Babesia Divergens Research Articles

Babesia divergens glycosylphosphatidylinositols modulate blood coagulation and induce Th2-biased cytokine profiles in antigen presenting cells

Glycosylphosphatidylinositols (GPIs) are glycolipids described as toxins of protozoan parasites due to their inflammatory properties in mammalian hosts characterized by the production of interleukin (IL)-1, IL-12 and tumor necrosis factor (TNF)-α. In the present work, we studied the cytokines produced by antigen presenting cells in response to ten different GPI species extracted from Babesia divergens, responsible for babesiosis. Interestingly, B. divergens GPIs induced the production of anti-inflammatory cytokines (IL-2, IL-5) and of the regulatory cytokine IL-10 by macrophages and dendritic cells. In contrast to all protozoan GPIs studied until now, GPIs from B. divergens did not stimulate the production of TNF-α and IL-12, leading to a unique Th1/Th2 profile. Analysis of the carbohydrate composition of the B. divergens GPIs indicated that the di-mannose structure was different from the evolutionary conserved tri-mannose structure, which might explain the particular cytokine profile they induce. Expression of major histocompatibility complex (MHC) molecules on dendritic cells and apoptosis of mouse peritoneal cells were also analysed. B. divergens GPIs did not change expression of MHC class I, but decreased expression of MHC class II at the cell surface, while GPIs slightly increased the percentages of apoptotic cells. During pathogenesis of babesiosis, the inflammation-coagulation auto-amplification loop can lead to thrombosis and the effect of GPIs on coagulation parameters was investigated. Incubation of B. divergens GPIs with rat plasma ex vivo led to increase of fibrinogen levels and to prolonged activated partial thromboplastin time, suggesting a direct modulation of the extrinsic coagulation pathway by GPIs.

Laboratory diagnostic of human babesiosis.

Human babesiosis caused by parasitic protozoan Babesia spp. is sporadic zoonotic vector-borne infection. The course of babesiosis and prognosis depend on the type of pathogen and on the patient's immunological status. Significance this disease is a severe, often fatal course with immunocompromissed patients resembling complicated falciparum malaria. In Europe to date, more than 50 cases of confirmed human babesiosis have been reported in most cases caused by Babesia divergens. Possible there are unrecognized cases. Pathogen is an obligate intraerythrocyte parasite of vertebrate animals. The organism is transmitted from animal to man through bite of Ixodidae tick. Asexual reproduction of the parasite occurs in a vertebrate host. The pathogenesis of babesiosis is caused by the destruction of host cells. Intensive haemolysis of red blood cells leads to the development of haemolytic anemia, haematuria, jaundice, and polyorgan failure may develop. The clinical manifestations of the disease are nonspecific. Detection of intraerythrocyte parasites in blood smears stained Gimsa-Romanovsky confirms the proposed diagnosis. Blood smears and some laboratory signs from fatal cases were analyzed in the Reference-centre of E. I. Martsinovskiĭ Institute. Original microphotographs B. divergens are shown. The main morphological forms of the parasite are shown. In addition to the well-known tetrades of parasites «Maltese Cross», for the first time, the parasites dividing into 6 interconnected trophozoites - "sextet" - were found. Originally, the invasion of Babesia in a normoblast is shown. An unusually high multiple invasion (14 parasites) of erythrocytes is noted. Because the patients, initially, were incorrectly diagnosed with malaria, the differential diagnosis of Babesia with Plasmodium is described step-by-step. It is important, since the treatment with antimalarial drugs is ineffective. Deviation laboratory signs are discussed. Complex morphological characteristics allowed us to speciated the parasites as B. divergens. DNA was detected in the sample with specific primers Bab di hsp70F/Bab di hsp70R and the probe Bab di hsp70P. The sequence demonstrated 99-100% and 98% similarity to the 18S rRNA gene fragment of B. divergence and Babesia venatorum, respectively. Molecular biological and serological methods of laboratory diagnosis of babesiosis are considered.

