Publications by authors named "Henri-Jean Boulouis"

73 Publications

Sensitivity of bovine tuberculosis surveillance through intradermal tests in cattle in France: An evaluation of different scenarios.

Prev Vet Med 2021 Apr 24;191:105364. Epub 2021 Apr 24.

INRAE, Oniris, BIOEPAR, 44300, Nantes, France. Electronic address:

The current situation regarding bovine tuberculosis (bTB) in Europe is spatially heterogeneous, with stagnating or increasing trends in bTB prevalence in many European regions, underlying the challenge in controlling this disease. In France, in spite of the implementation of two control programs in 2010-2012 to eradicate the disease and maintain the bTB-free status, bTB prevalence has continued to increase, underlying the need to reinforce and adapt surveillance measures. The goal of this study was to evaluate the effectiveness of bTB surveillance in high-risk areas in metropolitan France, with an emphasis on the criteria to select herds and animals within herds in the context of programmed surveillance and movement testing. The fraction of bTB-infected herds detected by the surveillance was quantified using a stochastic scenario tree modelling approach, with input parameter values based on surveillance and cattle traceability data and literature. The detection fraction was assessed for the current surveillance system and for alternative scenarios. The model predicted that the median detection fraction of infected herds by the current programmed surveillance in high-risk areas, which consists in annual testing of herds with a minimum age of testing of 24 months, was 71.5 % (interquartile interval: 47.4-89.4). The results showed a significant gain of the detection fraction with a decrease from 24 to 12 months old (83.5 % [60.6-95.9]) or to six weeks old (91.3 % [71.6-99.0]). Regarding pre-movement surveillance, tests are currently mandatory for bovines that originate from a previously infected herd or from a herd epidemiologically linked to a bTB-infected herd. The median detection fraction predicted by the model for this surveillance scenario was 1.2 % [0.7-1.8]. For the alternative scenario, where surveillance would be extended to all herds in high-risk areas, the model predicted a significant increase of the detection fraction to 26.5 % [18.1-37.9]. The results were sensitive to the following input values: the number of infected bovines within herds and, to a lower extent, the comparative intradermal tuberculin test sensitivity for both models, and surveillance coverage for the model on pre-movement surveillance. Our study underlines several complementary ways to improve the detection of infected herds, which is critical for implementing control measures and epidemiological investigations as early as possible. These necessary changes in surveillance must be accompanied by a global reflexion on surveillance financing.
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http://dx.doi.org/10.1016/j.prevetmed.2021.105364DOI Listing
April 2021

High-Throughput Microfluidic Real-Time PCR for the Detection of Multiple Microorganisms in Ixodid Cattle Ticks in Northeast Algeria.

Pathogens 2021 Mar 18;10(3). Epub 2021 Mar 18.

Laboratoire de Santé Animale, Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR, ANSES, INRAE, F-94700 Maisons-Alfort, France.

Ixodid ticks are hematophagous arthropods considered to be prominent ectoparasite vectors that have a negative impact on cattle, either through direct injury or via the transmission of several pathogens. In this study, we investigated the molecular infection rates of numerous tick-borne pathogens in ticks sampled on cattle from the Kabylia region, northeastern Algeria, using a high-throughput microfluidic real-time PCR system. A total of 235 ticks belonging to seven species of the genera , , and were sampled on cattle and then screened for the presence of 36 different species of bacteria and protozoans. The most prevalent tick-borne microorganisms were spp. at 79.1%, followed by -like endosymbionts (62.9%), spp. (17.8%), spp. (14.4%), spp. (6.8%), spp. (6.8%), and spp. (2.5%). Among the 80.4% of ticks bearing microorganisms, 20%, 36.6%, 21.7%, and 2.1% were positive for one, two, three, and four different microorganisms, respectively. was detected in , , and ticks. was found in , and and were detected in . was found in all identified tick genera, but was detected exclusively in spp. ticks. The DNA of spp. and spp. was identified in several tick species. was found in , , , , and and was found in and . Our study highlights the importance of tick-borne pathogens in cattle in Algeria.
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http://dx.doi.org/10.3390/pathogens10030362DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8002991PMC
March 2021

Enlisting the Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick-Pathogen Interactions.

Pathogens 2021 Jan 14;10(1). Epub 2021 Jan 14.

UMR BIPAR, Laboratoire de Santé Animale, ANSES, INRAE, Ecole Nationale Vétérinaire d'Alfort, Paris-Est Sup, 94700 Maisons-Alfort, France.

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like embryonic cell line, ISE6, is an effective tool frequently used for examining tick-pathogen interactions. We detected 37 neuropeptide transcripts in the ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, . The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in -infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick-pathogen interactions.
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http://dx.doi.org/10.3390/pathogens10010070DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7828734PMC
January 2021

Bovines Harbor a Diverse Array of Vector-Borne Pathogens in Northeast Algeria.

Pathogens 2020 Oct 25;9(11). Epub 2020 Oct 25.

UMR BIPAR, National Veterinary School of Alfort, Anses, INRAE, Paris-Est University, 7 Avenue du Général de Gaulle, 94700 Maisons-Alfort, France.

