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Philippe Jacquiet - One of the best experts on this subject based on the ideXlab platform.

  • stomoxys calcitrans mechanical vector of virulent besnoitia besnoiti from chronically infected cattle to susceptible rabbit
    Medical and Veterinary Entomology, 2019
    Co-Authors: Philippe Jacquiet, Isabelle Raymondletron, S Sharif, Francoise Prevot, Christelle Grisez, Marie Odile Semin
    Abstract:

    : Cattle Besnoitiosis caused by Besnoitia besnoiti (Eucoccidiorida: Sarcocystidae) is a re-emerging disease in Europe. Its mechanical transmission by biting flies has not been investigated since the 1960s. The aim of this study was to re-examine the ability of Stomoxys calcitrans (Diptera: Muscidae) to transmit virulent B. besnoiti bradyzoites from chronically infected cows to susceptible rabbits. Three batches of 300 stable flies were allowed to take an interrupted bloodmeal on chronically infected cows, followed by an immediate bloodmeal on three rabbits (Group B). A control group of rabbits and a group exposed to the bites of non-infected S. calcitrans were included in the study. Blood quantitative polymerase chain reaction (qPCR) analyses, and clinical, serological and haematological surveys were performed in the three groups over 152 days until the rabbits were killed. Quantitative PCR analyses and histological examinations were performed in 24 tissue samples per rabbit. Only one rabbit in Group B exhibited clinical signs of the acute phase of Besnoitiosis (hyperthermia, weight loss, regenerative anaemia and transient positive qPCR in blood) and was seroconverted. Parasite DNA was detected in four tissue samples from this rabbit, but no cysts were observed on histological examination. These findings indicate that S. calcitrans may act as a mechanical vector of B. besnoiti more efficiently than was previously considered.

  • Phenotypic susceptibility to pyrethroids and organophosphate of wild Stomoxys calcitrans (Diptera: Muscidae) populations in southwestern France
    Parasitology Research, 2018
    Co-Authors: Krajana Tainchum, Sharif Shukri, Gérard Duvallet, Lucas Etienne, Philippe Jacquiet
    Abstract:

    Stomoxys calcitrans (Diptera: Muscidae), is an important vector of lumpy skin disease and bovine Besnoitiosis in Europe. Control of this biting fly could represent a keystone in the containment of this emerging disease. Reports of insecticide resistance in S. calcitrans are scarce in Europe. Therefore, the purpose of this study was to evaluate the phenotypic susceptibility to deltamethrin, cypermethrin and phoxim of five wild S. calcitrans populations from southwestern France, where transmission of bovine Besnoitiosis is very prevalent. Adult S. calcitrans were caught at each study site and exposed to insecticide-impregnated filter papers under laboratory conditions. Quantities of active ingredients on filter papers corresponded to the recommended doses proposed by the manufacturers (37.5 mg a.i./m^2 of cattle’s skin, 125 mg a.i./m^2 and 750 mg a.i./m^2 for deltamethrin, cypermethrin and phoxim respectively) were tested. Knock-down effects (KD) (1 h after the onset of exposure) and mortality rates (24 h and 48 h after exposure) were evaluated. Phoxim showed a rapid and full efficacy in all populations. However, the KD effects (37.5 to 97.5%) and the mortality rates at 48 h (10 to 91.25%) induced by the exposure to pyrethroids varied greatly according to the study site but none of the populations showed full susceptibility. Therefore, the current recommended doses of these pyrethroids are probably less efficient than expected in the field and should be considered with caution in the control of bovine Besnoitiosis in France.

  • does bovine Besnoitiosis affect the sexual function of chronically infected bulls
    Theriogenology, 2016
    Co-Authors: Angel Estebangil, Philippe Jacquiet, S Florentin, A Decaudin, Xavier Berthelot, P Ronsin, J P Alzieu, Francoise Prevot, Christelle Grisez, M Marois
    Abstract:

