The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in ... more The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in 343 well maintained horses belonging to 11 horse stables located in northern Tunisia with indirect immunofluorescence test. Overall, 16.3% (56/343) of tested animals were positive. Anti-A. phagocytophilum antibodies were present in horses located in all studied governorates except the governorate of Ben Arous. Nine horse stables out of 11 contained positive animals, the seroprevalence of each one varied from nought to 50%. Seroprevalence varied according to gender, breed and type of activity, contrary to age. Seroprevalence was higher in females than males (21.4 and 9.5% respectively). Similarly, seropositive animals belonging to the English thoroughbred breed was significantly higher (30.2%) than other breeds. According to the utilization of horses, the highest positivity rate (21.1%) was observed in brood mares. Our results demonstrate that A. phagocytophilum infection is widespread in well maintained horses from northern Tunisia, so equine granulocytic anaplasmosis should be suspected by veterinarians in front of suggestive clinical signs like fever, ataxia and reluctance to move.
Additional file 8: Figure S3. Sensitivity of PCR-HRM targeting 7SL RNA and HSP70 genes. Detection... more Additional file 8: Figure S3. Sensitivity of PCR-HRM targeting 7SL RNA and HSP70 genes. Detection limit of PCR-HRM using 7SL RNA and HSP70 genes was determined using tenfold dilutions of DNA from each reference Leishmania isolate, representing L. infantum (MHOM/TN/94/LV50), L. major (MHOM/TN/2011/EMPA10), and L. tropica (MHOM/IQ/65/L75). a. 7SL PCR-HRM using L. infantum DNA b. HSP70 PCR-HRM using L. infantum DNA. c. 7SL PCR-HRM using L. major DNA d. HSP70 PCR-HRM using L. major DNA. e. 7SL PCR-HRM using L. tropica DNA f. HSP70 PCR-HRM using L. tropica DNA. The right sections of the figures show the tested samples, which are color-coded according to the amplification result, red when positive and green when negative. In all panels, sample codes correspond to the following DNA amounts: A: 20 ng; B: 10 ng; C: 1 ng; D: 0.1 ng; E: 0.01 ng; F: 0.001 ng; G: 0.0001 ng; H: negative control (no DNA).
Crimean-Congo hemorrhagic fever virus (CCHFV, Nairoviridae family) and Rift Valley fever virus (R... more Crimean-Congo hemorrhagic fever virus (CCHFV, Nairoviridae family) and Rift Valley fever virus (RVFV, Phenuiviridae family) are zoonotic vector-borne pathogens with clinical relevance worldwide. Our study aimed to determine seroprevalences of these viruses and potential risk factors among livestock (cattle, sheep, and goats) in Tunisia. Sera were tested for antibodies against CCHFV (n = 879) and RVFV (n = 699) using various enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence assays (IIFA). The overall seroprevalence of IgG antibodies was 8.6% (76/879) and 2.3% (16/699) against CCHFV and RVFV, respectively. For CCHF seropositivity bioclimatic zones and breed were potential risk factors for the three tested animal species; while the season was associated with cattle and sheep seropositivity, tick infestation was associated with cattle and goats seropositivity and age as a risk factor was only associated with cattle seropositivity. Age and season were significan...
Background Discovered by Nicolle and Comte in 1908 in Tunisia, Leishmania infantum is an intracel... more Background Discovered by Nicolle and Comte in 1908 in Tunisia, Leishmania infantum is an intracellular protozoan responsible for zoonotic canine leishmaniosis (CanL) and zoonotic human visceral leishmaniasis (HVL). It is endemic in several regions of the world, including Tunisia, with dogs considered as the main domestic reservoir. The geographic expansion of canine leishmaniosis (CanL) has been linked to global environmental changes that have affected the density and the distribution of its sand fly vectors. Methodology/Principal findings In this study, a cross-sectional epidemiological survey on CanL was carried out in 8 localities in 8 bioclimatic areas of Tunisia. Blood samples were taken from 317 dogs after clinical examination. Collected sera were tested by indirect fluorescent antibody test (IFAT; 1:80) for the presence of anti-Leishmania infantum antibodies. The overall seroprevalence was 58.3% (185/317). Among positive dogs, only 16.7% showed clinical signs suggestive of le...