Arthropod-borne hemoparasites represent a serious health problem in livestock, causing significant production losses. Currently, the evidence of spp., spp., spp., and hemotropic spp. in Algeria remains limited to a few scattered geographical regions. In this work, our objectives were to study the prevalence of these vector-borne pathogens and to search other agents not yet described in Algeria as well as the identification of statistical associations with various risk factors in cattle in the northeast of Algeria. Among the 205 cattle blood samples tested by PCR analysis, 42.4% positive results were obtained for at least one pathogen. The overall rates of spp., / spp., and spp. in the cattle sampled were respectively 30.7%, 18.5%, and 2.9%; co-infections with multiple species was also detected. spp. and spp. were detected at a higher rate in cattle under 3 years old, according to univariate analysis. spp. DNA was detected more frequently in our sample in cattle living in semi extensive farming. Our study provides additional data about spp., spp. and reveals for the first time that and ' Mycoplasma hemobos are present in cattle in Northeast Algeria.
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http://dx.doi.org/10.3390/pathogens9110883DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7692033PMC
October 2020

First report on Bartonella henselae in dromedary camels (Camelus dromedarius).

Infect Genet Evol 2020 11 7;85:104496. Epub 2020 Aug 7.

Service de Microbiologie et Immunologie, Ecole Nationale de Médecine Vétérinaire, Univ. Manouba, Sidi Thabet, Tunisie. Electronic address:

Bartonellosis is one of the clinically underdiagnosed emerging bacterial diseases among domestic livestock, particularly in camels. Until now, the natural infection of camels with Bartonella species was not investigated in Tunisia. In the attempt of filling this gap in knowledge, a total of 412 dromedary camels (Camelus dromedarius) as well as 300 associated ticks (Hyalomma dromedarii (160; 53.4%), H. impeltatum (131; 43.6%) and H. excavatum (9; 3%) were screened for the presence of Bartonella spp. by PCR followed by a sequencing step through the amplification of the rpoB gene. Positive samples were then tested and further characterized by the combined use of the ftsZ and gltA genes. Fifteen camels (3.6%) were found to be positive to Bartonella spp. However, there was no evidence of Bartonella DNA in any of the analyzed ticks. Risk factors' analysis shows that camels derived from arid and semi-arid bioclimatic areas were more infected than those originated from desert area. Molecular characterization and phylogenetic analysis revealed the occurrence of novel B. henselae genotypes closely related to those isolated from humans, cats, and lions. By combining the characteristics of each single gene with those of concatenated sequences, we report here the first molecular detection of B. henselae in the dromedary camel suggesting a possible involvement of camelids as hosts or reservoirs in the transmission cycle of this emerging bacterium in arid and saharan areas.
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http://dx.doi.org/10.1016/j.meegid.2020.104496DOI Listing
November 2020

Multiple locus variable number tandem repeat analysis for the characterization of wild feline Bartonella species and subspecies.

Vet Microbiol 2020 Aug 17;247:108788. Epub 2020 Jul 17.

UMR BIPAR 956, Ecole nationale vétérinaire d'Alfort, Anses, INRA, Université Paris-Est, Maisons-Alfort, France.

Bartonella genus includes an increasing number of species and subspecies, especially among wild felids, the positioning of which, with regards to the zoonotic species Bartonella henselae, is important to determine. The aim of this study was to test the ability of a molecular typing technique to distinguish between various Bartonella isolates obtained from four different species of free-ranging and captive wild felids and to identify key profiles or markers allowing differentiating them from each other and/or from B. henselae or B. koehlerae. A molecular typing technique for B. henselae based on the polymorphism of variable number tandem repeat units (VNTR) called MLVA (Multiple Locus VNTR Analysis) was applied to 24 Bartonella isolates from free-ranging or captive wild felids, 19 of which were obtained from California and five from three countries in Southern Africa, and compared with 49 B. henselae isolates from cats, dog or humans from the United States including the human ATCC (American Type Culture Collection) reference strain, B. henselae Houston 1. MLVA allowed distinguishing Bartonella isolates from wild felids from either B. henselae or B. koehlerae. We confirmed infection of semi-captive cheetahs with an isolate similar to a Californian bobcat isolate. MLVA also confirmed the unique profile of a free-ranging cheetah isolate from Namibia. Specific profiles were observed making MVLA a useful identification/classification tool of these wild felid isolates and suggesting that they are highly adapted to a specific feline reservoir. Finally, circulation of B. henselae isolates between domestic cats, wild felids and humans is likely occurring, based on the close allelic profiles of some isolates.
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http://dx.doi.org/10.1016/j.vetmic.2020.108788DOI Listing
August 2020

Bovine Organospecific Microvascular Endothelial Cell Lines as New and Relevant In Vitro Models to Study Viral Infections.

Int J Mol Sci 2020 Jul 24;21(15). Epub 2020 Jul 24.

Center for Molecular Biophysics UPR4301 CNRS, 45000 Orléans, France.

Microvascular endothelial cells constitute potential targets for exogenous microorganisms, in particular for vector-borne pathogens. Their phenotypic and functional variations according to the organs they are coming from provide an explanation of the organ selectivity expressed in vivo by pathogens. In order to make available relevant tools for in vitro studies of infection mechanisms, our aim was to immortalize bovine organospecific endothelial cells but also to assess their permissivity to viral infection. Using transfection with SV40 large T antigen, six bovine microvascular endothelial cell lines from various organs and one macrovascular cell line from an umbilical cord were established. They display their own panel of endothelial progenitor/mature markers, as assessed by flow cytometry and RT-qPCR, as well as the typical angiogenesis capacity. Using both Bluetongue and foot-and-mouth disease viruses, we demonstrate that some cell lines are preferentially infected. In addition, they can be transfected and are able to express viral proteins such as BTV8-NS3. Such microvascular endothelial cell lines bring innovative tools for in vitro studies of infection by viruses or bacteria, allowing for the study of host-pathogen interaction mechanisms with the actual in vivo target cells. They are also suitable for applications linked to microvascularization, such as anti-angiogenic and anti-tumor research, growing fields in veterinary medicine.
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http://dx.doi.org/10.3390/ijms21155249DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7432920PMC
July 2020

Pathogenic in Dogs Reveals the Predominance of ST372 and the Human-Associated ST73 Extra-Intestinal Lineages.

