M. bovis from lechwe antelopes are much conserved as only one spoligotype was detected. The study has shown that three loci differentiated fairly well. This option is cheap and less laborious, and hence a better option in resource-strained country like Zambia. The study further showed that some of the loci recommended by the European

  19. Toxicogenomic response of Mycobacterium bovis BCG to peracetic acid and a comparative analysis of the M. bovis BCG response to three oxidative disinfectants.

    PubMed

    Nde, Chantal W; Toghrol, Freshteh; Jang, Hyeung-Jin; Bentley, William E

    2011-04-01

    Tuberculosis is a leading cause of death worldwide and infects thousands of Americans annually. Mycobacterium bovis causes tuberculosis in humans and several animal species. Peracetic acid is an approved tuberculocide in hospital and domestic environments. This study presents for the first time the transcriptomic changes in M. bovis BCG after treatment with 0.1 mM peracetic acid for 10 and 20 min. This study also presents for the first time a comparison among the transcriptomic responses of M. bovis BCG to three oxidative disinfectants: peracetic acid, sodium hypochlorite, and hydrogen peroxide after 10 min of treatment. Results indicate that arginine biosynthesis, virulence, and oxidative stress response genes were upregulated after both peracetic acid treatment times. Three DNA repair genes were downregulated after 10 and 20 min and cell wall component genes were upregulated after 20 min. The devR-devS signal transduction system was upregulated after 10 min, suggesting a role in the protection against peracetic acid treatment. Results also suggest that peracetic acid and sodium hypochlorite both induce the expression of the ctpF gene which is upregulated in hypoxic environments. Further, this study reveals that in M. bovis BCG, hydrogen peroxide and peracetic acid both induce the expression of katG involved in oxidative stress response and the mbtD and mbtI genes involved in iron regulation/virulence. PMID:21152916

  20. Humoral Immune Responses of White-tailed Deer (Odocoileus virginianus) to Mycobacterium bovis BCG Vaccination and Experimental Challenge with M. bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Monitoring serum antibody production kinetics to multiple mycobacterial antigens can be useful as a diagnostic tool for the detection of Mycobacterium bovis infection as well as for the characterization of disease progression and efficacy of intervention strategies in several species. Humoral immun...

  1. T-cell mRNA Expression in Response to Mycobacterium bovis BCG Vaccination and Mycobacterium bovis Infection of White-tailed deer

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Understanding immune responses of white-tailed deer (WTD) to infection with Mycobacterium bovis provides insight into mechanisms of pathogen control and may provide clues to development of effective vaccine strategies. WTD were vaccinated with either BCG strain Pasteur or BCG Danish. Both vaccinates...

  2. Babesia infection in naturally exposed pet dogs from a north-eastern state (Assam) of India: detection by microscopy and polymerase chain reaction.

    PubMed

    Laha, R; Bhattacharjee, K; Sarmah, P C; Das, M; Goswami, A; Sarma, D; Sen, A

    2014-12-01

    The objective of the study was to detect Babesia infections in pet dogs of a north-eastern state of India. The diagnostic efficacy of Babesia infection by polymerase chain reaction (PCR) technique has been compared with microscopy examination. For this, a total of 111 blood samples of pet dogs presented at clinical complex of the College of Veterinary Science, Guwahati, Assam with clinical signs suspected for Babesia infection subjected to the study. A total of 44 (39.63 %) dogs were diagnosed as positive for Babesia infections after microscopic examination. Among these, Babesia canis infection was diagnosed in 5 dogs (4.50 %) and B. gibsoni infection in 39 (35.13 %) dogs microscopically in Giemsa stained blood smears. Molecular diagnosis using PCR detected 63 (56.75 %) dogs positive for Babesia infection. Single infection with B. canis was found in 9 (8.10 %) dogs while B. gibsoni alone was detected in 3 (2.70 %) dogs. Mixed infections by both these species were detected in 51 (45.94 %) dogs. Overall, PCR detected 54 (48.64 %) dogs as B. gibsoni and 60 (54.05 %) dogs as B. canis positive. PMID:25320489

  3. Detection of Babesia Sp. EU1 and members of spotted fever group rickettsiae in ticks collected from migratory birds at Curonian Spit, North-Western Russia.

    PubMed

    Movila, Alexandru; Reye, Anna L; Dubinina, Helen V; Tolstenkov, Oleg O; Toderas, Ion; Hübschen, Judith M; Muller, Claude P; Alekseev, Andrey N

    2011-01-01

    To reveal the prevalence of spotted fever group (SFG) rickettsiae and Babesia sp. in Ixodes ricinus (L.) ticks from migratory birds, 236 specimens represented 8 species of Passeriformes and were collected at Curonian Spit in Kaliningrad enclave of North-Western Russia. The ticks (total 126) being detached from four bird species, Turdus philomelos, Fringilla coelebs, Parus major, and Sturnus vulgaris, were investigated by PCR using the primers Rp CS.877p/Rp CS.1258n for the detection of Rickettsia and BJ1/BN2 for Babesia spp. Babesia spp. were detected in 2 of 126 (1.6%) ticks. The partial sequence of 18S rDNA had 100% similarity to human pathogenic Babesia sp. EU1. The SFG rickettsiae were detected in 19 of 126 (15.1%) ticks collected from the above-mentioned bird species. BLAST analysis of SFG rickettsia gltA assigned sequences to human pathogenic Rickettsia helvetica (10.3%), Rickettsia monacensis (3.9%), and Rickettsia japonica (0.8%) with 98%-100% sequence similarity. The SFG rickettsiae and Babesia sp. EU1 in ticks collected from the passerines in Russia were detected for the first time. The survey indicates that migratory birds may become a reservoir for Babesia spp. and SFG rickettsiae. Future investigations need to characterize the role of birds in the epidemiology of these human pathogens in the region.

  4. A Study of Naturally Acquired Canine Babesiosis Caused by Single and Mixed Babesia Species in Zambia: Clinicopathological Findings and Case Management

    PubMed Central

    Nalubamba, King Shimumbo; Mudenda, Ntombi Basimbi; Namwila, Mwaka Mwangala; Mulenga, Chilufya Susan; Bwalya, Eugene Chisela; M'kandawire, Ethel; Saasa, Ngonda; Hankanga, Careen; Oparaocha, Elizabeth; Simuunza, Martin

    2015-01-01

    A retrospective and prospective analysis of clinical records of dogs diagnosed with Babesia infections was carried out for the years 2000 to 2013 from practices in Lusaka, Zambia. Records of 363 dogs with confirmed Babesia infections were analysed using demographic factors including sex, breed, age, and clinical signs in relation to haematological findings and Babesia species. The clinical and laboratory findings observed are described as well as Babesia species identification. The study included 18 breeds and the highest proportion were mongrels (32.2%), males representing 64.5% of the population. The most common presenting problems were anorexia (65.3%) and lethargy/weakness (65.3%). The most common clinical signs were fever (87.3%), pallor (52.3%), lymphadenopathy (47.4%), and presence of ticks (44.9%). Anaemia (96.4%) and nucleated erythrocytes (42.2%) were the most common laboratory findings. A mixed infection of Babesia rossi and Babesia gibsoni was present in 59.7% of dogs, whilst 8% and 32.2% had B. rossi and B. gibsoni as a single infection, respectively. Case management mainly involved therapy with tetracyclines and imidocarb and was usually accompanied by clinical improvement. This study highlights, for the first time, the presence of B. gibsoni in natural dog populations in Zambia, where previously only B. rossi was reported. PMID:26682062

  5. Allele-Specific PCR Method Based on pncA and oxyR Sequences for Distinguishing Mycobacterium bovis from Mycobacterium tuberculosis: Intraspecific M. bovis pncA Sequence Polymorphism

    PubMed Central

    de los Monteros, Luz Elena Espinosa; Galán, Juan Carlos; Gutiérrez, Montserrat; Samper, Sofía; García Marín, Juan F.; Martín, Carlos; Domínguez, Lucas; de Rafael, Luis; Baquero, Fernando; Gómez-Mampaso, Enrique; Blázquez, Jesús

    1998-01-01

    An allele-specific amplification method based on two genetic polymorphisms to differentiate Mycobacterium tuberculosis from Mycobacterium bovis was tested. Based on the differences found at position 169 in the pncA genes from M. tuberculosis and M. bovis, a PCR system which was able to differentiate most of the 237 M. tuberculosis complex isolates tested in one of the two species was developed. All 121 M. tuberculosis strains showed the expected base (cytosine) at position 169. Most of the M. bovis isolates had a guanine at the cited position. Nevertheless, 18 of the 116 M. bovis isolates, all of them goat isolates, showed the pncA polymorphism specific to M. tuberculosis. These results suggest that goat M. bovis may be the nicotinamidase-missing link at the origin of the M. tuberculosis species. Based on the polymorphism found at position 285 in the oxyR gene, the same system was used to differentiate M. tuberculosis from M. bovis. In this case, DNAs from all 121 M. tuberculosis isolates had the expected base (guanine) at this position. In addition, all 116 M. bovis isolates, including those from goats, showed the identical polymorphism (adenine). The oxyR allele-specific amplification method can differentiate M. bovis from M. tuberculosis, is rapid (results can be obtained in less than 3 h), and is easy to perform. PMID:9431955

  6. Evaluation of Cocktails with Recombinant Proteins of Mycobacterium bovis for a Specific Diagnosis of Bovine Tuberculosis

    PubMed Central

    Mon, María Laura; Moyano, Roberto Damián; Viale, Mariana Noelia; Colombatti Olivieri, María Alejandra; Gamietea, Ignacio José; Montenegro, Valeria Noely; Alonso, Bernardo; Santangelo, María de la Paz; Singh, Mahavir; Duran, Rosario; Romano, María Isabel

    2014-01-01

    The Delayed type hypersensitivity skin test (DTH) and interferon-gamma assay are used for the diagnosis of bovine tuberculosis (TBB). The specificity of these diagnoses, however, is compromised because both are based on the response against purified protein derivative of Mycobacterium bovis (PPD-B). In this study, we assessed the potential of two cocktails containing M. bovis recombinant proteins: cocktail 1 (C1): ESAT-6, CFP-10 and MPB83 and cocktail 2 (C2): ESAT-6, CFP-10, MPB83, HspX, TB10.3, and MPB70. C1, C2, and PPD-B showed similar response by DTH in M. bovis-sensitized guinea pigs. Importantly, C1 induced a lower response than PPD-B in M. avium-sensitized guinea pigs. In cattle, C1 displayed better performance than PPD-B and C2; indeed, C1 showed the least detection of animals either vaccinated or Map-infected. To optimize the composition of the cocktails, we obtained protein fractions from PPD-B and tested their immunogenicity in experimentally M. bovis-infected cattle. In one highly reactive fraction, seven proteins were identified. The inclusion of FixB in C1 enhanced the recognition of M. bovis-infected cattle without compromising specificity. Our data provide a promising basis for the future development of a cocktail for TBB detection without interference by the presence of sensitized or infected animals with other mycobacteria. PMID:25110654

  7. In vitro antimicrobial susceptibility of Mycoplasma bovis clinical isolates recovered from bison (Bison bison).

    PubMed

    Suleman, Muhammad; Prysliak, Tracy; Windeyer, Claire; Perez-Casal, Jose

    2016-03-01

    Mycoplasma bovis is a pathogen globally affecting cattle and bison herds, causing pneumonia, arthritis, mastitis, abortions, and other symptoms, leading to huge economic losses. Many studies have been done regarding the antimicrobial susceptibility of M. bovis isolated from cattle, but no such study is available for isolates recovered from bison. For the first time, in vitro susceptibilities of 40 M. bovis clinical isolates collected from bison herds in Canada are reported here. Minimal inhibitory concentration (MIC) values were determined using Sensititre® plates. The most effective MIC50 and MIC90 were for spectinomycin (1 and >64 μg/mL), tiamulin (1 and >32 μg/mL), and tulathromycin (16 and 64 μg/mL), whereas tetracyclines, fluoroquinolones, and florfenicol failed to inhibit growth of M. bovis bison isolates. Isolates were nonsusceptible to tetracyclines (100%), fluoroquinolones (97.5%), and tilmicosin (100%), whereas the highest susceptibility of bison clinical isolates was seen with spectinomycin (95%) and tulathromycin (67.5%). Two lung isolates (Mb283 and 348) were found resistant to both spectinomycin and tulathromycin. These results show a marked difference in antimicrobial susceptibility of bison isolates as compared with previously reported and laboratory reference cattle isolates, emphasizing the necessity of testing antimicrobial susceptibility of M. bovis bison isolates and to generate better therapeutic regime for improved recovery chances for infected bison herds across North America. PMID:26854525

  8. [Mycobacterium bovis in wildlife of the dairy regions of Santa Fe (Argentina)].

    PubMed

    Abdala, Alejandro A; Garbaccio, Sergio; Zumárraga, Martín; Tarabla, Héctor D

    2015-01-01

    Control eradication campaigns of bovine tuberculosis based on the «test and slaughter» approach were successful in many countries and regions; however, in some areas the infection persists and one of the main reasons is Mycobacterium bovis infection in wild life species. Argentina has applied the same approach since 1999, achieving progress in dairy cattle herds. Nonetheless, the wildlife role has never been investigated. The objective of this study was to determine if wildlife from the Santa Fe dairy area is infected with M. bovis. Wildlife species having a positive tuberculin skin test were captured in five dairy farms. Ninety five wildlife mammals were captured; M. bovis was recovered from 7 possums (Didelphys albiventris), from one fox (Lycolapex gimnocercus) and from one rat (Rattus norvegicus). None of the animals exhibited macroscopic lesions. The most frequently isolated M. bovis spoligotypes were types 34 (4 isolates) and 12 (3 isolates). Spoligotype 34 is the most frequently isolated type in Argentine cattle. The role of D. albiventris as spillover host of M. bovis is discussed in this study. PMID:26376835

  9. Epidemiology of human Mycobacterium bovis disease, California, USA, 2003-2011.

    PubMed

    Gallivan, Mark; Shah, Neha; Flood, Jennifer

    2015-03-01

    We conducted a retrospective review of California tuberculosis (TB) registry and genotyping data to evaluate trends, analyze epidemiologic differences between adult and child case-patients with Mycobacterium bovis disease, and identify risk factors for M. bovis disease. The percentage of TB cases attributable to M. bovis increased from 3.4% (80/2,384) in 2003 to 5.4% (98/1,808) in 2011 (p = 0.002). All (6/6) child case-patients with M. bovis disease during 2010-2011 had >1 parent/guardian who was born in Mexico, compared with 38% (22/58) of child case-patients with M. tuberculosis disease (p = 0.005). Multivariate analysis of TB case-patients showed Hispanic ethnicity, extrapulmonary disease, diabetes, and immunosuppressive conditions, excluding HIV co-infection, were independently associated with M. bovis disease. Prevention efforts should focus on Hispanic binational families and adults with immunosuppressive conditions. Collection of additional risk factors in the national TB surveillance system and expansion of whole-genome sequencing should be considered. PMID:25693687

  10. Molecular epidemiology of Mycobacterium bovis isolates from free-ranging wildlife in South African game reserves.

    PubMed

    Michel, A L; Coetzee, M L; Keet, D F; Maré, L; Warren, R; Cooper, D; Bengis, R G; Kremer, K; van Helden, P

    2009-02-01

    Bovine tuberculosis is endemic in African buffalo and a number of other wildlife species in the Kruger National Park (KNP) and Hluhluwe-iMfolozi Park (HiP) in South Africa. It was thought that the infection had been introduced into the KNP ecosystem through direct contact between cattle and buffalo, a hypothesis which was confirmed in this study by IS6110 and PGRS restriction fragment length polymorphism (RFLP) typing. The molecular characterisation of 189 Mycobacterium bovis isolates from nine wildlife species in the HiP, including three smaller associated parks, and the Kruger National Park with adjacent areas showed that the respective epidemics were each caused by an infiltration of a single M. bovis genotype. The two M. bovis strains had different genetic profiles, as demonstrated by hybridisation with the IS6110 and PGRS RFLP probes, as well as with regard to evidence of evolutionary changes to the IS profile. While the M. bovis type in HiP was transmitted between buffaloes and to at least baboon, bushpig and lion without obvious genetic changes in the RFLP patterns, in the KNP a dominant strain was represented in 73% of the M. bovis isolates, whilst the remaining 27% were variants of this strain. No species-specific variants were observed, except for one IS6110 type which was found only in a group of five epidemiologically related greater kudu. This finding was attributed to species-specific behaviour patterns rather than an advanced host-pathogen interaction.

  11. [Mycobacterium bovis in wildlife of the dairy regions of Santa Fe (Argentina)].

    PubMed

    Abdala, Alejandro A; Garbaccio, Sergio; Zumárraga, Martín; Tarabla, Héctor D

    2015-01-01

    Control eradication campaigns of bovine tuberculosis based on the «test and slaughter» approach were successful in many countries and regions; however, in some areas the infection persists and one of the main reasons is Mycobacterium bovis infection in wild life species. Argentina has applied the same approach since 1999, achieving progress in dairy cattle herds. Nonetheless, the wildlife role has never been investigated. The objective of this study was to determine if wildlife from the Santa Fe dairy area is infected with M. bovis. Wildlife species having a positive tuberculin skin test were captured in five dairy farms. Ninety five wildlife mammals were captured; M. bovis was recovered from 7 possums (Didelphys albiventris), from one fox (Lycolapex gimnocercus) and from one rat (Rattus norvegicus). None of the animals exhibited macroscopic lesions. The most frequently isolated M. bovis spoligotypes were types 34 (4 isolates) and 12 (3 isolates). Spoligotype 34 is the most frequently isolated type in Argentine cattle. The role of D. albiventris as spillover host of M. bovis is discussed in this study.

  12. Necrotic pharyngitis associated with Mycoplasma bovis infections in American bison (Bison bison).

    PubMed

    Dyer, Neil; Register, Karen B; Miskimins, Dale; Newell, Teresa

    2013-03-01

    Mycoplasma bovis has emerged as a significant and costly infectious disease problem in bison, generally presenting as severe, caseonecrotic pneumonia. Three diagnostic cases in which M. bovis is associated with necrotic pharyngitis in bison are described in the current study. The bacterium was isolated from lesions of the pharynx or lung of 3 American bison (Bison bison), at 2 different locations in the upper Midwestern United States, with severe, necrotic pharyngeal abscesses. Chronic caseonecrotic inflammation typical of M. bovis infection in bovines was observed microscopically in the pharynxes of affected bison. A mixed population of bacteria was recovered from the pharyngeal lesions, and Trueperella pyogenes, a frequent secondary pathogen in ruminant respiratory disease, was consistently isolated from the affected animals. Distinctive histopathological features of the pharyngeal lesions favor causation by M. bovis, although a role for T. pyogenes in the clinical presentation cannot be excluded. Veterinarians and producers working with bison should be aware that M. bovis may be associated with pharyngitis in bison.

  13. Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison.

    PubMed

    Register, Karen B; Thole, Luke; Rosenbush, Ricardo F; Minion, F Chris

    2015-01-30

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and the bacterium most frequently isolated from the polymicrobial syndrome known as bovine respiratory disease complex. Recently, M. bovis has emerged as a significant health problem in bison, causing necrotic pharyngitis, pneumonia, dystocia and abortion. Whether isolates from cattle and bison comprise genetically distinct populations is unknown. This study describes the development of a highly discriminatory multilocus sequencing typing (MLST) method for M. bovis and its use to investigate the population structure of the bacterium. Genome sequences from six M. bovis isolates were used for selection of gene targets. Seven of 44 housekeeping genes initially evaluated were selected as targets on the basis of sequence variability and distribution within the genome. For each gene target sequence, four to seven alleles could be distinguished that collectively define 32 sequence types (STs) from a collection of 94 cattle isolates and 42 bison isolates. A phylogeny based on concatenated target gene sequences of each isolate revealed that bison isolates are genetically distinct from strains that infect cattle, suggesting recent disease outbreaks in bison may be due to the emergence of unique genetic variants. No correlation was found between ST and disease presentation or geographic origin. MLST data reported here were used to populate a newly created and publicly available, curated database to which researchers can contribute. The MLST scheme and database provide novel tools for exploring the population structure of M. bovis and tracking the evolution and spread of strains.

  14. An experimental vaccine composed of two adjuvants gives protection against Mycoplasma bovis in calves.

    PubMed

    Dudek, Katarzyna; Bednarek, Dariusz; Ayling, Roger D; Kycko, Anna; Szacawa, Ewelina; Karpińska, Teresa A

    2016-06-01

    Mycoplasma bovis is a major pathogen affecting cattle causing bronchopneumonia, mastitis, and other disorders. Only autogenous vaccines made specifically for individual farms are available in parts of Europe and the United States. A novel experimental vaccine composed of a field M. bovis isolate combined with saponin and Emulsigen(®) adjuvants was evaluated. Eighteen 3-4 week old calves were placed in three equal groups: vaccinated (Vac), positive control (PC) and negative control (NC). The Vac calves were subcutaneously injected with 8ml of the vaccine; the PC and NC calves received phosphate buffered saline (PBS). Three weeks later the Vac and PC calves were challenged with a virulent M. bovis strain, the NC group received PBS. Throughout the study clinical observations, microbiology and immunological tests were carried out. Three weeks post challenge two calves from each group were euthanased for necropsy and histopathological examination. The vaccine effectively stimulated the humoral immune response, with high titres of anti-M. bovis specific antibodies and total Ig concentration. This vaccine also intensified the IgA response. A clinically protective effect of the vaccine was demonstrated as it also reduced the gross pathological lung lesions and nasal shedding of M. bovis. PMID:27156637

  15. Molecular epidemiology of Mycobacterium bovis isolates from free-ranging wildlife in South African game reserves.

    PubMed

    Michel, A L; Coetzee, M L; Keet, D F; Maré, L; Warren, R; Cooper, D; Bengis, R G; Kremer, K; van Helden, P

    2009-02-01

    Bovine tuberculosis is endemic in African buffalo and a number of other wildlife species in the Kruger National Park (KNP) and Hluhluwe-iMfolozi Park (HiP) in South Africa. It was thought that the infection had been introduced into the KNP ecosystem through direct contact between cattle and buffalo, a hypothesis which was confirmed in this study by IS6110 and PGRS restriction fragment length polymorphism (RFLP) typing. The molecular characterisation of 189 Mycobacterium bovis isolates from nine wildlife species in the HiP, including three smaller associated parks, and the Kruger National Park with adjacent areas showed that the respective epidemics were each caused by an infiltration of a single M. bovis genotype. The two M. bovis strains had different genetic profiles, as demonstrated by hybridisation with the IS6110 and PGRS RFLP probes, as well as with regard to evidence of evolutionary changes to the IS profile. While the M. bovis type in HiP was transmitted between buffaloes and to at least baboon, bushpig and lion without obvious genetic changes in the RFLP patterns, in the KNP a dominant strain was represented in 73% of the M. bovis isolates, whilst the remaining 27% were variants of this strain. No species-specific variants were observed, except for one IS6110 type which was found only in a group of five epidemiologically related greater kudu. This finding was attributed to species-specific behaviour patterns rather than an advanced host-pathogen interaction. PMID:18786785

  16. Epidemiology of Human Mycobacterium bovis Disease, California, USA, 2003–2011

    PubMed Central

    Shah, Neha; Flood, Jennifer

    2015-01-01

    We conducted a retrospective review of California tuberculosis (TB) registry and genotyping data to evaluate trends, analyze epidemiologic differences between adult and child case-patients with Mycobacterium bovis disease, and identify risk factors for M. bovis disease. The percentage of TB cases attributable to M. bovis increased from 3.4% (80/2,384) in 2003 to 5.4% (98/1,808) in 2011 (p = 0.002). All (6/6) child case-patients with M. bovis disease during 2010–2011 had >1 parent/guardian who was born in Mexico, compared with 38% (22/58) of child case-patients with M. tuberculosis disease (p = 0.005). Multivariate analysis of TB case-patients showed Hispanic ethnicity, extrapulmonary disease, diabetes, and immunosuppressive conditions, excluding HIV co-infection, were independently associated with M. bovis disease. Prevention efforts should focus on Hispanic binational families and adults with immunosuppressive conditions. Collection of additional risk factors in the national TB surveillance system and expansion of whole-genome sequencing should be considered. PMID:25693687

  17. Characterization of Mycobacterium bovis from Humans and Cattle in Namwala District, Zambia

    PubMed Central

    Johansen, Tone Bjordal; Muma, John Bwalya; Munyeme, Musso; Mbulo, Grace; Muwonge, Adrian; Djønne, Berit

    2014-01-01

    Tuberculosis remains a major public health problem in Zambia. While human to human transmission of Mycobacterium tuberculosis is of major importance in driving the tuberculosis epidemic, the impact of Mycobacterium bovis transmission from infected cattle is largely unknown. This cross-sectional study aimed at molecular characterization of M. bovis in humans and cattle. A total of 100 human sputum samples and 67 bovine tissues were collected and analyzed for the presence of mycobacteria. Of 65 human samples that harbored acid fast bacteria (AFB), 55 isolates were obtained of which 34 were identified as M. tuberculosis and 2 as M. bovis. AFB-positive bovine samples (n = 67) yielded 47 mycobacterial isolates among which 25 were identified as M. bovis and no M. tuberculosis was found. Among the M. bovis isolates, spoligotyping revealed a high homogeneity in genotypes circulating in Namwala district. Human and cattle isolates shared identical MIRU-VNTR genotypes, suggesting that transmission between the two hosts may occur. Therefore, this study has documented zoonotic TB in human patients in Namwala district of Zambia. However, further molecular epidemiological studies in the study area are recommended. PMID:24847441

  18. Evaluation of Mycobacterium bovis double knockout mce2-phoP as candidate vaccine against bovine tuberculosis.

    PubMed

    García, Elizabeth; Bianco, María V; Gravisaco, María José; Rocha, Rosana V; Blanco, Federico C; Bigi, Fabiana

    2015-03-01

    In this study, a Mycobacterium bovis knockout strain in phoP-phoR and mce2 operons was tested as an antituberculosis experimental vaccine in animal models. The double mutant strain was significantly more attenuated than the wild type strain in inmunocompetent and inmunodeficient mice. Vaccination with the double mutant protected mice against challenge with a virulent M. bovis strain.

  19. Oral vaccination of white-tailed deer (Odocoileus virginianus) with Mycobacterium bovis Bacillus Calmette-Guerin (BCG)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis from livestock, particularly cattle. In Michigan, USA tuberculous white-tailed deer transmit M. bovis to other deer and cattle. One approach in dealing with this wildlife reservoir is to vaccina...

