Genetic characterization of the causative agent of besnoitiosis in goats in Iran on the basis of internal transcribed spacer rDNA and its comparison with Besnoitia species of other hosts

Namazi, Fatemeh; Oryan, Ahmad; Sharifiyazdi, Hassan

doi:10.1007/s00436-010-2107-4

Cited by 12 publications

(9 citation statements)

References 20 publications

(29 reference statements)

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“…The ITS1 sequence was 100% identical or almost 100% identical to those deposited for B. besnoiti and B. bennetti isolates in GenBank™ including isolates from Germany, Israel, Italy, Portugal, Spain, South Africa and USA. This shows the close phylogenetic relationship between B. besnoiti, B. tarandi and B. bennetti , which had already been observed in a number of studies ( Dubey et al, 2004 ; Madubata et al, 2012 ; Schares et al, 2011 ); this very close relationship was also noted, when the ribosomal DNAs of B. besnoiti and B. caprae were compared ( Namazi et al, 2011 ). Consequently, rDNA based PCRs established for B. besnoiti ( Cortes et al, 2007 ; Schares et al, 2011 ) may also be suitable for a PCR diagnosis of B. tarandi .…”

Section: Discussionsupporting

confidence: 80%

Besnoitia tarandi in Canadian woodland caribou – Isolation, characterization and suitability for serological tests

Schares

Jutras

Bärwald

et al. 2019

International Journal for Parasitology: Parasites and Wildlife

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In the present study, we report the first in vitro isolation of Besnoitia tarandi from North America and the second of B. tarandi at all. The parasite was isolated directly from the skin of a Canadian woodland caribou from the migratory ecotype. The animal belonged to the Leaf River Herd, in Northern Quebec, Canada. The isolate was designated Bt-CA-Quebec1.Sequencing of the 3’-end of the 18S rRNA gene, the complete sequence of the ITS1 and the 5’-end of the 5.8S rRNA gene of Bt-CA-Quebec1 revealed only minor differences to rDNA gene fragments of B. besnoiti. In contrast, the patterns for the microsatellite loci Bt-20 and Bt-21 varied substantially from those reported for B. besnoiti and B. bennetti. Surprisingly, the typing results in the loci Bt-6 and Bt-7 differed between Bt-CA-Quebec1 and results obtained for skin samples from caribou of the Canadian regions of Nunavut and the Northwest Territories reported by other investigators. This indicates that differences might exist among B. tarandi in caribou from different regions in Canada.Mice (γ-interferon knockout) intraperitoneally inoculated with 1.2 × 106 or 1.5 × 106 bradyzoites mechanically released from skin tissue cysts fell ill 8, 9 or 18 days post inoculation. GKO mice inoculated with 3.0 × 104 tachyzoites isolated from the peritoneal cavity of a bradyzoites-inoculated mouse became ill earlier, i.e. 5 days post inoculation. Lung was the predilection site in all mice.Bt-CA-Quebec1 tachyzoites rapidly grew in MARC-145 cells and were used for antigen production. Comparative Western blot analyses revealed only a few differences between B. tarandi Bt-CA-Quebec1 and B. besnoiti Evora antigen when probed with sera collected from chronically infected caribou.Due to its fast growth in vitro, the Bt-CA-Quebec1 isolate may represent an interesting antigen source to establish B. tarandi-specific serological tools and to study the biology of this parasite species further.

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Section: Discussionsupporting

confidence: 80%

Besnoitia tarandi in Canadian woodland caribou – Isolation, characterization and suitability for serological tests

Schares

Jutras

Bärwald

et al. 2019

International Journal for Parasitology: Parasites and Wildlife

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“…Following qPCR results, only the positive Besnoitia DNA sample was used for specific identification. A portion of ribosomal DNA cistron containing 5 genes [ 18S , ITS1, 5.8S , internal transcribed spacer 2 (ITS2) and 28S ] was amplified using the primers Bes-F and Bes-R as designed by others [ 7 ] and synthesized by Eurogentec (Angers, France). The PCR reaction was performed in a total volume of 50 μl per sample with 5 μl DNA template, 10 pmol of each primer and 25 μl of the Taq PCR Master Mix® (Qiagen) containing (final concentrations) 1.5 mM MgCl 2 , 200 μM of each dNTP, 1.25 units of Taq polymerase and Qiagen PCR Buffer 1× (pH = 8.7).…”

Section: Case Presentationmentioning

confidence: 99%

“…The ITS1 sequence (244 bp) was completely identical with B. bennetti ITS1 sequences already deposited in the GenBank database (AY827839, AY665399 and JQ013812). ITS1 sequence comparisons between B. bennetti , B. besnoiti , B. caprae and B. tarandi have been previously performed [ 18 ] and emphasized the close relationship between besnoitiosis agents infecting ungulates [ 7 ]. Moreover, as 5.8S (164 bp), ITS2 (393 bp) and partial 18S (56 bp) and 28S (81 bp) sequences are not currently available in the GenBank database for B. bennetti , phylogenetic tree reconstructions were performed using this partial 938 bp rDNA sequence with B. besnoiti and B. caprae rDNA sequences.…”

