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J S Gray - One of the best experts on this subject based on the ideXlab platform.
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four dimensional characterization of the Babesia divergens asexual life cycle from the trophozoite to the multiparasite stage
mSphere, 2020Co-Authors: Jose Javier Conesa, J S Gray, Elena Sevilla, Luis Miguel Gonzalez, Maria C Terron, Ana J Perezberna, Jose L Carrascosa, Eva Pereiro, Francisco Javier Chichon, Daniel LuqueAbstract:ABSTRACT Babesia is an apicomplexan parasite of significance that causes the disease known as babesiosis in domestic and wild animals and in humans worldwide. Babesia infects vertebrate hosts and reproduces asexually by a form of binary fission within erythrocytes/red blood cells (RBCs), yielding a complex pleomorphic population of intraerythrocytic parasites. Seven of them, clearly visible in human RBCs infected with Babesia divergens, are considered the main forms and named single, double, and quadruple trophozoites, paired and double paired pyriforms, tetrad or Maltese Cross, and multiparasite stage. However, these main intraerythrocytic forms coexist with RBCs infected with transient parasite combinations of unclear origin and development. In fact, little is understood about how Babesia builds this complex population during its asexual life cycle. By combining cryo-soft X-ray tomography and video microscopy, main and transitory parasites were characterized in a native whole cellular context and at nanometric resolution. The architecture and kinetics of the parasite population was observed in detail and provide additional data to the previous B. divergens asexual life cycle model that was built on light microscopy. Importantly, the process of multiplication by binary fission, involving budding, was visualized in live parasites for the first time, revealing that fundamental changes in cell shape and continuous rounds of multiplication occur as the parasites go through their asexual multiplication cycle. A four-dimensional asexual life cycle model was built highlighting the origin of several transient morphological forms that, surprisingly, intersperse in a chronological order between one main stage and the next in the cycle. IMPORTANCE Babesiosis is a disease caused by intraerythrocytic Babesia parasites, which possess many clinical features that are similar to those of malaria. This worldwide disease is increasing in frequency and geographical range and has a significant impact on human and animal health. Babesia divergens is one of the species responsible for human and cattle babesiosis causing death unless treated promptly. When B. divergens infects its vertebrate hosts, it reproduces asexually within red blood cells. During its asexual life cycle, B. divergens builds a population of numerous intraerythrocytic (IE) parasites of difficult interpretation. This complex population is largely unexplored, and we have therefore combined three- and four-dimensional imaging techniques to elucidate the origin, architecture, and kinetics of IE parasites. Unveiling the nature of these parasites has provided a vision of the B. divergens asexual cycle in unprecedented detail and is a key step to develop control strategies against babesiosis.
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four dimensional characterization of the Babesia divergens asexual life cycle from the trophozoite to the multiparasite stage
bioRxiv, 2020Co-Authors: Jose Javier Conesa, J S Gray, Elena Sevilla, Luis Miguel Gonzalez, Maria C Terron, Ana J Perezberna, Jose L Carrascosa, Eva Pereiro, Javier Chichon, Daniel LuqueAbstract:Babesia is an apicomplexan parasite of significance that causes the disease known as babesiosis in domestic and wild animals and in humans worldwide. Babesia infects vertebrate hosts and reproduces asexually by a form of binary fission within erythrocytes/red blood cells (RBCs), yielding a complex pleomorphic population of intraerythrocytic parasites. Seven of them, clearly visible in human RBCs infected with Babesia divergens, are considered the main forms and named single, double and quadruple trophozoites, paired and double paired-pyriforms, tetrad or Maltese Cross, and multiparasite stage. However, these main intraerythrocytic forms coexist with RBCs infected with transient parasite combinations of unclear origin and development. In fact, little is understood about how Babesia builds this complex population during its asexual life cycle. By combining the emerging technique cryo soft X-ray tomography and video microscopy, main and transitory parasites were characterized in a native whole cellular context and at nanometric resolution. As a result, the architecture and kinetic of the parasite population has been elucidated. Importantly, the process of multiplication by binary fission, involving budding, was visualized in live parasites for the first time, revealing that fundamental changes in cell shape and continuous rounds of multiplication occur as the parasites go through their asexual multiplication cycle. Based on these observations, a four-dimensional (4D) asexual life cycle model has been designed highlighting the origin of the tetrad, double paired-pyriform and multiparasite stages and the transient morphological forms that, surprisingly, intersperse in a chronological order between one main stage and the next along the cycle.