  20. In-depth analysis of the genome sequence of a clinical, extensively drug-resistant Mycobacterium bovis strain.

    PubMed

    Sagasti, Sara; Millán-Lou, María Isabel; Soledad Jiménez, María; Martín, Carlos; Samper, Sofía

    2016-09-01

    Although human-to-human transmission of Mycobacterium bovis strains and other members of the animal lineage of the tubercle bacilli is a rare event, an extensively drug resistant (XDR) strain, named M. bovis B strain, caused a lethal outbreak in the nineties in Spain. The genome of M. bovis B strain was re-sequenced by SOLiD platform and mapped to the reference M. bovis AF2122/97. The genetic polymorphisms detected have been analysed in depth. One hundred and fifty-eight specific non-synonymous SNPs were detected; ninety-two of these were non-conservative. In addition, one specific 3195-bp insertion could be identified as an ABC transporter gene by homology with tbd2 gene, which was found to be present in other clinical M. bovis strains. Its peculiar phenotype profile of resistance was explained by molecular characteristics, including a 5685-bp specific deletion that revealed a novel polymorphism associated with resistance to paraminosalicilic acid. From a phylogenetical point of view, according to the SNPs detected, M. bovis B could be included into the clonal complex M. bovis European 2. This is the first time that a deep analysis of the whole-genome sequencing of an extensively drug-resistant M. bovis strain is detailed. It offers the explanation for the resistance and found several data to be incorporated for future research. PMID:27553409

  1. Evaluation of enzyme-linked immunosorbent assays for detection of Mycoplasma bovis-Specific antibody in bison sera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis has recently emerged as a significant and costly infectious disease problem in bison. This report demonstrates that ELISAs for detection of M. bovis-specific antibody in cattle are not optimal for identification of seropositive bison. An ELISA optimized for use with bison sera is ...

  2. Animal-side Serologic Assay for Rapid Detection of Mycobacterium bovis Infection in Multiple Species of Free-ranging Wildlife

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous species of wild mammals are susceptible to Mycobacterium bovis, a cause of bovine tuberculosis (TB). Eurasian badgers, white-tailed deer, brushtail possums, and wild boar are implicated in the maintenance of wildlife reservoirs of M. bovis infection in different countries, fueling bovine TB...

  3. Crystalline inclusions in erythrocytes parasitized with Babesia equi following treatment of ponies with imidocarb.

    PubMed

    Simpson, C F; Taylor, W J; Kitchen, H

    1980-08-01

    Four splenectomized Welsh ponies were infected with Babesia equi. Two ponies were treated with imidocarb dipropionate, and two were not treated. By light microscopic examination, 1% to 2% of the parasitized erythrocytes of treated ponies contained crystalline inclusions. The crystals were rectangular, diamond, or burr shaped. They occupied most of the erythrocytic cytoplasm, and, as a result, the remainder of the pale staining cytoplasm was inconspicuous in Wright-Giemsa-stained blood smears. The size and shape of intraerythrocytic inclusions varied when examined by electron microscopy, but in most instances they were either adhered to or were located close to the parasite. The sides of crystals were either smooth or serrated, and corners were either sharp or notched. Fractures or faults were common in large crystals. Parasitized erythrocytes of nontreated ponies and nonparasitized erythrocytes of treated ponies did not contain crystals. Four hemoglobulin types were identified in five noninfected, nontreated Welsh ponies from the same herd. PMID:6255836

  4. Molecular identification of different Theileria and Babesia species infecting sheep in Sudan.

    PubMed

    El Imam, Ahmed H; Hassan, Shawgi M; Gameel, Ahmed A; El Hussein, Abdelrahim M; Taha, Khalid M; Oosthuizen, Marinda C

    2016-01-01

    The epidemiological aspects of sheep piroplasmosis in Sudan are poorly studied, and further investigations using sensitive and precise techniques are required. In this study, the Reverse Line Blot (RLB) hybridization assay was used to detect and simultaneously differentiate between Theileria and Babesia species. DNA was extracted from blood collected on filter paper (n=219) from apparently healthy sheep from six different geographical localities in Sudan. Results indicated that Theileria ovis (88.6%), T. separata (20.1%), T. lestoquardi (16.4%) and T. annulata (16.4%) DNA could be detected in the blood samples. Single and mixed Theileria infections were detected in 74 (33.8%) and 124 (56.6%) respectively and T. ovis being the most prevalent species in the country. T. ovis and T. separata were reported for the first time in sheep in Sudan. PMID:27262957

  5. Sequencing of the smallest Apicomplexan genome from the human pathogen Babesia microti†

    PubMed Central

    Cornillot, Emmanuel; Hadj-Kaddour, Kamel; Dassouli, Amina; Noel, Benjamin; Ranwez, Vincent; Vacherie, Benoît; Augagneur, Yoann; Brès, Virginie; Duclos, Aurelie; Randazzo, Sylvie; Carcy, Bernard; Debierre-Grockiego, Françoise; Delbecq, Stéphane; Moubri-Ménage, Karina; Shams-Eldin, Hosam; Usmani-Brown, Sahar; Bringaud, Frédéric; Wincker, Patrick; Vivarès, Christian P.; Schwarz, Ralph T.; Schetters, Theo P.; Krause, Peter J.; Gorenflot, André; Berry, Vincent; Barbe, Valérie; Ben Mamoun, Choukri

    2012-01-01

    We have sequenced the genome of the emerging human pathogen Babesia microti and compared it with that of other protozoa. B. microti has the smallest nuclear genome among all Apicomplexan parasites sequenced to date with three chromosomes encoding ∼3500 polypeptides, several of which are species specific. Genome-wide phylogenetic analyses indicate that B. microti is significantly distant from all species of Babesidae and Theileridae and defines a new clade in the phylum Apicomplexa. Furthermore, unlike all other Apicomplexa, its mitochondrial genome is circular. Genome-scale reconstruction of functional networks revealed that B. microti has the minimal metabolic requirement for intraerythrocytic protozoan parasitism. B. microti multigene families differ from those of other protozoa in both the copy number and organization. Two lateral transfer events with significant metabolic implications occurred during the evolution of this parasite. The genomic sequencing of B. microti identified several targets suitable for the development of diagnostic assays and novel therapies for human babesiosis. PMID:22833609

  6. Babesia gibsoni internal transcribed spacer 1 region is highly conserved amongst isolates from dogs across Japan

    PubMed Central

    LIU, Mingming; CAO, Shinuo; VUDRIKO, Patrick; SUZUKI, Hiroshi; SOMA, Takehisa; XUAN, Xuenan

    2016-01-01

    Babesia gibsoni is a tick-borne apicomplexan parasite of dogs that often causes fever and hemolytic anemia with highly variable clinical outcome. In this study, we sequenced the 254bp Internal Transcribed Spacer 1 region (ITS1) of 54 B. gibsoni isolates from 14 different geographical regions of Japan. The 54 isolates shared high sequence identity with each other and with B. gibsoni isolates reported in GenBank database (97.2–100%). Consistent with previous reports, phylogenetic analysis showed that B. gibsoni isolates from Japan formed the same clade with those from U.S.A., Australia, India and Taiwan. Our finding indicates that B. gibsoni ITS1 region is highly conserved among isolates from dogs in Japan, making it a useful genetic marker for molecular epidemiology of the parasite. PMID:26806537

  7. The epidemiological survey for atovaquone resistant related gene of Babesia gibsoni in Japan

    PubMed Central

    IGUCHI, Aiko; SOMA, Takehisa; SUZUKI, Hiroshi; XUAN, Xuenan

    2015-01-01

    In 73 gDNA samples from Babesia gibsoni-infected dogs, the M121I variant population was measured by using allele-specific real-time PCR. Although the mechanism of atovaquone against B. gibsoni has not been clearly identified, it is reported that the mitochondria cytochrome b gene of the atovaquone-resistant B. gibsoni had a single-nucleotide substitution at nt363 (G to T), which resulted in the substitution of methionine with isoleucine (M121I). In this study, 3/73 samples showed over 5% M121I variant population. Although the M121I variant population is a low percentage, it runs the risk of spreading drug-resistant parasites. It is important to prevent the spread of drug-resistance, so we need to gather information about this at regular intervals. PMID:26549436

  8. Molecular identification of different Theileria and Babesia species infecting sheep in Sudan.

    PubMed

    El Imam, Ahmed H; Hassan, Shawgi M; Gameel, Ahmed A; El Hussein, Abdelrahim M; Taha, Khalid M; Oosthuizen, Marinda C

    2016-01-01

    The epidemiological aspects of sheep piroplasmosis in Sudan are poorly studied, and further investigations using sensitive and precise techniques are required. In this study, the Reverse Line Blot (RLB) hybridization assay was used to detect and simultaneously differentiate between Theileria and Babesia species. DNA was extracted from blood collected on filter paper (n=219) from apparently healthy sheep from six different geographical localities in Sudan. Results indicated that Theileria ovis (88.6%), T. separata (20.1%), T. lestoquardi (16.4%) and T. annulata (16.4%) DNA could be detected in the blood samples. Single and mixed Theileria infections were detected in 74 (33.8%) and 124 (56.6%) respectively and T. ovis being the most prevalent species in the country. T. ovis and T. separata were reported for the first time in sheep in Sudan.

  9. First report of two asymptomatic cases of human infection with Babesia microti (Franca, 1910) in Poland.

    PubMed

    Welc-Falęciak, Renata; Pawełczyk, Agnieszka; Radkowski, Marek; Pancewicz, Sławomir A; Zajkowska, Joanna; Siński, Edward

    2015-01-01

    Human infection by Babesia microti has been recognized as an emerging zoonosis with important public health implications worldwide. In Europe the reported cases of human babesiosis have been attributed mostly to B. divergens infection, with only sporadic cases of the disease caused by B. microti or B. venatorum. This study, based on molecular methods (PCR, R-T PCR, DNA sequencing and phylogenetic analysis), reveals for the first time in Poland, asymptomatic infection with . microti in immunocompetent healthy individuals working in forest ecosystems. Of the 58 professional foresters examined, two (3.4%) were identified as B. microti-positive by specific PCR. The results of this study also provide strong evidence that in eastern Poland, where tick-borne diseases (TBDs) are endemic, there is a potential risk of acquiring human babesiosis due to zoonotic B. microti parasites commonly found in rodents and I. ricinus ticks. The potential public health importance of this finding is discussed. PMID:25780828

  10. Molecular characteristics of phosphoenolpyruvate: mannose phosphotransferase system in Streptococcus bovis.

    PubMed

    Asanuma, Narito; Yoshii, Takahiro; Hino, Tsuneo

    2004-07-01

    To elucidate the regulatory mechanism of catabolite control in Streptococcus bovis, we investigated the molecular properties and gene expression of the mannose-specific phosphoenolpyruvate (PEP)-dependent sugar: phosphotransferase system (PTS). The mannose PTS gene cluster (man) was found to comprise a gene encoding enzyme (E) II AB (manL) and genes encoding EIIC (manM), EIID (manN), and a putative regulator (manO). The gene cluster (man operon) was transcribed from one transcriptional start site, which was located 40 bp upstream of the manL start codon. However, two transcriptional start sites were found between manN and manO in primer extension analysis, and the manO may be transcribed independently from the man operon. The man operon and manO were constitutively transcribed without being affected by culture conditions, such as the sugar supplied (glucose, galactose, fructose, maltose, lactose, sucrose, or mannose), growth rate, or pH. PMID:15297922

  11. Antibiotic sensitivity of an Argentine strain collection of Moraxella bovis.

    PubMed

    Zielinski, G; Piscitelli, H; Perez-Monti, H; Stobbs, L A

    2000-01-01

    The antimicrobial susceptibility of 88 isolates of Moraxella bovis of Argentine origin was evaluated for 12 antimicrobials by broth microdilution procedures. The isolates had a minimum inhibitory concentration (MIC90) of < or = 0.06 microg/mL to enrofloxacin; < or = 0.12 microg/mL to ceftiofur; < or = 0.25 microg/mL to ampicillin; < or = 0.5 microg/mL to florfenicol and gentamicin; < or = 1.0 microg/mL to tilmicosin, erythromycin, and oxytetracycline; < or = 4.0 microg/mL to tylosin; < or = 8.0 microg/mL to spectinomycin; < or = 0.25/4.75 microg/mL to trimethoprim/sulfamethoxazole; and > or = 32 microg/mL to lincomycin. Modal MIC values for these antimicrobials were as follows: enrofloxacin, 0.03 microg/mL; ceftiofur, 0.06 pg/mL; ampicillin, 0.25 microg/mL; florfenicol, gentamicin, erythromycin, and oxytetracycline, 0.5 microg/mL; tilmicosin, 1.0 microg/mL; tylosin and spectinomycin, 4.0 microg/mL; lincomycin and erythromycin, 16 microg/mL; and trimethoprim/ sulfamethoxazole, < or = 0.25/4.75 microg/mL. These data show that all antimicrobials except lincomycin have MICs suggestive of sensitivity in vitro, though confirmation of clinical efficacy can only be properly assessed based on pharmacologic and/or clinical data to support the MIC values. PMID:19757583

  12. Serum concentrations of eicosanoids and lipids in dogs naturally infected with Babesia canis.

    PubMed

    Mrljak, Vladimir; Kučer, Nada; Kuleš, Josipa; Tvarijonaviciute, Asta; Brkljačić, Mirna; Crnogaj, Martina; Zivičnjak, Tatjana; Smit, Iva; Ceron, Jose Joaquin; Rafaj, Renata Barić

    2014-03-17

    Canine babesiosis is a tick-borne disease with world-wide significance caused by intraerythrocytic protozoa of the genus Babesia. The eicosanoids, as inflammatory mediators, are involved in the regulation of the immune response and inflammatory reaction. Metabolism of lipids is of great importance in babesiosis. In this study it was aimed to investigate the dynamics of serum concentration of prostaglandin E2 (PGE2), thromboxane B2 (TxB2), leukotriene B4 (LTB4), triglycerides, total cholesterol (Chol), HDL- and LDL-cholesterol in dogs naturally infected with Babesia canis and healthy dogs. Both groups were measured for all parameters on the admission day and on the first, second and seventh day of the disease. Dogs that were included in this study had systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS). It was demonstrated that the level of LTB4, PGE2, TxB2 in dogs naturally infected with B. canis significantly changed during the disease. The level of LTB4 was significantly higher during the study, while the concentration of PGE2 was significantly higher second, third and seventh day of disease in relation with healthy dogs. The level of TxB2 was significantly lower at the beginning of the disease, but after seven days concentration was significantly higher. Both group of patients with SIRS and MODS had significantly higher level of LTB4. Substained high concentrations of PGE2 were observed in dogs with MODS after therapy but not in dogs with SIRS, and LTB4 followed a similar tendency. On the other hand, increases in TxB2 were only significant in dogs with SIRS. The lipid profile in naturally infected dogs with B. canis infection was significantly changed. Further studies are needed to assess the prognostic values of lipid mediators in dogs with B. canis infection, and the ability of these markers to predict the progress of SIRS and MODS.

  13. Mycobacterium bovis BCG Causing Vertebral Osteomyelitis (Pott’s Disease) Following Intravesical BCG Therapy

    PubMed Central

    Aljada, Ibrahim S.; Crane, John K.; Corriere, Nancy; Wagle, Datta G.; Amsterdam, Daniel

    1999-01-01

    We report a case of Mycobacterium bovis BCG vertebral osteomyelitis in a 79-year-old man 2.5 years after intravesical BCG therapy for bladder cancer. The recovered isolate resembled M. tuberculosis biochemically, but resistance to pyrazinamide (PZA) rendered that diagnosis suspect. High-pressure liquid chromatographic studies confirmed the diagnosis of M. bovis BCG infection. The patient was originally started on a four-drug antituberculous regimen of isoniazid, rifampin, ethambutol, and PZA. When susceptibility studies were reported, the regimen was changed to isoniazid and rifampin for 12 months. Subsequently, the patient was transferred to a skilled nursing facility for 3 months, where he underwent intensive physical therapy. Although extravesical adverse reactions are rare, clinicians and clinical microbiologists need to be aware of the possibility of disseminated infection by M. bovis BCG in the appropriate setting of clinical history, physical examination, and laboratory investigation. PMID:10325395

  14. Infection of Eurasian badgers (Meles meles) with Mycobacterium bovis and Mycobacterium avium complex in Spain.

    PubMed

    Balseiro, Ana; Rodríguez, Oscar; González-Quirós, Pablo; Merediz, Isabel; Sevilla, Iker A; Davé, Dipesh; Dalley, Deanna J; Lesellier, Sandrine; Chambers, Mark A; Bezos, Javier; Muñoz, Marta; Delahay, Richard J; Gortázar, Christian; Prieto, José M

    2011-11-01

    The prevalence, distribution and pathology related to infection with Mycobacterium bovis and other mycobacteria were determined in trapped (n=36) and road-killed (n=121) badgers in Spain from 2006 to 2010. The prevalence of M. bovis based on bacteriological culture from road-killed badgers was 8/121 (6.6%) and from trapped badgers was 0/36 (0%). Tuberculosis/M. bovis infection was evident in 15/121 (12.4%) road-killed badgers when bacteriology and histopathology were combined. Mycobacterium avium complex was isolated by culture from the tracheal aspirate of 1/36 (2.8%) trapped badgers and from tissue pools from 8/121 (6.6%) road-killed badgers.

  15. Menadione-catalyzed luminol chemiluminescent assay for viability of Mycobacterium bovis.

    PubMed

    Yamashoji, Shiro

    2002-01-01

    Stable luminol chemiluminescence was observed 10 min after the addition of menadione to a suspension of Mycobacterium bovis homogenized in Middlebrook 7H9 broth base including OADC enrichment. The chemiluminescence intensity was proportional to the absorbance of the bacterial suspension at 600 nm in a range of 0.005 to 0.15. Luminol chemiluminescence disappeared after 10 min incubation of M. bovis at over 60% of ethanol or 4 days of cultivation of M. bovis in the presence of 40 microg/ml of streptomycin. The bacterium showing the disappearance of chemiluminescence could not grow after being washed, suggesting that the inhibition concentration of the antimicrobials can be estimated on the basis of the disappearance of chemiluminescence. Menadione-catalyzed luminol chemiluminescent assay was rapid and sensitive in comparison to turbidimetry, tetrazolium (WST-8) reduction assay, and the assay using the Mycobacteria growth indicator tube (MGIT).

  16. Development of a loop-mediated isothermal amplification (LAMP) assay for rapid diagnosis of Babesia canis infections.

    PubMed

    Müller, H; Aysul, N; Liu, Z; Salih, D A; Karagenc, T; Beyer, D; Kullmann, B; Ahmed, J S; Seitzer, U

    2010-04-01

    Vector-borne diseases are rising in interest due to global warming, which is believed to impact on the distribution of vectors into new areas thus influencing the occurrence and epidemiology of vector-borne pathogens. Babesia canis belongs to the Piroplasmidae and there are three described subspecies, namely B. canis canis, B. canis rossi and B. canis vogeli. They are each transmitted by a different tick-species, Dermacentor reticulatus, Haemaphysalis leachi and Rhipicephalus sanguineus, respectively. There are also differences in the geographical distribution and pathogenicity to dogs of each subspecies. In this study, we aimed to establish a rapid and easy to perform DNA-based test using loop-mediated isothermal amplification to detect all three Babesia canis subspecies in one assay.

  17. Identification and phylogenetic analysis of Japanese Macaque Babesia-1 (JM-1) detected from a Japanese Macaque (Macaca fuscata fuscata).

    PubMed

    Hirata, Haruyuki; Kawai, Satoru; Maeda, Mari; Jinnai, Michio; Fujisawa, Kohei; Katakai, Yuko; Hikosaka, Kenji; Tanabe, Kazuyuki; Yasutomi, Yasuhiro; Ishihara, Chiaki

    2011-10-01

    We demonstrate here the identification and phylogenetic characterization of Babesia microti (B. microti)-like parasite detected from a splenectomized Japanese macaque (Macaca fuscata fuscata) at a facility for laboratory animal science. On Day 133 after splenectomy, intra-erythrocytic parasites were found on light microscopic examination, and the level of parasitemia reached 0.3% on blood smear. Molecular characterization of the parasite using nested-polymerization chain reactions targeting the 18S rRNA, β-tubulin, and subunit 7 (eta) of the chaperonin-containing t-complex polypeptide 1 (CCT7) genes were identified as a B. microti-like parasite, designated the Japanese Macaque Babesia-1 (JM-1).

  18. Identification of new 18S rRNA strains of Babesia canis isolated from dogs with subclinical babesiosis.

    PubMed

    Łyp, P; Adaszek, Ł; Furmaga, B; Winiarczyk, S

    2015-01-01

    In this study, we used PCR to detect and characterize B. canis from naturally infected dogs in Poland with subclinical babesiosis by amplifying and sequencing a portion of the 18S ribosomal RNA (rRNA) gene. Venous blood samples were collected from ten dogs with subclinical babesiosis. A 559-bp fragment of the B. canis 18S rRNA gene was amplified by PCR. Sequencing of the PCR products led to the identification of a new variant of Babesia canis, differing from the previously detected protozoa genotypes (18S rRNA-A and 18S rRNA-B) with nucleotide substitutions in positions 150 and 151 of the tested gene fragment. The results indicate the emergence within the Polish territory of a new, previously unencountered Babesia canis genotype responsible for the development of subclinical babesiosis. PMID:26618590

  19. Identification of new 18S rRNA strains of Babesia canis isolated from dogs with subclinical babesiosis.

    PubMed

    Łyp, P; Adaszek, Ł; Furmaga, B; Winiarczyk, S

    2015-01-01

    In this study, we used PCR to detect and characterize B. canis from naturally infected dogs in Poland with subclinical babesiosis by amplifying and sequencing a portion of the 18S ribosomal RNA (rRNA) gene. Venous blood samples were collected from ten dogs with subclinical babesiosis. A 559-bp fragment of the B. canis 18S rRNA gene was amplified by PCR. Sequencing of the PCR products led to the identification of a new variant of Babesia canis, differing from the previously detected protozoa genotypes (18S rRNA-A and 18S rRNA-B) with nucleotide substitutions in positions 150 and 151 of the tested gene fragment. The results indicate the emergence within the Polish territory of a new, previously unencountered Babesia canis genotype responsible for the development of subclinical babesiosis.

  20. Nitric oxide not apoptosis mediates differential killing of Mycobacterium bovis in bovine macrophages.

    PubMed

    Esquivel-Solís, Hugo; Vallecillo, Antonio J; Benítez-Guzmán, Alejandro; Adams, L Garry; López-Vidal, Yolanda; Gutiérrez-Pabello, José A

    2013-01-01

    To identify the resistance phenotype against Mycobacterium bovis in cattle, we used a bactericidal assay that has been considered a marker of this trait. Three of 24 cows (12.5%) were phenotyped as resistant and 21 as susceptible. Resistance of bovine macrophages (MΦ) to BCG challenge was evaluated for its association with SLC11A1 GT microsatellite polymorphisms within 3'UTR region. Twenty-three cows (95.8%) had a GT13 genotype, reported as resistant, consequently the SLC11A1 polymorphism was not in agreement with our bactericidal assay results. MΦ of cows with resistant or susceptible phenotype were challenged in vitro with virulent M. bovis field strain or BCG, and nitric oxide production, bacterial killing and apoptosis induction were measured in resting and LPS-primed states. M. bovis field strain induced more apoptosis than BCG, although the difference was not significant. Resistant MΦ controlled better the replication of M. bovis (P<0.01), produced more nitric oxide (P<0.05) and were slightly more prone to undergo apoptosis than susceptible cells. LPS pretreatment of MΦ enhanced all the functional parameters analyzed. Inhibition of nitric oxide production with n (G)-monomethyl-L-arginine monoacetate enhanced replication of M. bovis but did not modify apoptosis rates in both resistant and susceptible MΦ. We conclude that nitric oxide production not apoptosis is a major determinant of macrophage resistance to M. bovis infection in cattle and that the influence of SLC11A1 gene 3'UTR polymorphism is not associated with this event.

  1. Development of a Recombinant Antigen for Antibody-Based Diagnosis of Mycoplasma bovis Infection in Cattle

    PubMed Central

    Brank, Marion; Le Grand, Dominique; Poumarat, François; Bezille, Pierre; Rosengarten, Renate; Citti, Christine

    1999-01-01

    Mycoplasma bovis induces various clinical manifestations in cattle, such as mastitis, arthritis, and pneumonia. We have evaluated the immunoreactivity of three variable surface proteins (Vsps) of M. bovis, namely VspA, VspB, and VspC, with sera collected from herds with mycoplasmosis or from cattle experimentally infected with M. bovis. Western blot analysis revealed that the Vsps are the predominant antigens recognized by the host humoral response during M. bovis infection. The immunoreactivity of VspA, VspB, and VspC with host antibodies was independent of the clinical manifestations, the geographical origin of the M. bovis isolates, the mode of infection, and the animal’s history. Moreover, the results showed that Vsp-specific host antibodies can be detected about 10 days after experimental infection and for up to several months. The full-length or truncated versions of the VspA product were overexpressed in Escherichia coli as fusion proteins (FP-VspA). Recombinant products showed strong immunoreactivity with the Vsp-specific monoclonal antibodies 1A1 and 1E5, with the corresponding epitopes localized at the VspA N-terminal and C-terminal ends, respectively. Anti-M. bovis sera of cattle naturally or experimentally infected also strongly recognized the full-length FP-VspA. The seroreactivity of sera collected from cattle between 6 and 10 days after experimental infection was weaker with truncated versions of VspA lacking the 1E5 epitope than with the full-length VspA or the truncated versions lacking the 1A1 epitope. Overall, the results indicate that the Vsps, despite their inter- and intraclonal variability, may be applied as target antigens in serodiagnostic assays for epidemiological studies. PMID:10548577

  2. Asymptomatic Cattle Naturally Infected with Mycobacterium bovis Present Exacerbated Tissue Pathology and Bacterial Dissemination

    PubMed Central

    Menin, Álvaro; Fleith, Renata; Reck, Carolina; Marlow, Mariel; Fernandes, Paula; Pilati, Célso; Báfica, André

    2013-01-01

    Rational discovery of novel immunodiagnostic and vaccine candidate antigens to control bovine tuberculosis (bTB) requires knowledge of disease immunopathogenesis. However, there remains a paucity of information on the Mycobacterium bovis-host immune interactions during the natural infection. Analysis of 247 naturally PPD+ M. bovis-infected cattle revealed that 92% (n = 228) of these animals were found to display no clinical signs, but presented severe as well as disseminated bTB-lesions at post-mortem examination. Moreover, dissemination of bTB-lesions positively correlated with both pathology severity score (Spearman r = 0.48; p<0.0001) and viable tissue bacterial loads (Spearman r = 0.58; p = 0.0001). Additionally, granuloma encapsulation negatively correlated with M. bovis growth as well as pathology severity, suggesting that encapsulation is an effective mechanism to control bacterial proliferation during natural infection. Moreover, multinucleated giant cell numbers were found to negatively correlate with bacterial counts (Spearman r = 0.25; p = 0.03) in lung granulomas. In contrast, neutrophil numbers in the granuloma were associated with increased M. bovis proliferation (Spearman r = 0.27; p = 0.021). Together, our findings suggest that encapsulation and multinucleated giant cells control M. bovis viability, whereas neutrophils may serve as a cellular biomarker of bacterial proliferation during natural infection. These data integrate host granuloma responses with mycobacterial dissemination and could provide useful immunopathological-based biomarkers of disease severity in natural infection with M. bovis, an important cattle pathogen. PMID:23326525

  3. Mycobacterium bovis infection in cattle induces differential expression of prolactin receptor isoforms in macrophages.