Section: Case Presentationmentioning

confidence: 99%

“…Its propagation from historic endemic areas in Europe (Spain, Portugal and southern France) took place during the last two decades and currently at least eight European countries have reported clinical cases in imported or autochthonous animals: Switzerland, Italy, Germany, Croatia, Greece, Hungary, Belgium and Ireland [ 2 – 6 ]. For instance, a western blot revealed infection in a six-year-old Blonde d’Aquitaine bull which was imported in Belgium in 2012 from the Pyrenees Mountain endemic region in southern France [ 7 ]. The first stage of infection is the acute systemic phase (tachyzoites are present in the blood) characterized by fever, lymphadenopathy and oedematous swelling, sometimes associated with mortality, abortion and male transient or definitive sterility [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

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First evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium

Liénard

Nabuco²,

Vandenabeele

et al. 2018

Parasites Vectors

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BackgroundBesnoitiosis 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 presentationA 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.ConclusionThis is the first evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium.Electronic supplementary materialThe online version of this article (10.1186/s13071-018-2993-3) contains supplementary material, which is available to authorized users.

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“…B. caprae , a parasite very closely related to B. besnoiti [ 17 , 18 ], causes similar clinical signs in goats [ 19 , 20 ] and data on altered laboratory parameters in caprine besnoitiosis are already available. In naturally acquired caprine besnoitiosis in Iran, alterations of various hematological and biochemical parameters as well as enzyme activities have been observed [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Natural Besnoitia besnoiti infections in cattle: hematological alterations and changes in serum chemistry and enzyme activities

Langenmayer¹,

Scharr²,

Sauter‐Louis

et al. 2015

BMC Vet Res

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BackgroundThe emerging disease bovine besnoitiosis is caused by the apicomplexan parasite Besnoitia besnoiti. Clinical signs of acute besnoitiosis are pyrexia, anorexia and subcutaneous edema. In subacute and chronic besnoitiosis parasitic cysts arise in a variety of tissues and affected cattle display skin lesions and weight loss. In all stages of bovine besnoitiosis, lesions can be found in many organ systems and therefore presumably alter a variety of laboratory parameters. In this study, the impact of naturally acquired acute, subacute and chronic bovine besnoitiosis on hematologic parameters, serum chemistry, and enzyme activities was investigated. Laboratory parameters of two Simmental heifers and two Limousin cows were monitored during acute, subacute and chronic besnoitiosis and in another Simmental heifer during subclinical besnoitiosis. To determine aberrations of laboratory parameters, values were compared with reference ranges obtained from B. besnoiti negative Simmentals (224 samples of nine animals) and Limousins (41 animals). Further, laboratory parameters of B. besnoiti seropositive Limousin cows (54 animals; 32 of these showing clinical signs) and healthy B. besnoiti seronegative Limousin cows (41 animals) were compared.ResultsDuring acute and subacute besnoitiosis, a reduction of leukocyte and erythrocyte concentrations, hematocrit, serum albumin, urea, magnesium, and calcium concentrations were observed. Serum total protein, globulin, total bilirubin and creatinine concentrations were increased and aspartate transaminase (AST) and creatine kinase (CK) activities were elevated. In chronic besnoitiosis, erythrocyte parameters were statistically significantly lower, and total protein and globulin concentrations were significantly higher in B. besnoiti seropositive compared with B. besnoiti seronegative Limousin cows.ConclusionsIn this study, altered laboratory parameters during the course of naturally acquired acute, subacute and chronic bovine besnoitiosis are described for the first time. Only a few animals were examined in acute and subacute besnoitiosis, however the alterations of laboratory parameters during these stages reflected i) the acute inflammatory state (e.g. high levels of serum globulin fractions), ii) clinical findings such as disturbed condition (e.g. bilirubin concentrations), and iii) lesions such as muscle necroses described in the literature (e.g. AST or CK activities). Chronic besnoitiosis led to typical alterations of chronic inflammatory diseases like hyper-(gamma)-globulinemia or reduced erythrocyte concentrations.

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Genetic characterization of the causative agent of besnoitiosis in goats in Iran on the basis of internal transcribed spacer rDNA and its comparison with Besnoitia species of other hosts

Cited by 12 publications

References 20 publications

Besnoitia tarandi in Canadian woodland caribou – Isolation, characterization and suitability for serological tests

Besnoitia tarandi in Canadian woodland caribou – Isolation, characterization and suitability for serological tests

First evidence of Besnoitia bennetti infection (Protozoa: Apicomplexa) in donkeys (Equus asinus) in Belgium

Natural Besnoitia besnoiti infections in cattle: hematological alterations and changes in serum chemistry and enzyme activities

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