Front Microbiol 2020 21;11:580. Epub 2020 Apr 21.

Unité Antibiorésistance et Virulence Bactériennes, ANSES Laboratoire de Lyon - Université de Lyon, Lyon, France.

is a ubiquitous commensal and pathogen that has also been recognized as a multi-sectoral indicator of antimicrobial resistance (AMR). Given that latter focus, such as on resistances to extended-spectrum cephalosporins (ESC) and carbapenems, the reported population structure of is generally biased toward resistant isolates, with sequence type (ST)131 being widely reported in humans, and ST410 and ST648 being reported in animals. In this study, we characterized 618 non-duplicate isolates collected throughout France independently of their resistance phenotype. The B2 phylogroup was over-represented (79.6%) and positively associated with the presence of numerous virulence factors (VFs), including those defining the extra-intestinal pathogenic isolates (presence of ≥2 VFs: , , , , and ) and those more specifically related to uropathogenic (, ). The major STs associated with clinical isolates from dogs were by far the dog-associated ST372 (20.7%) and ST73 (20.1%), a lineage that had commonly been considered until now as human-associated. Resistance to ESC was found in 33 isolates (5.3%), along with one carbapenemase-producing isolate, and was mostly restricted to non-B2 isolates. In conclusion, the presence of virulent lineages may be the issue, rather than the presence of ESC-resistant isolates, and the risk of transmission of such virulent isolates to humans needs to be further studied.
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http://dx.doi.org/10.3389/fmicb.2020.00580DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7186358PMC
April 2020

Experimental -Sheep Cycle of NV2Os Propagated in Tick Cell Cultures.

Front Vet Sci 2020 6;7:40. Epub 2020 Feb 6.

UMR BIPAR, INRAE, Ecole Nationale Vétérinaire d'Alfort, ANSES, Université Paris-Est, Maisons-Alfort, France.

The causative agent of tick-borne fever and human granulocytic anaplasmosis, , is transmitted by , and is currently considered an emerging disease throughout Europe. In this study, we established a model of sheep infection and transmission using the European Norway variant 2 ovine strain (NV2Os) propagated in both IDE8 and ISE6 tick cells. Two sheep were inoculated with IDE8 tick cells infected with NV2Os. Both sheep developed infection as determined by qPCR and PCR, the presence of fever 4 days post inoculation (dpi), the observation of morulae in granulocytes at 6 dpi, and the detection of antibodies at 14 dpi. was detected by PCR in skin, lung, small intestine, liver, spleen, uterus, bone marrow, and mesenteric lymph node from necropsies performed at 14 and 15 dpi. One sheep was infested during the acute phase of infection with nymphs from a pathogen-free colony. After molting, transstadial transmission in ticks was validated with qPCR positive bacterial detection in 80% of salivary glands and 90% of midguts from female adults. Infected sheep blood collected at 14 dpi was demonstrated to be able to infect ISE6 tick cells, thus enabling the infection of two additional naive sheep, which then went on to develop similar clinical signs to the sheep infected previously. One of the sheep remained persistently infected until 115 dpi when it was euthanized, and transmitted bacteria to 70 and 2.7% of nymphs engorged as larvae during the acute and persistent infection stages, respectively. We then demonstrated that these infected nymphs were able to transmit the bacteria to one of two other naive infested sheep. As expected, when females were engorged during the acute phase of infection, no transovarial transmission was detected. The development of this new experimental model will facilitate future research on this tick-borne bacterium of increasing importance, and enable the evaluation of any new tick/transmission control strategies.
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http://dx.doi.org/10.3389/fvets.2020.00040DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7015893PMC
February 2020

Infection in Stray Dogs from Central and Southern Chile (Linares and Puerto Montt).

Vector Borne Zoonotic Dis 2020 03 29;20(3):187-192. Epub 2019 Oct 29.

Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, Davis, California.

Bartonellae are emerging zoonotic vector-borne pathogens causing a broad spectrum of clinical symptoms in humans and animals, including life-threatening endocarditis. Dogs are infected with a wide range of species and infection has been reported in free-roaming dogs from various South American countries. We report a high seroprevalence in 82 Chilean stray dogs. More than half of the dogs from Linares (72.7%,  = 66) and Puerto Montt (56.2%,  = 16) were seropositive for , ssp. , or with antibody titers ranging from 1:64 to 1:512. Three dogs (3.6%) were PCR positive for sp. Partial sequencing of the A gene indicated that two dogs were infected with , and one with a strain close to ssp. . Exposure to species was common in stray Chilean dogs, as for other South American countries, likely associated with heavy ectoparasite infestation.
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http://dx.doi.org/10.1089/vbz.2019.2505DOI Listing
March 2020

Genomic characterisation of a multidrug-resistant TEM-52b extended-spectrum β-lactamase-positive Escherichia coli ST219 isolated from a cat in France.

J Glob Antimicrob Resist 2019 09 17;18:223-224. Epub 2019 Jul 17.