    PubMed

    López-Rincón, Gonzalo; Gutiérrez-Pabello, José Ángel; Díaz-Otero, Fernando; Muñoz-Valle, José Francisco; Pereira-Suárez, Ana Laura; Estrada-Chávez, Ciro

    2013-12-01

    Prolactin receptor (PRLr) is a member of the cytokine receptor superfamily 1 showing tissue specific structural diversity. Expression of PRLr isoforms in lymphoid tissues has been associated with immunomodulatory function of prolactin. Bovine tuberculosis (bTB) is characterized by chronic inflammation caused by the persistent infection of lymphoid tissues with Mycobacterium bovis. To test the hypothesis of the influence of PRLr in the pathogenesis of bTB, the aim of this study was to identify PRLr isoforms expressed during bTB in different tissues and to analyze their association with the pathogenesis of bTB. We examined lymphoid and non-lymphoid tissues ex vivo from experimentally and naturally infected cattle, as well as from bTB-free cattle, by Western blot (WB) and immunohistochemistry (IH). In vitro, monocytes from exposed, infected, and healthy cattle were stimulated with M. bovis antigens and then analyzed by WB. To detect transcriptional levels of PRLr in macrophages (MØ) exposed to M. bovis, real time PCR was performed. WB revealed diversity of PRLr isoforms in tissues from infected cattle but not in tissues from bTB-free cattle. PRLr isoforms 100 kDa 75, 50 and 40 were found expressed in tissues of animals infected with M. bovis, while only the short isoform of 40 kDa correlated with the immunopathology and ability to infect MØ. We confirmed the synthesis of PRLr mRNA in MØ after M. bovis exposure and propose that molecular pathogen patterns of M. bovis might modulate inflammation during bTB through expression of the PRLr isoform in MØ.

  4. Nitric Oxide Not Apoptosis Mediates Differential Killing of Mycobacterium bovis in Bovine Macrophages

    PubMed Central

    Esquivel-Solís, Hugo; Vallecillo, Antonio J.; Benítez-Guzmán, Alejandro; Adams, L. Garry; López-Vidal, Yolanda; Gutiérrez-Pabello, José A.

    2013-01-01

    To identify the resistance phenotype against Mycobacterium bovis in cattle, we used a bactericidal assay that has been considered a marker of this trait. Three of 24 cows (12.5%) were phenotyped as resistant and 21 as susceptible. Resistance of bovine macrophages (MΦ) to BCG challenge was evaluated for its association with SLC11A1 GT microsatellite polymorphisms within 3′UTR region. Twenty-three cows (95.8%) had a GT13 genotype, reported as resistant, consequently the SLC11A1polymorphism was not in agreement with our bactericidal assay results. MΦ of cows with resistant or susceptible phenotype were challenged in vitro with virulent M. bovis field strain or BCG, and nitric oxide production, bacterial killing and apoptosis induction were measured in resting and LPS-primed states. M. bovis field strain induced more apoptosis than BCG, although the difference was not significant. Resistant MΦ controlled better the replication of M. bovis (P<0.01), produced more nitric oxide (P<0.05) and were slightly more prone to undergo apoptosis than susceptible cells. LPS pretreatment of MΦ enhanced all the functional parameters analyzed. Inhibition of nitric oxide production with nG-monomethyl-L-arginine monoacetate enhanced replication of M. bovis but did not modify apoptosis rates in both resistant and susceptible MΦ. We conclude that nitric oxide production not apoptosis is a major determinant of macrophage resistance to M. bovis infection in cattle and that the influence of SLC11A1 gene 3′UTR polymorphism is not associated with this event. PMID:23691050

  5. Cost-Effectiveness of Blood Donor Screening for Babesia microti in Endemic Regions of the United States

    PubMed Central

    Simon, Matthew S.; Leff, Jared A.; Pandya, Ankur; Cushing, Melissa; Shaz, Beth H.; Calfee, David P.; Schackman, Bruce R.; Mushlin, Alvin I.

    2014-01-01

    Background Babesia microti is the leading reported cause of red blood cell (RBC) transfusion-transmitted infection in the United States (US). Donor screening assays are in development. Study Design and Methods A decision analytic model estimated the cost-effectiveness of screening strategies for preventing transfusion-transmitted babesiosis (TTB) in a hypothetical cohort of transfusion recipients in Babesia-endemic areas of the US. Strategies included: (1) No screening, (2) Uniform Donor Health History Questionnaire (UDHQ), “status quo”, (3) Recipient risk-targeting using donor antibody (Ab) and polymerase chain reaction (PCR) screening, (4) Universal endemic donor Ab screening, (5) Universal endemic donor Ab and PCR screening. Outcome measures were TTB cases averted, costs, quality-adjusted life years (QALYs) and incremental cost-effectiveness ratios ($/QALY). We assumed a societal willingness to pay of $1 million/QALY based on screening for other transfusion-transmitted infections. Results Compared to no screening, the UDHQ avoids 0.02 TTB cases per 100,000 RBC transfusions at an incremental cost effectiveness ratio (ICER) of $160,000/QALY whereas recipient risk-targeted strategy using Ab/PCR avoids 1.62 TTB cases per 100,000 RBC transfusions at an ICER of $713,000/QALY compared to the UDHQ. Universal endemic Ab screening avoids 3.39 cases at an ICER of $760,000/QALY compared to the recipient-risk targeted strategy. Universal endemic Ab/PCR screening avoids 3.60 cases and has an ICER of $8.8 million/QALY compared to universal endemic Ab screening. Results are sensitive to blood donor Babesia prevalence, TTB transmission probability, screening test costs, risk and severity of TTB complications, and impact of babesiosis diagnosis on donor quality of life. Conclusion Antibody screening for Babesia in endemic regions is appropriate from an economic perspective based on the societal willingness to pay for preventing infectious threats to blood safety. PMID

  6. A molecular survey of Babesia spp. and Theileria spp. in red foxes (Vulpes vulpes) and their ticks from Thuringia, Germany.

    PubMed

    Najm, Nour-Addeen; Meyer-Kayser, Elisabeth; Hoffmann, Lothar; Herb, Ingrid; Fensterer, Veronika; Pfister, Kurt; Silaghi, Cornelia

    2014-06-01

    Wild canines which are closely related to dogs constitute a potential reservoir for haemoparasites by both hosting tick species that infest dogs and harbouring tick-transmitted canine haemoparasites. In this study, the prevalence of Babesia spp. and Theileria spp. was investigated in German red foxes (Vulpes vulpes) and their ticks. DNA extracts of 261 spleen samples and 1953 ticks included 4 tick species: Ixodes ricinus (n=870), I. canisuga (n=585), I. hexagonus (n=485), and Dermacentor reticulatus (n=13) were examined for the presence of Babesia/Theileria spp. by a conventional PCR targeting the 18S rRNA gene. One hundred twenty-one out of 261 foxes (46.4%) were PCR-positive. Out of them, 44 samples were sequenced, and all sequences had 100% similarity to Theileria annae. Similarly, sequencing was carried out for 65 out of 118 PCR-positive ticks. Theileria annae DNA was detected in 61.5% of the sequenced samples, Babesia microti DNA was found in 9.2%, and Babesia venatorum in 7.6% of the sequenced samples. The foxes were most positive in June and October, whereas the peak of tick positivity was in October. Furthermore, the positivity of the ticks was higher for I. canisuga in comparison to the other tick species and for nymphs in comparison to adults. The high prevalence of T. annae DNA in red foxes in this study suggests a reservoir function of those animals for T. annae. To our knowledge, this is the first report of T. annae in foxes from Germany as well as the first detection of T. annae and B. microti in the fox tick I. canisuga. Detection of DNA of T. annae and B. microti in three tick species collected from foxes adds new potential vectors for these two pathogens and suggests a potential role of the red fox in their natural endemic cycles.

  7. A molecular study of tick-borne haemoprotozoan parasites (Theileria and Babesia) in small ruminants in Northern Tunisia.

    PubMed

    M'ghirbi, Youmna; Ros-García, Amaia; Iribar, Pilar; Rhaim, Adel; Hurtado, Ana; Bouattour, Ali

    2013-11-15

    In this study, the frequency of Theileria and Babesia species in sheep and goats was assessed via reverse line blotting (RLB). A total of 263 apparently healthy sheep and goats, from 16 randomly selected flocks located in 9 localities situated in 3 bioclimatic zones in Tunisia, were investigated for the blood protozoans. RLB hybridization with polymerase chain reaction detected only Theileria ovis in sheep and goats, accounting for 22.4% (95% confidence interval [CI]: 17.6-27.1%) positive samples. The infection rate in sheep (28.1%; 95% CI: 23.8-32.3%) was higher than in goats (4.7%; 95% CI: -10.9 to 20.4%). Neither Babesia nor mixed infections were detected. Only two Ixodid tick species (Rhipicephalus turanicus and Rhipicephalus bursa) were collected from the examined sheep and goats in 5 localities. R. turanicus was the dominant species (95.5%) collected mainly in the humid zone, while apparently rare in the sub-humid zone. R. bursa was the only species collected in the semi-arid area. RLB analysis identified six different piroplasms in ticks, with an overall prevalence of 31.5% (95% CI: 28.1-34.9%). Twenty percent (95% CI: 14.4-25.5%) of the collected ticks tested positive for Theileria spp., 3% (95% CI: -5.6 to 11.6%) for Babesia spp. and 0.9% (95% CI: -8.1 to 9.9%) of the ticks harbored both genera; several of these species are not known to occur in small ruminants. This is the first report on the detection of Theileria and Babesia species DNA in small ruminants and ticks in Tunisia.

  8. Prevalence of Borrelia burgdorferi and Babesia microti in mice on islands inhabited by white-tailed deer.

    PubMed Central

    Anderson, J F; Johnson, R C; Magnarelli, L A; Hyde, F W; Myers, J E

    1987-01-01

    Borrelia burgdorferi and Babesia microti were isolated from 35 of 51 white-footed mice (Peromyscus leucopus) and meadow voles (Microtus pennsylvanicus) captured on two Narragansett Bay, R.I., islands inhabited by deer, the principal host for the adult stages of the vector tick, Ixodes dammini. Immature ticks parasitized mice from both islands. From 105 mice captured on four other islands not inhabited by deer neither pathogen was isolated, nor were I. dammini found. PMID:3555339

  9. Chemical Studies on the Structure of Mucopeptide Isolated from Streptococcus bovis

    PubMed Central

    Kane, Judith; Lackland, Henry; Karakawa, W. W.; Krause, R. M.

    1969-01-01

    Mucopeptides isolated from Streptococcus bovis cell walls were found to be composed of alanine, glutamic acid, lysine, and threonine in a mole ratio of 3:1:1:1. A dipeptide, Nε-lysylthreonine, was isolated from S. bovis mucopeptide by ion-exchange chromatography. This finding suggests that threonine is associated with the bridge which cross-links adjacent tetrapeptides by connecting the ε-amino group of lysine of one tetrapeptide to the carboxyl group of d-alanine of another to form the mucopeptide matrix. PMID:5802603

  10. Report of a case of bronchopneumonia associated with Moraxella bovis isolation in a chamois (Rupicapra pyrenaica).

    PubMed

    Lavin, S; Lastras, M E; Marco, I; Cabañes, F X

    2000-04-01

    A case of fibrinopurulent bronchopneumonia associated with Moraxella bovis infection in a chamois (Rupicapra pyrenaica) is described. The animal, a 4-month-old female, was referred by the staff warden of the National Game Reserve of Freser-Setcases (Catalonia, north-eastern Spain). The animal was in good general condition and was found 4 h before death. On necropsy the lungs were congested and oedematous, with haemorrhagic areas in the cranial and middle lobes. The microscopic lesions were those of a fibrinopurulent bronchopneumonia. Microbiological study of the samples obtained showed numerous small beta-haemolytic colonies in pure culture, identified as Moraxella (Moraxella) bovis.

  11. The bactericidal effect of microwaves on Mycobacterium bovis dried on scalpel blades.

    PubMed

    Rosaspina, S; Salvatorelli, G; Anzanel, D

    1994-01-01

    The action of microwaves on stainless steel scalpel blades contaminated with Mycobacterium bovis was investigated. The complete destruction of M. bovis was obtained with 4 min of microwave exposure. When the preparations were subjected to scanning electron microscopy, the bacteria had undergone a progressive series of alterations consisting, initially, of the formation of deep pits in the bacterial body and eventually the complete disintegration of the microorganisms. Such phenomena are less evident when this mycobacterium is exposed to other sterilization methods such as dry heat or autoclaving. PMID:7910182

  12. [Rifampicin-resistant Mycobacterium bovis BCG strain isolated from an infant with NEMO mutation].

    PubMed

    Çavuşoğlu, Cengiz; Edeer Karaca, Neslihan; Azarsız, Elif; Ulusoy, Ezgi; Kütükçüler, Necil

    2015-04-01

    It is well known that disseminated Mycobacterium bovis BCG infection is developed after BCG vaccination in infants with congenital cellular immune deficiencies such as mutations in genes along the interleukin (IL)-12/interferon (IFN)-γ pathway and mutations in nuclear factor-kB essential modulator (NEMO). In this report, a rifampicin-resistant M.bovis BCG strain isolated from an infant with NEMO defect was presented. An 8-month-old male infant with NEMO defect admitted to the pediatric outpatient clinic of our hospital with fever, generalized lymphadenopathy and hepatosplenomegaly. Microscopic examination of the smears prepared from lymph node and liver biopsy specimens revealed abundant amount (3+) of acid-fast bacilli (AFB). Rifampicin-susceptible Mycobacterium tuberculosis complex (MTC) was detected by real-time PCR (GeneXpert MTB/RIF; Cepheid, USA) in the samples. The growth of mycobacteria was determined on the 20th day of culture performed in MGIT960 system (Becton Dickinson, USA). The isolate was identified as M.bovis BCG by GenoType MTBC kit (Hain Lifescience, Germany) and defined as M.bovis BCG [SIT 482 (BOV_1)] by spoligotyping. In the primary anti-tuberculosis drug susceptibility test performed by MGIT960 system, the isolate was found susceptible to rifampicin (RIF), isoniazid (INH), streptomycin (STM) and ethambutol (EMB). Then anti-tuberculosis treatment was started to the patient. However, the patient at the age of 2 years, re-admitted to the hospital with the complaint of hepatosplenomegaly. Smear of spontaneously draining abscess material obtained from subcutaneous nodules revealed intensive AFB positivity (3+) once again. In the present instance RIF-resistant MTC was detected with GeneXpert system in the specimen. The growth of mycobacteria was determined on the 13th day of culture and isolate was identified as M.bovis BCG. The present isolate was found susceptible to INH, STM and EMB but resistant to RIF. A mutation in the rpoB gene (codon 531, S

  13. Differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis

    PubMed Central

    Maboni, Grazieli; Gressler, Leticia T.; Espindola, Julia P.; Schwab, Marcelo; Tasca, Caiane; Potter, Luciana; de Vargas, Agueda Castagna

    2015-01-01

    The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented. PMID:26273272

  14. Detection of Babesia spp. in free-ranging Pukus, Kobus vardonii, on a game ranch in Zambia.

    PubMed

    Munang'andu, Hetron Mweemba; Munyeme, Musso; Nambota, Andrew Mubila; Nalubamba, King Shimumbo; Siamudaala, Victor M

    2011-12-01

    Babesia spp. were detected from 4 asymptomatic pukus captured on a game ranch in central Zambia in October 2008. Blood smears were examined in 4 species of aymptomatic free-ranging antelopes, namely the puku (Kobus vordanii), reedbuck (Redunca arundinum), bushbuck (Tragelaphus sylvaticus), and kudu (Tragelaphus strepsiceros), and showed the presence of Babesia parasites only in the puku. In the puku, the prevalence of babesiosis was estimated at 33.3% (n = 12), while the overall prevalence in all examined animals was 8.5% (n = 47). The parasites showed morphological characteristics of paired ring-like stages with the length varying between 1.61 µm and 3.02 µm (mean = 2.12 µm, n = 27; SD = 0.76 µm). Both the infected and non-infected pukus showed good body condition scores (BCS), while the dominant tick species detected from all animals were Rhipicephalus appendiculatus, Rhipicephalus spp., and Boophilus spp. To our knowledge this is the first report of Babesia spp. infection in pukus in Zambia. These findings suggest that wildlife could play an important role in the epidemiology of babesiosis in Zambia.

  15. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil.

    PubMed

    Rotondano, Tereza Emmanuelle de Farias; Almeida, Herta Karyanne Araújo; Krawczak, Felipe da Silva; Santana, Vanessa Lira; Vidal, Ivana Fernandes; Labruna, Marcelo Bahia; de Azevedo, Sérgio Santos; Ade lmeida, Alzira Maria Paiva; de Melo, Marcia Almeida

    2015-01-01

    This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State. PMID:25909253

  16. Discrimination between ovine Babesia and Theileria species in China based on the ribosomal protein S8 (RPS8) gene.

    PubMed

    Tian, Zhancheng; Liu, Guangyuan; Yin, Hong; Luo, Jianxun; Guan, Guiquan; Luo, Jin; Xie, Junren; Zheng, Jinfeng; Yuan, Xiaosong; Wang, Fangfang; Shen, Hui; Tian, Meiyuan

    2013-10-18

    Ovine babesiosis and theileriosis are important hemoprotozoal diseases of sheep and goats in tropical and subtropical regions that lead to economic losses in these animals. PCR-restriction fragment length polymorphism (PCR-RFLP) is a reliable molecular diagnostic tool for discriminating Theileria or Babesia species in the same host. In this study, the DNA sequences of a ribosomal protein S8 (RPS8) gene from four species of piroplasms in China were used to develop a species-specific PCR-RFLP diagnostic tool. The sensitivity of the PCR assays was 0.1 pg DNA for B. motasi and 1 pg DNA for T. uilenbergi and 10 pg DNA for Babesia sp. Xinjiang-2005 and T. luwenshuni. The clear size difference of the PCR products allowed for a direct discrimination for B. motasi, Babesia sp. Xinjiang-2005 and ovine Theileria species (T. uilenbergi and T. luwenshuni), except that the mixed infection between T. uilenbergi and T. luwenshuni may be difficult to distinguish, simply after the electrophoretic separation of the amplification products. Further T. uilenbergi and T. luwenshuni diagnoses were made by digesting the PCR product with SacI. The established method could be applicable for the survey of parasite dynamics, and epidemiological studies as well as prevention and control of the disease.

  17. Evaluation of pathogenesis caused in cattle and guinea pig by a Mycobacterium bovis strain isolated from wild boar

    PubMed Central

    2011-01-01

    Background In many regions of the world, wild mammals act as reservoir of Mycobacterium bovis, a situation that prevents the eradication of bovine tuberculosis. In order to observe whether a strain isolated from a wild boar, previously tested as highly virulent in a mice model, is also virulent in cattle, we performed cattle experimental inoculation with this strain Results Groups of Friesian calves were either infected with the wild boar strain M. bovis 04-303 or with the bovine strain NCTC10772 as a control. We found that antigen-specific IFN-γ release in whole blood samples occurred earlier in animals infected with M. bovis 04-303. Both M. bovis strains resulted in a positive skin test, with animals infected with the wild boar isolate showing a stronger response. These results and the presence of more severe organ lesions, with granuloma and pneumonic areas in cattle demonstrate that the wild boar isolate is more virulent than the NCTC10772 strain. Additionally, we tested the infectivity of the M. bovis strains in guinea pigs and found that M. bovis 04-303 had the highest pathogenicity. Conclusions M. bovis strains isolated from wild boars may be pathogenic for cattle, producing TB lesions. PMID:21745408

  18. Chemical composition and nutrient degradability in elephant grass silage inoculated with Streptococcus bovis isolated from the rumen.

    PubMed

    Ferreira, Daniele J; Zanine, Anderson M; Lana, Rogério P; Ribeiro, Marinaldo D; Alves, Guilherme R; Mantovani, Hilário C

    2014-03-01

    The objective of the present study was to assess the chemical and bromatological composition and in situ degradability of elephant grass silages inoculated with Streptococcus bovis isolated from cattle rumen. A complete randomized design was used with four treatments and six replications: elephant grass silage, elephant grass silage inoculated with 10(6) CFU/g Streptococcus bovis JB1 strains; elephant grass silage inoculated with 106 CFU/g Streptococcus bovis HC5 strains; elephant grass silage inoculated with 106 CFU/g Enterococcus faecium with six replications each. The pH and ammoniacal nitrogen values were lower (P<0.05) for the silages inoculated with Streptococcus bovis JB1 and HC5, respectively. The silage inoculated with Streptococcus bovis had a higher crude protein content (P<0.05) and there were no differences for the fiber contents in the silage. The (a)soluble fraction degradability, especially in the silages inoculated with Streptococcus bovis JB1 and HC5, had higher values, 30.77 and 29.97%, for dry matter and 31.01 and 36.66% for crude protein, respectively. Inoculation with Streptococcus bovis improved the fermentation profile, protein value and rumen degradability of the nutrients.

  19. Evaluation of enzyme-linked immunosorbent assays for detection of Mycoplasma bovis-specific antibody in bison sera.

    PubMed

    Register, Karen B; Sacco, Randy E; Olsen, Steven C

    2013-09-01

    Mycoplasma bovis has recently emerged as a significant and costly infectious disease problem in bison. A method for the detection of M. bovis-specific serum antibodies is needed in order to establish prevalence and transmission patterns. Enzyme-linked immunosorbent assays (ELISAs) validated for the detection of M. bovis-specific serum IgG in cattle are commercially available, but their suitability for bison sera has not been determined. A collection of bison sera, most from animals with a known history of infection or vaccination with M. bovis, was tested for M. bovis-specific IgG using commercially available kits as well as an in-house ELISA in which either cattle or bison M. bovis isolates were used as a source of antigen. Comparison of the results demonstrates that ELISAs optimized for cattle sera may not be optimal for the identification of bison seropositive for M. bovis, particularly those with low to moderate antibody levels. The reagent used for the detection of bison IgG and the source of the antigen affect the sensitivity of the assay. Optimal performance was obtained when the capture antigen was derived from bison isolates rather than cattle isolates and when a protein G conjugate rather than an anti-bovine IgG conjugate was used for the detection of bison IgG.

  20. Sex-related heterogeneity in the life-history correlates of Mycobacterium bovis infection in European badgers (Meles meles).

    PubMed

    Tomlinson, A J; Chambers, M A; Wilson, G J; McDonald, R A; Delahay, R J

    2013-11-01

    Heterogeneity in the progression of disease amongst individual wild animals may impact on both pathogen and host dynamics at the population level, through differential effects on transmission, mortality and reproductive output. The role of the European badger (Meles meles) as a reservoir host for Mycobacterium bovis infection in the UK and Ireland has been the focus of intense research for many years. Here, we investigate life-history correlates of infection in a high-density undisturbed badger population naturally infected with M. bovis. We found no evidence of a significant impact of M. bovis infection on female reproductive activity or success, with evidence of reproduction continuing successfully for several years in the face of M. bovis excretion. We also found evidence to support the hypothesis that female badgers are more resilient to established M. bovis infection than male badgers, with longer survival times following the detection of bacterial excretion. We discuss the importance of infectious breeding females in the persistence of M. bovis in badger populations, and how our findings in male badgers are consistent with testosterone-induced immunosuppression. In addition, we found significant weight loss in badgers with evidence of disseminated infection, based on the culture of M. bovis from body systems other than the respiratory tract. For females, there was a gradual loss of weight as infection progressed, whereas males only experienced substantial weight loss when infection had progressed to the point of dissemination. We discuss how these differences may be explained in terms of resource allocation and physiological trade-offs.

  1. Assessment of Bdellovibrio bacteriovorus 109J killing of Moraxella bovis in an in vitro model of infectious bovine keratoconjunctivitis.

    PubMed

    Boileau, Mélanie J; Clinkenbeard, Kenneth D; Iandolo, John J

    2011-10-01

    The objective of this study was to determine the potential of Bdellovibrio bacteriovorus 109J as an alternative non-chemotherapeutic treatment of infectious bovine keratoconjunctivitis (IBK). To accomplish this, various parameters of B. bacteriovorus predation of Moraxella bovis were determined in vitro. Initial passage of B. bacteriovorus using M. bovis as prey required 10 d for active cultures to develop compared with 2 d for culture on normal Escherichia coli prey; however by the 5th passage, time to active predatory morphology was reduced to 2 d. This high passage B. bacteriovorus culture [1 × 10(10) plaque forming units (PFU)/mL] killed 76% of M. bovis [1 × 10(7) colony forming units (CFU)/mL] present in suspension broth in a 4 h assay. The minimal level of M. bovis supporting B. bacteriovorus predation was 1 × 10(4) CFU/mL. To assess the ability of B. bacteriovorus to kill M. bovis on an epithelial surface mimicking IBK, an in vitro assay with Madin-Darby bovine kidney (MDBK) cells inoculated with 4 × 10(7) CFU/mL M. bovis was used. Treatment with a B. bacteriovorus suspension (1.6 × 10(11) PFU/mL) decreased adherence of M. bovis to MDBK cells by 6-fold at 12 h of treatment, as well as decreased the number of unattached M. bovis cells by 1.4-fold. This study demonstrates that B. bacteriovorus has potential as an effective biological control of M. bovis at levels likely present in IBK-infected corneal epithelia and ocular secretions.

  2. Clinical characteristics and significance of Streptococcus salivarius bacteremia and Streptococcus bovis bacteremia: a prospective 16-year study.

    PubMed

    Corredoira, J C; Alonso, M P; García, J F; Casariego, E; Coira, A; Rodriguez, A; Pita, J; Louzao, C; Pombo, B; López, M J; Varela, J

    2005-04-01

    The aim of this study was to determine the clinical significance of Streptococcus salivarius isolates recovered from blood cultures and compare them with isolates of Streptococcus bovis biotypes I and II. Seventeen of the 52 (32%) S. salivarius isolates recovered were considered clinically significant, compared with 62 of the 64 (97%) S. bovis isolates (p<0.0001). Bacteremia caused by S. salivarius occurred mostly in patients who showed relevant disruption of the mucous membranes and/or serious underlying diseases. Patients with S. salivarius bacteremia were younger than those with S. bovis bacteremia (57 vs. 67 years; p<0.01). Patients with S. salivarius bacteremia and patients with S. bovis II bacteremia had similar rates of endocarditis, colon tumors, and non-colon cancer. On the other hand, when compared with S. bovis I bacteremia, S. salivarius bacteremia was associated with lower rates of endocarditis (18% vs. 74%, respectively) (p<0.01) and colon tumors (0% vs. 57%, respectively) (p<0.005) and higher rates of non-colon cancer (53% vs. 9.5%, respectively) (p<0.01). Bacteremia caused by S. bovis II had a hepatobiliary origin in 50% of the patients, while, in contrast, that due to S. salivarius or S. bovis I was less frequently associated with a hepatobiliary origin (12% and 5%, respectively) (p<0.00001). The rate of penicillin resistance was 31% among S. salivarius isolates and 0% among S. bovis isolates (p<0.0001). In conclusion, the clinical characteristics of S. salivarius bacteremia and S. bovis II bacteremia are similar, and the isolation of S. salivarius in blood should not be systematically regarded as contamination. PMID:15902530

  3. Mycobacterium bovis infection in the lion (Panthera leo): Current knowledge, conundrums and research challenges.

    PubMed

    Viljoen, Ignatius M; van Helden, Paul D; Millar, Robert P

    2015-06-12

    Mycobacterium bovis has global public-health and socio-economic significance and can infect a wide range of species including the lion (Panthera leo) resulting in tuberculosis. Lions are classified as vulnerable under the IUCN Red List of Threatened Species and have experienced a 30% population decline in the past two decades. However, no attempt has been made to collate and critically evaluate the available knowledge of M. bovis infections in lions and potential effects on population. In this review we set out to redress this. Arguments suggesting that ingestion of infected prey animals are the main route of infection for lions have not been scientifically proven and research is needed into other possible sources and routes of infection. The paucity of knowledge on host susceptibility, transmission directions and therefore host status, manifestation of pathology, and epidemiology of the disease in lions also needs to be addressed. Advances have been made in diagnosing the presence of M. bovis in lions. However, these diagnostic tests are unable to differentiate between exposure, presence of infection, or stage of disease. Furthermore, there are contradictory reports on the effects of M. bovis on lion populations with more data needed on disease dynamics versus the lion population's reproductive dynamics. Knowledge on disease effects on the lion reproduction and how additional stressors such as drought or co-morbidities may interact with tuberculosis is also lacking. Filling these knowledge gaps will contribute to the understanding of mycobacterial infections and disease in captive and wild lions and assist in lion conservation endeavours.