    Abstract Bovine Besnoitiosis is a reemerging disease in Europe. The clinically Besnoitia besnoiti infection in bulls is characterized by fever, nasal discharge, and orchitis in the acute phase and by scleroderma in the chronic phase. However, in many bulls, B besnoiti infection remains at a subclinical stage. Bull infertility is an economically relevant consequence of Besnoitiosis infection. It is not clear, however, if semen quality returns to normal levels when infected animals have clinically recovered. The aim of this study was to examine the relationship between chronic Besnoitiosis and bull sexual function in a region of eastern France, where the disease is reemerging, by comparing semen quality and genital lesions in 11 uninfected, 17 subclinically infected, and 12 clinically infected bulls. The presence of anti- B besnoiti antibodies was detected by Western blot test. Semen was collected by electroejaculation. Bulls clinically infected with B besnoiti showed significantly more genital tract alterations than uninfected or subclinically infected bulls. No relationship was evidenced between Besnoitiosis infectious status and semen quality, whereas a significant relationship was noted between genital lesions and semen score. This means that in the absence of moderate to severe genital lesions, chronic bovine Besnoitiosis is unlikely to alter semen quality. However, as the presence of infected animals could lead to spread of the disease, culling or separation of clinically infected bulls from the remaining healthy animals is strongly recommended.

  • besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates
    Parasites & Vectors, 2016
    Co-Authors: Paula Garcialunar, Javier Regidorcerrillo, D Gutierrezexposito, Caroline F Frey, Nelson Marreros, Arcangelo Gentile, J P Dubey, Gereon Schares, Philippe Jacquiet
    Abstract:

    Background Bovine Besnoitiosis, caused by the protozoan Besnoitia besnoiti, reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of Besnoitiosis.

  • Besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates
    Parasites & Vectors, 2016
    Co-Authors: Caroline F Frey, Paula García-lunar, Daniel Gutiérrez-expósito, Nelson Marreros, Arcangelo Gentile, Philippe Jacquiet, Gereon Schares, Javier Regidor-cerrillo, Jitender P. Dubey, Varda Shkap
    Abstract:

    Background Bovine Besnoitiosis, caused by the protozoan Besnoitia besnoiti , reduces productivity and fertility of affected herds. Besnoitiosis continues to expand in Europe and no effective control tools are currently available. Experimental models are urgently needed. Herein, we describe for the first time the kinetics of standardised in vitro models for the B. besnoiti lytic cycle. This will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful for the treatment of Besnoitiosis. Methods We compared invasion and proliferation of one B. tarandi (from Finland) and seven B. besnoiti isolates (Bb-Spain1, Bb-Spain2, Bb-Israel, Bb-Evora03, Bb-Ger1, Bb-France, Bb-Italy2) in MARC-145 cell culture. Host cell invasion was studied at 4, 6, 8 and 24 h post infection (hpi), and proliferation characteristics were compared at 24, 48, 72, 96, 120, and 144 hpi. Results In Besnoitia spp., the key parameters that determine the sequential adhesion-invasion, proliferation and egress steps are clearly distinct from those in the related apicomplexans Toxoplasma gondii and Neospora caninum. Besnoitia spp. host cell invasion is a rather slow process, since only 50 % of parasites were found intracellular after 3–6 h of exposure to host cells, and invasion still took place after 24 h. Invasion efficacy was significantly higher for Bb-France, Bb-Evora03 and Bb-Israel. In addition, the time span for endodyogeny to take place was as long as 18–35 h. Bb-Israel and B. tarandi isolates were most prolific, as determined by the tachyzoite yield at 72 hpi. The total tachyzoite yield could not be predicted neither by invasion-related parameters (velocity and half time invasion) nor by proliferation parameters (lag phase and doubling time (dT)). The lytic cycle of Besnoitia was asynchronous as evidenced by the presence of three different plaque-forming tachyzoite categories (lysis plaques, large and small parasitophorous vacuoles). Conclusions This study provides first insights into the lytic cycle of B. besnoiti isolates and a standardised in vitro model that allows screening of drug candidates for the treatment of Besnoitiosis.

Francoise Prevot - One of the best experts on this subject based on the ideXlab platform.

  • Real-time PCR on skin biopsies for super-spreaders’ detection in bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background: Bovine Besnoitiosis, an emerging disease in Europe that can be transmitted by vectors, is caused by the Apicomplexa Besnoitia besnoiti. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods: Previous real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled at the tail base and their skin sample analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time.Results: Among the 518 seropositive animals, a low proportion of individuals (14.5%) showed Ct values below 36, 17.8% had doubtful results (36 < Ct ≤ 40) and 67.8% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions: Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • Super-spreaders: detection and culling in the control of bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background Bovine Besnoitiosis is an emerging vector-borne disease in Europe caused by the Apicomplexa Besnoitia besnoiti. The mechanical transmission from infected to naïve hosts is permitted by horse flies and stable flies. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods Real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled and their skin analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time. Results Among the 518 seropositive animals, a low proportion of individuals (13%) showed Ct values below 36, 17% had doubtful results (36 < Ct ≤ 40) and 70% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • stomoxys calcitrans mechanical vector of virulent besnoitia besnoiti from chronically infected cattle to susceptible rabbit
    Medical and Veterinary Entomology, 2019
    Co-Authors: Philippe Jacquiet, Isabelle Raymondletron, S Sharif, Francoise Prevot, Christelle Grisez, Marie Odile Semin
    Abstract:

    : Cattle Besnoitiosis caused by Besnoitia besnoiti (Eucoccidiorida: Sarcocystidae) is a re-emerging disease in Europe. Its mechanical transmission by biting flies has not been investigated since the 1960s. The aim of this study was to re-examine the ability of Stomoxys calcitrans (Diptera: Muscidae) to transmit virulent B. besnoiti bradyzoites from chronically infected cows to susceptible rabbits. Three batches of 300 stable flies were allowed to take an interrupted bloodmeal on chronically infected cows, followed by an immediate bloodmeal on three rabbits (Group B). A control group of rabbits and a group exposed to the bites of non-infected S. calcitrans were included in the study. Blood quantitative polymerase chain reaction (qPCR) analyses, and clinical, serological and haematological surveys were performed in the three groups over 152 days until the rabbits were killed. Quantitative PCR analyses and histological examinations were performed in 24 tissue samples per rabbit. Only one rabbit in Group B exhibited clinical signs of the acute phase of Besnoitiosis (hyperthermia, weight loss, regenerative anaemia and transient positive qPCR in blood) and was seroconverted. Parasite DNA was detected in four tissue samples from this rabbit, but no cysts were observed on histological examination. These findings indicate that S. calcitrans may act as a mechanical vector of B. besnoiti more efficiently than was previously considered.

  • first evidence of besnoitia bennetti infection protozoa apicomplexa in donkeys equus asinus in belgium
    Parasites & Vectors, 2018
    Co-Authors: Emmanuel Lienard, Adriana Nabuco, Sophie Vandenabeele, Irene Tosi, Emilie Bouhsira, Shukri Sharif, Michel Franc, Bertrand Losson, Francoise Prevot, Caroline Vanvinckenroye
    Abstract:

    Background Besnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti, ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey Besnoitiosis is considered as an emerging disease.

  • First evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium
    Parasites & Vectors, 2018
    Co-Authors: Emmanuel Lienard, Adriana Nabuco, Sophie Vandenabeele, Irene Tosi, Emilie Bouhsira, Shukri Sharif, Michel Franc, Bertrand Losson, Francoise Prevot, Caroline Vanvinckenroye
    Abstract:

    Background Besnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti , ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey Besnoitiosis is considered as an emerging disease. Case presentation A two-year-old male donkey was purchased in May 2016 in poor body condition (cachexia, alopetic areas and pruritus mainly on neck and head) by the present owner in Le Roeulx (Belgium) from a milk producing donkey farm in Frasnes-lez-Buissenal (Belgium). Shortly after its purchase and shearing, the donkey presented with crusts, hyperkeratosis (both flanks and neck) anorexia and cachexia. A treatment with phoxim was given with no improvement. A cutaneous biopsy of hyperkeratotic skin was performed in July. It showed a perivascular eosinophilic infiltrate with a large thick walled cyst located in the dermis containing numerous bradyzoites. This was highly suggestive of Besnoitiosis. Several skin biopsy samples were obtained for qPCR analysis and confirmed the presence of Besnoitia spp. DNA. Further laboratory diagnosis tests were performed (western blot and rDNA sequencing) confirming Besnoitia bennetti aetiology for the male. For the female, the punch-biopsy, haematology and qPCR were negatives but the western blot showed the presence of antibodies directed to Besnoitia spp. Further clinical examination performed in August highlighted scleral pinhead sized cysts (pearl) in the right eye and between nares. Another ten-year-old female donkey purchased in France and sharing the same accommodation showed a good clinical condition, but a thorough clinical examination showed the presence of numerous cysts on the inner face of upper labial mucosa. A daily treatment based on sulfamethaxzole and trimethoprim (Emdotrim 60% Mix®, 30 mg/kg) was given orally and some improvement was noticed. Conclusion This is the first evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys ( Equus asinus ) in Belgium.

Emmanuel Lienard - One of the best experts on this subject based on the ideXlab platform.