Department of Microbiology, Institute of Biomedical Sciences, Universidade de São Paulo, São Paulo, Brazil; Department of Clinical Analysis, School of Pharmacy, Universidade de São Paulo, São Paulo, Brazil. Electronic address:

Objectives: TEM-52 extended-spectrum β-lactamases (ESBLs) have been detected in members of the Enterobacteriaceae isolated from human and non-human reservoirs, mainly in European countries. Here we report the first draft genome of a multidrug-resistant TEM-52b-positive Escherichia coli isolated from a companion animal in France.

Methods: Whole genomic DNA from E. coli 39590 was extracted and was sequenced using an Illumina NextSeq platform. De novo genome assembly was performed using Velvet v.1.2.10 and the draft genome was annotated using the NCBI Prokaryotic Genome Annotation Pipeline v.3.2. Genomic analyses were performed through bioinformatics tools from the Center for Genomic Epidemiology.

Results: The genome size was calculated as 5362108bp, with 5268 protein-coding sequences and a GC content of 50.5%. E. coli strain 39590 belonged to ST219, serotype O4:H34 and phylogroup E. The antimicrobial resistome consisted of genes encoding resistance to β-lactams (bla), aminoglycosides [aph(3″)-Ib, aph(6)-Id, aadA2, aadA24], phenicols (catA1), sulfonamides (sul1, sul2), trimethoprim (dfrA1, dfrA14), lincosamides (lnuG) and tetracycline (tetA) as well as mutations in gyrA (Ser83Leu, Asp87Asn) and parC (Ser80Ile) conferring resistance to quinolones. Virulome analysis revealed iss, astA and eilA genes, and IncQ1, IncX4, IncX1, IncFIB and IncFIC plasmid incompatibility groups were identified.

Conclusion: This draft genome can be used as a reference sequence for comparative studies using human and non-human E. coli isolates to identify genetic events that have contributed to pathogenicity and adaptation of TEM-52-producing E. coli clones at the human-animal interface as well as to elucidate dynamics of the spread of bla ESBL genes.
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http://dx.doi.org/10.1016/j.jgar.2019.07.012DOI Listing
September 2019

Supposed endogenous endophthalmitis caused by in a cat.

Open Vet J 2019 04 23;9(1):13-17. Epub 2019 Jan 23.

Unité d'Ophtalmologie, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.

An 8-year-old male neutered domestic shorthair cat was presented for evaluation of acute respiratory distress. Respiratory auscultation revealed a diffuse and symmetric increase in bronchovesicular sounds. Thoracic radiographs showed a diffuse unstructured interstitial pulmonary pattern with multifocal alveolar foci. Despite an aggressive treatment with supportive care, including oxygenotherapy and systemic antibiotics, progressive respiratory distress increased. Three days after the presentation, acute anterior uveitis was noticed on left eye. Ophthalmic examination and ocular ultrasonography revealed unilateral panuveitis with ocular hypertension. The right eye examination was unremarkable. Cytological examination of aqueous humor revealed a suppurative inflammation. was identified from aqueous humor culture. Primary pulmonary infection was suspected but was not confirmed as owners declined bronchoalveolar lavage. Active uveitis resolved and cat's pulmonary status improved after appropriate systemic antibacterial therapy. Vision loss was permanent due to secondary mature cataract. To the best of authors' knowledge, this is the first report of endogenous bacterial endophthalmitis secondary to infection in a cat.
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http://dx.doi.org/10.4314/ovj.v9i1.3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6500867PMC
April 2019

Frequency of bacteriuria in dogs with chronic kidney disease: A retrospective study of 201 cases.

J Vet Intern Med 2019 Mar 14;33(2):640-647. Epub 2019 Feb 14.

Unité de Médecine Interne, Ecole Nationale Vétérinaire d'Alfort, Maisons Alfort, France.

Background: Studies have shown an increased prevalence of positive urine culture (PUC) in cats with chronic kidney disease (CKD); no information is available in dogs.

Objectives: To document the PUC frequency in a cohort of dogs with CKD, determine risk factors for PUC, and identify associations between clinicopathologic data and PUC.

Animals: Two hundred one client-owned dogs with CKD.

Methods: Retrospective, observational study. Dogs recruited from 2 veterinary teaching hospitals were included if they were diagnosed with CKD and had a culture performed on urine collected by cystocentesis. The PUC frequency was calculated, multivariate analysis was performed to identify risk factors, and associations with clinicopathologic data were investigated.

Results: Sixty-five dogs (32%) with CKD had PUC, including 8 (28%) in International Renal Interest Society (IRIS) stage 1; only 8% showed signs of a urinary tract infection. Escherichia coli was the most common isolate (67%). A PUC was more likely in females (odds ratio [OR], 3.22; 95% confidence interval [CI], 1.67-6.37; P < .001) than males and in dogs with isosthenuria (OR, 2.48; 95% CI, 1.24-5.03; P = .01) than in dogs with urine-specific gravity 1.013-1.024. A positive leukocyte esterase test and microorganisms found by urine sediment analysis were significantly associated with PUC (both P < .001).

Conclusions And Clinical Importance: Dogs with CKD, even IRIS stage 1, have a high frequency of PUC and most cases are asymptomatic. A urine culture could be considered in the routine evaluation of dogs with CKD, but the clinical relevance of a PUC remains unknown and needs further evaluation.
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http://dx.doi.org/10.1111/jvim.15434DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6430954PMC
March 2019

Management of Polyarthritis Affecting Sea Turtles at Kélonia, the Reunion Island Sea Turtle Observatory (2013-17).

J Wildl Dis 2019 04 5;55(2):455-461. Epub 2018 Oct 5.