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kinetics of the invasion and egress processes of Babesia divergens observed by time lapse video microscopy
Scientific Reports, 2018Co-Authors: Elena Sevilla, J S Gray, Luis Miguel Gonzalez, Daniel Luque, Estrella MonteroAbstract:Based on confocal fluorescence and bright field video microscopy, we present detailed observations on the processes of invasion and egress of erythrocytes by the apicomplexan parasite Babesia divergens. Time-lapse images reveal numerous unexpected findings associated with the dynamics of B. divergens and its ability to manipulate the erythrocyte during both processes in its asexual cycle under in vitro conditions. Despite the speed at which these processes occur and the small size of the parasite, we capture infective merozoites moving vigorously and causing striking deformations in the erythrocyte’s plasma membrane during an active invasion. We also observed intraerythrocytic dynamic stages as paired pyriforms, double paired pyriforms, tetrads, unattached pyriform sister cells and multiple parasite stages resulting in the release of large numbers of merozoites over a short period. Of considerable interest is that time-lapse images reveal a novel mechanism of egress used by B. divergens to exit the human erythrocyte. The release occurs when B. divergens parasites establish contacts with the plasma membrane of the erythrocyte from within, before exiting the cell. Visualization and analysis of the images enabled us to obtain useful information and broaden our knowledge of complex and crucial events involved with parasitisation of human erythrocytes by B. divergens.
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splenic dysfunction from celiac disease resulting in severe babesiosis
Ticks and Tick-borne Diseases, 2017Co-Authors: Sarah Oconnell, Annetta Zintl, Craig Lyons, Moustafa Abdou, Rittick Patowary, Salman Aslam, Nora Kinsella, Klauspeter Hunfeld, Gary P Wormser, J S GrayAbstract:We describe a 79-year-old Irish man who, because he had hyposplenism and splenic atrophy due to adult celiac disease, became critically ill from a severe Babesia divergens infection. Greater awareness of the possible consequences of splenic dysfunction from adult celiac disease, such as serious pneumococcal infections and babesiosis, is warranted.
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identification and characterization of the rhoptry neck protein 2 in Babesia divergens and b microti
Infection and Immunity, 2016Co-Authors: Rosalynn Ord, J S Gray, Marilis Rodriguez, Jeny R Cursinosantos, Hyunryung Hong, Manpreet Singh, Cheryl A LoboAbstract: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.
Annetta Zintl - One of the best experts on this subject based on the ideXlab platform.
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splenic dysfunction from celiac disease resulting in severe babesiosis
Ticks and Tick-borne Diseases, 2017Co-Authors: Sarah Oconnell, Annetta Zintl, Craig Lyons, Moustafa Abdou, Rittick Patowary, Salman Aslam, Nora Kinsella, Klauspeter Hunfeld, Gary P Wormser, J S GrayAbstract:We describe a 79-year-old Irish man who, because he had hyposplenism and splenic atrophy due to adult celiac disease, became critically ill from a severe Babesia divergens infection. Greater awareness of the possible consequences of splenic dysfunction from adult celiac disease, such as serious pneumococcal infections and babesiosis, is warranted.
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Babesias of red deer cervus elaphus in ireland
Veterinary Research, 2011Co-Authors: Annetta Zintl, Eugene J Finnerty, Thomas M Murphy, Theo De Waal, J S GrayAbstract: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.
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Babesia divergens a bovine blood parasite of veterinary and zoonotic importance
Clinical Microbiology Reviews, 2003Co-Authors: Annetta Zintl, H E Skerrett, Stuart M Taylor, Grace Mulcahy, J S GrayAbstract:Babesia divergens is an intraerythrocytic protozoan parasite, transmitted by the tick Ixodes ricinus, and is the main agent of bovine babesiosis in Europe. It is not only a cause of significant loss to the cattle industry; it can also infect immunocompromised humans, causing medical emergencies characterized by rapid fulmination and parasitemias that may exceed 70%. The current emphasis in Europe on sustainable agriculture and extensification is likely to lead to an increase in vector tick populations with increased risk of infection. Despite the veterinary and zoonotic importance of this parasite, relatively little research has been carried out on B. divergens, and many questions regarding the parasite's epidemiology and the host's response remain unanswered. A better understanding of the species' biology and host-parasite interactions may lead to improved control mechanisms and new trends in vaccine and antiBabesial drug development. This review provides the first comprehensive summary of B. divergens biology, including its morphology, life cycle, and host specificity, and the current state of knowledge of both human and bovine infections.