Table S2. Classification of proteins identified in the salivary glands of Hyalomma dromedarii mal... more Table S2. Classification of proteins identified in the salivary glands of Hyalomma dromedarii males. Description of additional files. Accession: protein ID in the database. %coverage: percentage Coverage_ The percentage of all the amino acids in the protein sequence that were covered by identified peptides detected in the sample. -10logP: the P-value is converted to -10log10 (P-value). In PEAKS, this value is denoted by -10lgP as lg is the ISO reserved notation for log10. By this conversion, a more significant match will have a higher -10lgP value. Additionally, a P-value of 1% is equivalent to -10lgP of 20. #Peptides: number of peptides identified following LC-MS/MS analysis for a single protein. #Unique: Unique peptides_peptides with the same amino acid sequence but different charges or with different modifications are grouped together and counted only once. Avg.Mass: the average mass of a single protein obtained by summing the average atomic masses of the constituent elements. De...
Results of the relationship between percentage of autogeny of Cx. pipiens forms and the type of b... more Results of the relationship between percentage of autogeny of Cx. pipiens forms and the type of breeding site, based on a Generalized Linear Model with Poisson distribution. (PDF 313Â kb)
Relationship between the different types of habitat and breeding site and the proportion of Cx. p... more Relationship between the different types of habitat and breeding site and the proportion of Cx. pipiens based on a Generalized Linear Model (GLM) with Poisson distribution. (PDF 98Â kb)
Relationship between the bioclimatic region and the proportion of Cx. pipiens forms based on a Ge... more Relationship between the bioclimatic region and the proportion of Cx. pipiens forms based on a Generalized Linear Model (GLM) with Poisson distribution. (PDF 98Â kb)
Additional file 1. Presence of Aedes aegypti and Aedes albopictus and case data of Dengue, chikun... more Additional file 1. Presence of Aedes aegypti and Aedes albopictus and case data of Dengue, chikungunya and yellow fever in the EMR.
Distribution of Bartonella species by bioclimatic zones, site, flea species and animal host and p... more Distribution of Bartonella species by bioclimatic zones, site, flea species and animal host and partial sequencing analysis of gltA gene and the ITS region. (PDF 23 kb)
Hemostatic disorders are caused either by platelet-related dysfunctions, defective blood coagulat... more Hemostatic disorders are caused either by platelet-related dysfunctions, defective blood coagulation, or by a combination of both, leading to an increased susceptibility to cardiovascular diseases (CVD) and other related illnesses. The unique specificity of anticoagulants from hematophagous arthropods, such as ticks, suggests that tick saliva holds great promise for discovering new treatments for these life-threatening diseases. In this study, we combined in silico and in vitro analyses to characterize the first recombinant serpin, herein called Dromaserpin, from the sialotranscriptome of the Hyalomma dromedarii tick. Our in silico data described Dromaserpin as a secreted protein of ~43 kDa with high similarities to previously characterized inhibitory serpins. The recombinant protein (rDromaserpin) was obtained as a well-structured monomer, which was tested using global blood coagulation and platelet aggregation assays. With this approach, we confirmed rDromaserpin anticoagulant act...
Small wild mammals are an important element in the emergence and transmission of vector-borne pat... more Small wild mammals are an important element in the emergence and transmission of vector-borne pathogens (VBPs). Among these species, hedgehogs have been found to be a reservoir of VBPs and host of arthropod vectors. Surveillance of VBPs in wildlife and their arthropods are crucial in a one health context. We conducted an exploratory study to screen Atelerix algirus hedgehogs and their infesting ticks and fleas for VBPs using a high throughput microfluidic real-time PCR system. Tested biopsies from hedgehogs were found to be naturally infected by Theileria youngi, Hepatozoon sp., Ehrlichia ewingii, Coxiella burnetii, and Candidatus Ehrlichia shimanensis. Similarly, Haemaphysalis erinacei and Rhipicephalus sanguineus tick species were infected by Ehrlichia ewingii, Rickettsia spp., Rickettsia massiliae, Borrelia sp., Coxiella burnetii, Rickettsia lusitaniae and Anaplasma sp. Archaeopsylla erinacei fleas were infected by Rickettsia asembonensis, Coxiella burnetii, and Rickettsia massil...
Veterinary Parasitology: Regional Studies and Reports, 2021
A paucity of studies is available on haemoparasites in dogs in Tunisia. In this study, we used mo... more A paucity of studies is available on haemoparasites in dogs in Tunisia. In this study, we used molecular techniques (PCR/sequencing) to detect and characterize haemoprotozoa in sick dogs from Tunisia. A total of 99 dogs displaying such clinical symptoms as fever, anorexia, and depression were presented for treatment to the hospital of the Veterinary School of Sidi Thabet (Tunisia). Among dogs screened by PCR, five (5%) proved to be infected with a hemoprotozoa species. An analysis of all the sequences that were obtained enabled us to identify two species of Protozoa: Babesia vogeli (in three dogs) and Hepatozoon canis (in two other dogs). This is the first time that an infection of dogs by Hepatozoon canis in Tunisia has been reported. Veterinary practitioners should be aware that these two haemoparasites can infect dogs and should include them in any differential diagnosis of clinical illnesses with manifestations compatible with tick-borne diseases.