1 Faculté de Médecine Vétérinaire, Université de Montréal, 3200 Rue Sicotte, Saint-Hyacinthe, Québec, J2S 2M2, Canada.

All sea turtle species are listed on the Red List of the International Union for the Conservation of Nature due to multiple threats. Among these, disease is a frequent cause of stranded sea turtles encountered in rehabilitation centers. Since 2013, we found joint swelling in 13 sea turtles belonging to all four sea turtle species submitted to the Kélonia Sea Turtle Observatory of Reunion Island, France. Affected sea turtles presented with lameness, anorexia, and lethargy. Polyarthritis was radiographically confirmed and lesions were characterized by progressive osteolysis of bones surrounding joints. Anterior flippers were affected in all cases and posterior flippers were also involved in some cases. We isolated several bacterial agents from blood and synovial fluid. We attempted a collective treatment with injectable florfenicol, based on sensitivity results, which was continued for 4 wk and then as needed based on radiographic evolution of the lesions. Radiographic stabilization of the lesions occurred in nine of 13 cases. We reviewed environmental conditions and optimized them to minimize stress that could predispose these rehabilitated sea turtles to opportunistic infections. Handling techniques used to move sea turtles were also improved. While we can make no conclusion regarding the cause of polyarthritis in this population of sea turtles, we successfully managed this poorly described problem in a rehabilitation setting.
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http://dx.doi.org/10.7589/2018-04-103DOI Listing
April 2019

Bilateral bullous keratopathy secondary to melting keratitis in a Suri alpaca ().

Clin Case Rep 2018 04 15;6(4):626-630. Epub 2018 Feb 15.

CHUVA, Unité d'Opthalmologie Ecole Nationale Vétérinaire d'Alfort Université Paris - Est Maisons-Alfort F-94700 France.

An young alpaca was evaluated for bilateral progressive melting corneal ulcers and developped secondary bullous keratopathy during hospitalization. The tragic progression of melting ulcers in both eyes observed in our case leads us to recommend a rapid intensive medical therapy in young and debilitated alpacas presenting a corneal ulcer.
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http://dx.doi.org/10.1002/ccr3.1389DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5889221PMC
April 2018

Co-circulation of different A. phagocytophilum variants within cattle herds and possible reservoir role for cattle.

Parasit Vectors 2018 03 9;11(1):163. Epub 2018 Mar 9.

UMR BIPAR, Ecole Nationale Vétérinaire d'Alfort, Anses, INRA, Université Paris-Est, Maisons-Alfort, France.

Background: Anaplasma phagocytophilum is a zoonotic tick-borne intracellular alpha-proteobacterium causing tick-borne fever, which leads to significant economic losses in domestic ruminants in Europe. Its epidemiological cycles are complex and reservoir host species of bovine strains have not yet been identified. Given that little genetic information is available on strains circulating within a defined bovine environment, our objective was to assess the genetic diversity of A. phagocytophilum obtained from the same farms over time.

Methods: Blood samplings were performed several times in two European herds. In the French herd, 169 EDTA-blood samples were obtained from 115 cows (32 were sampled two to four times). In the German herd, 20 cows were sampled six times (120 EDTA-blood samples). The presence of A. phagocytophilum DNA was assessed using a qPCR targeting msp2. The positive DNA samples underwent MLST at nine genetic markers (typA, ctrA, msp4, pleD, recG, polA, groEL, gyrA, and ankA). For each locus, sequences were aligned with available bacterial sequences derived from cattle, horse, dog, and roe deer hosts, and concatenated neighbor joining trees were constructed using three to six loci.

Results: Around 20% (57/289) of samples were positive. Forty positive samples from 23 French and six German cows (11 of them being positive at two time points) were sequenced. Six loci (typA, ctrA, msp4, pleD, recG, and polA) allowed to build concatenated phylogenetic trees, which led to two distinct groups of bovine variants in the French herd (hereafter called A and B), whereas only group A was detected in the German herd. In 42% of French samples, double chromatogram peaks were encountered in up to four loci. Eleven cows were found infected three weeks to 17 months after first sampling and harboured a new variant belonging to one or the other group.

Conclusions: Our results demonstrate the occurrence of two major bovine strain groups and the simultaneous infection of single cows by more than one A. phagocytophilum strain. This challenges the role of cattle as reservoirs for A. phagocytophilum. This role may be facilitated via long-term bacterial persistence in individual cows and active circulation at the herd scale.
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http://dx.doi.org/10.1186/s13071-018-2661-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5845262PMC
March 2018

Bartonella, bats and bugs: A review.

Comp Immunol Microbiol Infect Dis 2017 Dec 18;55:20-29. Epub 2017 Sep 18.

Graduate Institute of Microbiology and Public Health, National Chung Hsing University, Taichung, Taiwan.

Ecological, immunological, and epidemiological factors enable bats to transmit an increasingly recognized spectrum of zoonotic agents, and bartonellae are among those emerging pathogens identified in bats and their arthropod ectoparasites. Current data reveal a multifaceted disease ecology where diverse host species distributed around the world interact with a number of Bartonella spp. and several potential vectors. This review summarizes the methods and findings of studies conducted since 2005 to illustrate that Bartonella bacteremia varies by bat species, location, and other potential variables, such as diet with a very high prevalence in hematophagous bats. Among bat families, Bartonella prevalence ranged from 7.3% among Nycteridae to 54.4% in Miniopteridae. Further research can build on these current data to better determine risk factors associated with Bartonella infection in bat populations and the role of their ectoparasites in transmission.
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http://dx.doi.org/10.1016/j.cimid.2017.09.001DOI Listing
December 2017

One particular Anaplasma phagocytophilum ecotype infects cattle in the Camargue, France.