Laurence Malandrin - One of the best experts on this subject based on the ideXlab platform.
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Stimulation and quantification of Babesia divergens gametocytogenesis
Parasites and Vectors, 2016Co-Authors: Marie Jalovecka, Ondrej Hajdusek, Claire Bonsergent-guillou, Petr Kopacek, Laurence MalandrinAbstract:Background Babesia divergens is the most common blood parasite in Europe causing babesiosis, a tick-borne malaria-like disease. Despite an increasing focus on B. divergens, especially regarding veterinary and human medicine, the sexual development of Babesia is poorly understood. Development of Babesia sexual stages in the host blood (gametocytes) plays a decisive role in parasite acquisition by the tick vector. However, the exact mechanism of gametocytogenesis is still unexplained. Methods Babesia divergens gametocytes are characterized by expression of bdccp1, bdccp2 and bdccp3 genes. Using previously described sequences of bdccp1, bdccp2 and bdccp3, we have established a quantitative real-time PCR (qRT-PCR) assay for detection and assessment of the efficiency of B. divergens gametocytes production in bovine blood. We analysed fluctuations in expression of bdccp genes during cultivation in vitro, as well as in cultures treated with different drugs and stimuli. Results We demonstrated that all B. divergens clonal lines tested, originally derived from naturally infected cows, exhibited sexual stages. Furthermore, sexual commitment was stimulated during continuous growth of the cultures, by addition of specific stress-inducing drugs or by alternating cultivation conditions. Expression of bdccp genes was greatly reduced or even lost after long-term cultivation, suggesting possible problems in the artificial infections of ticks in feeding assays in vitro. Conclusions Our research provides insight into sexual development of B. divergens and may facilitate the development of transmission models in vitro, enabling a more detailed understanding of Babesia-tick interactions.
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A retrospective serological survey on human babesiosis in Belgium
Epidemiology and Infection, 2015Co-Authors: Laetitia Lempereur, Emmanuelle Moreau, Brian Shiels, Paul Heyman, Claude Saegerman, Bertrand Losson, Laurence MalandrinAbstract:In Europe, most clinical babesiosis cases in humans have been attributed to Babesia divergens and Babesia sp. EU1. Babesia microti infection of humans occurs mainly in the United States; although a case of autochthonous B. microti infection and serological evidence of infection have been reported in Europe. The Indirect Fluorescent Antibody Test was used to screen sera from 199 anonymous Belgian patients with history of tick bite and clinical symptoms compatible with a tick-borne disease. The serological screen detected positive reactivity in 9% (n = 18), 33.2% (n = 66), and 39.7% (n = 79) of the samples against B. microti, B. divergens, and Babesia sp. EU1, respectively. Thus, evidence of contact among three potentially zoonotic species of Babesia and humans has been confirmed in Belgium. Preventive action and development of better diagnostic tools should help in prevention of clinical cases and to clarify the true burden of such infection for individuals and public health.
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Redescription of Babesia capreoli (Enigk and Friedhoff, 1962) from roe deer (Capreolus capreolus): Isolation, cultivation, host specificity, molecular characterisation and differentiation from Babesia divergens
International Journal for Parasitology, 2010Co-Authors: Laurence Malandrin, Maggy Jouglin, Yi Sun, Nadine Brisseau, Alain ChauvinAbstract:The recent use of the sole molecular identification of Babesia infecting European cervids has led to confusion between the closely related Babesia divergens and Babesia capreoli, and to their grouping together as “B. divergens-like”. In order to clarify this taxonomic confusion, Babesia from roe deer, cattle and human blood were isolated, cultured and their biological as well as molecular characteristics compared. On this basis, we conclude that: (i) the parasites isolated from roe deer blood are B. capreoli; (ii) there are no intraspecific variations in the 18S rDNA within B. capreoli and B. divergens spp.; (iii) these two species are closely related as demonstrated by their morphology, serological cross-reactions and 99.83% identity in their 18S rDNA; (iv) these two species are distinct as demonstrated by their different abilities to grow in vitro in cattle, human and sheep erythrocytes, by their infectivity for gerbils, and by a conserved three bases difference at positions 631, 663 and 1637 of their 18S rDNA; (v) B. capreoli does not pose a threat to either humans or livestock. An integrated description is given of the host range, geographical distribution, biological and molecular characterisation of B. capreoli, and reference materials have been deposited at the Museum d’Histoire Naturelle de Paris.