  4. Inhibitory effects of bezoar bovis on intimal formation and vascular smooth muscle cell proliferation in rat.

    PubMed

    Mizuno, Masaki; Chung, Hwa-Jin; Maruyama, Ikuro; Tani, Tadato

    2005-01-01

    Intimal formation of animal carotid arteries induced by balloon endothelial denudation has been considered to be an "accelerated atherosclerosis" model and used in primary screening methods to evaluate natural drugs and chemical candidates. The aim of the present study was to examine whether intimal formation is prevented by Bezoar Bovis (dried cattle gallbladder stones: Niuhuang in Chinese and Go-o in Japanese), which has been used to prevent heart palpitation in patients with hypertension. The intimal-to-medial area ratio in rat carotid arteries 7 days after balloon endothelial denudation was significantly reduced by oral administration of Bezoar Bovis. Bezoar Bovis also suppressed vascular smooth muscle cells (VSMCs) proliferation, which is thought to play important roles in the intimal formation after endothelial damage and also atherosclerosis resulting from long-term inappropriate lifestyle. The present findings suggest that Bezoar Bovis may be useful for preventing atherosclerosis and for protection against restenosis after percutaneous coronary intervention, for which effective reduction method is not currently available.

  5. Identification of Mycobacterium bovis in bovine clinical samples by PCR species-specific primers.

    PubMed Central

    Romero, R E; Garzón, D L; Mejía, G A; Monroy, W; Patarroyo, M E; Murillo, L A

    1999-01-01

    Tuberculosis, caused by Mycobacterium bovis is emerging as the most important disease affecting cattle. Furthermore, it results in a major public health problem when transmitted to humans. Due to its difficult and non-specific diagnosis, M. bovis has been declared to be one of the etiologic agents causing significant economic loss in the cattle industry. Our group evaluated a more rapid and specific method, based on a new polymerase chain reaction species-specific primers, which amplifies a 470-base pair fragment of the M. bovis genome. A total of 275 milk-producing cows were studied by intradermal tuberculin test (ITT) which gave 184 positive and 91 negative cases. From them, 50 animals were taken from a cattle ranch free of tuberculosis. Three different samples were collected from each animal (blood, nasal mucus, and milk). Positive results were obtained from 26 animals by PCR (11.4%), 1 by bacteriological culturing (0.4%) and 1 by bacilloscopy (0.4%). This finding suggests, as in previous reports, that ITT, normally used for bovine tuberculosis detection, has the inconvenience of having a broad range of specificity and sensitivity, and the PCR technique is a more specific and sensitive test to detect infection associated with M. bovis. Therefore, we propose this PCR assay as a useful tool in the epidemiological characterization of infected animals in areas considered to be at high risk of transmission. Images Figure 1. PMID:10369566

  6. Effects of inulin chain length on fermentation by equine fecal bacteria and Streptococcus bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fructans from pasture can be fermented by Gram-positive bacteria (e.g., Streptococcus bovis) in the equine hindgut, increasing production of lactic acid and decreasing pH. The degree of polymerization (DP) of fructans has been suggested to influence fermentation rates. The objective of the current ...

  7. Viral Booster Vaccines Improve Mycobacterium bovis BCG-Induced Protection Against Bovine Tuberculosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous work in small animal laboratory models of tuberculosis have shown that vaccination strategies based on heterologous prime-boost protocols using Mycobacterium bovis bacille Calmette-Guerin (BCG) to prime and Modified Vaccinia Ankara strain (MVA85A) or recombinant attenuated adenoviruses (Ad8...

  8. Update on vaccination of white-tailed deer with Mycobacterium bovis BCG: Safety and Efficacy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 1994, white-tailed deer in northeast Michigan were found to be harboring Mycobacterium bovis, the causative agent of tuberculosis in most animals including humans. Although deer likely contracted tuberculosis from cattle in the early 20th century, when the disease was present in Michigan cattle, ...

  9. Measuring bovine gamma delta T cell function at the site of Mycobacterium bovis infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The causative agent of tuberculosis (TB) in cattle is Mycobacterium bovis. The characteristic lesions of bovine TB are well-organized pulmonary granulomas. Gamma delta T cells are a unique subset of nonconventional T cells that play major roles in both the innate and adaptive arms of the immune sys...

  10. Draft Genome Sequence of the Vaccination Strain Mycobacterium bovis BCG S4-Jena

    PubMed Central

    Wibberg, Daniel; Winkler, Anika; Straube, Eberhard; Karrasch, Matthias; Keller, Peter M.

    2016-01-01

    Here, we present the draft genome sequence of Mycobacterium bovis BCG S4-Jena, a tuberculosis vaccine strain. The genome of S4-Jena is represented by 48 scaffolds, consisting of 132 scaffolded contigs and amounting to a size of about 4.2 Mb. New genes potentially encoding a phage fragment were identified in the genome. PMID:27103721

  11. Association of microRNAs with antibody response to mycoplasma bovis in beef cattle

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to identify microRNAs associated with a serum antibody response to Mycoplasma bovis in beef cattle. Serum from sixteen beef calves was collected at three points: in summer after calves were born, in fall at weaning, and in the following spring. All sera collected in t...

  12. Global Multilocus Sequence Typing Analysis of Mycoplasma bovis Isolates Reveals Two Main Population Clusters

    PubMed Central

    Churchward, C. P.; Schnee, C.; Sachse, K.; Lysnyansky, I.; Catania, S.; Iob, L.; Ayling, R. D.; Nicholas, R. A. J.

    2014-01-01

    Mycoplasma bovis is a major bovine pathogen associated with bovine respiratory disease complex and is responsible for substantial economic losses worldwide. M. bovis is also associated with other clinical presentations in cattle, including mastitis, otitis, arthritis, and reproductive disorders. To gain a better understanding of the genetic diversity of this pathogen, a multilocus sequence typing (MLST) scheme was developed and applied to the characterization of 137 M. bovis isolates from diverse geographical origins, obtained from healthy or clinically infected cattle. After in silico analysis, a final set of 7 housekeeping genes was selected (dnaA, metS, recA, tufA, atpA, rpoD, and tkt). MLST analysis demonstrated the presence of 35 different sequence types (STs) distributed in two main clonal complexes (CCs), defined at the double-locus variant level, namely, CC1, which included most of the British and German isolates, and CC2, which was a more heterogeneous and geographically distant group of isolates, including European, Asian, and Australian samples. Index of association analysis confirmed the clonal nature of the investigated M. bovis population, based on MLST data. This scheme has demonstrated high discriminatory power, with the analysis showing the presence of genetically distant and divergent clusters of isolates predominantly associated with geographical origins. PMID:25540400

  13. Key Hub and Bottleneck Genes Differentiate the Macrophage Response to Virulent and Attenuated Mycobacterium bovis

    PubMed Central

    Killick, Kate E.; Magee, David A.; Park, Stephen D. E.; Taraktsoglou, Maria; Browne, John A.; Conlon, Kevin M.; Nalpas, Nicolas C.; Gormley, Eamonn; Gordon, Stephen V.; MacHugh, David E.; Hokamp, Karsten

    2014-01-01

    Mycobacterium bovis is an intracellular pathogen that causes tuberculosis in cattle. Following infection, the pathogen resides and persists inside host macrophages by subverting host immune responses via a diverse range of mechanisms. Here, a high-density bovine microarray platform was used to examine the bovine monocyte-derived macrophage transcriptome response to M. bovis infection relative to infection with the attenuated vaccine strain, M. bovis Bacille Calmette–Guérin. Differentially expressed genes were identified (adjusted P-value ≤0.01) and interaction networks generated across an infection time course of 2, 6, and 24 h. The largest number of biological interactions was observed in the 24-h network, which exhibited scale-free network properties. The 24-h network featured a small number of key hub and bottleneck gene nodes, including IKBKE, MYC, NFKB1, and EGR1 that differentiated the macrophage response to virulent and attenuated M. bovis strains, possibly via the modulation of host cell death mechanisms. These hub and bottleneck genes represent possible targets for immuno-modulation of host macrophages by virulent mycobacterial species that enable their survival within a hostile environment. PMID:25324841

  14. Draft genome sequences of Streptococcus bovis strains ATCC 33317 and JB1

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We report the draft genome sequences of Streptococcus bovis type strain ATTC 33317 (CVM42251) isolated from cow dung and strain JB1 (CVM42252) isolated from a cow rumen in 1977. Strains were subjected to Next Generation sequencing and the genome sizes are approximately 2 MB and 2.2 MB, respectively....

  15. Antigen-Specific IP-10 Release Is a Sensitive Biomarker of Mycobacterium bovis Infection in Cattle.

    PubMed

    Parsons, Sven D C; McGill, Kevina; Doyle, Mairead B; Goosen, Wynand J; van Helden, Paul D; Gormley, Eamonn

    2016-01-01

    The most widely used ante-mortem diagnostic tests for tuberculosis in cattle are the tuberculin skin test and the interferon-gamma (IFN-γ) release assay, both of which measure cell-mediated immune responses to Mycobacterium bovis infection. However, limitations in the performance of these tests results in a failure to identify all infected animals. In attempting to increase the range of diagnostic tests for tuberculosis, measurement of the cytokine IP-10 in antigen-stimulated blood has previously been shown to improve the detection of M. tuberculosis and M. bovis infection, in humans and African buffaloes (Syncerus caffer), respectively. In the present study, 60 cattle were identified by the single intradermal comparative tuberculin test as tuberculosis reactors (n = 24) or non-reactors (n = 36) and the release of IFN-γ and IP-10 in antigen-stimulated whole blood from these animals was measured using bovine specific ELISAs. There was a strong correlation between IP-10 and IFN-γ production in these samples. Moreover, measurement of the differential release of IP-10 in response to stimulation with M. bovis purified protein derivative (PPD) and M. avium PPD distinguished between reactor and non-reactor cattle with a sensitivity of 100% (95% CI, 86%-100%) and a specificity of 97% (95% CI, 85%-100%). These results suggest that IP-10 might prove valuable as a diagnostic biomarker of M. bovis infection in cattle. PMID:27167122

  16. 16S rRNA Gene Mutations Associated with Decreased Susceptibility to Tetracycline in Mycoplasma bovis

    PubMed Central

    Amram, E.; Mikula, I.; Schnee, C.; Ayling, R. D.; Nicholas, R. A. J.; Rosales, R. S.; Harrus, S.

    2014-01-01

    Mycoplasma bovis isolates with decreased susceptibilities to tetracyclines are increasingly reported worldwide. The acquired molecular mechanisms associated with this phenomenon were investigated in 70 clinical isolates of M. bovis. Sequence analysis of the two 16S rRNA-encoding genes (rrs3 and rrs4 alleles) containing the primary binding pocket for tetracycline (Tet-1 site) was performed on isolates with tetracycline hydrochloride MICs of 0.125 to 16 μg/ml. Mutations at positions A965T, A967T/C (Escherichia coli numbering) of helix 31, U1199C of helix 34, and G1058A/C were identified. Decreased susceptibilities to tetracycline (MICs, ≥2 μg/ml) were associated with mutations present at two (A965 and A967) or three positions (A965, A967, and G1058) of the two rrs alleles. No tet(M), tet(O), or tet(L) determinants were found in the genome of any of the 70 M. bovis isolates. The data presented correlate (P < 0.0001) the mutations identified in the Tet-1 site of clinical isolates of M. bovis with decreased susceptibility to tetracycline. PMID:25403668

  17. Multiple sampling and discriminatory fingerprinting reveals clonally complex and compartmentalized infections by M. bovis in cattle.

    PubMed

    Navarro, Yurena; Romero, Beatriz; Copano, María Francisca; Bouza, Emilio; Domínguez, Lucas; de Juan, Lucía; García-de-Viedma, Darío

    2015-01-30

    The combination of new genotyping tools and a more exhaustive sampling policy in the analysis of infection by Mycobacterium tuberculosis has shown that infection by this pathogen is more complex than initially expected. Mixed infections, coexistence of clonal variants from a parental strain, and compartmentalized infections are all different modalities of this clonal complexity. Until recently, genotyping of Mycobacterium bovis in animal populations was based on spoligotyping and analysis of a single isolate per infection; therefore, clonal complexity is probably underdetected. We used multiple sampling combined with highly discriminatory MIRU-VNTR to study compartmentalized infections by M. bovis in a low-tuberculosis prevalence setting. We spoligotyped the M. bovis isolates from two or more anatomic locations sampled from 55 animals on 39 independent farms. Compartmentalized infections, with two different strains infecting independent lymph nodes in the same animal, were found in six cases (10.9%). MIRU-VNTR analysis confirmed that the compartmentalization was strict and that only one strain was present in each infected node. MIRU-VNTR analysis of additional infected animals on one of the farms confirmed that the compartmentalized infection was a consequence of superinfection, since the two strains were independently infecting other animals. This same analysis revealed the emergence of a microevolved clonal variant in one of the lymph nodes of the compartmentalized animal. Clonal complexity must also be taken into consideration in M. bovis infection, even in low-prevalence settings, and analyses must be adapted to detect it and increase the accuracy of molecular epidemiology studies.

  18. Mycobacterium bovis: a model pathogen at the interface of domestic livestock, wildlife, and humans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Complex and dynamic interactions involving domestic animals, wildlife and humans create environments favorable to the emergence of new diseases, or re-emergence of diseases in new host species. Today, reservoirs of Mycobacterium bovis, the causative agent of tuberculosis in animals and a serious zoo...

  19. Observations on natural and experimental interactions between Schistosoma bovis and S. curassoni from West Africa.

    PubMed

    Rollinson, D; Southgate, V R; Vercruysse, J; Moore, P J

    1990-02-01

    Surveys of 332 naturally infected bovines at eight abattoirs in Senegal, The Gambia and Mali were carried out to determine the prevalence of infection with Schistosoma bovis and S. curassoni and to pinpoint areas where the distribution of the species overlap. S. bovis was the commonest schistosome of cattle in Senegal and Mali being found in animals at seven abattoirs, the highest prevalence of 85.1% occurred at Mopti in Mali. S. bovis was the only bovine schistosome observed in The Gambia. S. curassoni was isolated from a cow at Bamako and shown to have similar glucose-6-phosphate dehydrogenase, phosphoglucomutase and acid phosphatase profiles to those described for a Senegalese isolate. Evidence of interaction of S. bovis with S. curassoni was found in cattle from Senegal, at Tambacounda and Kolda, and from Mali, at Bamako and Mopti. A mixed experimental infection of both species in a sheep showed the lack of any specific mate recognition system: identification of the worms was facilitated by analysis of acid phosphatase by isoelectric focusing in polyacrylamide gels. Viable hybrid parasites were produced in the laboratory and were maintained up until the F4 generation. Comparisons of egg morphology, surface structure of adult male worms and enzyme profiles have been made between experimental hybrid lines and field isolates. Possible mechanisms maintaining species integrity are discussed. PMID:1969699

  20. Spillover of Mycobacterium bovis from Wildlife to Livestock, South 
Africa

    PubMed Central

    Hlokwe, Tiny; Marcotty, Tanguy; du Plessis, Ben J.A.; Michel, Anita L.

    2015-01-01

    During August 2012–February 2013, bovine tuberculosis was detected in communal livestock bordering the Greater Kruger National Park Complex (GKNPC) in South Africa. Using spacer oligonucleotide and variable number tandem repeat typing, we identified the Mycobacterium bovis strain endemic in GKNPC wildlife. Our findings indicate bovine tuberculosis spillover from GKNPC wildlife to neighboring livestock. PMID:25695846

  1. Relative virulence in bison and cattle of bison-associated genotypes of Mycoplasma bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background. Mycoplasma bovis is a cause of respiratory disease in cattle and the bacterium most frequently isolated from bovine respiratory disease complex. It has recently emerged as a major health problem in bison, causing pharyngitis, pneumonia, arthritis, dystocia and abortion. In cattle, M. b...

  2. Virulence of two strains of Mycobacterium bovis in cattle following aerosol infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background Over the past two decades, highly virulent strains of Mycobacterium tuberculosis have emerged and spread rapidly in humans, suggesting a selective advantage based upon virulence. A similar scenario has not been described for Mycobacterium bovis infection in cattle (i.e., Bovine Tuberculos...

  3. A new predilection site of Mycoplasma bovis: Postsurgical seromas in beef cattle.

    PubMed

    Gille, L; Pilo, P; Valgaeren, B R; Van Driessche, L; Van Loo, H; Bodmer, M; Bürki, S; Boyen, F; Haesebrouck, F; Deprez, P; Pardon, B

    2016-04-15

    Mycoplasma bovis is a highly contagious bacterium, which predominantly causes chronic pneumonia, otitis and arthritis in calves and mastitis in adult cattle. In humans, Mycoplasma species have been associated with post-surgical infections. The present study aimed to identify the bacteria associated with three outbreaks of infected seromas after caesarian section in Belgian Blue beef cattle. A total of 10 cases occurred in three herds which were in close proximity of each other and shared the same veterinary practice. M. bovis could be cultured from seroma fluid in five of the six referred animals, mostly in pure culture and was isolated from multiple chronic sites of infection (arthritis and mastitis) as well. DNA fingerprinting of the isolates targeting two insertion sequence elements suggested spread of M. bovis from chronic sites of infection (udder and joints) to the postsurgical seromas. Identical genetic profiles were demonstrated in two animals from two separate farms, suggesting spread between farms. Mortality rate in the referred animals positive for M. bovis in a seroma was 80% (4/5), despite intensive treatment. A massive increase in antimicrobial use was observed in every affected farm. These observations demonstrate involvement of mycoplasmas in outbreaks of postsurgical seromas in cattle. PMID:27016759

  4. Immunological responses following experimental endobronchial infection of badgers (Meles meles) with different doses of Mycobacterium bovis.

    PubMed

    Lesellier, Sandrine; Corner, Leigh; Costello, Eamon; Sleeman, Paddy; Lyashchenko, Konstantin P; Greenwald, Rena; Esfandiari, Javan; Glyn Hewinson, R; Chambers, Mark; Gormley, Eamonn

    2009-01-15

    The Eurasian badger (Meles meles) is a wildlife reservoir for Mycobacterium bovis infection in Ireland and Great Britain and has been implicated in the transmission of tuberculosis to cattle. Vaccination of badgers is an option that could be used as part of a strategy to control the disease. In this study we used an endobronchial infection procedure to inoculate groups of badgers with three different doses (3x10(3), 2x10(2) and <10 Colony Forming Units (CFUs)) of M. bovis. After 17 weeks the disease status of each animal was determined by post-mortem pathology and culture for M. bovis. Each of the inoculum doses resulted in establishment of infection in the badgers. The cell-mediated immune (CMI) responses were measured by lymphocyte transformation assay (LTA) of peripheral blood mononuclear cells (PBMCs) cultured with bovine tuberculin (PPD-B). In each infected group the CMI responses increased with a kinetic profile corresponding to the delivered dose and the post-mortem pathology. The serological responses were measured by ELISA and a multi-antigen print immunoassay (MAPIA) in order to investigate any changes in the antigenic repertoire associated with different infective doses. In contrast to the CMI responses, the ELISA and MAPIA showed that the recognition of antigens by the badgers was intermittent and not strongly influenced by the dose of M. bovis.

  5. Virulent Mycobacterium bovis Beijing Strain Activates the NLRP7 Inflammasome in THP-1 Macrophages

    PubMed Central

    Zhou, Yang; Shah, Syed Zahid Ali; Yang, Lifeng; Zhang, Zhongqiu; Zhou, Xiangmei; Zhao, Deming

    2016-01-01

    Mycobacterium bovis is the causative agent of tuberculosis in a wide range of mammals, including humans. Macrophages are the first line of host defense. They secrete proinflammatory cytokines, such as interleukin-1 beta (IL-1β), in response to mycobacterial infection, but the underlying mechanisms by which human macrophages are activated and release IL-1β following M. bovis infection are poorly understood. Here we show that the ‘nucleotide binding and oligomerization of domain-like receptor (NLR) family pyrin domain containing 7 protein’ (NLRP7) inflammasome is involved in IL-1β secretion and caspase-1 activation induced by M. bovis infection in THP-1 macrophages. NLRP7 inflammasome activation promotes the induction of pyroptosis as well as the expression of tumor necrosis factor alpha (TNF-α), Chemokine (C-C motif) ligand 3 (CCL3) and IL-1β mRNAs. Thus, the NLRP7 inflammasome contributes to IL-1β secretion and induction of pyroptosis in response to M. bovis infection in THP-1 macrophages. PMID:27043315

  6. Spatial relationships between Eurasian badgers (Meles meles) and cattle infected with Mycobacterium bovis in Northern Spain.

    PubMed

    Balseiro, Ana; González-Quirós, Pablo; Rodríguez, Óscar; Francisca Copano, M; Merediz, Isabel; de Juan, Lucía; Chambers, Mark A; Delahay, Richard J; Marreros, Nelson; Royo, Luis J; Bezos, Javier; Prieto, José M; Gortázar, Christian

    2013-09-01

    Recent studies suggest that badgers may be a potential reservoir of Mycobacterium bovis infection for cattle in Northern Spain. The objective of this study was to investigate potential epidemiological links between cattle and badgers. Culture and molecular typing data were available for cattle culled during the national tuberculosis (TB) eradication campaigns between 2008 and 2012, as well as from 171 necropsied badgers and 60 live animals trapped and examined over the same time period. Mycobacterium tuberculosis complex strains were isolated from pooled tissues of 14 (8.2%) necropsied badgers, of which 11 were identified as M. bovis: six different spoligotypes of M. bovis were subsequently identified. In two geographical locations where these isolates were shared between cattle and badgers, infected cattle herds and badgers lived in close contact. Although it remains unclear if badgers are a maintenance or spill-over host of M. bovis in this setting, it would appear prudent to have precautionary measures in place to reduce contact between cattle and badgers.

  7. The phylogeny and population structure of Mycobacterium bovis in the British Isles.

    PubMed

    Allen, A R; Dale, J; McCormick, C; Mallon, T R; Costello, E; Gordon, S V; Hewinson, R G; Skuce, R A; Smith, N H

    2013-12-01

    To further understand the epidemic of bovine tuberculosis in Great Britain, Northern Ireland and the Republic of Ireland, we identified 16 mutations that are phylogenetically informative for Mycobacterium bovis strains from these regions. We determined the status of these mutations among a collection of 501 strains representing the molecular diversity found in these three regions of the British Isles. The resulting linear phylogenies from each region were concordant, showing that the same lineage of M. bovis was present. The dominance of this lineage is unique within Europe, and suggests that in the past the populations were homogenous. Comparison of approximately 500 strains isolated in 2005 from each region by spoligotype and 5 locus VNTR profiling, revealed distinct differences in the genotype frequencies and sub-lineage makeup between each region. We concluded that whilst each region shared the same major phylogenetic lineage of M. bovis, more recent evolution had resulted in the development of region-specific populations. Regional differences in the M. bovis populations suggest that it may be possible to identify the movement of strains from one region to another. PMID:23933404

  8. Performance of a Noninvasive Test for Detecting Mycobacterium bovis Shedding in European Badger (Meles meles) Populations

    PubMed Central

    Murphy, Andrew; James, Phillip; Travis, Emma; Porter, David; Sawyer, Jason; Cork, Jennifer; Delahay, Richard J.; Gaze, William; Courtenay, Orin; Wellington, Elizabeth M.

    2015-01-01

    The incidence of Mycobacterium bovis, the causative agent of bovine tuberculosis, in cattle herds in the United Kingdom is increasing, resulting in substantial economic losses. The European badger (Meles meles) is implicated as a wildlife reservoir and is the subject of control measures aimed at reducing the incidence of infection in cattle populations. Understanding the epidemiology of M. bovis in badger populations is essential for directing control interventions and understanding disease spread; however, accurate diagnosis in live animals is challenging and currently uses invasive methods. Here we present a noninvasive diagnostic procedure and sampling regimen using field sampling of latrines and detection of M. bovis with quantitative PCR tests, the results of which strongly correlate with the results of immunoassays in the field at the social group level. This method allows M. bovis infections in badger populations to be monitored without trapping and provides additional information on the quantities of bacterial DNA shed. Therefore, our approach may provide valuable insights into the epidemiology of bovine tuberculosis in badger populations and inform disease control interventions. PMID:26041891

  9. Risk of Mycoplasma bovis transmission from contaminated sand bedding to naive dairy calves.

    PubMed

    Wilson, D J; Justice-Allen, A; Goodell, G; Baldwin, T J; Skirpstunas, R T; Cavender, K B

    2011-03-01

    The objective of this study was to evaluate the possible transmission of Mycoplasma bovis from positive sand bedding to naïve dairy calves. Twelve preweaned Holstein bull calves were blocked in pairs and randomly assigned as unexposed controls (n=6) bedded with control sand, or exposed calves (n=6) bedded with sand previously positive for M. bovis at a dairy farm. Bedding sand was cultured weekly. Nasal and ear swabs and sera were collected weekly, tracheal swabs were collected monthly, and by the end of the 105-d study, all calves were euthanized (n=10) or died (n=2). Sera were tested for M. bovis-specific antibody. Mycoplasma spp. culture was performed on nasal and ear swabs; culture and a PCR differentiating multiple Mycoplasma spp. were performed on postmortem samples of lung, retropharyngeal lymph node, and trachea from each calf. A complete necropsy also was performed. During 6 wk, mycoplasma concentration in exposed group sand was between 200 and 32,000 cfu/g. All 166 tracheal swabs, nasal and ear swabs, and postmortem tests from all calves were negative for mycoplasma. All 94 sera were negative for M. bovis-specific antibody. No gross pathology suggestive of mycoplasma disease was detected. The probability of mycoplasma detection, if an exposed calf had become infected 4 wk after exposure, ranged between 97 and 99% depending on time of exposure for individual calves. There was no evidence that sand bedding contaminated with M. bovis might serve as a source of transmission to naïve dairy calves.