Parasit Vectors 2017 Aug 2;10(1):371. Epub 2017 Aug 2.

UMR BIPAR, Université Paris-Est, Ecole Nationale Vétérinaire d'Alfort, Maisons-Alfort, France.

Background: Anaplasma phagocytophilum is a zoonotic tick-borne pathogen responsible for granulocytic anaplasmosis, a mild to a severe febrile disease that affects man and several animal species, including cows and horses. In Europe, I. ricinus is the only proven vector for this pathogen, but studies suggest that other tick genera and species could be involved in its transmission. Our objective was to assess the presence and genetic diversity of A. phagocytophilum in domestic animals and different tick species from the Camargue region, located in the south of France.

Methods: A total of 140 ticks and blood samples from 998 cattle and 337 horses were collected in Camargue and tested for the presence of A. phagocytophilum DNA by msp2 quantitative real-time PCR. Molecular typing with four markers was performed on positive samples.

Results: Anaplasma phagocytophilum DNA was detected in 6/993 (0.6%) cows, 1/20 (5%) Haemaphysalis punctata, 1/57 (1.75%) Rhipicephalus pusillus, and was absent in horses (0%). All cattle A. phagocytophilum presented a profile identical to an A. phagocytophilum variant previously detected in Dermacentor marginatus, Hyalomma marginatum, and Rhipicephalus spp. in Camargue.

Conclusions: Our results demonstrate that one particular A. phagocytophilum variant infects cattle in Camargue, where I. ricinus is supposed to be rare or even absent. Dermacentor marginatus, Rhipicephalus spp. and Hyalomma spp., and possibly other tick species could be involved in the transmission of this variant in this region.
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http://dx.doi.org/10.1186/s13071-017-2305-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5540577PMC
August 2017

Draft Genome Sequences of 12 Feline Isolates.

Genome Announc 2017 Mar 30;5(13). Epub 2017 Mar 30.

Food Safety Laboratory, Platform IdentyPath, Université Paris-Est, Anses, Maisons-Alfort, France

is the main causative agent of cat scratch disease. In this report, we present the draft genome sequences of 12 strains of originating from the United States, Denmark, and France. These strains were isolated from cats and belonged to either 16S rRNA genotype I or 16S rRNA genotype II.
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http://dx.doi.org/10.1128/genomeA.00075-17DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374228PMC
March 2017

Prevalence and Potential Risk Factors for Bartonella Infection in Tunisian Stray Dogs.

Vector Borne Zoonotic Dis 2017 06 27;17(6):388-397. Epub 2017 Mar 27.

1 Department of Population Health and Reproduction, University of California Davis School of Veterinary Medicine , Davis, California.

Bartonellae are blood-borne and vector-transmitted pathogens, some are zoonotic, which have been reported in several Mediterranean countries. Transmission from dogs to humans is suspected, but has not been clearly demonstrated. Our objectives were to determine the seroprevalence of Bartonella henselae, Bartonella vinsonii subsp. berkhoffii, Bartonella clarridgeiae, and Bartonella bovis (as a proxy for Candidatus Bartonella merieuxii) in stray dogs from Tunisia, identify the Bartonella species infecting the dogs and evaluate potential risk factors for canine infection. Blood samples were collected between January and November 2013 from 149 dogs in 10 Tunisian governorates covering several climatic zones. Dog-specific and geographic variables were analyzed as potential risk factors for Bartonella spp. seropositivity and PCR-positivity. DNA was extracted from the blood of all dogs and tested by PCR for Bartonella, targeting the ftsZ and rpoB genes. Partial sequencing was performed on PCR-positive dogs. Twenty-nine dogs (19.5%, 95% confidence interval: 14-27.4) were seropositive for one or more Bartonella species, including 17 (11.4%) for B. vinsonii subsp. berkhoffii, 14 (9.4%) for B. henselae, 13 (8.4%) for B. clarridgeiae, and 7 (4.7%) for B. bovis. Statistical analysis revealed a few potential risk factors, mainly dog's age and breed, latitude and average winter temperature. Twenty-two (14.8%) dogs, including 8 of the 29 seropositive dogs, were PCR-positive for Bartonella based on the ftsZ gene, with 18 (81.8%) of these 22 dogs also positive for the rpoB gene. Partial sequencing showed that all PCR-positive dogs were infected with Candidatus B. merieuxii. Dogs from arid regions and regions with cold average winter temperatures were less likely to be PCR-positive than dogs from other climatic zones. The widespread presence of Bartonella spp. infection in Tunisian dogs suggests a role for stray dogs as potential reservoirs of Bartonella species in Tunisia.
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http://dx.doi.org/10.1089/vbz.2016.2039DOI Listing
June 2017

Potentially Zoonotic Bartonella in Bats from France and Spain.

Emerg Infect Dis 2017 03;23(3):539-541

We detected Bartonella in 11 of 109 insectivorous bats from France and 1 of 26 bats from Spain. These genetic variants are closely related to bat-associated Bartonella described in Finland and the United Kingdom and to B. mayotimonensis, the agent of a human endocarditis case in the United States.
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http://dx.doi.org/10.3201/eid2303.160934DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5382759PMC
March 2017

"Candidatus anaplasma camelii" in one-humped camels (Camelus dromedarius) in Morocco: a novel and emerging anaplasma species?

Infect Dis Poverty 2017 Feb 5;6(1). Epub 2017 Feb 5.