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

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

Honey inhibits the in vitro growth of four Babesia species and Theileria equi

Honey has antioxidant, immunostimulant, antibacterial, and antileishmanial activities. In this study, we evaluated the in vitro babesicidal and theilericidal effects of honey on Babesia bovis, Babesia bigemina, Babesia divergens, Babesia caballi, and Theileria equi. There was noteworthy suppression of growth at a concentration of 0.5% (V/V) for B. bovis, B. bigemina, B. divergens, and T. equi and 1% (V/V) for B. caballi. The IC50 values of honey were 1.98, 1.82, 0.42, 1.7, and 1.43% (V/V) for B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi, respectively. The growth was entirely repressed at 1% (V/V) for B. divergens,2.5% (V/V) for T. equi, and 5% (V/V) for B. bovis, B. bigemina, and B. caballi. The regrowth was repressed in the viability test at a concentration of 1% (V/V) for B. divergens,2.5% (V/V) for T. equi,and5% (V/V) for B. caballi, B. bovis,and B. bigemina. These results indicate honey as a natural killer of Babesia species and T. equi. Its use in the treatment of clinical cases requires further in vivo evaluation. Honey has antioxidant, immunostimulant, antibacterial, and antileishmanial activities. In this study, we evaluated the in vitro babesicidal and theilericidal effects of honey on Babesia bovis, Babesia bigemina, Babesia divergens, Babesia caballi, and Theileria equi. There was noteworthy suppression of growth at a concentration of 0.5% (V/V) for B. bovis, B. bigemina, B. divergens, and T. equi and 1% (V/V) for B. caballi. The IC50 values of honey were 1.98, 1.82, 0.42, 1.7, and 1.43% (V/V) for B. bovis, B. bigemina, B. divergens, B. caballi, and T. equi, respectively. The growth was entirely repressed at 1% (V/V) for B. divergens,2.5% (V/V) for T. equi, and 5% (V/V) for B. bovis, B. bigemina, and B. caballi. The regrowth was repressed in the viability test at a concentration of 1% (V/V) for B. divergens,2.5% (V/V) for T. equi,and5% (V/V) for B. caballi, B. bovis,and B. bigemina. These results indicate honey as a natural killer of Babesia species and T. equi. Its use in the treatment of clinical cases requires further in vivo evaluation.

Comparative and functional genomics of the protozoan parasite Babesia divergens highlighting the invasion and egress processes.

Babesiosis is considered an emerging disease because its incidence has significantly increased in the last 30 years, providing evidence of the expanding range of this rare but potentially life-threatening zoonotic disease. Babesia divergens is a causative agent of babesiosis in humans and cattle in Europe. The recently sequenced genome of B. divergens revealed over 3,741 protein coding-genes and the 10.7-Mb high-quality draft become the first reference tool to study the genome structure of B. divergens. Now, by exploiting this sequence data and using new computational tools and assembly strategies, we have significantly improved the quality of the B. divergens genome. The new assembly shows better continuity and has a higher correspondence to B. bovis chromosomes. Moreover, we present a differential expression analysis using RNA sequencing of the two different stages of the asexual lifecycle of B. divergens: the free merozoite capable of invading erythrocytes and the intraerythrocytic parasite stage that remains within the erythrocyte until egress. Comparison of mRNA levels of both stages identified 1,441 differentially expressed genes. From these, around half were upregulated and the other half downregulated in the intraerythrocytic stage. Orthogonal validation by real-time quantitative reverse transcription PCR confirmed the differential expression. A moderately increased expression level of genes, putatively involved in the invasion and egress processes, were revealed in the intraerythrocytic stage compared with the free merozoite. On the basis of these results and in the absence of molecular models of invasion and egress for B. divergens, we have proposed the identified genes as putative molecular players in the invasion and egress processes. Our results contribute to an understanding of key parasitic strategies and pathogenesis and could be a valuable genomic resource to exploit for the design of diagnostic methods, drugs and vaccines to improve the control of babesiosis.