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individual heterogeneity in erythrocyte susceptibility to Babesia divergens is a critical factor for the outcome of experimental spleen intact sheep infections
Veterinary Research, 2009Co-Authors: Laurence Malandrin, Maggy Jouglin, Emmanuelle Moreau, Alain ChauvinAbstract:Susceptibility of sheep erythrocytes to Babesia divergens was investigated in vitro and a high inter-individual variability in their ability to support parasite population development was demonstrated, with some individuals having refractory red blood cells (RBC). As neither changes in growth conditions nor the use of different B. divergens strains influenced the level of susceptibility, the main factor postulated for this variability is the erythrocyte itself. Sheep therefore represent an excellent in vitro model to study the parasite-erythrocyte interaction. In addition, the existence of refractory RBC should help in the identification of the erythrocyte components required for B. divergens development. Experimental infections were carried out on spleen-intact sheep charac- terized by refractory or fully susceptible erythrocyte types. These differences translated into the successful infection of only those animals with susceptible erythrocytes: infected animals showed no clinical signs, but maintained an asymptomatic persistent infection, as usually observed in the natural bovine host. Sheep therefore represent model organisms that can allow us to study interactions between B. divergens and its vertebrate host at different levels of biological organisation, from the target cell to the intact animal, and represent an experimental infection model of concomitant immunity. Only a low percentage (13%) of the sheep population tested possessed susceptible erythrocytes and the potential role of sheep as a natural host or reservoir of B. divergens is discussed. Babesia divergens / sheep / erythrocyte / red blood cell / experimental animal model
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Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique
Parasitology, 2007Co-Authors: Sarah Bonnet, Laurence Malandrin, Maggy Jouglin, C. Becker, Albert Agoulon, Monique L'hostis, Alain ChauvinAbstract:Although Babesia divergens is the the principal confirmed zoonotic Babesia sp. in Europe, there are gaps in our knowledge of its biology and transmission by the tick Ixodes ricinus. In order to reproduce the part of the parasite cycle that occurs in the vector, an in vitro animal skin feeding technique on blood containing in vitro cultivated B. divergens was developed. Parasite DNA was detected in all samples of salivary glands of nymphs and adults that had fed on parasitized blood as larvae and nymphs, respectively, indicating acquisition as well as a transtadial persistence of B. divergens. PCR performed on eggs and larvae produced by females that had fed on parasitized blood demonstrated the existence of a transovarial transmission of the parasite. Gorging B. divergens infected larvae on non-infected gerbils showed persistance of the parasite over moulting into the resulting nymphs. These results indicate that the parasitic stages infective for the vector (i.e. the sexual stages) can be produced in vitro. To our knowledge, this is the first report of artificial feeding of I. ricinus via membrane as well as in vitro transmission of B. divergens to its vector. The opportunities offered by the use of such a transmission model of a pathogen by I. ricinus are discussed.
Alain Chauvin - One of the best experts on this subject based on the ideXlab platform.
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Redescription of Babesia capreoli (Enigk and Friedhoff, 1962) from roe deer (Capreolus capreolus): Isolation, cultivation, host specificity, molecular characterisation and differentiation from Babesia divergens
International Journal for Parasitology, 2010Co-Authors: Laurence Malandrin, Maggy Jouglin, Yi Sun, Nadine Brisseau, Alain ChauvinAbstract:The recent use of the sole molecular identification of Babesia infecting European cervids has led to confusion between the closely related Babesia divergens and Babesia capreoli, and to their grouping together as “B. divergens-like”. In order to clarify this taxonomic confusion, Babesia from roe deer, cattle and human blood were isolated, cultured and their biological as well as molecular characteristics compared. On this basis, we conclude that: (i) the parasites isolated from roe deer blood are B. capreoli; (ii) there are no intraspecific variations in the 18S rDNA within B. capreoli and B. divergens spp.; (iii) these two species are closely related as demonstrated by their morphology, serological cross-reactions and 99.83% identity in their 18S rDNA; (iv) these two species are distinct as demonstrated by their different abilities to grow in vitro in cattle, human and sheep erythrocytes, by their infectivity for gerbils, and by a conserved three bases difference at positions 631, 663 and 1637 of their 18S rDNA; (v) B. capreoli does not pose a threat to either humans or livestock. An integrated description is given of the host range, geographical distribution, biological and molecular characterisation of B. capreoli, and reference materials have been deposited at the Museum d’Histoire Naturelle de Paris.