  • Real-time PCR on skin biopsies for super-spreaders’ detection in bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background: Bovine Besnoitiosis, an emerging disease in Europe that can be transmitted by vectors, is caused by the Apicomplexa Besnoitia besnoiti. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods: Previous real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled at the tail base and their skin sample analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time.Results: Among the 518 seropositive animals, a low proportion of individuals (14.5%) showed Ct values below 36, 17.8% had doubtful results (36 < Ct ≤ 40) and 67.8% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions: Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • Super-spreaders: detection and culling in the control of bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background Bovine Besnoitiosis is an emerging vector-borne disease in Europe caused by the Apicomplexa Besnoitia besnoiti. The mechanical transmission from infected to naïve hosts is permitted by horse flies and stable flies. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods Real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled and their skin analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time. Results Among the 518 seropositive animals, a low proportion of individuals (13%) showed Ct values below 36, 17% had doubtful results (36 < Ct ≤ 40) and 70% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • first evidence of besnoitia bennetti infection protozoa apicomplexa in donkeys equus asinus in belgium
    Parasites & Vectors, 2018
    Co-Authors: Emmanuel Lienard, Adriana Nabuco, Sophie Vandenabeele, Irene Tosi, Emilie Bouhsira, Shukri Sharif, Michel Franc, Bertrand Losson, Francoise Prevot, Caroline Vanvinckenroye
    Abstract:

    Background Besnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti, ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey Besnoitiosis is considered as an emerging disease.

  • First evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium
    Parasites & Vectors, 2018
    Co-Authors: Emmanuel Lienard, Adriana Nabuco, Sophie Vandenabeele, Irene Tosi, Emilie Bouhsira, Shukri Sharif, Michel Franc, Bertrand Losson, Francoise Prevot, Caroline Vanvinckenroye
    Abstract:

    Background Besnoitiosis is caused by different species of intracellular protozoan parasites belonging to the family Sarcocystidae and affecting multiple host species worldwide. Including B. besnoiti , ten species are described infecting animals. Among ungulates, Besnoitia bennetti infects horses, donkeys and zebras and was described in Africa and in the USA where donkey Besnoitiosis is considered as an emerging disease. Case presentation A two-year-old male donkey was purchased in May 2016 in poor body condition (cachexia, alopetic areas and pruritus mainly on neck and head) by the present owner in Le Roeulx (Belgium) from a milk producing donkey farm in Frasnes-lez-Buissenal (Belgium). Shortly after its purchase and shearing, the donkey presented with crusts, hyperkeratosis (both flanks and neck) anorexia and cachexia. A treatment with phoxim was given with no improvement. A cutaneous biopsy of hyperkeratotic skin was performed in July. It showed a perivascular eosinophilic infiltrate with a large thick walled cyst located in the dermis containing numerous bradyzoites. This was highly suggestive of Besnoitiosis. Several skin biopsy samples were obtained for qPCR analysis and confirmed the presence of Besnoitia spp. DNA. Further laboratory diagnosis tests were performed (western blot and rDNA sequencing) confirming Besnoitia bennetti aetiology for the male. For the female, the punch-biopsy, haematology and qPCR were negatives but the western blot showed the presence of antibodies directed to Besnoitia spp. Further clinical examination performed in August highlighted scleral pinhead sized cysts (pearl) in the right eye and between nares. Another ten-year-old female donkey purchased in France and sharing the same accommodation showed a good clinical condition, but a thorough clinical examination showed the presence of numerous cysts on the inner face of upper labial mucosa. A daily treatment based on sulfamethaxzole and trimethoprim (Emdotrim 60% Mix®, 30 mg/kg) was given orally and some improvement was noticed. Conclusion This is the first evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys ( Equus asinus ) in Belgium.

  • experimental infections of rabbits with proliferative and latent stages of besnoitia besnoiti
    Parasitology Research, 2015
    Co-Authors: Emmanuel Lienard, Emilie Bouhsira, Michel Franc, Isabelle Raymondletron, Virginie Mallet, Francoise Prevot, Christelle Grisez, Philippe Jacquiet
    Abstract:

    Cattle Besnoitiosis due to Besnoitia besnoiti is spreading across Europe and is responsible for severe economic losses in newly infected herds. Experimentally speaking, rabbits have been found to be susceptible to this parasite. The adaptation of B. besnoiti to rabbits may offer a new, easier and cheaper model of investigation for this disease. This study compared the virulence between tachyzoites and bradyzoites of B. besnoiti in rabbits. Eighteen New Zealand rabbits were allocated into three groups of six animals each. The rabbits from the control (group C), “tachyzoite” (group T) and “bradyzoite” (group B) groups were subcutaneously injected in the right flank with 66 μg of ovalbumin, 6.106 tachyzoites (125th passage on Vero cells) and 6.106 bradyzoites (collected from a natural infected cow) of B. besnoiti, respectively. Clinical follow-up and blood sampling for serological survey and qPCR were performed during 10 weeks until euthanasia. Molecular and immunohistochemistry examination was achieved on 25 samples of tissue per rabbit. Seroconversion occurred in group T without any clinical signs. Rabbits of group B exhibited a febrile condition (temperature above 40 °C from day 8 to day 11 following injection) with positive qPCR in blood. Cysts of B. besnoiti were found on skin samples and organs of rabbits from group B in tissue explored with threshold cycle (Ct) values below 30. These results suggest a higher virulence of bradyzoites in rabbits than Vero cell-cultivated tachyzoites. The proposed model could be used to assess the in vivo effectiveness of vaccine or drugs against cattle Besnoitiosis.

Christelle Grisez - One of the best experts on this subject based on the ideXlab platform.

  • Real-time PCR on skin biopsies for super-spreaders’ detection in bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background: Bovine Besnoitiosis, an emerging disease in Europe that can be transmitted by vectors, is caused by the Apicomplexa Besnoitia besnoiti. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods: Previous real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled at the tail base and their skin sample analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time.Results: Among the 518 seropositive animals, a low proportion of individuals (14.5%) showed Ct values below 36, 17.8% had doubtful results (36 < Ct ≤ 40) and 67.8% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions: Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • Super-spreaders: detection and culling in the control of bovine Besnoitiosis.
    2020
    Co-Authors: Christelle Grisez, Emmanuel Lienard, J P Alzieu, Francoise Prevot, Leslie Bottari, Fabien Corbière, Marie Rameil, Xavier Desclaux, Christophe Lacz, Christian Boulon
    Abstract:

    Abstract Background Bovine Besnoitiosis is an emerging vector-borne disease in Europe caused by the Apicomplexa Besnoitia besnoiti. The mechanical transmission from infected to naïve hosts is permitted by horse flies and stable flies. Bovine Besnoitiosis is difficult to control due to the complexity of its diagnosis in the acute stage of the disease, poor treatment success and chronically asymptomatic cattle acting as parasite reservoirs. When serological prevalence is low, detection and specific culling of seropositive cattle is feasible; however, economic considerations preclude this approach when serological prevalence is high. The aims of this study were to evaluate the accuracy of detection of super-spreaders in highly infected herds and to test their selective elimination as a new control strategy for bovine Besnoitiosis. Methods Real-time PCR analyses performed on skin tissues from 160 asymptomatic animals sampled at slaughterhouses showed that the tail base was the best location to evaluate the dermal parasite DNA load. All seropositive animals (N = 518) from eight dairy or beef cattle farms facing a high serological prevalence of Besnoitiosis were sampled and their skin analysed by real-time PCR. A recommendation of rapid and selective culling of super-spreaders was formulated and provided to the cattle breeders. Subsequent serological monitoring of naïve animals was used to evaluate the interest of this control strategy over time. Results Among the 518 seropositive animals, a low proportion of individuals (13%) showed Ct values below 36, 17% had doubtful results (36 < Ct ≤ 40) and 70% had negative PCR results. These proportions were grossly similar on the eight farms, regardless of their production type (beef or dairy cattle), size, geographic location or history of Besnoitiosis. Within two weeks of the biopsy, the rapid culling of super-spreaders was implemented on only three farms. The numbers of newly infected animals were lower on these farms compared to those where super-spreaders were maintained in the herd. Conclusions Real-time PCR analyses performed on skin biopsies of seropositive cattle showed huge individual variabilities in parasite DNA load. The rapid culling of individuals considered as super-spreaders seems to be a new and encouraging strategy for bovine Besnoitiosis control.