The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in ... more The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in 343 well maintained horses belonging to 11 horse stables located in northern Tunisia with indirect immunofluorescence test. Overall, 16.3% (56/343) of tested animals were positive. Anti-A. phagocytophilum antibodies were present in horses located in all studied governorates except the governorate of Ben Arous. Nine horse stables out of 11 contained positive animals, the seroprevalence of each one varied from nought to 50%. Seroprevalence varied according to gender, breed and type of activity, contrary to age. Seroprevalence was higher in females than males (21.4 and 9.5% respectively). Similarly, seropositive animals belonging to the English thoroughbred breed was significantly higher (30.2%) than other breeds. According to the utilization of horses, the highest positivity rate (21.1%) was observed in brood mares. Our results demonstrate that A. phagocytophilum infection is widespread in...
SUMMARYDescriptions of Babesia occultans have previously been restricted to sub-Saharan Africa. H... more SUMMARYDescriptions of Babesia occultans have previously been restricted to sub-Saharan Africa. Here, we report the finding, for the first time, of this low or non-pathogenic bovine Babesia species in Tunisia, northern Africa. B. occultans DNA was detected by molecular methods in Hyalomma marginatum unfed ticks collected in 3 bioclimatic regions of Tunisia. The near-full-length 18S rRNA gene was sequenced and compared with related sequences retrieved from GenBank. Phylogenetic analysis indicated that other sequences deposited as Babesia sp. could also correspond to B. occultans, suggesting that this species may have a wide distribution in Mediterranean and Asiatic regions, and not only in sub-Saharan Africa as previously described. A B. occultans-specific Reverse Line Blot (RLB) oligonucleotide probe was designed for future epidemiological studies that would help to clarify this possibility.
The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in ... more The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in 343 well maintained horses belonging to 11 horse stables located in northern Tunisia with indirect immunofluorescence test. Overall, 16.3% (56/343) of tested animals were positive. Anti-A. phagocytophilum antibodies were present in horses located in all studied governorates except the governorate of Ben Arous. Nine horse stables out of 11 contained positive animals, the seroprevalence of each one varied from nought to 50%. Seroprevalence varied according to gender, breed and type of activity, contrary to age. Seroprevalence was higher in females than males (21.4 and 9.5% respectively). Similarly, seropositive animals belonging to the English thoroughbred breed was significantly higher (30.2%) than other breeds. According to the utilization of horses, the highest positivity rate (21.1%) was observed in brood mares. Our results demonstrate that A. phagocytophilum infection is widespread in well maintained horses from northern Tunisia, so equine granulocytic anaplasmosis should be suspected by veterinarians in front of suggestive clinical signs like fever, ataxia and reluctance to move.
Additional file 8: Figure S3. Sensitivity of PCR-HRM targeting 7SL RNA and HSP70 genes. Detection... more Additional file 8: Figure S3. Sensitivity of PCR-HRM targeting 7SL RNA and HSP70 genes. Detection limit of PCR-HRM using 7SL RNA and HSP70 genes was determined using tenfold dilutions of DNA from each reference Leishmania isolate, representing L. infantum (MHOM/TN/94/LV50), L. major (MHOM/TN/2011/EMPA10), and L. tropica (MHOM/IQ/65/L75). a. 7SL PCR-HRM using L. infantum DNA b. HSP70 PCR-HRM using L. infantum DNA. c. 7SL PCR-HRM using L. major DNA d. HSP70 PCR-HRM using L. major DNA. e. 7SL PCR-HRM using L. tropica DNA f. HSP70 PCR-HRM using L. tropica DNA. The right sections of the figures show the tested samples, which are color-coded according to the amplification result, red when positive and green when negative. In all panels, sample codes correspond to the following DNA amounts: A: 20 ng; B: 10 ng; C: 1 ng; D: 0.1 ng; E: 0.01 ng; F: 0.001 ng; G: 0.0001 ng; H: negative control (no DNA).