  10. Acquired resistance to the 16-membered macrolides tylosin and tilmicosin by Mycoplasma bovis.

    PubMed

    Lerner, Uri; Amram, Eytan; Ayling, Roger D; Mikula, Inna; Gerchman, Irena; Harrus, Shimon; Teff, Dina; Yogev, David; Lysnyansky, Inna

    2014-01-31

    The molecular mechanism of acquired resistance to the 16-membered macrolides tylosin (Ty) and tilmicosin (Tm) was investigated in Mycoplasma bovis field isolates. Sequence analysis of domains II and V of the two 23S rRNA alleles and ribosomal proteins L4 and L22 was performed on 54 M. bovis isolates showing different minimal inhibitory concentrations (MIC). The presence of any one of the point mutations G748A, C752T, A2058G, A2059G or A2059C (Escherichia coli numbering) in one or both alleles of the 23S rRNAs was correlated with decreased susceptibility to Ty (8-1024 μg/ml) and to Tm (32 to >256 μg/ml) in 27/27 and 27/31 M. bovis isolates, respectively. Although a single mutation in domain II or V could be sufficient to cause decreased susceptibility to Ty, our data imply that a combination of mutations in two domains is necessary to achieve higher MICs (≥ 128 μg/ml). The influence of a combination of mutations in two domains II and V on enhancement of resistance to Tm was less clear. In addition, the amino acid (aa) substitution L22-Q90H was found in 24/32 representative M. bovis isolates with different MICs, but no correlation with decreased susceptibility to Ty or Tm was identified. Multiple aa substitutions were also identified in the L4 protein, including at positions 185-186 (positions 64 and 65 in E. coli) which are adjacent to the macrolide-binding site. This is the first description of the molecular mechanism of acquired resistance to the 16-membered macrolides in M. bovis. PMID:24393633

  11. Endometrial inflammation and abnormal expression of extracellular matrix proteins induced by Mycoplasma bovis in dairy cows.

    PubMed

    Guo, Mengyao; Wang, Guoqing; Lv, Tingting; Song, Xiaojing; Wang, Tiancheng; Xie, Guanghong; Cao, Yongguo; Zhang, Naisheng; Cao, Rongfeng

    2014-03-15

    Mycoplasma bovis infection can cause endometrial inflammation leading to infertility and involuntary culling in dairy cows. Because extracellular matrix (ECM) proteins affect the adherence of mycoplasma to eukaryotic cell surface, they may play a role in the pathogenesis of the bacteria. The objective of the present study was to evaluate the endometrial inflammatory response and ECM protein expression induced by M bovis. Endometrial concentrations of inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and mRNA and protein expression of collagen IV (CL-IV), fibronectin (FN), and laminin (LN) were evaluated 10, 20, and 30 days after M bovis intrauterine infusion in breed cows 18 days postpartum. The presence of the bacteria in the uterus was detected by nested polymerase chain reaction and denaturing gradient gel electrophoresis. Endometrial TNF-α, IL-1β, and IL-6 concentrations in the treatment group were greater (P < 0.05) than in the positive and negative control groups 20 and 30 days after infusion. Endometrial CL-IV, FN, and LN mRNA and protein expression increased (P < 0.01) 20 days after infusion in all groups. However, the increase was more pronounced in the treatment group and reactive expressions were greater (P < 0.05) than in the positive and negative control groups 10, 20, and 30 days after infusion. In conclusion, M bovis triggered endometrial inflammatory response and increased CL-IV, FN, and LN mRNA and protein expression. The abnormal expression of ECM these proteins may promote the pathogenic effects of M bovis that lead to endometrial tissue damage and infertility.

  12. Acquired resistance to the 16-membered macrolides tylosin and tilmicosin by Mycoplasma bovis.

    PubMed

    Lerner, Uri; Amram, Eytan; Ayling, Roger D; Mikula, Inna; Gerchman, Irena; Harrus, Shimon; Teff, Dina; Yogev, David; Lysnyansky, Inna

    2014-01-31

    The molecular mechanism of acquired resistance to the 16-membered macrolides tylosin (Ty) and tilmicosin (Tm) was investigated in Mycoplasma bovis field isolates. Sequence analysis of domains II and V of the two 23S rRNA alleles and ribosomal proteins L4 and L22 was performed on 54 M. bovis isolates showing different minimal inhibitory concentrations (MIC). The presence of any one of the point mutations G748A, C752T, A2058G, A2059G or A2059C (Escherichia coli numbering) in one or both alleles of the 23S rRNAs was correlated with decreased susceptibility to Ty (8-1024 μg/ml) and to Tm (32 to >256 μg/ml) in 27/27 and 27/31 M. bovis isolates, respectively. Although a single mutation in domain II or V could be sufficient to cause decreased susceptibility to Ty, our data imply that a combination of mutations in two domains is necessary to achieve higher MICs (≥ 128 μg/ml). The influence of a combination of mutations in two domains II and V on enhancement of resistance to Tm was less clear. In addition, the amino acid (aa) substitution L22-Q90H was found in 24/32 representative M. bovis isolates with different MICs, but no correlation with decreased susceptibility to Ty or Tm was identified. Multiple aa substitutions were also identified in the L4 protein, including at positions 185-186 (positions 64 and 65 in E. coli) which are adjacent to the macrolide-binding site. This is the first description of the molecular mechanism of acquired resistance to the 16-membered macrolides in M. bovis.

  13. Glutamate Dehydrogenase Is Required by Mycobacterium bovis BCG for Resistance to Cellular Stress

    PubMed Central

    Gallant, James L.; Viljoen, Albertus J.; van Helden, Paul D.; Wiid, Ian J. F.

    2016-01-01

    We recently reported on our success to generate deletion mutants of the genes encoding glutamate dehydrogenase (GDH) and glutamine oxoglutarate aminotransferase (GOGAT) in M. bovis BCG, despite their in vitro essentiality in M. tuberculosis. We could use these mutants to delineate the roles of GDH and GOGAT in mycobacterial nitrogen metabolism by using M. bovis BCG as a model for M. tuberculosis specifically. Here, we extended our investigation towards the involvement of GDH and GOGAT in other aspects of M. bovis BCG physiology, including the use of glutamate as a carbon source and resistance to known phagosomal stresses, as well as in survival inside macrophages. We find that gdh is indispensable for the utilization of glutamate as a major carbon source, in low pH environments and when challenged with nitric oxide. On the other hand, the gltBD mutant had increased viability under low pH conditions and was unaffected by a challenge with nitric oxide. Strikingly, GDH was required to sustain M. bovis BCG during infection of both murine RAW 264.7 and bone-marrow derived and macrophages, while GOGAT was not. We conclude that the catabolism of glutamate in slow growing mycobacteria may be a crucial function during infection of macrophage cells and demonstrate a novel requirement for M. bovis BCG GDH in the protection against acidic and nitrosative stress. These results provide strong clues on the role of GDH in intracellular survival of M. tuberculosis, in which the essentiality of the gdh gene complicates knock out studies making the study of the role of this enzyme in pathogenesis difficult. PMID:26824899

  14. Epizootiologic survey of Mycobacterium bovis in wildlife and farm environments in northern Michigan.

    PubMed

    Witmer, Gary; Fine, Amanda E; Gionfriddo, James; Pipas, Michael; Shively, Kirk; Piccolo, Kim; Burke, Patrick

    2010-04-01

    Bovine tuberculosis (bovine TB), caused by Mycobacterium bovis, has reemerged in northern Michigan, USA, with detections in white-tailed deer (Odocoileus virginianus) in 1994 and in cattle in 1998. Since then, significant efforts have been directed toward reducing deer densities in the area in the hopes of reducing the bovine TB prevalence rate in deer and eliminating spillover of the disease into cattle. Despite the success of the efforts to reduce deer densities, additional cattle herds have become infected. Other mammals can be infected with M. bovis, and some carnivores and omnivores had been found to be infected with the disease in northern Michigan, USA. We conducted a multiyear surveillance effort to detect bovine TB in wild species of mammals in the Michigan, USA, outbreak area. From 2002 to 2004, tissue samples from 1,031 individual animals of 32 species were collected, processed, and cultured for M. bovis. Only 10 (1.0%) were culture-positive for M. bovis (five raccoons [Procyon lotor], four opossums [Didelphis virginiana], and one grey fox [Urocyon cinereoargenteus]). We also found two raccoons and four opossums to be positive for Mycobacterium avium. We collected 503 environmental samples from cattle farms recently identified as bovine TB positive; none yielded positive M. bovis culture results. Finally, we used infrared cameras to document wildlife use of four barns in the area. Many avian and mammalian species of wildlife were observed, with raccoons being the most commonly observed species. This surveillance study identified no new wildlife species that should be considered significant reservoirs of bovine TB in the outbreak area in northern Michigan, USA. However, the relatively high, apparent bovine TB prevalence rates in some carnivorous and omnivorous species, their relatively long life spans, and their frequent use of barns, suggests that removal of raccoons, opossums, foxes, and coyotes (Canis latrans) should be considered when a newly infected

  15. Use of an electronic nose to diagnose Mycobacterium bovis infection in badgers and cattle.

    PubMed

    Fend, R; Geddes, R; Lesellier, S; Vordermeier, H-M; Corner, L A L; Gormley, E; Costello, E; Hewinson, R G; Marlin, D J; Woodman, A C; Chambers, M A

    2005-04-01

    It is estimated that more than 50 million cattle are infected with Mycobacterium bovis worldwide, resulting in severe economic losses. Current diagnosis of tuberculosis (TB) in cattle relies on tuberculin skin testing, and when combined with the slaughter of test-positive animals, it has significantly reduced the incidence of bovine TB. The failure to eradicate bovine TB in Great Britain has been attributed in part to a reservoir of the infection in badgers (Meles meles). Accurate and reliable diagnosis of infection is the cornerstone of TB control. Bacteriological diagnosis has these characteristics, but only with samples collected postmortem. Unlike significant wild animal reservoirs of M. bovis that are considered pests in other countries, such as the brushtail possum (Trichosurus vulpecula) in New Zealand, the badger and its sett are protected under United Kingdom legislation (The Protection of Badgers Act 1992). Therefore, an accurate in vitro test for badgers is needed urgently to determine the extent of the reservoir of infection cheaply and without destroying badgers. For cattle, a rapid on-farm test to complement the existing tests (the skin test and gamma interferon assay) would be highly desirable. To this end, we have investigated the potential of an electronic nose (EN) to diagnose infection of cattle or badgers with M. bovis, using a serum sample. Samples were obtained from both experimentally infected badgers and cattle, as well as naturally infected badgers. Without exception, the EN was able to discriminate infected animals from controls as early as 3 weeks after infection with M. bovis, the earliest time point examined postchallenge. The EN approach described here is a straightforward alternative to conventional methods of TB diagnosis, and it offers considerable potential as a sensitive, rapid, and cost-effective means of diagnosing M. bovis infection in cattle and badgers.

  16. Effect of milk fermentation by kefir grains and selected single strains of lactic acid bacteria on the survival of Mycobacterium bovis BCG.

    PubMed

    Macuamule, C L S; Wiid, I J; van Helden, P D; Tanner, M; Witthuhn, R C

    2016-01-18

    Mycobacterium bovis that causes Bovine tuberculosis (BTB) can be transmitted to humans thought consumption of raw and raw fermented milk products from diseased animals. Lactic acid bacteria (LAB) used in popular traditional milk products in Africa produce anti-microbial compounds that inhibit some pathogenic and spoilage bacteria. M. bovis BCG is an attenuated non-pathogenic vaccine strain of M. bovis and the aim of the study was to determine the effect of the fermentation process on the survival of M. bovis BCG in milk. M. bovis BCG at concentrations of 6 log CFU/ml was added to products of kefir fermentation. The survival of M. bovis BCG was monitored at 12-h intervals for 72 h by enumerating viable cells on Middlebrook 7H10 agar plates enriched with 2% BD BACTEC PANTA™. M. bovis BCG was increasingly reduced in sterile kefir that was fermented for a period of 24h and longer. In the milk fermented with kefir grains, Lactobacillus paracasei subsp. paracasei or Lactobacillus casei, the viability of M. bovis BCG was reduced by 0.4 logs after 24h and by 2 logs after 48 h of fermentation. No viable M. bovis BCG was detected after 60 h of fermentation. Results from this study show that long term fermentation under certain conditions may have the potential to inactivate M. bovis BCG present in the milk. However, to ensure safety of fermented milk in Africa, fermentation should be combined with other hurdle technologies such as boiling and milk pasteurisation.

  17. Recombinant methionine aminopeptidase protein of Babesia microti: immunobiochemical characterization as a vaccine candidate against human babesiosis.

    PubMed

    Munkhjargal, Tserendorj; Yokoyama, Naoaki; Igarashi, Ikuo

    2016-09-01

    Human babesiosis is the most important zoonotic protozoan infection in the world. This is the first report of the cloning, expression, purification, and immunobiochemical characterization of a methionine aminopeptidase 1 (MetAP1) protein from Babesia microti (B. microti). The gene encodes a MetAP1 protein of B. microti (BmMetAP1) of approximately 66.8 kDa that includes glutathione S-transferase (GST) tag and shows MetAP activity. BmMetAP1 was detected in a lysate of B. microti and further localized in cytoplasm of the B. microti merozoite. rBmMetAP1 was found to be immunogenic, eliciting a high antibody titer in mice. Moreover, rBmMetAP1 stimulated the production of IFN-γ and IL-12 but not IL-4. Finally, rBmMetAP1 was able to provide considerable protection to mice against a B. microti challenge infection based on a reduction in peak parasitemia levels and earlier clearance of the parasite as compared with control mice. Taken together, these results suggest that rBmMetAP1 confers significant protection against experimental B. microti infection and might be considered a potential vaccine target against human babesiosis. PMID:27306898

  18. Detection of Babesia divergens in southern Norway by using an immunofluorescence antibody test in cow sera

    PubMed Central

    2010-01-01

    Background The incidence of bovine babesiosis, caused by Babesia divergens (Apicomplexa: Piroplasmida) has decreased markedly since the 1930 s, but may re-emerge as a consequence of climate change and changes in legislation and pasturing practices. This is a potentially serious disease, with both economical and animal welfare consequences. Therefore, there is a need to survey the distribution of B. divergens. Methods We tested sera from 306 healthy pastured cows from 24 farms along the southern Norwegian coast by using an indirect immunofluorescence IgG antibody test (IFAT). Fractions of seropositive cows were compared by calculating 95% CI. Results The results of this test showed that 27% of the sera were positive for B. divergens antibodies. The fraction of antibody-positive sera that we detected showed a two-humped distribution, with a high fraction of positives being found in municipalities in the western and eastern parts of the study area, while the municipalities between these areas had few or no positive serum samples. Conclusions Neither the farmers' observations nor the Norwegian Dairy Herd Recording System give an adequate picture of the distribution of bovine babesiosis. Serological testing of cows by using IFAT is a convenient way of screening for the presence of B. divergens in an area. PMID:20925923

  19. Molecular detection and prevalence of Theileria equi and Babesia caballi in horses of central Balkan.

    PubMed

    Davitkov, Darko; Vucicevic, Milos; Stevanovic, Jevrosima; Krstic, Vanja; Slijepcevic, Dajana; Glavinic, Uros; Stanimirovic, Zoran

    2016-03-01

    Equine piroplasmosis is significant tick-borne disease with wide distribution. The prevalence of equine piroplasmosis in Serbia, Montenegro and Bosnia and Herzegovina is unknown. In aim to obtain a first insight into the prevalence we performed molecular epidemiological study which included 142 horses, on seven locations in these three countries. We first performed PCR for the detection of a 450bp long section of the 18S rRNA of piroplasma-specific region. For all positive samples we have done multiplex PCR for the species detection. Species determination was further confirmed by sequencing PCR products of 10 randomly selected Theileria equi and all Babesia caballi samples. The overall prevalence rates in analysed region for T. equi and B. caballi were 22.5% and 2.1%, respectively. Possible risk factors (such as location, age, sex and activity) associated with PCR positivity were evaluated. Marked differences were found in prevalence between geographic areas. There was no significant association between positivity and age group. T. equi was more prevalent in females and farming horses. This is the first report on the molecular survey of T. equi and B. caballi in central Balkan. Further prevalence studies on definitive host and vectors in this region are necessary. PMID:27078657

  20. Human Coinfection with Borrelia burgdorferi and Babesia microti in the United States

    PubMed Central

    Knapp, Kristen L.; Rice, Nancy A.

    2015-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease, and Babesia microti, a causative agent of babesiosis, are increasingly implicated in the growing tick-borne disease burden in the northeastern United States. These pathogens are transmitted via the bite of an infected tick vector, Ixodes scapularis, which is capable of harboring and inoculating a host with multiple pathogens simultaneously. Clinical presentation of the diseases is heterogeneous and ranges from mild flu-like symptoms to near-fatal cardiac arrhythmias. While the reason for the variability is not known, the possibility exists that concomitant infection with both B. burgdorferi and B. microti may synergistically increase disease severity. In an effort to clarify the current state of understanding regarding coinfection with B. burgdorferi and B. microti, in this review, we discuss the geographical distribution and pathogenesis of Lyme disease and babesiosis in the United States, the immunological response of humans to B. burgdorferi or B. microti infection, the existing knowledge regarding coinfection disease pathology, and critical factors that have led to ambiguity in the literature regarding coinfection, in order to eliminate confusion in future experimental design and investigation. PMID:26697208

  1. First report of Anaplasma phagocytophilum and Babesia microti in rodents in Finland.

    PubMed

    Kallio, Eva R; Begon, Michael; Birtles, Richard J; Bown, Kevin J; Koskela, Esa; Mappes, Tapio; Watts, Phillip C

    2014-06-01

    Tick-borne diseases pose an increasingly important public health problem in Europe. Rodents are the reservoir host for many tick-transmitted pathogens, including Anaplasma phagocytophilum and Babesia microti, which can cause human granulocytic anaplasmosis and babesiosis, respectively. To estimate the presence of these pathogens in rodents in Finland, we examined blood samples from 151 bank voles (Myodes glareolus) and demonstrate, for the first time, that A. phagocytophilum and B. microti commonly infect bank voles (in 22% and 40% of animals, respectively) in Finland. Sequence analysis of a fragment of 18S rRNA showed that the B. microti strain isolated was identical to the Munich strain, which is considered to be nonzoonotic. The A. phagocytophilum strain (based on a fragment of the msp4 gene) was identical to one found earlier in rodents in the United Kingdom that is transmitted by the tick Ixodes trianguliceps, all the life stages of which feed on small mammals. The infection probability of B. microti in the bank voles was the greater the older the individual was, and males were more often infected than females. A. phagocytophilum infection probability first increased and then decreased with the age of individual without any difference between sexes. While these pathogens presumably pose a limited zoonotic risk to humans in Finland, they might have important interactions with other rodent pathogens and therefore affect infection dynamics of, for example, zoonotic pathogens.

  2. Theileria equi and Babesia caballi infection of equids in Punjab, India: a serological and molecular survey.

    PubMed

    Sumbria, Deepak; Das Singla, Lachhman; Sharma, Amrita

    2016-01-01

    A cross-sectional study was conducted in Submountain undulating, Undulating plain, Western and Western plain agro-climatic zones of Punjab province, India, to determine the prevalence, agreement between diagnostic tests and associated related risk factors of Theileria equi and Babesia caballi infection in equids (horses, donkey, mules). An overall prevalence of 14.14 and 0.0% of T. equi and B. caballi was recorded by multiplex polymerase chain reaction targeting 18S ribosomal RNA (rRNA) for both the parasites and 75 and 1.11% by competitive enzyme-linked immunosorbent assay in a representative sample of 180 animals. Only two animals with positive antibody titre from B. caballi and none with PCR indicated T. equi as the predominant haemoprotozoan responsible for equine piroplasmosis in the study area. Among the PCR-positive animals, presence of tick vectors in farm vicinity was the most influential associated with T. equi infection (P = 0.002; odds ratio (OR) 9.30; 95% confidence interval (CI) = 3.32-27.10). For animals with higher anti-T. equi antibody titres, strong association of sero-prevalence for T. equi was recorded with age, sex, usage, tick infestation and deworming/vaccination status of host animals and farm management strategies. The study has demonstrated the possible absence of B. caballi in both conducive and non-conducive areas of Punjab and demonstrated T. equi as the potential agent of equine piroplasmosis in Punjab.

  3. The Effect of Babesia divergens Infection on the Spleen of Mongolian Gerbils

    PubMed Central

    Dkhil, Mohamed A.; Al-Khalifa, Mohamed S.

    2014-01-01

    Babesiosis is caused by intraerythrocytic protozoan parasites transmitted by ticks and affects a wide range of domestic and wild animals and occasionally humans. The current study aimed to investigate the effect of B. divergens infected erythrocytes on spleen histopathology, cell cycle alteration, and the presence of oxidative stress. Mongolian gerbils were challenged with 5 × 106  Babesia divergens infected erythrocytes. Parasitemia reached approximately 77% at day 5 postinfection. Infection also induced injury of the spleen. This was evidenced with (i) increases in cellular damage of the spleen, (ii) decrease in antioxidant capacity as indicated by decreased glutathione, catalase, and superoxide dismutase levels, (iii) increased production of malondialdehyde and nitric oxide derived products (nitrite/nitrate), and (iv) increased lactic acid dehydrogenase activity and protein carbonyl content in the spleen. Infection interfered with normal cell cycle of the spleen cells at G0/G1, S, and G2/M phases. On the basis of the above results it can be hypothesized that B. divergens infected erythrocytes could alter the spleen histopathology and cause cell cycle alteration and induce oxidative stress in splenic tissue. PMID:25136591

  4. Serial haematology results in transfused and non-transfused dogs naturally infected with Babesia rossi.

    PubMed

    Scheepers, E; Leisewitz, A L; Thompson, P N; Christopher, M M

    2011-09-01

    This prospective longitudinal study investigated the progression of haematological changes in 32 transfused and 54 non-transfused dogs naturally infected with Babesia rossi over the 1st 6 days following diagnosis and treatment. The effect of patient age on the results of complete blood counts was determined. Haematology data were analysed at presentation and at 24 hours, 3 days and 6 days after presentation. Dogs were treated with diminazene aceturate at diagnosis and a blood transfusion was given if deemed clinically required. Mildly to moderately regenerative normocytic normochromic anaemia was observed in all dogs throughout the study period. Transfused dogs more often had an inflammatory leukogram at presentation and at 24 hours, than dogs that were not transfused. In dogs with a left shift, a concurrent normal or decreased segmented neutrophil count was found more commonly than neutrophilia. Severe thrombocytopenia that resolved within a week was common. Blood transfusion alleviated the anaemia, but had no significant effect on white blood cell or platelet responses. Blood cell responses were not significantly influenced by age. In conclusion, the red blood cell and white blood cell responses were less than expected in dogs with babesiosis, given the degree of anaemia and inflammation present. The magnitude of thrombocytopenia and rapid return of the platelet count to normal suggested a possible immune-mediated mechanism for the thrombocytopenia.

  5. Molecular and serological detection of Theileria equi and Babesia caballi in donkeys (Equus asinus) in Brazil.

    PubMed

    Machado, R Z; Toledo, C Z P; Teixeira, M C A; André, M R; Freschi, C R; Sampaio, P H

    2012-05-25

    Piroplasmosis in donkeys has been recognized as a serious problem of major economic importance, since the affected animals manifest loss of appetite and decreased working capacity. The present work is aimed at detecting infection or exposure of donkeys in São Paulo, Brazil to Theileria (T.) equi and Babesia (B.) caballi using molecular and serological approaches. EDTA-blood and serum samples were collected from 88 donkeys (Equus asinus). From 88 sampled donkeys, 65 (73.86%; 95% confidence interval, PI=63.41, 82.65) and 82 (93.2%; 95% confidence interval, PI=85.75, 97.46) animals showed IgG antibodies to T. equi (by ELISA) and B. caballi (by IFAT), respectively. Twenty-eight (31.81%; 95% confidence interval, PI=22.3, 42.61) and 18 (20.45%; 95% confidence interval, PI=12.6, 30.39) donkeys were positive to T. equi and B. caballi nested PCR assays, respectively. The results indicated that T. equi and B. caballi are prevalent among donkeys in Brazil.

  6. Identification of Babesia bigemina infected erythrocyte surface antigens containing epitopes conserved among strains.

    PubMed

    Shompole, S; McElwain, T F; Jasmer, D P; Hines, S A; Katende, J; Musoke, A J; Rurangirwa, F R; McGuire, T C

    1994-03-01

    The presence of previously uncharacterized antigens (new antigens) on the surface of intact erythrocytes infected with three strains of Babesia bigemina from Kenya and one each from Puerto Rico, Mexico, St. Croix, and Texcoco-Mexico was demonstrated by indirect immunofluorescent antibody (IFA) reactions. These antigens were not strain specific because antibodies in bovine immune serum to either the Mexico or Kenya isolates reacted with all seven strains tested. Homologous and heterologous immune serum antibodies bound a maximum of 83% and 55%, respectively, of intact erythrocytes infected with the Kenya-Ngong strain but not uninfected erythrocytes. Both sera caused agglutination of only infected erythrocytes. Antibodies eluted from the surface of glutaraldehyde (0.25%) fixed infected erythrocytes had IFA reaction patterns among strains similar to those of immune sera before elution. Eluted antibodies were used to determine if these antigens were protein and encoded by B. bigemina. Eluted antibodies bound seven parasite-encoded proteins of 240, 220, 66, 62, 58, 52 and 38 kDa in an erythrocyte surface-specific immunoprecipitation reaction of 35S-methionine labelled proteins. It was concluded that the surface of B. bigemina infected erythrocytes had parasite-encoded proteins and that these proteins had surface exposed epitopes that were conserved among the seven strains examined which were from two continents.

  7. Longevity of Mycobacterium bovis in Raw and Traditional Souring Milk as a Function of Storage Temperature and Dose

    PubMed Central

    Hlokwe, Tiny; Raseleka, Keneilwe; Getz, Wayne M.; Marcotty, Tanguy

    2015-01-01

    Background Unpasteurised fresh and souring dairy products form an essential component of household diets throughout many rural communities in southern Africa. The presence of milk-borne zoonotic pathogens such as Mycobacterium bovis (M. bovis), the causative agent of bovine tuberculosis and zoonotic tuberculosis in humans, constitute a public health threat, especially in remote areas with poor disease surveillance in livestock and highly compromised human health due to HIV/AIDS. Methods In this study we used culture to determine the longevity of M. bovis in experimentally inoculated fresh and naturally souring milk obtained from communal cattle in the KwaZulu-Natal province of South Africa. The effect of bacterial load and storage temperature on the survival of M. bovis was evaluated by spiking mixtures of fresh milk and starter soured milk (aMasi) culture with three concentrations of bacteria (102, 104, 107 colony forming units/ml), followed by incubation under controlled laboratory conditions that mimicked ambient indoor (20°C) and outdoor (33°C) temperatures and periodic sampling and testing over time (0-56 days). Results M. bovis cultured from samples of the fresh and souring milk was identified by PCR analysis. At the highest spiking concentration (107cfu/ml), M. bovis survived for at least 2 weeks at 20°C; but, at all concentrations in the 33°C treatment, M. bovis was absent by three days after inoculation. Logistic regression analysis was used to assess the effects of bacterial concentration and time since inoculation, as well as determine the potential half-life of M. bovis in raw souring milk. Given the most favourable tested conditions for bacterial survival (20°C), approximately 25% of mycobacteria were alive after one day of storage (95% CI: 9-53%), giving an estimated half-life of M. bovis in raw souring milk of approximately 12 hours (95% CI: 7-27 hours). Conclusions This study demonstrates that M. bovis may survive in fresh and souring milk for

  8. Loss of diversity within Mycoplasma bovis isolates collected in France from bovines with respiratory diseases over the last 35 years.

