The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several... more The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several areas where Lyme disease is endemic in northern Spain. A low rate of infestation by Ixodes ricinus nymphs was found in the small mammal populations studied that correlated with the near-absence of B. burgdorferi sensu lato in 184 animals tested and with the lack of
Fifteen Borrelia burgdorferi sensu lato isolates from questing ticks and skin biopsy specimens fr... more Fifteen Borrelia burgdorferi sensu lato isolates from questing ticks and skin biopsy specimens from erythema migrans patients in three different areas of Spain were characterized. Four different genospecies were found (nine Borrelia garinii, including the two human isolates, three B. burgdorferi sensu stricto, two B. valaisiana, and one B. lusitaniae), showing a diverse spectrum of B. burgdorferi sensu lato species. B. garinii isolates were highly variable in terms of pulsed-field gel electrophoresis pattern and OspA serotype, with four of the seven serotypes described. One of the human isolates was OspA serotype 5, the same found in four of seven tick isolates. The second human isolate was OspA serotype 3, which was not present in ticks from the same area. Seven B. garinii isolates were able to disseminate through the skin of C3H/HeN mice and to cause severe inflammation of joints. One of the two B. valaisiana isolates also caused disease in mice. Only one B. burgdorferi sensu stri...
Revue scientifique et technique (International Office of Epizootics), 2004
The authors investigated the prevalence of Salmonella spp. in 205 wild birds and mammals belongin... more The authors investigated the prevalence of Salmonella spp. in 205 wild birds and mammals belonging to 45 species during the years 2001 and 2002 in the Basque Country (Spain). Salmonella was isolated from 16 (7.8%) animals. The prevalence was 8.5% (7/82) in birds, and 7.2% (9/123) in mammals. Nine serotypes, all of them belonging to the species Salmonella enterica, were identified: two isolates of Typhimurium (from 1/3 griffon vultures [Gyps fulvus], and 1/5 sparrowhawks [Accipiter nisus]); one of 6,14:z4, z23: (subsp. houtenae, 1/1 common kestrel [Falco tinnunculus]); one of Muenchen (1/1 captive Harris's hawk [Parabuteo unicinctus]); two of Enteritidis (1/5 tawny owls [Strix aluco], and 1/14 foxes [Vulpes vulpes]); one of Give, Newport and Umbilo and one untyped islolate (4/22 badgers [Meles meles]); two of Worthington and one of 38:IV:z35 (subsp. arizonae, 3/40 wild boars [Sus scrofa]); and three other untyped isolates (1/1 northern fulmar [Fulmarus glacialis], 1/11 buzzards [...
Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish ... more Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n = 53), 29% of foxes (n = 48), 29% of stone martens (n = 21) and in 25-50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.
An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic... more An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV) was carried out in red-legged partridges (Alectoris rufa) in order to study clinical signs, gross and microscopic lesions, and viral distribution in tissues and viral shedding. Birds were infected with a HPAIV subtype H7N1 (A/Chicken/Italy/5093/1999) and a LPAIV subtype H7N9 (A/Anas crecca/Spain/1460/2008). Uninoculated birds were included as contacts in both groups. In HPAIV infected birds, the first clinical signs were observed at 3 dpi, and mortality started at 4 dpi, reaching 100% at 8 dpi. The presence of viral antigen in tissues and viral shedding were confirmed by immunohistochemistry and quantitative real time RT-PCR (qRRT-PCR), respectively, in all birds infected with HPAIV. However, neither clinical signs nor histopathological findings were observed in LPAIV infected partridges. In addition, only short-term viral shedding together with seroconversion was detected in some LPAIV inoculated animals. The present study demonstrates that the red-legged partridge is highly susceptible to the H7N1 HPAIV strain, causing severe disease, mortality and abundant viral shedding and thus contributing to the spread of a potential local outbreak of this virus. In contrast, our results concerning H7N9 LPAIV suggest that the red-legged partridge is not a reservoir species for this virus.
We describe the distribution of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red... more We describe the distribution of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red deer (Cervus elaphus) in Spain. Animals with TBL were confirmed in 84.21% of mixed populations (n = 57) of red deer and wild boar and in 75% of populations of wild boar alone (n = 8) in central and southern Spain (core area). The prevalence of TBL declined towards the periphery of this region. In the core area, the prevalence ranged up to 100% in local populations of wild boar (mean estate prevalence 42.51%) and up to 50% in red deer (mean estate prevalence 13.70%). We carried out exploratory statistical analyses to describe the epidemiology of TBL in both species throughout the core area. Prevalence of TBL increased with age in both species. Wild boar and red deer mean TBL prevalence at the estate level were positively associated, and lesion scores were consistently higher in wild boars than in red deer. The wild boar prevalence of TBL in wild boar did not differ between populations that were or were not cohabiting with red deer. Amongst the wild boars with TBL, 61.19% presented generalized lesions, and the proportion of generalized cases was similar between sex and age classes. In red deer, 57.14% of TBL-positive individuals presented generalized lesions, and the percentage of generalized cases increased with age class, but did not differ between the sexes. These results highlight the potential importance of wild boar and red deer in the maintenance of tuberculosis in south central Spain.