Crimean-Congo hemorrhagic fever virus (CCHFV, Nairoviridae family) and Rift Valley fever virus (R... more Crimean-Congo hemorrhagic fever virus (CCHFV, Nairoviridae family) and Rift Valley fever virus (RVFV, Phenuiviridae family) are zoonotic vector-borne pathogens with clinical relevance worldwide. Our study aimed to determine seroprevalences of these viruses and potential risk factors among livestock (cattle, sheep, and goats) in Tunisia. Sera were tested for antibodies against CCHFV (n = 879) and RVFV (n = 699) using various enzyme-linked immunosorbent assays (ELISAs) and indirect immunofluorescence assays (IIFA). The overall seroprevalence of IgG antibodies was 8.6% (76/879) and 2.3% (16/699) against CCHFV and RVFV, respectively. For CCHF seropositivity bioclimatic zones and breed were potential risk factors for the three tested animal species; while the season was associated with cattle and sheep seropositivity, tick infestation was associated with cattle and goats seropositivity and age as a risk factor was only associated with cattle seropositivity. Age and season were significan...
Background Discovered by Nicolle and Comte in 1908 in Tunisia, Leishmania infantum is an intracel... more Background Discovered by Nicolle and Comte in 1908 in Tunisia, Leishmania infantum is an intracellular protozoan responsible for zoonotic canine leishmaniosis (CanL) and zoonotic human visceral leishmaniasis (HVL). It is endemic in several regions of the world, including Tunisia, with dogs considered as the main domestic reservoir. The geographic expansion of canine leishmaniosis (CanL) has been linked to global environmental changes that have affected the density and the distribution of its sand fly vectors. Methodology/Principal findings In this study, a cross-sectional epidemiological survey on CanL was carried out in 8 localities in 8 bioclimatic areas of Tunisia. Blood samples were taken from 317 dogs after clinical examination. Collected sera were tested by indirect fluorescent antibody test (IFAT; 1:80) for the presence of anti-Leishmania infantum antibodies. The overall seroprevalence was 58.3% (185/317). Among positive dogs, only 16.7% showed clinical signs suggestive of le...
Table S2. Classification of proteins identified in the salivary glands of Hyalomma dromedarii mal... more Table S2. Classification of proteins identified in the salivary glands of Hyalomma dromedarii males. Description of additional files. Accession: protein ID in the database. %coverage: percentage Coverage_ The percentage of all the amino acids in the protein sequence that were covered by identified peptides detected in the sample. -10logP: the P-value is converted to -10log10 (P-value). In PEAKS, this value is denoted by -10lgP as lg is the ISO reserved notation for log10. By this conversion, a more significant match will have a higher -10lgP value. Additionally, a P-value of 1% is equivalent to -10lgP of 20. #Peptides: number of peptides identified following LC-MS/MS analysis for a single protein. #Unique: Unique peptides_peptides with the same amino acid sequence but different charges or with different modifications are grouped together and counted only once. Avg.Mass: the average mass of a single protein obtained by summing the average atomic masses of the constituent elements. De...
Results of the relationship between percentage of autogeny of Cx. pipiens forms and the type of b... more Results of the relationship between percentage of autogeny of Cx. pipiens forms and the type of breeding site, based on a Generalized Linear Model with Poisson distribution. (PDF 313Â kb)
Relationship between the different types of habitat and breeding site and the proportion of Cx. p... more Relationship between the different types of habitat and breeding site and the proportion of Cx. pipiens based on a Generalized Linear Model (GLM) with Poisson distribution. (PDF 98Â kb)
Relationship between the bioclimatic region and the proportion of Cx. pipiens forms based on a Ge... more Relationship between the bioclimatic region and the proportion of Cx. pipiens forms based on a Generalized Linear Model (GLM) with Poisson distribution. (PDF 98Â kb)
Additional file 1. Presence of Aedes aegypti and Aedes albopictus and case data of Dengue, chikun... more Additional file 1. Presence of Aedes aegypti and Aedes albopictus and case data of Dengue, chikungunya and yellow fever in the EMR.
Distribution of Bartonella species by bioclimatic zones, site, flea species and animal host and p... more Distribution of Bartonella species by bioclimatic zones, site, flea species and animal host and partial sequencing analysis of gltA gene and the ITS region. (PDF 23 kb)
Hemostatic disorders are caused either by platelet-related dysfunctions, defective blood coagulat... more Hemostatic disorders are caused either by platelet-related dysfunctions, defective blood coagulation, or by a combination of both, leading to an increased susceptibility to cardiovascular diseases (CVD) and other related illnesses. The unique specificity of anticoagulants from hematophagous arthropods, such as ticks, suggests that tick saliva holds great promise for discovering new treatments for these life-threatening diseases. In this study, we combined in silico and in vitro analyses to characterize the first recombinant serpin, herein called Dromaserpin, from the sialotranscriptome of the Hyalomma dromedarii tick. Our in silico data described Dromaserpin as a secreted protein of ~43 kDa with high similarities to previously characterized inhibitory serpins. The recombinant protein (rDromaserpin) was obtained as a well-structured monomer, which was tested using global blood coagulation and platelet aggregation assays. With this approach, we confirmed rDromaserpin anticoagulant act...