  • stomoxys calcitrans mechanical vector of virulent besnoitia besnoiti from chronically infected cattle to susceptible rabbit
    Medical and Veterinary Entomology, 2019
    Co-Authors: Philippe Jacquiet, Isabelle Raymondletron, S Sharif, Francoise Prevot, Christelle Grisez, Marie Odile Semin
    Abstract:

    : Cattle Besnoitiosis caused by Besnoitia besnoiti (Eucoccidiorida: Sarcocystidae) is a re-emerging disease in Europe. Its mechanical transmission by biting flies has not been investigated since the 1960s. The aim of this study was to re-examine the ability of Stomoxys calcitrans (Diptera: Muscidae) to transmit virulent B. besnoiti bradyzoites from chronically infected cows to susceptible rabbits. Three batches of 300 stable flies were allowed to take an interrupted bloodmeal on chronically infected cows, followed by an immediate bloodmeal on three rabbits (Group B). A control group of rabbits and a group exposed to the bites of non-infected S. calcitrans were included in the study. Blood quantitative polymerase chain reaction (qPCR) analyses, and clinical, serological and haematological surveys were performed in the three groups over 152 days until the rabbits were killed. Quantitative PCR analyses and histological examinations were performed in 24 tissue samples per rabbit. Only one rabbit in Group B exhibited clinical signs of the acute phase of Besnoitiosis (hyperthermia, weight loss, regenerative anaemia and transient positive qPCR in blood) and was seroconverted. Parasite DNA was detected in four tissue samples from this rabbit, but no cysts were observed on histological examination. These findings indicate that S. calcitrans may act as a mechanical vector of B. besnoiti more efficiently than was previously considered.

  • does bovine Besnoitiosis affect the sexual function of chronically infected bulls
    Theriogenology, 2016
    Co-Authors: Angel Estebangil, Philippe Jacquiet, S Florentin, A Decaudin, Xavier Berthelot, P Ronsin, J P Alzieu, Francoise Prevot, Christelle Grisez, M Marois
    Abstract:

    Abstract Bovine Besnoitiosis is a reemerging disease in Europe. The clinically Besnoitia besnoiti infection in bulls is characterized by fever, nasal discharge, and orchitis in the acute phase and by scleroderma in the chronic phase. However, in many bulls, B besnoiti infection remains at a subclinical stage. Bull infertility is an economically relevant consequence of Besnoitiosis infection. It is not clear, however, if semen quality returns to normal levels when infected animals have clinically recovered. The aim of this study was to examine the relationship between chronic Besnoitiosis and bull sexual function in a region of eastern France, where the disease is reemerging, by comparing semen quality and genital lesions in 11 uninfected, 17 subclinically infected, and 12 clinically infected bulls. The presence of anti- B besnoiti antibodies was detected by Western blot test. Semen was collected by electroejaculation. Bulls clinically infected with B besnoiti showed significantly more genital tract alterations than uninfected or subclinically infected bulls. No relationship was evidenced between Besnoitiosis infectious status and semen quality, whereas a significant relationship was noted between genital lesions and semen score. This means that in the absence of moderate to severe genital lesions, chronic bovine Besnoitiosis is unlikely to alter semen quality. However, as the presence of infected animals could lead to spread of the disease, culling or separation of clinically infected bulls from the remaining healthy animals is strongly recommended.

  • experimental infections of rabbits with proliferative and latent stages of besnoitia besnoiti
    Parasitology Research, 2015
    Co-Authors: Emmanuel Lienard, Emilie Bouhsira, Michel Franc, Isabelle Raymondletron, Virginie Mallet, Francoise Prevot, Christelle Grisez, Philippe Jacquiet
    Abstract:

    Cattle Besnoitiosis due to Besnoitia besnoiti is spreading across Europe and is responsible for severe economic losses in newly infected herds. Experimentally speaking, rabbits have been found to be susceptible to this parasite. The adaptation of B. besnoiti to rabbits may offer a new, easier and cheaper model of investigation for this disease. This study compared the virulence between tachyzoites and bradyzoites of B. besnoiti in rabbits. Eighteen New Zealand rabbits were allocated into three groups of six animals each. The rabbits from the control (group C), “tachyzoite” (group T) and “bradyzoite” (group B) groups were subcutaneously injected in the right flank with 66 μg of ovalbumin, 6.106 tachyzoites (125th passage on Vero cells) and 6.106 bradyzoites (collected from a natural infected cow) of B. besnoiti, respectively. Clinical follow-up and blood sampling for serological survey and qPCR were performed during 10 weeks until euthanasia. Molecular and immunohistochemistry examination was achieved on 25 samples of tissue per rabbit. Seroconversion occurred in group T without any clinical signs. Rabbits of group B exhibited a febrile condition (temperature above 40 °C from day 8 to day 11 following injection) with positive qPCR in blood. Cysts of B. besnoiti were found on skin samples and organs of rabbits from group B in tissue explored with threshold cycle (Ct) values below 30. These results suggest a higher virulence of bradyzoites in rabbits than Vero cell-cultivated tachyzoites. The proposed model could be used to assess the in vivo effectiveness of vaccine or drugs against cattle Besnoitiosis.