    PubMed

    Becker, Claire A M; Thibault, François M; Arcangioli, Marie-Anne; Tardy, Florence

    2015-07-01

    Mycoplasma (M.) bovis has recently emerged as a major, worldwide etiological agent of bovine respiratory diseases leading to huge economic losses mainly due to high morbidity and mortality as well as poor growth rates. The spread of M. bovis infections between different animals, herds, regions or countries has been often reported to be connected to the movement of animals. However, despite recent considerable efforts, no universal subtyping method is yet available to trace M. bovis isolates circulation at an international scale. Moreover in France, the overall population diversity of M. bovis isolates has not been assessed since the early 1990s. This study was conducted to fill in these gaps. The genotypic diversity between sixty isolates collected in France over the last 35 years was assessed using two molecular subtyping methods that addressed either the long-term epidemiological relationships (Multi Locus Sequence Typing, MLST) or the genetic microvariations (Multiple Locus VNTR Analysis, MLVA) between isolates. Phenotypic diversity was also analyzed by using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) to compare the main protein patterns of isolates. All proposed subtyping approaches were optimized and led to the same pattern in the French M. bovis population that consisted of two clusters, the first one comprising isolates collected before 2000 and the second, those collected after 2000. Recent strains were further shown to be more homogeneous than older ones, which is consistent with the spread of a single clone throughout the country. Because this spread was concomitant with the emergence of multiresistant M. bovis isolates, several hypotheses are discussed to explain the homogeneity of M. bovis isolates in France, even though the M. bovis species is fully equipped to generate diversity. PMID:25913158

  9. Utility of a fecal real-time PCR protocol for detection of Mycobacterium bovis infection in African buffalo (Syncerus caffer).

    PubMed

    Roug, Annette; Geoghegan, Claire; Wellington, Elizabeth; Miller, Woutrina A; Travis, Emma; Porter, David; Cooper, David; Clifford, Deana L; Mazet, Jonna A K; Parsons, Sven

    2014-01-01

    A real-time PCR protocol for detecting Mycobacterium bovis in feces was evaluated in bovine tuberculosis-infected African buffalo (Syncerus caffer). Fecal samples spiked with 1.42 × 10(3) cells of M. bovis culture/g and Bacille Calmette-Guérin standards with 1.58 × 10(1) genome copies/well were positive by real-time PCR but all field samples were negative.

  10. Loss of diversity within Mycoplasma bovis isolates collected in France from bovines with respiratory diseases over the last 35 years.

    PubMed

    Becker, Claire A M; Thibault, François M; Arcangioli, Marie-Anne; Tardy, Florence

    2015-07-01

    Mycoplasma (M.) bovis has recently emerged as a major, worldwide etiological agent of bovine respiratory diseases leading to huge economic losses mainly due to high morbidity and mortality as well as poor growth rates. The spread of M. bovis infections between different animals, herds, regions or countries has been often reported to be connected to the movement of animals. However, despite recent considerable efforts, no universal subtyping method is yet available to trace M. bovis isolates circulation at an international scale. Moreover in France, the overall population diversity of M. bovis isolates has not been assessed since the early 1990s. This study was conducted to fill in these gaps. The genotypic diversity between sixty isolates collected in France over the last 35 years was assessed using two molecular subtyping methods that addressed either the long-term epidemiological relationships (Multi Locus Sequence Typing, MLST) or the genetic microvariations (Multiple Locus VNTR Analysis, MLVA) between isolates. Phenotypic diversity was also analyzed by using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) to compare the main protein patterns of isolates. All proposed subtyping approaches were optimized and led to the same pattern in the French M. bovis population that consisted of two clusters, the first one comprising isolates collected before 2000 and the second, those collected after 2000. Recent strains were further shown to be more homogeneous than older ones, which is consistent with the spread of a single clone throughout the country. Because this spread was concomitant with the emergence of multiresistant M. bovis isolates, several hypotheses are discussed to explain the homogeneity of M. bovis isolates in France, even though the M. bovis species is fully equipped to generate diversity.

  11. Clinical and morphological characteristics in Streptococcus bovis endocarditis: a comparison with other causative microorganisms in 177 cases

    PubMed Central

    Kupferwasser, I; Darius, H; Muller, A; Mohr-Kahaly, S; Westermeier, T; Oelert, H; Erbel, R; Meyer, J

    1998-01-01

    Aim—To compare the clinical and morphological characteristics of patients with Streptococcus bovis endocarditis with those of patients with endocarditis caused by other microorganisms.
Methods—177 consecutive patients (Streptococcus bovis, 22; other streptococci, 94; staphylococci, 44; other, 17) with definite infective endocarditis according to the Duke criteria were included. All patients underwent transthoracic and transoesophageal echocardiography. In 88 patients, findings from surgery/necropsy were obtained.
Results—S bovis endocarditis was associated with older patients, with a higher mortality (p = 0.04), and with a higher rate of cardiac surgery (p < 0.001) than other microorganisms, although embolic events were observed less often (p = 0.02). Pathological gastrointestinal lesions were detected in 45% of the patients. Multiple valves were affected in 68% of the patients with S bovis endocarditis and in 20% of those with other organisms (p < 0.001). Moderate or severe regurgitation occurred more often in S bovis endocarditis than with other microorganisms (p = 0.05). When surgery or necropsy was performed, infectious myocardial infiltration of the left ventricle was confirmed histopathologically in 36% of the patients with S bovis endocarditis and in 10% of those with other organisms (p = 0.002).
Conclusions—S bovis endocarditis is a severe illness because of the more common involvement of multiple valves, and of the frequent occurrence of haemodynamically relevant valvar regurgitation and infectious myocardial infiltration.

 Keywords: infective endocarditis;  Streptococcus bovis;  transoesophageal echocardiography;  valvar disease PMID:9875088

  12. Improved Detection of Mycobacterium bovis Infection in Bovine Lymph Node Tissue Using Immunomagnetic Separation (IMS)-Based Methods

    PubMed Central

    Stewart, Linda D.; McNair, James; McCallan, Lyanne; Gordon, Alan; Grant, Irene R.

    2013-01-01

    Immunomagnetic separation (IMS) can selectively isolate and concentrate Mycobacterium bovis cells from lymph node tissue to facilitate subsequent detection by PCR (IMS-PCR) or culture (IMS-MGIT). This study describes application of these novel IMS-based methods to test for M. bovis in a survey of 280 bovine lymph nodes (206 visibly lesioned (VL), 74 non-visibly lesioned (NVL)) collected at slaughter as part of the Northern Ireland bovine TB eradication programme. Their performance was evaluated relative to culture. Overall, 174 (62.1%) lymph node samples tested positive by culture, 162 (57.8%) by IMS-PCR (targeting IS6110), and 191 (68.2%) by IMS-MGIT culture. Twelve (6.9%) of the 174 culture positive lymph node samples were not detected by either of the IMS-based methods. However, an additional 79 M. bovis positive lymph node samples (27 (13.1%) VL and 52 (70.3%) NVL) were detected by the IMS-based methods and not by culture. When low numbers of viable M. bovis are present in lymph nodes (e.g. in NVLs of skin test reactor cattle) decontamination prior to culture may adversely affect viability, leading to false negative culture results. In contrast, IMS specifically captures whole M. bovis cells (live, dead or potentially dormant) which are not subject to any deleterious treatment before detection by PCR or MGIT culture. During this study only 2.7% of NVL lymph nodes tested culture positive, whereas 70.3% of the same samples tested M. bovis positive by the IMS-based tests. Results clearly demonstrate that not only are the IMS-based methods more rapid but they have greater detection sensitivity than the culture approach currently used for the detection of M. bovis infection in cattle. Adoption of the IMS-based methods for lymph node testing would have the potential to improve M. bovis detection in clinical samples. PMID:23469275

  13. Evaluation of antioxidant status, oxidative stress and serum trace mineral levels associated with Babesia ovis parasitemia in sheep.

    PubMed

    Esmaeilnejad, Bijan; Tavassoli, Mousa; Asri-Rezaei, Siamak; Dalir-Naghadeh, Bahram; Malekinejad, Hassan; Jalilzadeh-Amin, Ghader; Arjmand, Jafar; Golabi, Mostafa; Hajipour, Naser

    2014-09-15

    Ovine babesiosis is a fatal disease characterized by severe progressive hemolytic anemia. In order to clarify the causal mechanisms implicated in anemia, this study was aimed to assess the antioxidant status and erythrocyte oxidative stress in sheep suffering from ovine babesiosis. Babesia infection was confirmed both with Giemsa's staining blood smears and semi-nested PCR amplified region of 18S rRNA gene. Thirty-eight Iranian sheep, naturally infected with Babesia spp., were considered as the infected group and divided into four subgroups according to parasitemia rates (<1%, 1-2%, 2-3% and >3%), and the same number non-infected animals were selected as the control group. Blood samples were taken and hematological parameters, activities of antioxidant enzymes including erythrocyte glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT), glucose-6-phosphate dehydrogenase (G6PD), total antioxidant capacity (TAC), median corpuscular fragility (MCF), and serum concentrations of some trace minerals (copper, iron, zinc, manganese, and selenium) were measured. In addition, as an index of lipid peroxidation, the level of malondialdehyde (MDA) was measured. The results revealed a significant decrease (P<0.01) in RBC count, packed cell volume (PCV) and Hb concentration as well as the activities of erythrocyte GSH-Px, SOD, CAT, G6PD, TAC, MCF and serum concentrations of Cu, Zn, Mn and Se in the infected sheep. In contrast, significantly increased (P<0.01) levels of MDA and erythrocyte osmotic fragility as well as serum concentration of iron were recorded in the infected animals. Overall, the observed remarkable decrease in the antioxidant enzyme activities, median corpuscular fragility and substantial elevated levels of lipid peroxidation associated with the notable increase in parasitemia indicate high exposure of RBCs to oxidative damage in Babesia infected sheep. These results indicate that the disturbed antioxidant defense mechanisms in ovine

  14. Oral Vaccination of White-Tailed Deer (Odocoileus virginianus) with Mycobacterium bovis Bacillus Calmette-Guerin (BCG)

    PubMed Central

    Palmer, Mitchell V.; Thacker, Tyler C.; Waters, W. Ray; Robbe-Austerman, Suelee

    2014-01-01

    Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis from livestock, particularly cattle. In Michigan, USA tuberculous white-tailed deer transmit M. bovis to other deer and cattle. One approach in dealing with this wildlife reservoir is to vaccinate deer, thus interfering with the intraspecies and interspecies transmission cycles. Thirty-three white-tailed deer were assigned to one of two groups; oral vaccination with 1×108 colony-forming units of M. bovis BCG Danish (n = 17); and non-vaccinated (n = 16). One hundred eleven days after vaccination deer were infected intratonsilarly with 300 colony-forming units of virulent M. bovis. At examination, 150 days after challenge, BCG vaccinated deer had fewer gross and microscopic lesions, fewer tissues from which M. bovis could be isolated, and fewer late stage granulomas with extensive liquefactive necrosis. Fewer lesions, especially those of a highly necrotic nature should decrease the potential for dissemination of M. bovis within the host and transmission to other susceptible hosts. PMID:24804678

  15. Oral vaccination of white-tailed deer (Odocoileus virginianus) with Mycobacterium bovis Bacillus Calmette-Guerin (BCG).

    PubMed

    Palmer, Mitchell V; Thacker, Tyler C; Waters, W Ray; Robbe-Austerman, Suelee

    2014-01-01

    Wildlife reservoirs of Mycobacterium bovis represent serious obstacles to the eradication of tuberculosis from livestock, particularly cattle. In Michigan, USA tuberculous white-tailed deer transmit M. bovis to other deer and cattle. One approach in dealing with this wildlife reservoir is to vaccinate deer, thus interfering with the intraspecies and interspecies transmission cycles. Thirty-three white-tailed deer were assigned to one of two groups; oral vaccination with 1 × 10(8) colony-forming units of M. bovis BCG Danish (n = 17); and non-vaccinated (n = 16). One hundred eleven days after vaccination deer were infected intratonsilarly with 300 colony-forming units of virulent M. bovis. At examination, 150 days after challenge, BCG vaccinated deer had fewer gross and microscopic lesions, fewer tissues from which M. bovis could be isolated, and fewer late stage granulomas with extensive liquefactive necrosis. Fewer lesions, especially those of a highly necrotic nature should decrease the potential for dissemination of M. bovis within the host and transmission to other susceptible hosts.

  16. Effects of Streptococcus bovis Isolated from Bovine Rumen on the Fermentation Characteristics and Nutritive Value of Tanzania Grass Silage

    PubMed Central

    Zanine, Anderson de Moura; Bonelli, Emerson Alencar; de Souza, Alexandre Lima; Ferreira, Daniele de Jesus; Santos, Edson Mauro; Ribeiro, Marinaldo Divino; Geron, Luiz Juliano Valério; Pinho, Ricardo Martins Araujo

    2016-01-01

    This study aimed to evaluate the effects of Streptococcus bovis on the fermentation characteristics and nutritive value of Tanzania grass silage. Tanzania grass was chopped and left untreated (U) or treated with Streptococcus bovis JB1 at 1 × 106 colony-forming units per gram (cfu/g) of fresh forage or Streptococcus bovis HC5 at 1 × 106 cfu/g of fresh forage and packed into sixtuplicate laboratory silos. The largest number of enterobacteria, molds and yeast (M&Y) occurred in untreated silages and the smallest populations of enterobacteria and M&Y and the largest numbers of lactic acid bacteria (LAB), at 9.81 and 9.87 log cfu/g, were observed in Streptococcus bovis JB1 and HC5, respectively (P < 0.05). Silages treated with JB1 and HC5 had lower (P < 0.05) silage pHs and concentrations of ammoniacal nitrogen (NH3-N) than untreated silages. The application of Streptococcus bovis JB1 and HC5 resulted in fewer losses through gases and effluents (P < 0.05), which resulted in greater dry matter recovery (DMR) and crude protein recovery (CPR) (P < 0.05). Streptococcus bovis JB1 and HC5 improved the fermentative profile and increased the concentration of crude protein and DMR and CPR in Tanzania grass silage. PMID:27073806

  17. Effects of Streptococcus bovis Isolated from Bovine Rumen on the Fermentation Characteristics and Nutritive Value of Tanzania Grass Silage.

    PubMed

    Zanine, Anderson de Moura; Bonelli, Emerson Alencar; de Souza, Alexandre Lima; Ferreira, Daniele de Jesus; Santos, Edson Mauro; Ribeiro, Marinaldo Divino; Geron, Luiz Juliano Valério; Pinho, Ricardo Martins Araujo

    2016-01-01

    This study aimed to evaluate the effects of Streptococcus bovis on the fermentation characteristics and nutritive value of Tanzania grass silage. Tanzania grass was chopped and left untreated (U) or treated with Streptococcus bovis JB1 at 1 × 10(6) colony-forming units per gram (cfu/g) of fresh forage or Streptococcus bovis HC5 at 1 × 10(6) cfu/g of fresh forage and packed into sixtuplicate laboratory silos. The largest number of enterobacteria, molds and yeast (M&Y) occurred in untreated silages and the smallest populations of enterobacteria and M&Y and the largest numbers of lactic acid bacteria (LAB), at 9.81 and 9.87 log cfu/g, were observed in Streptococcus bovis JB1 and HC5, respectively (P < 0.05). Silages treated with JB1 and HC5 had lower (P < 0.05) silage pHs and concentrations of ammoniacal nitrogen (NH3-N) than untreated silages. The application of Streptococcus bovis JB1 and HC5 resulted in fewer losses through gases and effluents (P < 0.05), which resulted in greater dry matter recovery (DMR) and crude protein recovery (CPR) (P < 0.05). Streptococcus bovis JB1 and HC5 improved the fermentative profile and increased the concentration of crude protein and DMR and CPR in Tanzania grass silage.

  18. Experimental Aerosol Inoculation and Investigation of Potential Lateral Transmission of Mycobacterium bovis in Virginia Opossum (Didelphis virginiana).

    PubMed

    Fenton, Karla A; Fitzgerald, Scott D; Bolin, Steve; Kaneene, John; Sikarskie, James; Greenwald, Rena; Lyashchenko, Konstantin

    2012-01-01

    An endemic focus of Mycobacterium bovis (M. bovis) infection in the state of Michigan has contributed to a regional persistence in the animal population. The objective of this study was to determine if Virginia opossums (Didelphis virginiana) contribute to disease persistence by experimentally assessing intraspecies lateral transmission. One wild caught pregnant female opossum bearing 11 joeys (young opossum) and one age-matched joey were obtained for the study. Four joeys were aerosol inoculated with M. bovis (inoculated), four joeys were noninoculated (exposed), and four joeys plus the dam were controls. Four replicate groups of one inoculated and one exposed joey were housed together for 45 days commencing 7 days after experimental inoculation. At day 84 opossums were sacrificed. All four inoculated opossums had a positive test band via rapid test, culture positive, and gross/histologic lesions consistent with caseogranulomatous pneumonia. The exposed and control groups were unremarkable on gross, histology, rapid test, and culture. In conclusion, M. bovis infection within the inoculated opossums was confirmed by gross pathology, histopathology, bacterial culture, and antibody tests. However, M. bovis was not detected in the control and exposed opossums. There was no appreciable lateral transmission of M. bovis after aerosol inoculation and 45 days of cohabitation between infected and uninfected opossums.

  19. Generation of transgenic cattle expressing human β-defensin 3 as an approach to reducing susceptibility to Mycobacterium bovis infection.

    PubMed

    Su, Feng; Wang, Yongsheng; Liu, Guanghui; Ru, Kun; Liu, Xin; Yu, Yuan; Liu, Jun; Wu, Yongyan; Quan, Fusheng; Guo, Zekun; Zhang, Yong

    2016-03-01

    Bovine tuberculosis results from infection with Mycobacterium bovis, a member of the Mycobacterium tuberculosis family. Worldwide, M. bovis infections result in economic losses in the livestock industry; cattle production is especially hard-hit by this disease. Generating M. bovis-resistant cattle may potentially mitigate the impact of this disease by reducing M. bovis infections. In this study, we used transgenic somatic cell nuclear transfer to generate cattle expressing the gene encoding human β-defensin 3 (HBD3), which confers resistance to mycobacteria in vitro. We first generated alveolar epithelial cells expressing HBD3 under the control of the bovine MUC1 promoter, and confirmed that these cells secreted HBD3 and possessed anti-mycobacterial capacity. We then generated and identified transgenic cattle by somatic cell nuclear transfer. The cleavage and blastocyst formation rates of genetically modified embryos provided evidence that monoclonal transgenic bovine fetal fibroblast cells have an integral reprogramming ability that is similar to that of normal cells. Five genetically modified cows were generated, and their anti-mycobacterial capacities were evaluated. Alveolar epithelial cells and macrophages from these cattle expressed higher levels of HBD3 protein compared with non-transgenic cells and possessed effective anti-mycobacterial capacity. These results suggest that the overall risk of M. bovis infection in transgenic cattle is efficiently reduced, and support the development of genetically modified animals as an effective tool to reduce M. bovis infection.

  20. Effect of synthesized inhibitors on babesipain-1, a new cysteine protease from the bovine piroplasm Babesia bigemina.

    PubMed

    Martins, T M; Gonçalves, L M D; Capela, R; Moreira, R; do Rosário, V E; Domingos, A

    2010-04-01

    Papain-like cysteine proteases (CP) have been shown to have essential roles in parasitic protozoa and are under study as promising drug targets. One gene was identified by sequence similarity search to be homologous to the CP family in the ongoing Babesia bigemina genome sequencing project database. The newly identified CP gene, called babesipain-1, was cloned and expressed as a fusion protein, and the effect of different inhibitors on proteolytic activity was tested. A series of new artemisinin-vinyl sulfone hybrid molecules were tested as inhibitors being effective on the range of 0.3-30 microm, depending on the core-containing molecule. PMID:20537109

  1. Expression of OmpATb is dependent on small membrane proteins in Mycobacterium bovis BCG.

    PubMed

    Veyron-Churlet, Romain; Brust, Belinda; Kremer, Laurent; Blanc-Potard, Anne-Béatrice

    2011-11-01

    Small membrane proteins emerge as a novel class of regulatory molecules in bacteria. Experiments carried out in Mycobacterium bovis BCG indicate that the ompATb gene (Rv0899), encoding a major outer membrane protein, is organized in operon with Rv0900 and Rv0901, encoding two small proteins with a predicted transmembrane domain. Fractioning experiment confirmed the association of Rv0901 with the membrane fraction. To investigate the role of Rv0900 and Rv0901 in M. bovis BCG, we have constructed a strain deleted for the whole operon as well as complemented strains carrying a deletion of Rv0900 or a frameshift mutation in either Rv0900 or Rv0901. Importantly, mutations in Rv0900 and/or Rv0901 strongly altered OmpATb expression, demonstrating that Rv0900 and Rv0901 play a regulatory role, which appears to occur at a post-transcriptional level. PMID:21802366

  2. Natural genetic transformation in the rumen bacterium Streptococcus bovis JB1.

    PubMed

    Mercer, D K; Melville, C M; Scott, K P; Flint, H J

    1999-10-15

    Natural transformation of Streptococcus bovis JB1 was demonstrated after development of competence in normal culture medium. Transformation efficiencies were not significantly increased when heat-inactivated horse serum was added to the medium before growth. This is the first time that a resident rumen bacterial species has been shown to be naturally transformable. Transformation allowed the acquisition of plasmids or integration of sequences into the chromosome. No transformation was observed in the presence of undiluted autoclaved or filter-sterilised ovine rumen fluid or filter-sterilised ovine saliva, suggesting that transformation in the ruminant digestive tract is a rare event, although transformation was observed in the presence of 1% and 0.5% filter-sterilised rumen fluid. The use of natural transformation of S. bovis should facilitate further molecular biological studies on this species.

  3. [Cysticercus bovis in Turkey and its importance from the public health aspect].

    PubMed

    Kuş, Fatma Selcan; Sevimli, Feride Kırcalı; Miman, Özlem

    2014-01-01

    This study was conducted in order to compare the different regions according to the literature on the prevalence of bovine cysticercosis and T. saginata in Turkey. Bovine cysticercosis and T. saginata status were evaluated retrospectively. The distribution of the data obtained according to provinces and regions were showed in the Table and the minumum / maximum values of this data in different regions in the Figure. The data obtained through the literature showed that the prevalence of C. bovis and T. saginata infections are parallel in the same region. The higher prevalence of both C. bovis and T. saginata infections was determined in the Southeastern Anatolia, Eastern Anatolia and Central Anatolia regions respectively. PMID:24659701

  4. Streptococcus bovis bacteraemia requires rigorous exclusion of colonic neoplasia and endocarditis.

    PubMed

    Beeching, N J; Christmas, T I; Ellis-Pegler, R B; Nicholson, G I

    1985-08-01

    Twelve patients presented to the hospitals of the Auckland Hospital Board with bacteraemia caused by Streptococcus bovis in the years 1979-84. Ten had endocarditis, affecting homograft valves in two cases and the tricuspid valve in one case. Of nine patients who underwent investigation of the large bowel, only one did not have a colorectal tumour. Three had colonic adenocarcinoma and three had colorectal villous adenoma. Two, including a patient with acute hepatic failure from alcoholic cirrhosis, had colonic adenomata. Colonoscopy provided a tissue diagnosis of colorectal neoplasia despite negative radiological studies in three patients. Bacteraemia due to S. bovis should prompt rigorous investigation to exclude both endocarditis and tumours of the large bowel.

  5. SERUM ANTIBODIES TO BORRELIA BURGDORFERI, ANAPLASMA PHAGOCYTOPHILUM, AND BABESIA MICROTI IN RECAPTURED WHITE-FOOTED MICE

    PubMed Central

    Magnarelli, Louis A.; Williams, Scott C.; Norris, Steven J.; Fikrig, Erol

    2013-01-01

    A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ≥2 yr. Sera from 561 mice were analyzed for total antibodies to B. burgdorferi and A. phagocytophilum by using whole-cell or recombinant (VlsE or protein 44) antigens in a solid-phase enzyme-linked immunosorbent assay or to whole-cell B. microti by indirect fluorescent antibody staining. Antibody prevalences were highly variable for B. burgdorferi (from 56% to 98%), A. phagocytophilum (from 11% to 85%), and B. microti (from 11% to 84%) depending on the site and time of sampling. Of 463 mice with antibodies, 206 (45%) had antibodies to all three pathogens. Changes in antibody status for some mice from negative to positive (117 seroconversions) or from positive to negative (55 reversions) were observed. Seroconversions were observed in 10.1% of 417 mice for B. burgdorferi, 18.0% of 306 mice for A. phagocytophilum, and 6.6% of 304 mice for B. microti; reversion rates were 5.3, 5.9, and 4.9%, respectively. Antibodies to all pathogens persisted in some mice over several weeks while, in others, there were marked declines in titration end points to negative status. The latter may indicate elimination of a certain pathogen, such as A. phagocytophilum, or that mouse immune systems ceased to produce antibodies despite an existing patent infection. PMID:23568904

  6. Borrelia burgdorferi promotes the establishment of Babesia microti in the northeastern United States.

    PubMed

    Dunn, Jessica M; Krause, Peter J; Davis, Stephen; Vannier, Edouard G; Fitzpatrick, Meagan C; Rollend, Lindsay; Belperron, Alexia A; States, Sarah L; Stacey, Andrew; Bockenstedt, Linda K; Fish, Durland; Diuk-Wasser, Maria A

    2014-01-01

    Babesia microti and Borrelia burgdorferi, the respective causative agents of human babesiosis and Lyme disease, are maintained in their enzootic cycles by the blacklegged tick (Ixodes scapularis) and use the white-footed mouse (Peromyscus leucopus) as primary reservoir host. The geographic range of both pathogens has expanded in the United States, but the spread of babesiosis has lagged behind that of Lyme disease. Several studies have estimated the basic reproduction number (R0) for B. microti to be below the threshold for persistence (<1), a finding that is inconsistent with the persistence and geographic expansion of this pathogen. We tested the hypothesis that host coinfection with B. burgdorferi increases the likelihood of B. microti transmission and establishment in new areas. We fed I. scapularis larva on P. leucopus mice that had been infected in the laboratory with B. microti and/or B. burgdorferi. We observed that coinfection in mice increases the frequency of B. microti infected ticks. To identify the ecological variables that would increase the probability of B. microti establishment in the field, we integrated our laboratory data with field data on tick burden and feeding activity in an R0 model. Our model predicts that high prevalence of B. burgdorferi infected mice lowers the ecological threshold for B. microti establishment, especially at sites where larval burden on P. leucopus is lower and where larvae feed simultaneously or soon after nymphs infect mice, when most of the transmission enhancement due to coinfection occurs. Our studies suggest that B. burgdorferi contributes to the emergence and expansion of B. microti and provides a model to predict the ecological factors that are sufficient for emergence of B. microti in the wild. PMID:25545393

  7. Serum antibodies to Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in recaptured white-footed mice.

    PubMed

    Magnarelli, Louis A; Williams, Scott C; Norris, Steven J; Fikrig, Erol

    2013-04-01

    A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ≥2 yr. Sera from 561 mice were analyzed for total antibodies to B. burgdorferi and A. phagocytophilum by using whole-cell or recombinant (VlsE or protein 44) antigens in a solid-phase enzyme-linked immunosorbent assay or to whole-cell B. microti by indirect fluorescent antibody staining. Antibody prevalences were highly variable for B. burgdorferi (from 56% to 98%), A. phagocytophilum (from 11% to 85%), and B. microti (from 11% to 84%) depending on the site and time of sampling. Of 463 mice with antibodies, 206 (45%) had antibodies to all three pathogens. Changes in antibody status for some mice from negative to positive (117 seroconversions) or from positive to negative (55 reversions) were observed. Seroconversions were observed in 10.1% of 417 mice for B. burgdorferi, 18.0% of 306 mice for A. phagocytophilum, and 6.6% of 304 mice for B. microti; reversion rates were 5.3, 5.9, and 4.9%, respectively. Antibodies to all pathogens persisted in some mice over several weeks while, in others, there were marked declines in titration end points to negative status. The latter may indicate elimination of a certain pathogen, such as A. phagocytophilum, or that mouse immune systems ceased to produce antibodies despite an existing patent infection. PMID:23568904

  8. Seroprevalence of Babesia caballi and Theileria equi in five draught equine populated metropolises of Punjab, Pakistan.