Severe babesiosis caused by Babesia divergens in a host with intact spleen, Russia, 2018

We report a case of severe babesiosis caused by the bovine pathogen Babesia divergens with the development of multisystem failure in a splenic host. Immunosuppression other than splenectomy can also predispose people to B. divergens. There was heavy multiple invasion of up to 14 parasites inside the erythrocyte, which had not been previously observed even in asplenic hosts. The piroplasm 18S rRNA sequence from our patient was identical B. divergens EU lineage with identity 99.5–100%.

Babesia divergens: A Drive to Survive

Babesia divergens is an obligate intracellular protozoan parasite that causes zoonotic disease. Central to its pathogenesis is the ability of the parasite to invade host red blood cells of diverse species, and, once in the host blood stream, to manipulate the composition of its population to allow it to endure unfavorable conditions. Here we will review key in vitro studies relating to the survival strategies that B. divergens adopts during its intraerythrocytic development to persist and how proliferation is restored in the parasite population once optimum conditions return.

WITHDRAWN: Babesia divergens glycosylphosphatidylinositols modulate blood coagulation and induce Th2-biased cytokine profiles in antigen presenting cells

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In vitro cultivation of Babesia duncani (Apicomplexa: Babesiidae), a zoonotic hemoprotozoan, using infected blood from Syrian hamsters (Mesocricetus auratus).

Human babesiosis, a tick-borne disease similar to malaria, is most often caused by the hemoprotozoans Babesia divergens in Europe, and Babesia microti and Babesia duncani in North America. Babesia microti is the best documented and causes more cases of human babesiosis annually than all other agents combined. Although the agents that cause human babesiosis are considered high-risk pathogens in transfusion medicine, federally licensed diagnostics are lacking for B. duncani in both the USA and Canada. Thus, there has been a need to develop and validate diagnostics specifically for this pathogen. In this study, B. duncani (WA1 isolate) was cultivated in vitro from Syrian hamster (Mesocricetus auratus) infected blood. We hypothesized HL-1 media with supplements would result in B. duncani propagating at higher levels in culture than supplemented M199 similar to the medium the parasite was originally cultivated with in 1994. We were unable to recreate Thomford's cultivation results with the M199 medium but supplemented HL-1 medium was able to successfully establish continuous culture. We further hypothesized that RBC from species other than hamsters would support B. duncani in vitro. However, rat, mouse, horse, and cow RBC did not support continuous culture of the parasite. Culture stocks of B. duncani were deposited at BEI Resources and are now commercially available to the scientific community to further research. The cultured parasite developed in this study was instrumental in the adaptation of B. duncani continuous culture to human RBC.