INP-ENVT, Toulouse, France.

Background: There has been a growing interest in camel anaplasmosis due to its recent emergence in this reservoir species and concerns for its zoonotic potential. The epidemiology of anaplasmosis in camels therefore remains poorly understood mostly because camels belong to marginalised poor and often transhumant populations whose interests are largely neglected. Most studies of anaplasmosis in camels have relied on microscopy and serology for diagnosis and only three studies, undertaken in Tunisia, Saudia Arabia and China, have used molecular diagnostics. The present work characterises Anaplasmataceae strains circulating in the Camelus dromedarius reservoir in Morocco using PCR.

Methods: Camels (n = 106) were randomly sampled from 6 regions representing different agro-ecological areas in southern Morocco. Whole blood was collected and screened using PCR methods targeting the gene groEL. Anaplasmataceae strains were characterised by sequence analysis of the gene groEL.

Results: A total of 39.62% (42/106) camels screened were positive for Anaplasmataceae spp. GenBank BLAST analysis of five positive sequenced samples revealed that all strains were 100% identical to "Candidatus Anaplasma camelii". Phylogenetic investigation and genetic characterisation of the aligned segment (650 bp) of the gene groEL confirmed high similarity with A. platys.

Conclusion: This study demonstrates the circulation of a previously unidentified species of the genus Anaplasma in Morocco which is genetically close to the agent causing canine anaplasmosis but whose main reservoir is thought to be Camelus dromedarius.

Trial Registration Number: This study is not a clinical trial and therefore a trial registration number does not apply.
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http://dx.doi.org/10.1186/s40249-016-0216-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5292149PMC
February 2017

OXA-48-producing ST372 Escherichia coli in a French dog.

J Antimicrob Chemother 2017 04;72(4):1256-1258

Unité Antibiorésistance et Virulence Bactériennes, Université Lyon-ANSES Site de Lyon, 31 avenue Tony Garnier, Lyon 69364, France.

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http://dx.doi.org/10.1093/jac/dkw531DOI Listing
April 2017

Draft Anaplasma phagocytophilum Genome Sequences from Five Cows, Two Horses, and One Roe Deer Collected in Europe.

Genome Announc 2016 Dec 1;4(6). Epub 2016 Dec 1.

Université Paris-Est, École Nationale Vétérinaire d'Alfort, UMR BIPAR ENVA Anses UPEC USC INRA, Maisons-Alfort, France

Anaplasma phagocytophilum is a zoonotic tick-borne intracellular bacterium responsible for granulocytic anaplasmosis. As it is difficult to isolate and cultivate, only 20 A. phagocytophilum genomes have been sequenced to date. Here, we present eight A. phagocytophilum genome sequences obtained using alternative approaches based on sequence capture technology.
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http://dx.doi.org/10.1128/genomeA.00950-16DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5137398PMC
December 2016

Multiple-locus variable-number tandem repeat analysis potentially reveals the existence of two groups of Anaplasma phagocytophilum circulating in cattle in France with different wild reservoirs.

Parasit Vectors 2016 11 22;9(1):596. Epub 2016 Nov 22.

Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR, Université Paris-Est, Maisons-Alfort, France.

Background: Anaplasma phagocytophilum is the causative agent of tick-borne fever, a disease with high economic impact for domestic ruminants in Europe. Epidemiological cycles of this species are complex, and involve different ecotypes circulating in various host species. To date, these epidemiological cycles are poorly understood, especially in Europe, as European reservoir hosts (i.e. vertebrate hosts enabling long-term maintenance of the bacterium in the ecosystem), of the bacterium have not yet been clearly identified. In this study, our objective was to explore the presence, the prevalence, and the genetic diversity of A. phagocytophilum in wild animals, in order to better understand their implications as reservoir hosts of this pathogen.

Methods: The spleens of 101 wild animals were collected from central France and tested for the presence of A. phagocytophilum DNA by msp2 qPCR. Positive samples were then typed by multi-locus variable-number tandem repeat (VNTR) analysis (MLVA), and compared to 179 previously typed A. phagocytophilum samples.

Results: Anaplasma phagocytophilum DNA was detected in 82/101 (81.2%) animals including 48/49 red deer (98%), 20/21 roe deer (95.2%), 13/29 wild boars (44.8%), and 1/1 red fox. MLVA enabled the discrimination of two A. phagocytophilum groups: group A contained the majority of A. phagocytophilum from red deer and two thirds of those from cattle, while group B included a human strain and variants from diverse animal species, i.e. sheep, dogs, a horse, the majority of variants from roe deer, and the remaining variants from cattle and red deer.

Conclusions: Our results suggest that red deer and roe deer are promising A. phagocytophilum reservoir host candidates. Moreover, we also showed that A. phagocytophilum potentially circulates in at least two epidemiological cycles in French cattle. The first cycle may involve red deer as reservoir hosts and cattle as accidental hosts for Group A strains, whereas the second cycle could involve roe deer as reservoir hosts and at least domestic ruminants, dogs, horses, and humans as accidental hosts for Group B strains.
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http://dx.doi.org/10.1186/s13071-016-1888-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5120488PMC
November 2016

[Zoonoses (synthetic tables): animal reservoirs of pathogens and way of transmission to man].

Rev Francoph Lab 2015 Dec 10;2015(477):67-79. Epub 2015 Dec 10.

Unité de Parasitologie, Mycologie, Maladies parasitaires et fongiques, Dermatologie, École nationale vétérinaire d'Alfort, UPE, 7, avenue du Général de Gaulle, 94704 Maisons-Alfort cedex.