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individual heterogeneity in erythrocyte susceptibility to Babesia divergens is a critical factor for the outcome of experimental spleen intact sheep infections
Veterinary Research, 2009Co-Authors: Laurence Malandrin, Maggy Jouglin, Emmanuelle Moreau, Alain ChauvinAbstract:Susceptibility of sheep erythrocytes to Babesia divergens was investigated in vitro and a high inter-individual variability in their ability to support parasite population development was demonstrated, with some individuals having refractory red blood cells (RBC). As neither changes in growth conditions nor the use of different B. divergens strains influenced the level of susceptibility, the main factor postulated for this variability is the erythrocyte itself. Sheep therefore represent an excellent in vitro model to study the parasite-erythrocyte interaction. In addition, the existence of refractory RBC should help in the identification of the erythrocyte components required for B. divergens development. Experimental infections were carried out on spleen-intact sheep charac- terized by refractory or fully susceptible erythrocyte types. These differences translated into the successful infection of only those animals with susceptible erythrocytes: infected animals showed no clinical signs, but maintained an asymptomatic persistent infection, as usually observed in the natural bovine host. Sheep therefore represent model organisms that can allow us to study interactions between B. divergens and its vertebrate host at different levels of biological organisation, from the target cell to the intact animal, and represent an experimental infection model of concomitant immunity. Only a low percentage (13%) of the sheep population tested possessed susceptible erythrocytes and the potential role of sheep as a natural host or reservoir of B. divergens is discussed. Babesia divergens / sheep / erythrocyte / red blood cell / experimental animal model
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Transstadial and transovarial persistence of Babesia divergens DNA in Ixodes ricinus ticks fed on infected blood in a new skin-feeding technique
Parasitology, 2007Co-Authors: Sarah Bonnet, Laurence Malandrin, Maggy Jouglin, C. Becker, Albert Agoulon, Monique L'hostis, Alain ChauvinAbstract:Although Babesia divergens is the the principal confirmed zoonotic Babesia sp. in Europe, there are gaps in our knowledge of its biology and transmission by the tick Ixodes ricinus. In order to reproduce the part of the parasite cycle that occurs in the vector, an in vitro animal skin feeding technique on blood containing in vitro cultivated B. divergens was developed. Parasite DNA was detected in all samples of salivary glands of nymphs and adults that had fed on parasitized blood as larvae and nymphs, respectively, indicating acquisition as well as a transtadial persistence of B. divergens. PCR performed on eggs and larvae produced by females that had fed on parasitized blood demonstrated the existence of a transovarial transmission of the parasite. Gorging B. divergens infected larvae on non-infected gerbils showed persistance of the parasite over moulting into the resulting nymphs. These results indicate that the parasitic stages infective for the vector (i.e. the sexual stages) can be produced in vitro. To our knowledge, this is the first report of artificial feeding of I. ricinus via membrane as well as in vitro transmission of B. divergens to its vector. The opportunities offered by the use of such a transmission model of a pathogen by I. ricinus are discussed.
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sheep as a new experimental host for Babesia divergens
Veterinary Research, 2002Co-Authors: Alain Chauvin, Laurence Malandrin, Alexis Valentin, Monique LhostisAbstract:Babesia divergens was cultivated in sheep erythrocytes in RPMI 1640 supplemented with 10% Fetal Calf Serum (FCS) or sheep serum. In vitro cultures in sheep red blood cells were initiated with human erythrocytes infected in vitro with B. divergens Rouen 1987 or with gerbil blood infected with several isolates from bovine origin. After the first subcultures on sheep erythrocytes, a ten-fold multiplication of the parasites was obtained within 48 h. Erythrocytes from three splenectomized sheep were infected in vitro with B. divergens; when parasitaemia reached 10%, the animals were inoculated with homologous parasitized erythrocytes. All sheep expressed hyperthermia with a peak between the 6th and the 9th day post-infection (p-i) and a transitory parasitaemia 10 days p-i. In vitro primary cultures were performed on two of these sheep, demonstrating the parasite persistence at very low parasitaemia in the infected animals. Splenectomized sheep can be used as a new model for B. divergens chronic infection.
Estrella Montero - One of the best experts on this subject based on the ideXlab platform.