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  • histological findings in experimentally infected male calves with chronic Besnoitiosis
    Veterinary Parasitology, 2020
    Co-Authors: Carlos Diezmadiaz, Enrique Tabanera, Marta Gonzalezhuecas, M Pizarrodiaz, Javier Blancomurcia, Luis Miguel Ortegamora, Ignacio Ferre, Gema Alvarezgarcia
    Abstract:

    Abstract The histological findings associated to Besnoitia besnoiti infection were exhaustively studied in target tissues from experimentally and chronically infected calves. Calves were inoculated with 106 bradyzoites via intravenous, subcutaneous and intradermal route. Visible pathognomonic sclera cysts were observed in all infected animals. Tissue cysts were more abundant and lesions were more frequent in calves inoculated via intradermal. The most parasitized tissues were skin, including scrotum (40.81% of positive samples), nostril and nasal turbinate. Tissue cysts were already fully developed as the average tissue cyst diameter was 181.20 μm. Microscopic lesions were mainly detected in skin samples, followed by reproductive and upper respiratory tracts. Mild lesions compatible with both acute (thrombus, oedema and inflammation) and chronic Besnoitiosis (skin lesions, hyperkeratosis and dilated sweat glands) coexisted. Vascular damage and inflammation were more frequently observed in skin (including scrotum) followed by testicular parenchyma, epididymis and pampiniform plexus. Histological findings evidenced a subclinical chronic Besnoitiosis.

  • bovine chronic Besnoitiosis in a calf characterization of a novel b besnoiti isolate from an unusual case report
    Veterinary Parasitology, 2017
    Co-Authors: Carlos Diezmadiaz, Paula Garcialunar, D Gutierrezexposito, Alejandro Jimenezmelendez, Javier Blancomurcia, Luis Miguel Ortegamora, Ignacio Ferre, M Fernandez, J A Perezsalas, Gema Alvarezgarcia
    Abstract:

    Abstract Bovine Besnoitiosis, caused by the apicomplexan Besnoitia besnoiti , is a chronic and debilitating disease characterized by cutaneous and systemic manifestations that primarily affects adult beef cattle. Previous studies have reported that clinical Besnoitiosisis is rare in calves. However, we isolated B. besnoiti from a chronically infected calf for the first time. The identity of the Besnoitia species was determined after parasite isolation and molecular genotyping. According to the results obtained in vitro the new isolate, named as Bb-Spain3, was characterized in a reproducible in vitro model and was categorized as a low invader and low prolific isolate with a slower lytic cycle compared to Bb-Spain 1 isolate. Specific traits that differentiate isolates obtained from adult animals from those infecting calves were not found. Next, we described the first case report of chronic Besnoitiosis in a female calf less than 6 months-old with a low body condition. The disease was confirmed by the presence of specific anti- B. besnoiti antibodies and parasite detection in the skin. At post-mortem examination, tissue samples were collected for histological, immunohistochemical and molecular analyses. DNA-parasite was detected in 31 different calf’s tissues, being the most highly parasitized tissues the skin and the respiratory and reproductive tracts. In addition, the parasite was also present in heart, eyes, lymph nodes and brain. The high parasite load, a wide intra-organic parasite distribution and the presence of both viable and degenerated cysts, were indicative of a rapid progression of the disease. This case report underlines the need to include the inspection of young animals in Besnoitiosis control.

  • advances in the diagnosis of bovine Besnoitiosis current options and applications for control
    International Journal for Parasitology, 2017
    Co-Authors: D Gutierrezexposito, Luis Miguel Ortegamora, Ignacio Ferre, Gema Alvarezgarcia
    Abstract:

    Bovine Besnoitiosis, which is caused by the tissue cyst-forming intracellular parasite Besnoitia besnoiti, is a chronic and debilitating disease that is responsible for severe economic losses in the cattle raised under extensive husbandry systems. The absence of vaccines, treatments or a health scheme at local, national and international levels has led to a rapid spread of bovine Besnoitiosis from western Europe towards eastern countries and northwards. Moreover, this parasitic disease is widely present in many sub-Saharan countries. Thus, bovine Besnoitiosis should be included in the animal health scheme of beef cattle herds. Accurate diagnostic tools and common diagnostic procedures are mandatory in any control programme. Relevant advances have been made in this field during the last decade. Succeeding with accurate diagnosis relies on the technique employed and the antibody and parasite kinetics of the infection stage, which may notably influence control programmes and surveillance. Moreover, control programmes should be adapted to the epidemiological status of the disease, as the disease presentation in a herd has important implications for prospective control. Herein, we review the clinical disease presentation of bovine Besnoitiosis and the correlation between its clinical course and laboratory parameters. We also provide an update on the available diagnostic tools, discuss their strengths and pitfalls, and provide guidelines for their use in control, surveillance and epidemiological studies. A rational control strategy is also recommended.