The knowledge of animals which act as reservoirs for human pathogenic agents is essential for the understanding of the epidemiology of zoonotic diseases and for the application of relevant methods of control. In the present article, synthetic tables present the main zoonoses due to viruses, bacteria, protozoa, helminths, arthropods and fungi. A list of the main pathogenic agents, the animal reservoirs and the way of transmission to man is detailed in each table.
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http://dx.doi.org/10.1016/S1773-035X(15)30318-XDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7270411PMC
December 2015

Opening the black box of Anaplasma phagocytophilum diversity: current situation and future perspectives.

Front Cell Infect Microbiol 2015 14;5:61. Epub 2015 Aug 14.

UMR Biologie Moléculaire et Immunologie Parasitaires, Ecole Nationale Vétérinaire d'Alfort, Université Paris-Est Paris, France.

Anaplasma phagocytophilum is a zoonotic obligate intracellular bacterium known to be transmitted by ticks belonging to the Ixodes persulcatus complex. This bacterium can infect several mammalian species, and is known to cause diseases with variable symptoms in many domestic animals. Specifically, it is the causative agent of tick-borne fever (TBF), a disease of important economic impact in European domestic ruminants, and human granulocytic anaplasmosis (HGA), an emerging zoonotic disease in Asia, USA and Europe. A. phagocytophilum epidemiological cycles are complex and involve different ecotypes, vectors, and mammalian host species. Moreover, the epidemiology of A. phagocytophilum infection differs greatly between Europe and the USA. These different epidemiological contexts are associated with considerable variations in bacterial strains. Until recently, few A. phagocytophilum molecular typing tools were available, generating difficulties in completely elucidating the epidemiological cycles of this bacterium. Over the last few years, many A. phagocytophilum typing techniques have been developed, permitting in-depth epidemiological exploration. Here, we review the current knowledge and future perspectives regarding A. phagocytophilum epidemiology and phylogeny, and then focus on the molecular typing tools available for studying A. phagocytophilum genetic diversity.
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http://dx.doi.org/10.3389/fcimb.2015.00061DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4536383PMC
January 2016

Bartonella Infection among Cats Adopted from a San Francisco Shelter, Revisited.

Appl Environ Microbiol 2015 Sep 10;81(18):6446-50. Epub 2015 Jul 10.

Department of Medicine and Epidemiology and Center for Companion Animal Health, School of Veterinary Medicine, University of California, Davis, Davis, California, USA.

Bartonella infection among cats from shelters can pose a health risk to adopters. Bartonella henselae is the most common species, with B. clarridgeiae and B. koehlerae being less common. The lower rates of infection by the latter species may reflect their rarity or an inefficiency of culture techniques. To assess the incidence of infection, blood cultures, serology, and PCR testing were performed on 193 kittens (6 to 17 weeks old) and 158 young adult cats (5 to 12 months old) from a modern regional shelter. Classical B. henselae culture medium was compared to a medium supplemented with insect cell growth factors. Bartonella colonies were isolated from 115 (32.8%) animals, including 50 (25.9%) kittens and 65 (41.1%) young adults. Therefore, young adults were twice as likely to be culture positive as kittens. Enhanced culture methods did not improve either the isolation rate or species profile. B. henselae was isolated from 40 kittens and 55 young adults, while B. clarridgeiae was cultured from 10 animals in each group. B. koehlerae was detected in one young adult by PCR only. B. henselae genotype II was more commonly isolated from young adults, and genotype I was more frequently isolated from kittens. Kittens were 4.7 times more likely to have a very high bacterial load than young adults. A significantly higher incidence of bacteremia in the fall and winter than in the spring and summer was observed. Bartonella antibodies were detected in 10% (19/193) of kittens and 46.2% (73/158) of young adults, with culture-positive kittens being 9.4 times more likely to be seronegative than young adults.
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http://dx.doi.org/10.1128/AEM.01864-15DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4542261PMC
September 2015

Prevalence of select vector-borne pathogens in stray and client-owned dogs from Algiers.

Comp Immunol Microbiol Infect Dis 2015 Feb 14;38:1-7. Epub 2015 Jan 14.

Université Paris-Est, Ecole Nationale Vétérinaire d'Alfort, UMR BIPAR, ENVA, Anses, USC INRA, Avenue du général de Gaulle, F94704 Maisons-Alfort, France. Electronic address:

Data on the prevalence of vector-borne diseases agents infecting canines in Algeria is currently lacking. The purpose of this study is to assess by serological and molecular methods the prevalence of select arthropod borne-bacterial infections in client-owned and stray dogs. Antibodies to Anaplasma phagocytophilum were the most prevalent at 47.7%, followed by Borrelia burgdorferi s.l. at 37.6%, Ehrlichia canis at 30.0%, Bartonella henselae at 32.4% and Bartonella vinsonii subsp. berkhoffii at 27%. Seroprevalence was statistically significantly higher in stray dogs than those owned by clients. Seropositivity was not associated with health status, except for E. canis. Molecular evaluation indicates that 17.8% of the 213 analyzed dogs were positive for Ehrlichia and Anaplasma with a prevalence of 4.2% for E. canis, 14.1% for Anaplama platys and 0% for A. phagocytophilum. Seven (7.1%) of the tested dogs were positive for Bartonella spp. with two characterized as Bartonella rochalimae, four as B. henselae and one as B.v. subsp. berkhoffii.
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http://dx.doi.org/10.1016/j.cimid.2015.01.001DOI Listing
February 2015