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Babesiasp. YLG has been recently described in Yellow-legged gull (Larus michahellis) chicks and belongs to the Peircei clade in the new classification of Piroplasms. Here, we studiedBabesiasp. YLG vectorial transmission by ticks in the simplified environment of a single seabird breeding colony where the Yellow-legged gull is the sole vertebrate host,Ornithodoros maritimus(syn.Alectorobius maritimus, Mans et al. 2021) the sole tick species, andBabesiasp. YLG is the unique blood parasite species previously detected in gull chicks in the colony. We collected ticks over four years, maintained certain individuals through moulting or oviposition, and dissected fresh ticks to isolate different organs and test for the presence of the parasite using molecular assays. We report the first strong evidence of a Piroplasmidae transmitted by a soft tick. Indeed,Babesiasp. YLG DNA was detected in the salivary glands of nymphs, females and males, a necessary organ to infect for transmission to a new vertebrate host. Parasite DNA was also found in tick ovaries, which could indicate possible transovarial transmission. Our detection ofBabesiasp. YLG DNA in several male testes and in endospermatophores, and notably in a parasite-free female (uninfected ovaries and salivary glands), raise the interesting possibility of sexual transmission from infected males to uninfected females. Future work in this system will now need to focus on the degree to which the parasite can be maintained locally by ticks and the epidemiological consequences of infection for bothO. maritimusand the avian host.

International audience; Avian infecting piroplasms are largely under-studied compared to other hemoparasites, and this paucity of information has blurred our phylogenetic and biological comprehension of this important group as a whole. In the present study, we detected and characterized Babesia from yellow-legged gull (Larus michahellis) chicks from a colony in southern France. Based on morphological and molecular characterizations, a new Babesia species belonging to the Peircei group, a clade of avian-specific piroplasms, was identified. Due to the complexity of species delineations and the low number of parasites characterized in this clade to date, a species name was not yet attributed; we refer to it for now as Babesia sp. YLG (Yellow-Legged Gull). High prevalence (85% and 58% in 2019 and 2020, respectively) and high parasitemia (up to 20% of parasitized erythrocytes) were recorded in chicks, without any obvious clinical signs of infection. Although the 16 isolates had identical 18S rRNA gene sequences, 6 genetic variants were described based on partial cox1 sequencing, with evidence of chicks co-infected by two variants. Transmission of Babesia sp. YLG via the soft tick Ornithodoros maritimus is discussed.

Environmental niche models predict the presence of the invasive Argentine ant in north-western Europe, especially along all the French Atlantic coast. Yet, the species has never been observed North from the 45th parallel in Europe, suggesting either that current models are wrong or that Argentine ants are already spreading north inconspicuously. Here, we report a three-hectare wide colony of Argentine ants, detected in 2016 in Nantes, France, which is 300 km north of the former northern-most outdoor population of this species in Europe. COI sequencing revealed that the haplotype of this new colony is the same as the one found in the so-called Catalonian supercolony, which is distinct from the haplotype found over most of the species range in Europe. Our discovery confirms models’ predictions that Argentine ants can colonize north-western Europe and suggests that they might have already reached several other locations along the French Atlantic coast. Detection surveys should be conducted in order to assess Argentine ants’ invasion patterns in Western France, particularly in high introduction risk areas such as major cities and maritime ports.

Introduction La Babesiose, maladie transmise par les tiques, est due a Babesia, un parasite sanguin proche de Plasmodium. L’homme ne represente qu’un hote accidentel, les differentes especes de i identifiees etant toutes zoonotiques. En Europe, B. divergens est infeode aux bovins et Babesia sp. EU1 (B. venatorum) aux chevreuils, tandis que B. microti est infeode aux rongeurs, principalement aux USA. Le tableau clinique est tres different aux USA et en Europe : aux USA, la babesiose humaine est tres largement repandue sur des patients immunodeprimes ou non avec un tableau clinique modere ; elle est rare mais severe en Europe sur individus immunodeprimes uniquement. Quelques cas moderes sur individus immunocompetents sont rapportes. Materiels et methodes De 2005 a 2014, 4694 analyses serologiques de babesiose humaine a B. divergens ont ete realisees par immunofluorescence indirecte. La lecture etant delicate, elle est realisee en double aveugle, et tout resultat discordant est re-evalue. En 2014, la technique a ete transferee vers un laboratoire prive. La detection par PCR nichee et l’identification moleculaire sans a priori des Babesia presentes dans le sang ont ete realisees sur un echantillon de prelevements pour comparer resultats serologiques et detection moleculaire. Resultats Depuis 2005, le nombre d’analyses serologiques a augmente, passant de 1 a plus de 2000 en 2014. Les cas analyses ne correspondent pas a une population representative, car les analyses concernent des patients avec historique de morsure de tiques, ou diagnostiques positifs pour la maladie de Lyme ou souffrant de symptomes tels que fatigue chronique, migraine, fievre recurrente, vertige. L’echantillon comporte 2/3 de femmes, et la majeure partie des analyses concerne les tranches d’âge 20–40 et 40–60 ans. La seroprevalence moyenne sur 10 ans est de 36,7 % (1721/4694), en augmentation depuis 10 ans, avec des titres allant du 1/8 au 1/512. Des titres superieurs a 1/16 ont ete obtenus pour pres de la moitie des cas seropositifs. Par contre, les analyses moleculaires ne permettent que tres rarement de detecter le parasite. Dans les quelques cas positifs, seule l’espece courante en Europe, B. divergens, a ete mise en evidence. Conclusion Les analyses serologiques indiquent une forte prevalence de la babesiose humaine a B. divergens en France. Elle n’est pas correlee avec le taux de detection moleculaire du parasite chez les patients, qui demeure tres faible. La serologie apporte une indication de contact de B. divergens avec le patient, qui etant le plus souvent immunocompetent elimine la parasite. Si le parasite persiste, c’est souvent a bas bruit, avec une localisation preferentielle dans les capillaires sanguins, rendant sa detection difficile. La mise au point de techniques de detection rapides et sensibles des Babesia zoonotiques chez l’homme est une necessite.