ANAPLASMOSIS AND BABESIOSIS OF CATTLE IN CENTRAL REGION OF RUSSIAN FEDERATION

An epizootological monitoring of anaplasmosis and babesiosis of cattle of Aberdeen-Angus breed in the Bryansk region was carried out. Acarological and entomological studies made it possible to detect Dermacentor reticulatus ixodic ticks on animals in spring, and Tabanus bovinus, T. sudeticus, Chrysops spp., Haematopota spp. and blood-sucking zoophilic flies Stomoxys calcitrans, Haematobia spp. summer and early spring. The higher number of gadflies and bloodsucking flies (mechanical carriers of Anaplasma marginale), compared with ixodic ticks (Babesia divergens biological carriers) in a number of areas of the Bryansk region, explains the widespread anaplasmosis and high rates of parasitemia. The acute course of anaplasmosis and babesiosis in a mixed form is characterized by persistent fever, decreased or no appetite, ataxia, tachycardia, shortness of breath, mucous membrane anemia. According to the results of research, the maximum intensity of the epizootic process in anaplasmosis and babesiosis was found in the Trubchevskiy district of the Bryansk region, which is caused by the numerous biotopes of Ixodic ticks Dermacentor reticulatus.In the clinically expressed form of anaplasmosis and babesiosis, 1% oxytetracycline (LA 200) and Babezan 12% intramuscularly were administered to animals once. For the prevention of diseases in young stock, taking into account the absence of non-sterile immunity, during the period of activity of ixodic ticks and blood-sucking dipter, the above preparations were prescribed three times: in spring (March-April), in summer (late June-July) with high rainfall and in autumn (September-October).In cases of severe anaplasmosis and babesiosis Oxytetracycline 1% (LA 200) and Babesan, 12% are prescribed simultaneously in the mixed form with the means of pathogenetic and symptomatic therapy. With moderate and severe babesiosis accompanied by fever, depression, and lack of appetite, Babezan 12% is used fractionally, i.e. 50% of the therapeutic dose on the day of manifestation of the first clinical signs and the rest in 18–20 hours.

Altered parasite life-cycle processes characterize Babesia divergens infection in human sickle cell anemia.

Babesia divergens is an intra-erythrocytic parasite that causes malaria-like symptoms in infected people. As the erythrocyte provides the parasite with the infra-structure to grow and multiply, any perturbation to the cell should impact parasite viability. Support for this comes from the multitude of studies that have shown that the sickle trait has in fact been selected because of the protection it provides against a related Apicomplexan parasite, Plasmodium, that causes malaria. In this paper, we examine the impact of both the sickle cell anemia and sickle trait red blood cell (RBC) environment on different aspects of the B. divergens life-cycle, and reveal that multiple aspects of parasite biological processes are altered in the mutant sickle anemia RBC. Such processes include parasite population progression, caused potentially by defective merozoite infectivity and/or defective egress from the sickle cell, resulting in severely lowered parasitemia in these cells with sickle cell anemia. In contrast, the sickle trait RBC provide a supportive environment permitting in vitro infection rates comparable to those of wild-type RBC. The elucidation of these naturally occurring RBC resistance mechanisms is needed to shed light on host-parasite interaction, lend evolutionary insights into these related blood-borne parasites, and to provide new insights into the development of therapies against this disease.

Detection of pathogens in Dermacentor reticulatus in northwestern Europe: evaluation of a high-throughput array

BackgroundThe geographic distribution of Dermacentor reticulatus is expanding in Europe. Surveillance of this tick species and its pathogens is desirable, as it transmits pathogens of public and veterinary importance. A high-throughput real-time PCR-based array was used to screen 1.741 D. reticulatus ticks from Belgium, Germany, The Netherlands, and Great Britain for the presence of 28 tick-borne bacteria and twelve protozoan parasites. The presence of pathogen DNA was confirmed by conventional PCR followed by sequencing. ResultsThe array detected the presence of DNA from Borrelia spp. (7%), B. afzelii (0.1%), B. garinii (0.1%), B. spielmanii (0.1%), B. miyamotoi (0.2%), Anaplasma marginale (0.1%), A. phagocytophilum (0.1%), Ehrlichia canis (2%), Rickettsia helvetica (0.2%), spotted fever group Rickettsia (9.6%), Francisella tularensis or Francisella-like endosymbionts (95%), Coxiella burnettii (0.1%), Babesia divergens (0.2%), B. canis (0.9%) B. vogeli (5.6%), and Theileria equi (0.1%). Only the presence of B. canis and spotted fever group Rickettsia could be confirmed by conventional PCR and sequencing. The spotted fever Rickettsia-positive samples were all identified as R. raoultii. ConclusionsWe successfully detected and determined the prevalence of B. canis and R. raoultii in D. reticulatus. An high-throughput array that allows fast and comprehensive testing of tick-borne pathogens is advantageous for surveillance and future epidemiological studies. The importance of thorough validation of real-time PCR-based assays and careful interpretation is evident.