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kinetics of the invasion and egress processes of Babesia divergens observed by time lapse video microscopy
Scientific Reports, 2018Co-Authors: Elena Sevilla, J S Gray, Luis Miguel Gonzalez, Daniel Luque, Estrella MonteroAbstract:Based on confocal fluorescence and bright field video microscopy, we present detailed observations on the processes of invasion and egress of erythrocytes by the apicomplexan parasite Babesia divergens. Time-lapse images reveal numerous unexpected findings associated with the dynamics of B. divergens and its ability to manipulate the erythrocyte during both processes in its asexual cycle under in vitro conditions. Despite the speed at which these processes occur and the small size of the parasite, we capture infective merozoites moving vigorously and causing striking deformations in the erythrocyte’s plasma membrane during an active invasion. We also observed intraerythrocytic dynamic stages as paired pyriforms, double paired pyriforms, tetrads, unattached pyriform sister cells and multiple parasite stages resulting in the release of large numbers of merozoites over a short period. Of considerable interest is that time-lapse images reveal a novel mechanism of egress used by B. divergens to exit the human erythrocyte. The release occurs when B. divergens parasites establish contacts with the plasma membrane of the erythrocyte from within, before exiting the cell. Visualization and analysis of the images enabled us to obtain useful information and broaden our knowledge of complex and crucial events involved with parasitisation of human erythrocytes by B. divergens.
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the efficacy of the ultraviolet c pathogen inactivation system in the reduction of Babesia divergens in pooled buffy coat platelets
Transfusion, 2014Co-Authors: Emma Castro, Luis Miguel Gonzalez, Jose M Rubio, Raquel Ramiro, Nuria Girones, Estrella MonteroAbstract:Background Babesia spp. is an intraerythrocytic parasite that causes human babesiosis and its transmission by transfusion has been extensively demonstrated. The aim of this study was to ascertain the efficacy of an ultraviolet C (UVC)-based pathogen inactivation system in the reduction of Babesia divergens–infected platelet (PLT) concentrates and to determine the parasite's ability to survive in PLT concentrates stored under blood bank conditions. Study Design and Methods This study was conducted using in vitro cultures of B. divergens. The detection limit of the culture assay was established and, subsequently, 15 buffy coat–derived PLT concentrates (BC-PCs) were inoculated with 107 B. divergens–infected red blood cells. Infected BC-PCs were irradiated with 0.2 J/cm2 UVC light using the THERAFLEX UV-Platelets method (Macopharma). Viability and parasite growth were evaluated before and after inactivation. Culture growth kinetics were monitored by DNA incorporation of [3H]thymidine. The ability of B. divergens to survive in PLT concentrates was also analyzed. Results The limit of detection in cultures was established at 0.1 × 10−6% parasites. The THERAFLEX UV-Platelets system inactivated B. divergens to below the limit of detection in 12 of 15 BC-PCs (log reduction, >6.0) and to the limit of detection (log reduction, 5.0) in three of 15. It was also demonstrated that B. divergens remains viable in BC-PCs stored up to 7 days. Conclusion Since B. divergens can survive in PLT concentrates and given the performance of UVC, this system could be considered as an alternative to prevent B. divergens and other Babesia species from being transmitted through PLT transfusions.
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a conserved subtilisin protease identified in Babesia divergens merozoites
Journal of Biological Chemistry, 2006Co-Authors: Estrella Montero, Michael J Blackman, Luis Miguel Gonzalez, Marilis Rodriguez, Yelena Oksov, Cheryl A LoboAbstract:Invasion of erythrocytes is an integral part of the Babesia divergens life cycle. Serine proteases have been shown to play an important role in invasion by related Apicomplexan parasites such as the malaria parasite Plasmodium falciparum. Here we demonstrate the presence of two dominant serine proteases in asexual B. divergens using a biotinylated fluorophosphonate probe. One of these active serine proteases (p48) and its precursors were recognized by anti-PfSUB1 antibodies. These antibodies were used to clone the gene encoding a serine protease using a B. divergens cDNA library. BdSub-1 is a single copy gene with no introns. The deduced gene product (BdSUB-1) clearly belongs to the subtilisin superfamily and shows significant homology to Plasmodium subtilisins, with the highest degree of sequence identity around the four catalytic residues. Like subtilisin proteases in other Apicomplexan parasites, BdSUB-1 undergoes two steps of processing during activation in the secretory pathway being finally converted to an active form (p48). The mature protease is concentrated in merozoite dense granules, apical secretory organelles involved in erythrocyte invasion. Anti-PfSUB1 antibodies have a potent inhibitory effect on erythrocyte invasion by B. divergens merozoites in vitro. This report demonstrates conservation of the molecular machinery involved in erythrocyte invasion by these two Apicomplexan parasites and paves the way for a comparative analysis of other molecules that participate in this process in the two parasites.