    PubMed

    Hussain, Muhammad Hammad; Saqib, Muhammad; Raza, Fahad; Muhammad, Ghulam; Asi, Muhammad Nadeem; Mansoor, Muhammad Khalid; Saleem, Muhammad; Jabbar, Abdul

    2014-05-28

    Equine piroplasmosis (EP) caused by intraerythrocytic parasites (Theileria equi and Babesia caballi) is an emerging equine disease of world-wide distribution. In Pakistan, the prevalence and incidence of EP are unknown. In order to obtain the first insights into the prevalence of the disease, a total of 430 equids, including 33 mules, 65 horses and 332 donkeys, aging from ≤ 5 to ≥ 10 years of either sex, from five metropolises of Punjab, Pakistan, were serologically tested for the presence of antibodies directed against B. caballi and T. equi, using a competitive enzyme-linked immunosorbent assay (cELISA). Out of 430 equid serum samples tested, 226 (52.6%, 95% CI 47.7-57.4) were found cELISA positive for EP (T. equi and/or B. caballi infections). The overall seroprevalence of EP was 41.2% (95% CI 36.5-46.0) for T. equi and 21.6% (95% CI 17.8-25.8) for B. caballi. A small proportion of equids (10.2%, 95% CI 7.5-13.5) was seropositive for both T. equi and B. caballi. Seroprevalence of T. equi was significantly higher (P<0.01) in equines from the metropolis of Lahore (66.7%, 95% CI 54.3-77.6) and in horses (56.9%, 95% CI 44.0-69.2). Multivariable logistic regression model analysis indicated that factors associated with prevalence of EP were being an equine species kept in metropolis Lahore (OR=4.24, 95% CI 2.28-7.90), horse (OR=2.82, 95% CI 1.53-5.20) and male equids (OR=1.81, 95% CI 1.15-2.86).

  9. Cloning, Characterization, and Expression of a 200-Kilodalton Diagnostic Antigen of Babesia bigemina†

    PubMed Central

    Tebele, N.; Skilton, R. A.; Katende, J.; Wells, C. W.; Nene, V.; McElwain, T.; Morzaria, S. P.; Musoke, A. J.

    2000-01-01

    Current serological tests for Babesia bigemina use semipurified merozoite antigens derived from infected erythrocytes. One of the major drawbacks of these tests is that antigen quality can vary from batch to batch. Since the quality of the antigen contributes to the sensitivity and specificity of serological tests, the use of standardized recombinant antigens should ensure consistency in assay quality. Previously, a 200-kDa merozoite antigen (p200) was identified as a candidate diagnostic antigen for use in a serological assay for the detection of B. bigemina antibodies in infected cattle. In this study, we have cloned, characterized, and expressed p200. A 3.5-kbp cDNA clone encoding p200 was isolated and shown to be almost full length, lacking approximately 300 bp at the 5′ end. The predicted amino acid sequence shows that p200 consists of a long, highly charged central repeat region of an uninterrupted α helix, indicative of a fibrous protein. Immunoelectron microscopy localized p200 to the merozoite cytoplasm, suggesting that the antigen may be a structural protein involved in forming filament structures within the cytoskeleton. The 3.5-kbp cDNA was expressed in bacteria as a fusion protein with glutathione S-transferase (GST), but the yield was poor. To improve the yield, cDNA fragments encoding antigenic domains of p200 were expressed as fusions with GST. One of these fusion proteins, C1A-GST, is composed of a 7-kDa fragment of the p200 repeat region and contains epitopes that react strongly with sera from cattle experimentally infected with B. bigemina. Recombinant C1A-GST should permit the development of an improved enzyme-linked immunosorbent assay for the detection of antibodies against B. bigemina. PMID:10834983

  10. Borrelia burgdorferi promotes the establishment of Babesia microti in the northeastern United States.

    PubMed

    Dunn, Jessica M; Krause, Peter J; Davis, Stephen; Vannier, Edouard G; Fitzpatrick, Meagan C; Rollend, Lindsay; Belperron, Alexia A; States, Sarah L; Stacey, Andrew; Bockenstedt, Linda K; Fish, Durland; Diuk-Wasser, Maria A

    2014-01-01

    Babesia microti and Borrelia burgdorferi, the respective causative agents of human babesiosis and Lyme disease, are maintained in their enzootic cycles by the blacklegged tick (Ixodes scapularis) and use the white-footed mouse (Peromyscus leucopus) as primary reservoir host. The geographic range of both pathogens has expanded in the United States, but the spread of babesiosis has lagged behind that of Lyme disease. Several studies have estimated the basic reproduction number (R0) for B. microti to be below the threshold for persistence (<1), a finding that is inconsistent with the persistence and geographic expansion of this pathogen. We tested the hypothesis that host coinfection with B. burgdorferi increases the likelihood of B. microti transmission and establishment in new areas. We fed I. scapularis larva on P. leucopus mice that had been infected in the laboratory with B. microti and/or B. burgdorferi. We observed that coinfection in mice increases the frequency of B. microti infected ticks. To identify the ecological variables that would increase the probability of B. microti establishment in the field, we integrated our laboratory data with field data on tick burden and feeding activity in an R0 model. Our model predicts that high prevalence of B. burgdorferi infected mice lowers the ecological threshold for B. microti establishment, especially at sites where larval burden on P. leucopus is lower and where larvae feed simultaneously or soon after nymphs infect mice, when most of the transmission enhancement due to coinfection occurs. Our studies suggest that B. burgdorferi contributes to the emergence and expansion of B. microti and provides a model to predict the ecological factors that are sufficient for emergence of B. microti in the wild.

  11. Identification of serum biomarkers in dogs naturally infected with Babesia canis canis using a proteomic approach

    PubMed Central

    2014-01-01

    Background Canine babesiosis is a tick-borne disease that is caused by the haemoprotozoan parasites of the genus Babesia. There are limited data on serum proteomics in dogs, and none of the effect of babesiosis on the serum proteome. The aim of this study was to identify the potential serum biomarkers of babesiosis using proteomic techniques in order to increase our understanding about disease pathogenesis. Results Serum samples were collected from 25 dogs of various breeds and sex with naturally occurring babesiosis caused by B. canis canis. Blood was collected on the day of admission (day 0), and subsequently on the 1st and 6th day of treatment. Two-dimensional electrophoresis (2DE) of pooled serum samples of dogs with naturally occurring babesiosis (day 0, day 1 and day 6) and healthy dogs were run in triplicate. 2DE image analysis showed 64 differentially expressed spots with p ≤ 0.05 and 49 spots with fold change ≥2. Six selected spots were excised manually and subjected to trypsin digest prior to identification by electrospray ionisation mass spectrometry on an Amazon ion trap tandem mass spectrometry (MS/MS). Mass spectrometry data was processed using Data Analysis software and the automated Matrix Science Mascot Daemon server. Protein identifications were assigned using the Mascot search engine to interrogate protein sequences in the NCBI Genbank database. A number of differentially expressed serum proteins involved in inflammation mediated acute phase response, complement and coagulation cascades, apolipoproteins and vitamin D metabolism pathway were identified in dogs with babesiosis. Conclusions Our findings confirmed two dominant pathogenic mechanisms of babesiosis, haemolysis and acute phase response. These results may provide possible serum biomarker candidates for clinical monitoring of babesiosis and this study could serve as the basis for further proteomic investigations in canine babesiosis. PMID:24885808

  12. Identification of a thioredoxin reductase from Babesia microti during mammalian infection.

    PubMed

    Zhao, Shaoruo; Gong, Haiyan; Zhou, Yongzhi; Zhang, Houshuang; Cao, Jie; Zhou, Jinlin

    2016-08-01

    Babesia microti is the primary causative agent of human babesiosis worldwide and associated with increased human health risks and the safety of blood supply. The parasite replicates in the host's red blood cells, thus, in order to counteract the oxidative stress and toxic effects, parasites employ a thioredoxin (Trx) system to maintain a redox balance. Since thioredoxin reductase (TrxR) plays a critical role in the system, in this study, we report the cloning, expression, and functional characterization of a novel TrxR from B. microti (BmiTrxR). The complete gene BmiTrxR was obtained by amplifying the 5' and 3' regions of messenger RNA (mRNA) by RACE. The full-length complementary DNA (cDNA) of BmiTrxR was 1766 bp and contained an intact open reading frame with 1662 bp that encoded a polypeptide with 553 amino acids. Molecular weight of the predicted protein was 58.4 kDa with an isoelectric point of 6.95, similar to high molecular weight TrxR. The recombinant protein of BmiTrxR was expressed in a His-fused soluble form in Escherichia coli. The native protein BmiTrxR was identified with the mouse anti-BmiTrxR polyclonal serum by western blotting and IFAT. Moreover, the enzyme showed a disulfide reductase activity using DTNB as substrate and catalyzed the NADPH-dependent reduction of Trx. Auranofin, a known inhibitor of TrxR, completely abrogated the activity of the recombinant enzyme in vitro. These results not only contribute to the understanding of redox pathway in this parasite but also suggest that BmiTrxR could be a potential target for the development of novel strategies to control B. microti thus reducing the incidence of babesiosis. PMID:27164832

  13. Molecular characterization of the Babesia caballi rap-1 gene and epidemiological survey in horses in Israel.

    PubMed

    Rapoport, Adi; Aharonson-Raz, Karin; Berlin, Dalia; Tal, Saar; Gottlieb, Yuval; Klement, Eyal; Steinman, Amir

    2014-04-01

    Equine piroplasmosis imposes great concerns for the equine industry regarding international horse movement, and therefore requires reliable diagnostic tools. Recent studies from South Africa and Jordan, including a preliminary study in Israel, reported extremely low seroprevalence to Babesia caballi (B. caballi) (0-1%) using the acceptable rhoptry-associated protein-1 (RAP-1) cELISA. In accordance with the study from South Africa demonstrating a significant heterogeneity in the rap-1 gene sequence of South African B. caballi isolates, the objectives of this study were to phylogenetically characterize the rap-1 gene of the Israeli isolates and determine the prevalence of B. caballi in horses in Israel. Out of 273 horses tested using the RAP-1 cELISA, only one was sero-positive, while 9.3% were positive on PCR performed on the rap-1 gene. Phylogenetic analysis of the rap-1 gene grouped the Israeli isolates in a cluster together with the South African strains (99% nt identity), but in a separate cluster from the American/Caribbean strains (81-82% nt identity). These findings support the existence of heterogeneity in the RAP-1 amino-acid sequences of the Israeli and South African isolates as compared to that used in the cELISA commercial kit and raise doubts as to the ability of this assay to serve as a sole regulatory test for international horse movement. Risk factor analysis found management and age to significantly associate with prevalence of B. caballi, as higher prevalence was noted in horses held out on pasture and a negative association was recorded with age. In addition, B. caballi was not detected in horses in the steppe-arid and extreme-arid climatic regions as compared to the wetter regions. Findings of this study emphasize the need to combine several detection methods to ameliorate the control and spread of the disease.

  14. Borrelia burgdorferi Promotes the Establishment of Babesia microti in the Northeastern United States

    PubMed Central

    Dunn, Jessica M.; Krause, Peter J.; Davis, Stephen; Vannier, Edouard G.; Fitzpatrick, Meagan C.; Rollend, Lindsay; Belperron, Alexia A.; States, Sarah L.; Stacey, Andrew; Bockenstedt, Linda K.; Fish, Durland; Diuk-Wasser, Maria A.

    2014-01-01

    Babesia microti and Borrelia burgdorferi, the respective causative agents of human babesiosis and Lyme disease, are maintained in their enzootic cycles by the blacklegged tick (Ixodes scapularis) and use the white-footed mouse (Peromyscus leucopus) as primary reservoir host. The geographic range of both pathogens has expanded in the United States, but the spread of babesiosis has lagged behind that of Lyme disease. Several studies have estimated the basic reproduction number (R0) for B. microti to be below the threshold for persistence (<1), a finding that is inconsistent with the persistence and geographic expansion of this pathogen. We tested the hypothesis that host coinfection with B. burgdorferi increases the likelihood of B. microti transmission and establishment in new areas. We fed I. scapularis larva on P. leucopus mice that had been infected in the laboratory with B. microti and/or B. burgdorferi. We observed that coinfection in mice increases the frequency of B. microti infected ticks. To identify the ecological variables that would increase the probability of B. microti establishment in the field, we integrated our laboratory data with field data on tick burden and feeding activity in an R0 model. Our model predicts that high prevalence of B. burgdorferi infected mice lowers the ecological threshold for B. microti establishment, especially at sites where larval burden on P. leucopus is lower and where larvae feed simultaneously or soon after nymphs infect mice, when most of the transmission enhancement due to coinfection occurs. Our studies suggest that B. burgdorferi contributes to the emergence and expansion of B. microti and provides a model to predict the ecological factors that are sufficient for emergence of B. microti in the wild. PMID:25545393

  15. Characterisation of a Babesia orientalis apical membrane antigen, and comparison of its orthologues among selected apicomplexans.

    PubMed

    He, Lan; Fan, Lizhe; Hu, Jinfang; Miao, Xiaoyan; Huang, Yuan; Zhou, Yanqin; Hu, Min; Zhao, Junlong

    2015-04-01

    In the present study, we identified and characterised the complete coding sequence of Babesia orientalis apical membrane antigen 1 (designated Bo-ama1); it is 1803bp in length and encodes a polypeptide of 601 amino acids (aa). The Bo-ama-1 gene product (Bo-AMA1) is predicted to be 67kDa in size and contains a signal peptide. Mature Bo-AMA1 is predicted to have one transmembrane region and a short cytoplasmic tail (C-terminal domain). The extracellular part of Bo-AMA1 has three functional domains (DI, DII and DIII) with 14 conserved cysteine residues. A Bo-AMA1 fragment containing all three of these domains (designated Bo-AMA1-DI/II/III) was cloned into the plasmid vector pET-28a and expressed as a recombinant (His-fusion) protein of 53kDa. Antibodies in the serum from a B. orientalis-infected water buffalo specifically recognised this protein in immunoblotting analysis. Rabbit antibodies raised against the recombinant protein were able to detect native Bo-AMA1 (67kDa) from erythrocytes of B. orientalis-infected water buffalo. Bo-AMA1 is a new member of the AMA1 family and might be a good antigen for the specific detection of antibodies produced in B. orientalis infected cattle. This protein is likely to play critical roles during host cell adherence and invasion by B. orientalis, as the AMA1s reported in other organisms such as Plasmodium falciparum and Toxoplasma gondii. Further research is required to explore the biological functions of this protein and to determine whether its immunisation can induce protective effects in water buffalo against B. orientalis infection.

  16. Babesia bigemina: identification of B cell epitopes associated with parasitized erythrocytes.

    PubMed

    Vidotto, O; McElwain, T F; Machado, R Z; Perryman, L E; Suarez, C E; Palmer, G H

    1995-12-01

    Rhoptries are involved in host cell invasion and rhoptry polypeptides, including the Babesia bigemina rhoptry-associated protein-1 (RAP-1), are targets for protective immune responses. Polyclonal antisera produced against isolated rhoptries is directed predominantly against RAP-1 and reacts with both the merozoite and the membrane of parasitized erythrocytes. To determine whether these B cell epitopes associated with the parasitized erythrocyte are derived from RAP-1 or, alternatively, from previously undetected merozoite polypeptides, monoclonal antibodies (mAbs) were generated from mice immunized with rhoptries isolated from the JG-29 clone of the Mexico strain. The anti-RAP-1 mAbs bound only merozoites in a punctate immunofluorescence pattern. A second group of four mAbs, none of which were reactive with RAP-1, bound the parasitized erythrocyte. Two of these latter mAbs, 64/44.17.3 and 64/05.7.2, reacted only with parasitized erythrocytes that had been permeabilized. MAb 64/44.17.3 bound a 54-kDa merozoite polypeptide while 64/05.7.2 bound a > or = 225-kDa merozoite polypeptide. MAbs 64/32.8.5 and 64/38.5.3 recognized epitopes on 17.5- and 76-kDa polypeptides exposed on the external surface of intact parasitized erythrocytes. The results indicate that the identified RAP-1 epitopes are not associated with the erythrocyte cytoskeleton or membrane and that anti-RAP-1 immunity is most likely generated against the free merozoite. All new mAbs reacted with every B. bigemina strain tested (Mexico, Puerto Rico, St. Croix, Texcoco, Jaboticabal). The conservation of RAP-1 epitopes among these strains supports the continued testing of RAP-1 as a vaccine component. In addition, the identification of epitopes expressed on the surface of erythrocytes infected with all five strains provides new candidate immunogens.

  17. Seroprevalence and risk factors associated with Babesia caballi and Theileria equi infection in equids.

    PubMed

    García-Bocanegra, Ignacio; Arenas-Montes, Antonio; Hernández, Esther; Adaszek, Lukasz; Carbonero, Alfonso; Almería, Sonia; Jaén-Téllez, Juan Antonio; Gutiérrez-Palomino, Pedro; Arenas, Antonio

    2013-02-01

    A cross-sectional study was carried out on equids (horses, mules and donkeys) in Andalusia, Southern Spain, to assess the level of exposure to equine piroplasmosis and to investigate risk factors associated with these infections. At least one animal seropositive for Theileria equi and/or Babesia caballi was detected in 222/380 (58.4%) herds sampled by competitive inhibition ELISAs. The seroprevalences for B. caballi and T. equi were 13.2% and 56.1%, respectively; there was serological evidence of co-circulation of both piroplasms in 10.8% of herds. Antibodies against equine piroplasms were detected in 286/537 (53.3%) animals; 61 (11.4%) were seropositive for B. caballi, 270 (50.3%) were seropositive for T. equi and 24 (8.4%) were seropositive for both T. equi and B. caballi. There was a significantly higher seroprevalence of B. caballi in mules (32.1%) compared with donkeys (17.0%) and horses (7.9%), and a significantly higher seroprevalence of T. equi in mules (66.1%) in comparison with horses (48.6%), but not donkeys (47.2%). There were significant differences in prevalence of both piroplasms among locations; the seroprevalence of B. caballi ranged from 0 to 22.5%, while the seropositivity to T. equi ranged from 26.7 to 63.3%. A multiple logistic regression model indicated that the risk factors associated with a higher T. equi seroprevalence were increased age, presence of ticks and vaccination against other diseases. Risk factors associated with a higher seroprevalence of B. caballi were species (mules compared to horses), entry of horses in the last 6months, presence of ticks and presence of shelter. The findings indicate widespread exposure to equine piroplasmosis in Southern Spain.

  18. Mycobacterium bovis infection in the lion (Panthera leo): Current knowledge, conundrums and research challenges.

    PubMed

    Viljoen, Ignatius M; van Helden, Paul D; Millar, Robert P

    2015-06-12

    Mycobacterium bovis has global public-health and socio-economic significance and can infect a wide range of species including the lion (Panthera leo) resulting in tuberculosis. Lions are classified as vulnerable under the IUCN Red List of Threatened Species and have experienced a 30% population decline in the past two decades. However, no attempt has been made to collate and critically evaluate the available knowledge of M. bovis infections in lions and potential effects on population. In this review we set out to redress this. Arguments suggesting that ingestion of infected prey animals are the main route of infection for lions have not been scientifically proven and research is needed into other possible sources and routes of infection. The paucity of knowledge on host susceptibility, transmission directions and therefore host status, manifestation of pathology, and epidemiology of the disease in lions also needs to be addressed. Advances have been made in diagnosing the presence of M. bovis in lions. However, these diagnostic tests are unable to differentiate between exposure, presence of infection, or stage of disease. Furthermore, there are contradictory reports on the effects of M. bovis on lion populations with more data needed on disease dynamics versus the lion population's reproductive dynamics. Knowledge on disease effects on the lion reproduction and how additional stressors such as drought or co-morbidities may interact with tuberculosis is also lacking. Filling these knowledge gaps will contribute to the understanding of mycobacterial infections and disease in captive and wild lions and assist in lion conservation endeavours. PMID:25891424

  19. Transcriptional Response of Peripheral Blood Mononuclear Cells from Cattle Infected with Mycobacterium bovis

    PubMed Central

    Blanco, Federico Carlos; Soria, Marcelo; Bianco, María Verónica; Bigi, Fabiana

    2012-01-01

    Mycobacterium bovis is the causative agent of most cases of bovine tuberculosis. The identification of bTB biomarkers in specific stages of the disease will contribute to a better understanding of the immunopathology associated with tuberculosis and will enable their use in disease diagnosis and prognosis. The aim of this study was to evaluate the gene expression profile induced after specific stimulation of bovine peripheral blood mononuclear cells from cattle infected with M. bovis using the Affymetrix® GeneChip® Bovine Genome Array. A total of 172 genes showed differential expression profile that was statistically significant with log2-fold change >2.5 and <−2.5. Twenty-four out of these genes were upregulated and 148 were downregulated in bovine peripheral blood mononuclear cells of M. bovis-infected cattle. The highest differentially-expressed genes were related to immune and inflammatory responses, apoptosis, endocytosis, cellular trafficking and genes encoding proteins involved in cellular matrix degradation. Microarray results were confirmed in another group of infected cattle by RT-qPCR for the CD14, IL-1R, THBS1, MMP9 and FYVE genes. This study confirms previous findings that have shown that M. bovis infection in cattle results in the downregulation of immune response-related genes. Moreover, it validates the use of microarray platforms in combination with RT-qPCR to identify biomarkers of bovine tuberculosis. In addition, we propose CD14, IL-1R, THBS1, MMP9 and FYVE as potential biomarkers of bovine tuberculosis. PMID:22815916

  20. Properties of two sugar phosphate phosphatases from Streptococcus bovis and their potential involvement in inducer expulsion.

    PubMed Central

    Cook, G M; Ye, J J; Russell, J B; Saier, M H

    1995-01-01

    Streptococcus bovis possesses two sugar phosphate phosphatases (Pases). Pase I is a soluble enzyme that is inhibited by the membrane fractions from lactose-grown cells and is insensitive to activation by S46D HPr, an analog of HPr(ser-P) of the sugar phosphotransferase system. Pase II is a membrane-associated enzyme that can be activated 10-fold by S46D HPr, and it appears to play a role in inducer expulsion. PMID:7592500

  1. RNA-seq Transcriptional Profiling of Peripheral Blood Leukocytes from Cattle Infected with Mycobacterium bovis

    PubMed Central

    McLoughlin, Kirsten E.; Nalpas, Nicolas C.; Rue-Albrecht, Kévin; Browne, John A.; Magee, David A.; Killick, Kate E.; Park, Stephen D. E.; Hokamp, Karsten; Meade, Kieran G.; O’Farrelly, Cliona; Gormley, Eamonn; Gordon, Stephen V.; MacHugh, David E.

    2014-01-01

    Bovine tuberculosis, caused by infection with Mycobacterium bovis, is a major endemic disease affecting cattle populations worldwide, despite the implementation of stringent surveillance and control programs in many countries. The development of high-throughput functional genomics technologies, including gene expression microarrays and RNA-sequencing (RNA-seq), has enabled detailed analysis of the host transcriptome to M. bovis infection, particularly at the macrophage and peripheral blood level. In the present study, we have analyzed the peripheral blood leukocyte (PBL) transcriptome of eight natural M. bovis-infected and eight age- and sex-matched non-infected control Holstein-Friesian animals using RNA-seq. In addition, we compared gene expression profiles generated using RNA-seq with those previously generated using the high-density Affymetrix® GeneChip® Bovine Genome Array platform from the same PBL-extracted RNA. A total of 3,250 differentially expressed (DE) annotated genes were detected in the M. bovis-infected samples relative to the controls (adjusted P-value ≤0.05), with the number of genes displaying decreased relative expression (1,671) exceeding those with increased relative expression (1,579). Ingenuity® Systems Pathway Analysis (IPA) of all DE genes revealed enrichment for genes with immune function. Notably, transcriptional suppression was observed among several of the top-ranking canonical pathways including Leukocyte Extravasation Signaling. Comparative platform analysis demonstrated that RNA-seq detected a larger number of annotated DE genes (3,250) relative to the microarray (1,398), of which 917 genes were common to both technologies and displayed the same direction of expression. Finally, we show that RNA-seq had an increased dynamic range compared to the microarray for estimating differential gene expression. PMID:25206354

  2. Association between spoligotype-VNTR types and virulence of Mycobacterium bovis in cattle

    PubMed Central

    Garbaccio, Sergio; Macias, Analía; Shimizu, Ernesto; Paolicchi, Fernando; Pezzone, Natalia; Magnano, Gabriel; Zapata, Laura; Abdala, Alejandro; Tarabla, Hector; Peyru, Maite; Caimi, Karina; Zumárraga, Martín; Canal, Ana; Cataldi, Angel

    2014-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis, a disease that affects approximately 5% of Argentine cattle. The aim of this research was to study if it is possible to infer the degree of virulence of different M. bovis genotypes based on scorified observations of tuberculosis lesions in cattle. In this study, we performed association analyses between several parameters with tuberculosis lesions: M. bovis genotype, degree of progression of tuberculosis, and animal age. For this purpose, the genotype was determined by spoligotyping and the degree of bovine tuberculosis gross lesion was quantified with a score based on clinical observations (number, size, and location of granulomas along with histopathologic features). This study was performed with naturally infected cattle of slaughterhouses from three provinces in Argentina. A total of 265 M. bovis isolates were obtained from 378 pathological lesion samples and 192 spoligotyping and VNTR (based on ETR sequences) typing patterns were obtained. SB0140 was the most predominant spoligotype, followed by SB0145. The spoligotype with the highest lesion score was SB0273 (median score of 27 ± 4.46), followed by SB0520 (18 ± 5.8). Furthermore, the most common spoligotype, SB0140, had a median score of 11 ± 0.74. Finally, the spoligotype with the lowest score was SB0145 (8 ± 1.0). ETR typing of SB0140, SB0145, SB0273, and SB0520 did not subdivide the lesion scores in those spoligotypes. In conclusion, SB0273 and SB0520 were the spoligotypes with the strongest association with hypervirulence and both spoligotypes were only found in Río Cuarto at the south of Córdoba province. Interestingly, there is no other report of any of these spoligotyes in Latin America. PMID:24398919

  3. Immunological responses of Eurasian badgers (Meles meles) vaccinated with Mycobacterium bovis BCG (bacillus calmette guerin).