Small wild mammals are an important element in the emergence and transmission of vector-borne pat... more Small wild mammals are an important element in the emergence and transmission of vector-borne pathogens (VBPs). Among these species, hedgehogs have been found to be a reservoir of VBPs and host of arthropod vectors. Surveillance of VBPs in wildlife and their arthropods are crucial in a one health context. We conducted an exploratory study to screen Atelerix algirus hedgehogs and their infesting ticks and fleas for VBPs using a high throughput microfluidic real-time PCR system. Tested biopsies from hedgehogs were found to be naturally infected by Theileria youngi, Hepatozoon sp., Ehrlichia ewingii, Coxiella burnetii, and Candidatus Ehrlichia shimanensis. Similarly, Haemaphysalis erinacei and Rhipicephalus sanguineus tick species were infected by Ehrlichia ewingii, Rickettsia spp., Rickettsia massiliae, Borrelia sp., Coxiella burnetii, Rickettsia lusitaniae and Anaplasma sp. Archaeopsylla erinacei fleas were infected by Rickettsia asembonensis, Coxiella burnetii, and Rickettsia massil...
Veterinary Parasitology: Regional Studies and Reports, 2021
A paucity of studies is available on haemoparasites in dogs in Tunisia. In this study, we used mo... more A paucity of studies is available on haemoparasites in dogs in Tunisia. In this study, we used molecular techniques (PCR/sequencing) to detect and characterize haemoprotozoa in sick dogs from Tunisia. A total of 99 dogs displaying such clinical symptoms as fever, anorexia, and depression were presented for treatment to the hospital of the Veterinary School of Sidi Thabet (Tunisia). Among dogs screened by PCR, five (5%) proved to be infected with a hemoprotozoa species. An analysis of all the sequences that were obtained enabled us to identify two species of Protozoa: Babesia vogeli (in three dogs) and Hepatozoon canis (in two other dogs). This is the first time that an infection of dogs by Hepatozoon canis in Tunisia has been reported. Veterinary practitioners should be aware that these two haemoparasites can infect dogs and should include them in any differential diagnosis of clinical illnesses with manifestations compatible with tick-borne diseases.
The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in ... more The aim of the present study was to determine the seroprevalence of Anaplasma phagocytophilum in 343 well maintained horses belonging to 11 horse stables located in northern Tunisia with indirect immunofluorescence test. Overall, 16.3% (56/343) of tested animals were positive. Anti-A. phagocytophilum antibodies were present in horses located in all studied governorates except the governorate of Ben Arous. Nine horse stables out of 11 contained positive animals, the seroprevalence of each one varied from nought to 50%. Seroprevalence varied according to gender, breed and type of activity, contrary to age. Seroprevalence was higher in females than males (21.4 and 9.5% respectively). Similarly, seropositive animals belonging to the English thoroughbred breed was significantly higher (30.2%) than other breeds. According to the utilization of horses, the highest positivity rate (21.1%) was observed in brood mares. Our results demonstrate that A. phagocytophilum infection is widespread in...
SUMMARYDescriptions of Babesia occultans have previously been restricted to sub-Saharan Africa. H... more SUMMARYDescriptions of Babesia occultans have previously been restricted to sub-Saharan Africa. Here, we report the finding, for the first time, of this low or non-pathogenic bovine Babesia species in Tunisia, northern Africa. B. occultans DNA was detected by molecular methods in Hyalomma marginatum unfed ticks collected in 3 bioclimatic regions of Tunisia. The near-full-length 18S rRNA gene was sequenced and compared with related sequences retrieved from GenBank. Phylogenetic analysis indicated that other sequences deposited as Babesia sp. could also correspond to B. occultans, suggesting that this species may have a wide distribution in Mediterranean and Asiatic regions, and not only in sub-Saharan Africa as previously described. A B. occultans-specific Reverse Line Blot (RLB) oligonucleotide probe was designed for future epidemiological studies that would help to clarify this possibility.
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