A preliminary molecular survey of Babesia divergens and first evidence of Theileria annulata in cattle from Saudi Arabia.

Background and Aim:Babesia divergens causes human babesiosis in Europe where the parasite utilizes cattle as animal reservoir and Ixodes ricinus as tick vector. Importation of infected animals and passive carriage of infected ticks through migratory birds can lead to tick/pathogen geographic expansion and emergence of diseases in naïve land. Given the information that Saudi Arabia imports cattle from the European countries and that two global bird flyways pass through the country geographic coordinates, we speculate that B. divergens might be introduced into the Kingdom. Therefore, the aim of this preliminary study was to molecularly detect and characterize B. divergens and other piroplasms (including Theileria spp.) in cattle from Taif district, Kingdom of Saudi Arabia.Materials and Methods:Blood samples from 20 cattle residing Taif district were collected, and polymerase chain reaction tested using wide and species-specific primers. Amplicons from a positive genus-wide reaction were purified, sequenced, and analyzed. Phylogenetic trees were constructed, and similarity to existing GenBank zoonotic piroplasms was also assessed.Results:All samples were negative for B. divergens, and only one sample proved positive for Theileria annulata in a wide reaction. Phylogeny clustered our strain with T. annulata from Spanish dog and another one detected in a cow from France. BLAST analysis showed genetic distance from zoonotic piroplasms with identity ranged from 88% to 91%.Conclusion:Although B. divergens was not detected, we are not able to rule out or affirm the existence of the pathogen in the country. On the other hand, identifying T. annulata strain with a southern European origin strongly supports our speculation that bovine zoonotic Babesia might be introduced into KSA. This study is not only the first molecular survey of B. divergens but also the first report of the molecular identity of T. annulata in Saudi Arabia. A national-wide bovine and tick surveillance are needed to further prove our speculation.

Evaluation of the in vitro and in vivo inhibitory effect of thymoquinone on piroplasm parasites

BackgroundDeveloping new antibabesial drugs with a low toxic effect to the animal and with no resistance from Babesia parasites is in urgent demand. In this concern, the antimalarial, anticancer and antioxidant effect of thymoquinone (TQ), a phytochemical compound found in the plant Nigella sativa, has been reported. Therefore, in the present study, the antibabesial effect of this compound was evaluated on the growth of piroplasm parasites.ResultsSignificant inhibition (P < 0.05) of the in vitro growth of piroplasm parasites were observed after treatment by TQ with IC50 values of 35.41 ± 3.60, 7.35 ± 0.17, 0.28 ± 0.016, 74.05 ± 4.55 and 67.33 ± 0.94 μM for Babesia bovis, Babesia bigemina, Babesia divergens, Theileria equi and Babesia caballi, respectively. The in vitro inhibitory effect of TQ was significantly enhanced (P < 0.05) when used in combination with either diminazene aceturate on bovine Babesia and equine Babesia and Theileria cultures. In B. microti-infected mice, oral and intraperitoneal administrations of TQ showed significant (P < 0.05) inhibition of parasite growth at a dose of 70 mg/kg and 50 mg/kg, respectively, compared to the control group.ConclusionsThe obtained results indicate that thymoquinone might be a promising medicinal compound for use in the treatment of animal piroplasmosis.

Tick-borne pathogens in Finland: comparison of Ixodes ricinus and I. persulcatus in sympatric and parapatric areas