    PubMed

    Southey, A; Sleeman, D P; Lloyd, K; Dalley, D; Chambers, M A; Hewinson, R G; Gormley, E

    2001-05-30

    Wildlife species, such as the badger (Meles meles), may act as maintenance hosts for Mycobacterium bovis and contribute to the spread and persistence of tuberculosis in associated cattle populations. Targeted vaccination of badgers against tuberculosis is an option that, if successfully employed, could directly facilitate the advancement of bovine tuberculosis eradication in affected areas. In this study, the immunological responses of a group of badgers vaccinated subcutaneously with low doses of Mycobacterium bovis bacillus calmette guerin (BCG) were measured in vitro and compared with non-vaccinated control animals over a period of 42 weeks. Peripheral blood mononuclear cells (PBMC) from badgers which had received repeated booster injections of BCG proliferated in response to culture with PPD-bovine (purified protein derivative of tuberculin). The proliferation was significantly greater than that seen in the non-vaccinated control group. In contrast, the proliferative response of PBMC from vaccinated badgers to PPD-avian declined relative to the control group. These results demonstrate that repeated vaccination of badgers with M. bovis BCG induced a population of T-lymphocytes responsive to specific antigens in PPD-bovine. Throughout the course of the study, the sera from all animals were tested (BrockTest) by an enzyme-linked immunosorbent assay (ELISA) system for the presence of antibodies to MPB83, a serodominant antigen whose expression is high in M. bovis, but very low in BCG (Pasteur). No animals at any stage showed seroconversion to the antigen, consistent with the tuberculosis-free status of the badgers under study. PMID:11389955

  4. Investigating Transmission of Mycobacterium bovis in the United Kingdom in 2005 to 2008▿

    PubMed Central

    Mandal, Sema; Bradshaw, Louise; Anderson, Laura F.; Brown, Tim; Evans, Jason T.; Drobniewski, Francis; Smith, Grace; Magee, John G.; Barrett, Anne; Blatchford, Oliver; Laurenson, Ian F.; Seagar, Amie-Louise; Ruddy, Michael; White, P. Lewis; Myers, Richard; Hawkey, Peter; Abubakar, Ibrahim

    2011-01-01

    Due to an increase in bovine tuberculosis in cattle in the United Kingdom, we investigated the characteristics of Mycobacterium bovis infection in humans and assessed whether extensive transmission of M. bovis between humans has occurred. A cross-sectional study linking demographic, clinical, and DNA fingerprinting (using 15-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat [MIRU-VNTR] typing) data on cases reported between 2005 and 2008 was undertaken. A total of 129 cases of M. bovis infection in humans were reported over the period, with a decrease in annual incidence from 0.065 to 0.047 cases per 100,000 persons. Most patients were born pre-1960, before widespread pasteurization was introduced (73%), were of white ethnicity (83%), and were born in the United Kingdom (76%). A total of 102 patients (79%) had MIRU-VNTR typing data. A total of 31 of 69 complete MIRU-VNTR profiles formed eight distinct clusters. The overall clustering proportion determined using the n − 1 method was 33%. The largest cluster, comprising 12 cases, was indistinguishable from a previously reported West Midlands outbreak strain cluster and included those cases. This cluster was heterogeneous, having characteristics supporting recent zoonotic and human-to-human transmission as well as reactivation of latent disease. Seven other, smaller clusters identified had demographics supporting recrudescence rather than recent infection. A total of 33 patients had incomplete MIRU-VNTR profiles, of which 11 may have yielded 2 to 6 further small clusters if typed to completion. The incidence of M. bovis in humans in the United Kingdom remains low, and the epidemiology is predominantly that of reactivated disease. PMID:21430093

  5. Possible Airborne Person-to-Person Transmission of Mycobacterium bovis - Nebraska 2014-2015.

    PubMed

    Buss, Bryan F; Keyser-Metobo, Alison; Rother, Julie; Holtz, Laura; Gall, Kristin; Jereb, John; Murphy, Caitlin N; Iwen, Peter C; Robbe-Austerman, Suelee; Holcomb, Melissa A; Infield, Pat

    2016-03-01

    Mycobacterium bovis, one of several mycobacteria of the M. tuberculosis complex, is a global zoonotic pathogen that primarily infects cattle. Humans become infected by consuming unpasteurized dairy products from infected cows; possible person-to-person airborne transmission has also been reported. In April 2014, a man in Nebraska who was born in Mexico was determined to have extensive pulmonary tuberculosis (TB) caused by M. bovis after experiencing approximately 3 months of cough and fever. Four months later, a U.S.-born Hispanic girl from a nearby town who had been ill for 4-5 months was also determined to have pulmonary TB caused by M. bovis. The only social connection between the two patients was attendance at the same church, and no common dietary exposure was identified. Both patients had pulmonary cavities on radiography and acid-fast bacilli (AFB) on sputum-smear microscopy, indicators of being contagious. Whole-genome sequencing results of the isolates were nearly indistinguishable. Initial examination of 181 contacts determined that 39 (22%) had latent infection: 10 (42%) of 24 who had close exposure to either patient, 28 (28%) of 100 who were exposed to one or both patients in church, and one (2%) of 57 exposed to the second patient at a school. Latent infection was diagnosed in six contacts on follow-up examination, 2 months after an initial negative test result, for an overall latent infection rate of 25%. No infected contacts recalled consuming unpasteurized dairy products, and none had active TB disease at the initial or secondary examination. Persons who have M. bovis TB should be asked about consumption of unpasteurized dairy products, and contact investigations should follow the same guidance as for M. tuberculosis TB. PMID:26938831

  6. Immunization with a Streptococcus bovis vaccine administered by different routes against lactic acidosis in sheep.

    PubMed

    Shu, Q; Gill, H S; Leng, R A; Rowe, J B

    2000-05-01

    Streptococcus bovis is an important lactic acid bacterium in the rumen, which contributes to the development of lactic acidosis. This study was designed to test the efficacy of immunization with S. bovis primed either intramuscularly (i.m.) or intraperitoneally (i.p. ) against lactic acidosis. Forty-five wethers were allocated to three treatment groups. Two groups were injected with a S. bovis vaccine by either the i.m. or i.p. route for primary immunization; both groups were further immunized by the same route(s) (oral and/or i.m.) for boosters. The third group was not immunized (control). Antibody concentrations were measured in saliva prior to and following animals being fed a grain diet, and also in the rumen fluid, before the animals were suddenly introduced to a grain diet. The average antibody concentration in the animals of the i.m. group was higher than the i.p. group (P< 0.05). The antibody concentration in the rumen fluid of immunized sheep was higher than the control animals (P< 0.01). The difference in the rumen fluid antibody concentration between the i.m. and i.p. groups was not statistically significant (P> 0.05). In the i.m. group, there was a significantly greater feed intake, higher rumen pH, lower diarrhoea scores, and less increase in blood packed cell volume following grain feeding than in the animals of the control group. The severity of diarrhoea and the increase of blood packed cell volume in the animals of the i. p. group were also less than in the animals of the control group. The results suggest that the risk of lactic acidosis can be reduced by immunization against S. bovis, and that the immunization primed i. m. is more effective than the immunization primed i.p.

  7. A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan).

    PubMed

    Iqbal, F; Fatima, M; Shahnawaz, S; Naeem, M; Shaikh, Rs; Ali, M; Shaikh, As; Aktas, M; Ali, M

    2011-08-01

    Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total) produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009) and young animals under one year of age (P = 0.01) were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007), composed of mixed species of small ruminants (P = 0.022), associated with dogs (P = 0.003) and dogs having ticks on their bodies (P = 0.011) were among the major risk factors for the spread of babesiosis in small ruminants. PMID:21894263

  8. A study on the determination of risk factors associated with babesiosis and prevalence of Babesia sp., by PCR amplification, in small ruminants from Southern Punjab (Pakistan)

    PubMed Central

    Iqbal, F.; Fatima, M.; Shahnawaz, S.; Naeem, M.; Shaikh, R.S.; Ali, M.; Shaikh, A.S.; Aktas, M.; Ali, M.

    2011-01-01

    Babesiosis is a parasitic infection due to the multiplication of tick borne parasite, Babesia sp., in erythrocytes of host, which includes a wide variety of vertebrates including small ruminants causing decreased livestock output and hence economic losses. The objective of the present study was to establish a PCR based method for the detection of Babesia sp. in small ruminant population in Southern Punjab and to determine the risk factors involve in the spread of babesiosis. A total of 107 blood samples were collected from 40 sheep and 67 goats in seven districts of Southern Punjab from randomly selected herds. Data on the characteristics of the animals and the herd were collected through questionnaires. 36 blood samples (34% of total) produced the DNA fragment specific for 18S rRNA gene of Babesia sp., by PCR amplification, of which 20 were sheep and 16 were goats. Samples from all seven district contained Babesia positive samples and prevalence varied between 18 to 68%. It was observed that male animals (P = 0.009) and young animals under one year of age (P = 0.01) were more prone to the parasite. It was observed that herds consist of more than 15 animals (P = 0.007), composed of mixed species of small ruminants (P = 0.022), associated with dogs (P = 0.003) and dogs having ticks on their bodies (P = 0.011) were among the major risk factors for the spread of babesiosis in small ruminants. PMID:21894263

  9. In vitro antimicrobial susceptibility of Mycoplasma bovis isolated in Israel from local and imported cattle.

    PubMed

    Gerchman, Irena; Levisohn, Sharon; Mikula, Inna; Lysnyansky, Inna

    2009-06-12

    Monitoring of susceptibility to antibiotics in field isolates of pathogenic bovine mycoplasmas is important for appropriate choice of treatment. Our study compared in vitro susceptibility profiles of Mycoplasma bovis clinical strains, isolated during 2005-2007 from Israeli and imported calves. Minimal inhibitory concentration (MIC) values were determined for macrolides by the microbroth dilution test, for aminoglycosides by commercial Etest, and for fluoroquinolones and tetracyclines by both methods. Notably, although correlation between the methods was generally good, it was not possible to determine the MIC endpoint for enrofloxacin-resistant strains (MIC > or =2.5 microg/ml in the microtest) by Etest. Comparison of antibiotic susceptibility profiles between local and imported M. bovis strains revealed that local strains were significantly more resistant to macrolides than most isolates from imported animals, with MIC(50) of 128 microg/ml vs. 2 microg/ml for tilmicosin and 8 microg/ml vs. 1 microg/ml for tylosin, respectively. However, local strains were more susceptible than most imported strains to fluoroquinolones and spectinomycin. Difference in susceptibility to tetracycline, doxycycline and oxytetracycline between local and imported strains was expressed in MIC(90) values for imported strains in the susceptible range compared to intermediate susceptibility for local strains. The marked difference in susceptibility profiles of M. bovis strains isolated from different geographical regions seen in this study emphasizes the necessity for performing of the antimicrobial susceptibility testing periodically and on a regional basis. PMID:19250777

  10. Genotyping Mycobacterium bovis from cattle in the Central Pampas of Argentina: temporal and regional trends

    PubMed Central

    Shimizu, Ernesto; Macías, Analía; Paolicchi, Fernando; Magnano, Gabriel; Zapata, Laura; Fernández, Analía; Canal, Ana; Garbaccio, Sergio; Cataldi, Angel; Caimi, Karina; Zumárraga, Martín

    2014-01-01

    Mycobacterium bovis is the causative agent of bovine tuberculosis (TB), a disease that affects approximately 5% of Argentinean cattle. Among the molecular methods for genotyping, the most convenient are spoligotyping and variable number of tandem repeats (VNTR). A total of 378 samples from bovines with visible lesions consistent with TB were collected at slaughterhouses in three provinces, yielding 265 M. bovis spoligotyped isolates, which were distributed into 35 spoligotypes. In addition, 197 isolates were also typed by the VNTR method and 54 combined VNTR types were detected. There were 24 clusters and 27 orphan types. When both typing methods were combined, 98 spoligotypes and VNTR types were observed with 27 clusters and 71 orphan types. By performing a meta-analysis with previous spoligotyping results, we identified regional and temporal trends in the population structure of M. bovis. For SB0140, the most predominant spoligotype in Argentina, the prevalence percentage remained high during different periods, varying from 25.5-57.8% (1994-2011). By contrast, the second and third most prevalent spoligotypes exhibited important fluctuations. This study shows that there has been an expansion in ancestral lineages as demonstrated by spoligotyping. However, exact tandem repeat typing suggests dynamic changes in the clonal population of this microorganism. PMID:24676658

  11. Mycobacterium bovis: A Model Pathogen at the Interface of Livestock, Wildlife, and Humans

    PubMed Central

    Palmer, Mitchell V.; Thacker, Tyler C.; Waters, W. Ray; Gortázar, Christian; Corner, Leigh A. L.

    2012-01-01

    Complex and dynamic interactions involving domestic animals, wildlife, and humans create environments favorable to the emergence of new diseases, or reemergence of diseases in new host species. Today, reservoirs of Mycobacterium bovis, the causative agent of tuberculosis in animals, and sometimes humans, exist in a range of countries and wild animal populations. Free-ranging populations of white-tailed deer in the US, brushtail possum in New Zealand, badger in the Republic of Ireland and the United Kingdom, and wild boar in Spain exemplify established reservoirs of M. bovis. Establishment of these reservoirs is the result of factors such as spillover from livestock, translocation of wildlife, supplemental feeding of wildlife, and wildlife population densities beyond normal habitat carrying capacities. As many countries attempt to eradicate M. bovis from livestock, efforts are impeded by spillback from wildlife reservoirs. It will not be possible to eradicate this important zoonosis from livestock unless transmission between wildlife and domestic animals is halted. Such an endeavor will require a collaborative effort between agricultural, wildlife, environmental, and political interests. PMID:22737588

  12. Genetic diversity based on MIRU-VNTR profile of isolates of Mycobacterium bovis from Mexican cattle.

    PubMed

    Nava Vargas, Alejandro; Milián Suazo, Feliciano; Cantó Alarcón, Germinal Jorge; Rubio Venegas, Yezenia; Guerrero Solorio, Roberto; Rodríguez Hernández, Elba; Pizano Martìnez, Oscar

    2016-09-01

    Bovine tuberculosis (bTB) is a disease caused by Mycobacterium bovis (M. bovis), which affects cattle, animal species and humans. To determinate the genetic structure of strains of M. bovis in mexican cattle, 467 isolates obtained from 2009 to 2010 from different regions of Mexico with known spoligotype were included in the study. The isolates were genotyped by interspersed repeated mycobacterial units-variable number tandem repeats (MIRU-VNTR) obtaining 13 MIRU-VNTR groups. When combining MIRU-VNTR patterns with its spolygotypes, the Hunter genetic discrimination index (HGDI), we obtained 421 genetic patterns distributed in 17 groups. The HGDI for the total loci was 0.99. The locus that presented the higher HGDI was 2461 (0.857), while the locus with the lowest HGDI was 2686 (0.239). When we analyzed our results, using just 6 or 8 MIRU-VNTR we obtained an discriminatory power of 0.8499 and 0.8875 respectively indicating lower HGDI than 12 MIRU-VNTR locus. PMID:27544255

  13. Visual format for detection of Mycobacterium tuberculosis and M. bovis in clinical samples using molecular beacons.

    PubMed

    Kumar, Parameet; Nath, Kapili; Rath, Bimba; Sen, Manas K; Vishalakshi, Potharuju; Chauhan, Devender S; Katoch, Vishwa M; Singh, Sarman; Tyagi, Sanjay; Sreenivas, Vishnubhatla; Prasad, Hanumanthappa K

    2009-09-01

    A real-time polymerase chain reaction (PCR) assay for the direct identification of Mycobacterium tuberculosis and M. bovis using molecular beacons was developed. The assay was modified for use in regular thermal cyclers. Molecular beacons that were specific for M. tuberculosis (Tb-B) and M. bovis (Bo-B) were designed. The fluorescence of the target PCR product-molecular beacon probe complex was detected visually using a transilluminator. The results were then compared with those of conventional multiplex PCR (CM-PCR) assays and biochemical identification. The detection limit of Tb-B and Bo-B beacons was 500 fg and 50 fg by the visual format and real-time PCR assay, respectively, compared with 5 pg by CM-PCR assay. Pulmonary and extrapulmonary samples were examined. The agreement between culture and the two assays was very good in sputum samples and fair in extrapulmonary samples. The agreement between clinical diagnoses with the two assays was moderate in extrapulmonary samples. There was very good agreement between CM-PCR and visual format assays for all samples used in the study. Concordance in the identification of isolates by the visual, CM-PCR assay, and biochemical identification was seen. Hence, the use of molecular beacon detection of M. tuberculosis and M. bovis in clinical samples is feasible by setting up two asymmetric PCRs concurrently. The assay is sensitive, specific, simple to interpret, and takes less than 3 hours to complete.

  14. Identification of proteins from Mycobacterium tuberculosis missing in attenuated Mycobacterium bovis BCG strains.

    PubMed

    Mattow, J; Jungblut, P R; Schaible, U E; Mollenkopf, H J; Lamer, S; Zimny-Arndt, U; Hagens, K; Müller, E C; Kaufmann, S H

    2001-08-01

    A proteome approach, combining high-resolution two-dimensional electrophoresis (2-DE) with mass spectrometry, was used to compare the cellular protein composition of two virulent strains of Mycobacterium tuberculosis with two attenuated strains of Mycobacterium bovis Bacillus Calmette-Guerin (BCG), in order to identify unique proteins of these strains. Emphasis was given to the identification of M. tuberculosis specific proteins, because we consider these proteins to represent putative virulence factors and interesting candidates for vaccination and diagnosis of tuberculosis. The genome of M. tuberculosis strain H37Rv comprises nearly 4000 predicted open reading frames. In contrast, the separation of proteins from whole mycobacterial cells by 2-DE resulted in silver-stained patterns comprising about 1800 distinct protein spots. Amongst these, 96 spots were exclusively detected either in the virulent (56 spots) or in the attenuated (40 spots) mycobacterial strains. Fifty-three of these spots were analyzed by mass spectrometry, of which 41 were identified, including 32 M. tuberculosis specific spots. Twelve M. tuberculosis specific spots were identified as proteins, encoded by genes previously reported to be deleted in M. bovis BCG. The remaining 20 spots unique for M. tuberculosis were identified as proteins encoded by genes that are not known to be missing in M. bovis BCG.

  15. Antigen specific immunological responses of badgers (Meles meles) experimentally infected with Mycobacterium bovis.

    PubMed

    Lesellier, Sandrine; Corner, Leigh; Costello, Eamon; Sleeman, Paddy; Lyashchenko, Konstantin; Greenwald, Rena; Esfandiari, Javan; Singh, Mahavir; Hewinson, R Glyn; Chambers, Mark; Gormley, Eamonn

    2008-03-15

    European badgers (Meles meles) are considered to be an important reservoir of infection for Mycobacterium bovis and are implicated in the transmission of tuberculosis to cattle in Ireland and Great Britain. Accurate tests are required for tuberculosis surveillance in badger populations and to provide a basis for the development of strategies, including vaccination, to reduce the incidence of the infection. In this study, we have developed an endobronchial M. bovis infection model in badgers in which we measured cell-mediated immune and serological responses for up to 24 weeks post-infection. Groups of badgers were subjected to necropsy at 6-week intervals and the gross lesion severity status compared with immune responses measured in blood samples taken throughout the course of the study. The panel of antigens included bovine and avian tuberculins (PPD) as well as single antigens, ESAT-6, CFP-10, MPB70, Rv3019c, Rv3873, Rv3878 and Rv3879, all known to be recognised by the immune system in other animal models of tuberculosis infection. Our results demonstrated that M. bovis infected badgers responded to specific antigens as early as 6 weeks post-infection, consistent with the presence of visible lesions. The data also revealed unique patterns of antigen recognition with high levels of PBMC proliferation in the presence of CFP-10 but low proliferation levels with ESAT-6. Using a multi-antigen print immunoassay (MAPIA), we were able to confirm that MPB83 is the dominant antigen recognised by serum antibodies in infected badgers.

  16. Immunological responses and protective immunity in BCG vaccinated badgers following endobronchial infection with Mycobacterium bovis.

    PubMed

    Lesellier, Sandrine; Corner, Leigh; Costello, Eamon; Lyashchenko, Konstantin; Greenwald, Rena; Esfandiari, Javan; Singh, Mahavir; Hewinson, R Glyn; Chambers, Mark; Gormley, Eamonn

    2009-01-14

    European badgers (Meles meles) are a reservoir host of Mycobacterium bovis and are implicated in the transmission of tuberculosis to cattle in Ireland and Great Britain. The development of a vaccine for use in badgers is considered a key element of any campaign to eradicate the disease in livestock in both countries. In this study we have vaccinated groups of badgers with approximately 5 x 10(5)cfu of the BCG vaccine delivered via two alternative routes, subcutaneous and mucosal (intranasal/conjunctival). Following experimental endobronchial infection with approximately 10(4)cfu of M. bovis, all badgers were euthanised at 12 weeks post-infection. At post-mortem examination both vaccinated groups had significantly reduced severity of disease compared with the non-vaccinated controls. The analysis of immune responses throughout the study showed that vaccination with BCG did not generate any detectable immunological responses as measured by IFN-gamma production in antigen-stimulated peripheral blood mononuclear cells (PBMC) and IgG serological responses. However, the levels of the responses increased following M. bovis infection, and the kinetic profiles corresponded to the severity of lesions recorded post-mortem. Significant differences were observed in the timing of development of the immune responses between vaccinates and controls. The results suggest that the immunological responses are associated with the levels of protective immunity and could be used as markers to monitor control of disease in badgers following vaccination.

  17. The variability and seasonality of the environmental reservoir of Mycobacterium bovis shed by wild European badgers.

    PubMed

    King, Hayley C; Murphy, Andrew; James, Phillip; Travis, Emma; Porter, David; Hung, Yu-Jiun; Sawyer, Jason; Cork, Jennifer; Delahay, Richard J; Gaze, William; Courtenay, Orin; Wellington, Elizabeth M

    2015-08-06

    The incidence of Mycobacterium bovis, the causative agent of bovine tuberculosis, has been increasing in UK cattle herds resulting in substantial economic losses. The European badger (Meles meles) is implicated as a wildlife reservoir of infection. One likely route of transmission to cattle is through exposure to infected badger urine and faeces. The relative importance of the environment in transmission remains unknown, in part due to the lack of information on the distribution and magnitude of environmental reservoirs. Here we identify potential infection hotspots in the badger population and quantify the heterogeneity in bacterial load; with infected badgers shedding between 1 × 10(3)- 4 × 10(5) M. bovis cells g(-1) of faeces, creating a substantial and seasonally variable environmental reservoir. Our findings highlight the potential importance of monitoring environmental reservoirs of M. bovis which may constitute a component of disease spread that is currently overlooked and yet may be responsible for a proportion of transmission amongst badgers and onwards to cattle.

  18. The pathophysiological effects of Moraxella bovis toxins on cattle, mice and guinea pigs.

    PubMed

    Pugh, G W; Hughes, D E; Schulz, V D

    1973-01-01

    In three experiments, cattle, mice and guinea pigs were inoculated with viable cultures of Moraxella bovis or fractions of this organism. Fractions were obtained by disruption of cells with a fractionator at 20,000 pounds per square inch, and separating the cell wall and cell sap fractions by differential centrifugation. Cell sap fractions were further separated by ultra-centrifugation, heating and precipitation with (NH(4))(2) SO(4). Different fractions induced different pathophysiological manifestations. The cell wall fractions caused localized lesions (necrosis) at the site of injection, and emphysema and congestion of the lungs. Cell sap fractions induced a "shock syndrome," as well as hemorrhage and inflammation of the intestines, hemorrhage and congestion of lymph nodes, liver, adrenal and spleen. Cell sap also induced conjunctivitis in mice and guinea pigs, and periocular edema, myosis, ocular pruritus and lacrimation in cattle. The authors suggest that M. bovis probably produces endotoxins and exotoxins as well as possibly a specific oculopathic substance, but more definitive work is needed to confirm this. They caution that consideration of these toxins should be made in any application of M. bovis for vaccines or other immunological studies.

  19. The variability and seasonality of the environmental reservoir of Mycobacterium bovis shed by wild European badgers

    PubMed Central

    King, Hayley C.; Murphy, Andrew; James, Phillip; Travis, Emma; Porter, David; Hung, Yu-Jiun; Sawyer, Jason; Cork, Jennifer; Delahay, Richard J.; Gaze, William; Courtenay, Orin; Wellington, Elizabeth M.

    2015-01-01

    The incidence of Mycobacterium bovis, the causative agent of bovine tuberculosis, has been increasing in UK cattle herds resulting in substantial economic losses. The European badger (Meles meles) is implicated as a wildlife reservoir of infection. One likely route of transmission to cattle is through exposure to infected badger urine and faeces. The relative importance of the environment in transmission remains unknown, in part due to the lack of information on the distribution and magnitude of environmental reservoirs. Here we identify potential infection hotspots in the badger population and quantify the heterogeneity in bacterial load; with infected badgers shedding between 1 × 103 − 4 × 105 M. bovis cells g−1 of faeces, creating a substantial and seasonally variable environmental reservoir. Our findings highlight the potential importance of monitoring environmental reservoirs of M. bovis which may constitute a component of disease spread that is currently overlooked and yet may be responsible for a proportion of transmission amongst badgers and onwards to cattle. PMID:26247348

  20. A presumptive case of cerebral babesiosis in a dog in Poland caused by a virulent Babesia canis strain.

    PubMed

    Adaszek, L; Górna, M; Klimiuk, P; Kalinowski, M; Winiarczyk, S

    2012-10-17

    The aim of this paper was to present the first case of cerebral canine babesiosis due to infection by Babesia canis in a dog in Poland. A 5-year-old American Staffordshire Terrier was presented with an unusual clinical manifestation of acute babesiosis that included neurological signs and pancytopenia. Despite treatment the dog died. Diagnosis was based on microscopic examination of Giemsa-stained blood smears (detection of piroplasms in red blood cells) and post mortem examination of the brain by histopathology and PCR method. The amplified segment of the Babesia 18S RNA gene was sequenced. This enabled to determine that the cause of the disease had been the strain 18S RNA-B EU622793. This is one of two B. canis strains found endemically in Poland, which reveals a greater virulence than the strain 18S RNA-A EU622792. The described case indicates that this form of canine babesiosis should be taken into account in differential diagnosis in dogs exhibiting neurological symptoms, especially in the tick activity season.