International audience; Babesia sp. BQ1 (Lintan) is one of the parasites isolated from infected sheep in China that belongs to the B. motasi-like phylogenetic group. The rhoptry-associated-protein 1 (rap-1) locus in this group consists of a complex organization of 12 genes of three main types: 6 rap-1a variants intercalated with 5 identical copies of rap-1b and a single 3′ ending rap-1c gene. In the present study, transcription analysis performed by standard RT-PCR demonstrated that the three different rap-1 gene types and the four rap-1a variants were transcribed by the parasite cultivated in vitro. Peptides, specific for each rap-1 type gene, were selected in putative linear B-epitopes and used to raise polyclonal rabbit antisera. Using these sera, the same expression pattern of RAP-1 proteins was found in parasites cultivated in vitro or collected from acute infection whereas only RAP-1a67 was detectable in merozoite extracts. However, ELISA performed with recombinant RAP-1a67, RAP-1b or RAP-1c and sera from infected sheep demonstrated that RAP-1a67 is the main RAP-1 recognized during infection, even if some infected sheep also recognized RAP-1b and/or RAP-1c

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. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1731-y) contains supplementary material, which is available to authorized users.

International audience; Sheep babesiosis occurs mainly in tropical and subtropical areas. The sheep parasite Babesia sp. Xinjiang is widespread in China, and our goal is to characterize rap-1 (rhoptry-associated protein 1) gene diversity and expression as a first step of a long term goal aiming at developing a recombinant subunit vaccine. Seven different rap-1a genes were amplified in Babesia sp. Xinjiang, using degenerate primers designed from conserved motifs. Rap-1b and rap-1c gene types could not be identified. In all seven rap-1a genes, the 5′ regions exhibited identical sequences over 936 nt, and the 3′ regions differed at 28 positions over 147 nt, defining two types of genes designated α and β. The remaining 3′ part varied from 72 to 360 nt in length, depending on the gene. This region consists of a succession of two to ten 36 nt repeats, which explains the size differences. Even if the nucleotide sequences varied, 6 repeats encoded the same stretch of amino acids. Transcription of at least four α and two β genes was demonstrated by standard RT-PCR

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 31Kb, 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.

Chantier qualité GA; Avian influenza virus (AIV) infections of the chicken occur via the respiratory route. Unlike ducks which are considered as a natural AIV reservoir, chickens are highly susceptible to AIV infections and do not possess the RIG-I pattern recognition receptor involved in triggering the antiviral interferon response. To study the chicken innate immune response to AIV in the respiratory tract, we established an epithelial cell line (CLEC213) from lung explants of white leghorn chickens. CLEC213 cells exhibited a polyhedral morphology and formed cohesive clusters bound through tight junctions as assessed by electron microscopy. Expression of E-cadherin but not vimentin could be detected as expected for cells of epithelial origin. In addition, CLEC213 cells showed characteristics similar to those of mammalian type II pneumocytes, including the presence of intracytoplasmic vacuoles filled with a mucopolysaccharide material, alkaline phosphatase activity, transcription of chicken lung collectins genes (cLL and SPA), and some intracytoplasmic lamellar-like bodies. CLEC213 cells showed a constitutive expression level of TLR3 and TLR4 and were responsive to stimulation with the respective agonists, poly (I:C) and LPS: between 4 h and 24 h after treatment, a strong increase in the expression of IFN-α, IFN-β and IL-8 genes could be detected. Furthermore, CLEC213 cells supported efficient growth of the low pathogenicity avian influenza virus H6N2 (A/duck/France/05057a/2005) in the presence or the absence of trypsin in the culture media. At 4 h post-infection, the H6N2 virus induced highly elevated levels of expression of IFN-α and IL-8, moderately elevated levels of LITAF, TGF-β4 and CCL5. However, an increase of IFN-β gene expression could not be detected in response to AIV infection. In conclusion, like mammalian type II pneumocytes, CLEC213 are able to mount a robust cytokine and chemokine immune response to microbial patterns and viral infection. We hypothesize that they could derive from lung atrial granular cells. The involvement of such type of lung epithelial cells in the respiratory tract defence of the chicken can thus be further studied.

International audience; Chemokines are key molecules that drive migration of lymphoid and myeloid cells toward organs in basal as well as inflammatory conditions. By recruiting immature dendritic cells to the mucosal surfaces, CCL20 acts in the very early events leading to the development of a specific immune response. In order to characterize dendritic cells in birds and better understand their role in the initiation of immune responses against pathogens of economic as well as human health relevance, we have cloned and expressed chicken CCL20 (chCCL20) and its specific receptor chCCR6. chCCL20 has 51% identity (60% similarity) with human CCL20, while the chicken receptor and its human counterpart display nearly 55% identity (and up to 70% similarity). chCCL20 and its specific receptor chCCR6 mRNAs are mainly expressed in bone marrow, secondary lymphoid organs and in the mucosal surfaces, in particular lungs and intestine. Both receptor and chemokine are functionally active when expressed as genuine or tagged proteins in mammalian expression systems, that is chCCR6 is mainly located at the cell surface within lipid rafts like its human counterpart. And secondly, both human and chicken chemokines were able to drive the migration of either chicken or human CCR6-transfected cells. (C) 2009 Elsevier Ltd. All rights reserved.