BackgroundAlmost 3500 tick samples, originally collected via a nationwide citizen science campaign in 2015, were screened to reveal the prevalence and distribution of a wide spectrum of established and putative tick-borne pathogens vectored by Ixodes ricinus and I. persulcatus in Finland. The unique geographical distribution of these two tick species in Finland allowed us to compare pathogen occurrence between an I. ricinus-dominated area (southern Finland), an I. persulcatus-dominated area (northern Finland), and a sympatric area (central Finland).ResultsOf the analysed ticks, almost 30% carried at least one pathogen and 2% carried more than one pathogen. A higher overall prevalence of tick-borne pathogens was observed in I. ricinus than in I. persulcatus: 30.0% (604/2014) versus 24.0% (348/1451), respectively. In addition, I. ricinus were more frequently co-infected than I. persulcatus: 2.4% (49/2014) versus 0.8% (12/1451), respectively. Causative agents of Lyme borreliosis, i.e. bacterial genospecies in Borrelia burgdorferi (sensu lato) group, were the most prevalent pathogens (overall 17%). “Candidatus Rickettsia tarasevichiae” was found for the first time in I. ricinus ticks and in Finnish ticks in general. Moreover, Babesia divergens, B. venatorum and “Candidatus Neoehrlichia mikurensis” were reported for the first time from the Finnish mainland.ConclusionsThe present study provides valuable information on the prevalence and geographical distribution of various tick-borne pathogens in I. ricinus and I. persulcatus ticks in Finland. Moreover, this comprehensive subset of ticks revealed the presence of rare and potentially dangerous pathogens. The highest prevalence of infected ticks was in the I. ricinus-dominated area in southern Finland, while the prevalence was essentially equal in sympatric and I. persulcatus-dominated areas. However, the highest infection rates for both species were in areas of their dominance, either in south or north Finland.

Optimizing exchange transfusion for patients with severe Babesia divergens babesiosis: Therapeutically-Rational Exchange (T-REX) of M antigen-negative and/or S antigen-negative red blood cells should be evaluated now

Babesia divergens is an intraerythrocytic parasite, which is the major cause of babesiosis in Europe. For years, clinicians have been publishing stunning case reports that describe how some – but not all – conventional red blood cell (RBC) exchange transfusions have saved the lives of severely ill babesiosis patients. Due to markedly different patient outcomes, clinicians agree that new treatments and additional studies are needed. Here we argue that we should evaluate “therapeutically-rational exchange” (T-REX) in which the RBCs used to replace Babesia-parasitized RBCs are special disease-resistant RBC genetic variants (instead of the nondescript, “standard issue” RBCs used in conventional exchanges). T-REX seems prudent because with conventional exchange only some units of “standard issue” RBCs may be disease-resistant, while other units may not protect or may even promote disease. The random selection of RBCs for conventional RBC exchange may explain why clinical outcomes can vary dramatically. Fortunately, researchers have found that M antigen-negative (M−) and S antigen-negative (S−) RBCs resist invasion by B. divergens. Thus, we recommend evaluating T-REX of RBC variants that are B. divergens invasion-resistant: RBCs that are (1) M−, (2) S−, or (3) both M− and S−. By using only Babesia-resistant RBCs, T-REX eliminates the risk of unintentionally infusing Babesia-susceptible RBCs that might increase the severity of babesiosis. Because the T-REX variation of the conventional RBC exchange procedure is feasible, safe, and biologically plausible, we feel T-REX of Babesia-resistant RBCs should now be evaluated.

Kinetics of the invasion and egress processes of Babesia divergens, observed by time-lapse video microscopy

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.

Inhibitory effects of fluoroquinolone antibiotics on Babesia divergens and Babesia microti, blood parasites of veterinary and zoonotic importance.