  • systemic Besnoitiosis in a juvenile roe deer capreolus capreolus
    Transboundary and Emerging Diseases, 2017
    Co-Authors: Maria C Arnal, Javier Regidorcerrillo, D Gutierrezexposito, David Martinezduran, Alejandro Jimenezmelendez, Luis Miguel Ortegamora, Miguel Revilla, Fernandez D De Luco, Gema Alvarezgarcia
    Abstract:

    Summary Herein, we report the first incidence of systemic Besnoitiosis in a male juvenile roe deer Capreolus capreolus. The animal was found dead in an area where bovine Besnoitiosis is endemic and showed cachexia and multiple skin erosions in the metacarpal and metatarsal areas. Moreover, round and elevated white structures suggestive of Besnoitia spp. tissue cysts were also present. Twenty-eight tissue samples from different anatomical locations were collected for microscopic lesion and parasite detection through histopathology and PCR. Immunohistochemistry was performed to confirm Besnoitia-positive reaction in the tissue cysts. In addition, the identity of Besnoitia spp. in PCR-positive tissue samples was also investigated using microsatellite (MS) markers, and the comparison of protein disulphide isomerase gene sequences (BbPDI) of B. besnoiti and B. tarandi isolated from cattle and reindeer, respectively. Besnoitia cysts were detected in the skin (several parts), respiratory and upper digestive tracts, eyes, kidney, liver, testicle, cardiac muscle and lymphoid tissue. Remarkably, the presence of tissue cysts in the brain confirmed the capacity of Besnoitia spp. to form tissue cysts in the central nervous system (CNS). Finally, the Besnoitia species detected showed the same MS genotype as B. besnoiti, and BbPDI sequences from roe deer and two B. besnoiti isolates were genetically identical throughout multiple sequence alignment. Thus, for the first time, there is evidence that roe deer might act as an intermediate host of B. besnoiti. Further molecular analyses and parasite isolations are needed to corroborate these findings.

  • absence of antibodies specific to besnoitia spp in european sheep and goats from areas in spain where bovine Besnoitiosis is endemic
    Parasitology Research, 2017
    Co-Authors: D Gutierrezexposito, Javier Carvajalvalilla, Angel Morales, Santiago Lavín, Luis Miguel Ortegamora, Ignasi Marco, Gema Alvarezgarcia
    Abstract:

    Besnoitia besnoiti and B. caprae, which infect bovids (cattle and antelopes) and goats, respectively, are responsible for Besnoitiosis, a chronic and debilitating disease. Bovine Besnoitiosis is considered to be a reemerging disease in Central and Western Europe. In addition, infection by Besnoitia spp. has been reported in reindeer from Sweden and Finland. Recently, the parasite was also detected in roe deer and red deer from Spain, where an interconnection between the domestic and sylvatic cycles of B. besnoiti has been presumed. In contrast, caprine Besnoitiosis seems to be enzootic to Kenya and Iran. The presence of Besnoitia spp. in small domestic ruminants has never been explored in Europe, and the role that these species might play in the epidemiology of bovine Besnoitiosis, as intermediate hosts or reservoirs of B. besnoiti, remains unknown. Herein, the first serosurvey conducted in European sheep and goats from areas in Spain where bovine Besnoitiosis is endemic is described. Convenience sampling was conducted of 1943 sheep and 342 goats close to cattle from the Pyrenees and Central Spain that were infected with endemic Besnoitia spp. Serum samples were first analyzed by ELISA and then by confirmatory Western blot. Specific antibodies were not found in any sampled animal. Thus, sheep are unlikely to play a role in the epidemiology of bovine Besnoitiosis, at least in the sampled areas. A larger serosurvey is necessary to determine whether goats might be a putative reservoir. To confirm the results of this study, sheep and goats should be further studied in other European countries and regions where their numbers are high and where bovine Besnoitiosis is spreading.