AimThis study aimed to evaluate the inhibitory effects of fluoroquinolone antibiotics, including enrofloxacin, enoxacin, trovafloxacin, norfloxacin, and ofloxacin, on the in vitro and in vivo growth of Babesia divergens and Babesia microti parasites, respectively.Materials and methodsThe in vitro and in vivo inhibitory effects of fluoroquinolone antibiotics against B. divergens and B. microti, respectively were evaluated using fluorescence-based assay. Additionally, combination therapies of highly effective fluoroquinolone antibiotics (enrofloxacin, enoxacin, and trovafloxacin) with diminazene aceturate, luteolin, or pyronaridine tetraphosphate were tested on the in vitro cultures of B. divergens.ResultsEnrofloxacin, trovafloxacin, and enoxacin were the most effective fluoroquinolones against the in vitro growth of B. divergens, followed by norfloxacin and ofloxacin. Furthermore, a combination of enoxacin or trovafloxacin with either diminazene aceturate, luteolin, or pyronaridine tetraphosphate significantly enhanced the inhibitory effect on the growth of B. divergens in in vitro cultures. In mice infected by B. microti, enoxacin and diminazene aceturate combination therapy exhibited a potential antibabesial effect.ConclusionThese results suggest that safe and cheap fluoroquinolone, such as enoxacin, might be used for the treatment of clinical cases caused by Babesia spp. in animals or humans.

Validation of Babesia proteasome as a drug target

Babesiosis is a tick-transmitted zoonosis caused by apicomplexan parasites of the genus Babesia. Treatment of this emerging malaria-related disease has relied on antimalarial drugs and antibiotics. The proteasome of Plasmodium, the causative agent of malaria, has recently been validated as a target for anti-malarial drug development and therefore, in this study, we investigated the effect of epoxyketone (carfilzomib, ONX-0914 and epoxomicin) and boronic acid (bortezomib and ixazomib) proteasome inhibitors on the growth and survival of Babesia. Testing the compounds against Babesia divergens ex vivo revealed suppressive effects on parasite growth with activity that was higher than the cytotoxic effects on a non-transformed mouse macrophage cell line. Furthermore, we showed that the most-effective compound, carfilzomib, significantly reduces parasite multiplication in a Babesia microti infected mouse model without noticeable adverse effects. In addition, treatment with carfilzomib lead to an ex vivo and in vivo decrease in proteasome activity and accumulation of polyubiquitinated proteins compared to untreated control. Overall, our results demonstrate that the Babesia proteasome is a valid target for drug development and warrants the design of potent and selective B. divergens proteasome inhibitors for the treatment of babesiosis.

Manifestations ostéo-articulaires secondaires à une piqûre, griffure, morsure ou à germes inhabituels ou émergents

Osteoarticular manifestations after a sting, a bite or a scratch are rare but the diagnosis should not be neglected. Targeted antibiotic therapy is mandatory. Surgery might help the diagnosis with adequate biopsy and is sometimes essential for recovery. A detective work is needed to identify a history of wound. Pantoea agglomerans (P. agglomerans) and nocardiosis are seen after a telluric wound, Capnocytophaga canimorsus (C. canimorsus) and Pasteurella multocida (P. multocida) after a dog bite, Bartonella henselae (B. henselae) and P. multocida after a cat scratch, Erysipelothrix rhusopathiae (E. rhusopathiae) when the wound is caused by a pig bone or a fishbone. A contact with water soiled with rat urine during work or leisure activity could indicate infection with Streptobacillus moniliformis (S. moniliformis) Spirillum minus (S. minus), Leptospira interrogans (L. interrogans). Bacteriological documentation might be difficult but is often mandatory to ascertain diagnosis and to adapt antibiotics. A sting can be responsible for a non-infectious arthritis. Inflammation is directly mediated by foreign body and is called “thorn arthritis”. Foreign body is often not seen on conventional radiography, hardly seen in ultrasonography, and surgery is needed to extract it and sometimes to perform synovectomy. Atypical mycobacteria are seen in patients with immunodepression but not only. Contact with fish or fishing material brings Mycobacterium marinum (M. marinum) to mind and a history of joint injection evoke Mycobacterium kansasii (M. kansasii). Surgery is frequently needed for diagnosis and treatment of tenosynovitis or arthritis caused by atypical mycobacteria. Tick-borne arthritis are mainly caused by Borrelia burgdorferi but can also involve Babesia divergens or Rickettsia conorii. Among arthropod-borne viruses, chikungunya provides a model of infection-triggered inflammatory joint disease.