At present few studies have been carried out on the distribution and incidence of Coxiella burnet... more At present few studies have been carried out on the distribution and incidence of Coxiella burnetii infection in wildlife. Therefore, the aim of this study was to investigate the distribution of C. burnetii in the main wild species in the Basque Country (Northern Spain), such as carnivores, cervids, wild boar, lagomorphs and several species of birds. Tissues from a total of 601 animals and 340 adult ticks collected from them were analyzed by PCR. DNA of C. burnetii was detected in 5.1% of roe deer (Capreolus capreolus), 4.3% of wild boar (Sus scrofa), 9.1% of European hare (Lepus europaeus), and among wild birds, in 11% of vultures (Gyps fulvus) and 14% of black kites (Milvus migrans). These results showed that C. burnetii circulates in wildlife in Spain participating in the cycle of Q fever in nature. All of the adult ticks analyzed were negative for C. burnetii, suggesting that ticks do not play an important role in the transmission of C. burnetii in this area.
Information on lesion distribution and characteristics is essential to determine the significance... more Information on lesion distribution and characteristics is essential to determine the significance of a species as a reservoir host for tuberculosis (TB). Herein, we describe the extension and distribution of lesions in 127 Mycobacterium tuberculosis Complex culture positive European wild boars (Sus scrofa), and use this information to discuss the role of this wildlife species in TB epidemiology in Mediterranean Spain. Macroscopic TB-compatible lesions were detected in 105 of 127 wild boars (82.68%). Only microscopic lesions were found in 11 wild boars (8.66%). Lesions were not evident in 11 wild boars (8.66%). A total of 49 wild boars had lesions confined to one anatomical region (42.2%, localized TB), while 67 animals had lesions in more than one anatomical region (57.8%, generalized TB). Head lymph nodes (LNs), particularly the mandibular LNs, were most frequently affected (107/116, 92.24%), and 43 wild boar had only mandibular LN lesions. Histopathology evidenced TB lesions in 38.1% of the lungs, 23% of the livers and 13% of the spleens examined. Mammary gland lesions were observed in three cases. When TB lesions were localized, granulomas characterized by a mixed inflammatory cell population were more predominant, whereas strongly necrotic-calcified granulomas were more prevalent in generalized cases of TB infection. Large lesions in more than one anatomical region were more frequent among juveniles. The histopathological characteristics of the tuberculous reaction and the associated tissue damage in various organs, together with the gross pathology, indicate that at least those wild boar with large lesions and generalized infections have the potential to excrete mycobacteria by several routes. This finding, in the context of unusually high densities of wild boar and fencing and feeding, reinforces the suggestion that wild boar can act as a true TB reservoir under the particular circumstances of Mediterranean Spain. Further studies on the routes of excretion as well as the effect of altering management methods would be of interest to confirm the role of wild boar in TB epidemiology in Spain.
This study was aimed at investigating risk factors associated with prevalence of tuberculosis (TB... more This study was aimed at investigating risk factors associated with prevalence of tuberculosis (TB)-like lesions and associated mycobacteria in Ethiopian cattle slaughtered. The study was carried out during 2006-2007 in five selected municipal and export abattoirs. Methods of investigation involved detailed necropsy examination of carcasses and isolation of mycobacteria from pathologic tissue specimens. Factors of animal and environment origin were recorded and examined as explanatory variables in relation to the presence or absence of TB-like lesions and the presence of viable mycobacteria. Multivariable logistic regression analysis was used to identify risk factors associated with prevalence of TB-like lesions and mycobacteria. Out of 3,322 carcasses inspected, 10.2% (95% confidence interval (CI) [9.2-11.3]) and 3.2% (95% CI [2.6-3.8]) were positive, respectively, based on necropsy and bacteriologic examinations. The highest and lowest lesion prevalence was recorded in Adama (24.7%) and Yabello (4.2%), respectively. Multivariable logistic regression analysis identified age, breed, abattoir location, geographic origin and management system as being risk factors for prevalence of TB-like lesions and occurrence of viable mycobacteria in Ethiopian cattle. The study demonstrated mycobacterial infection as important infectious disease of Ethiopian cattle. The reported confirmed cases of the disease in different management and geographic settings in Ethiopia disproved the earlier held opinion of its occurrence as a low sporadic profile. In view of a dietary proclivity of Ethiopian communities (milk and meat are predominantly consumed raw) and lifestyle (close contact of people with animals), the risk of bovine tuberculosis as a public health threat is eminent.
The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whethe... more The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whether sympatric carnivores are reservoirs of pathogens posing a disease risk for the lynx, evidence of exposure to 17 viral, bacterial and protozoan agents was investigated in 176 carnivores comprising 26 free-living lynx, 53 domestic cats, 28 dogs, 33 red foxes (Vulpes vulpes), 24 Egyptian mongooses (Herpestes ichneumon), 10 common genets (Genetta genetta) and 2 Eurasian badgers (Meles meles) in the areas inhabited by the last two populations of Iberian lynx, both in Andalusia (South-Western Spain).
Methods available for the diagnosis of Ehrlichia phagocytophila infection are limited and of low ... more Methods available for the diagnosis of Ehrlichia phagocytophila infection are limited and of low sensitivity. Since polymerase chain reaction (PCR) is a highly sensitive and specific method for the detection of infectious agents in animals, it was considered useful to develop a PCR method specific for this agent. An experimental protocol in which blood from 10 different flocks was inoculated into 10 naive 1 month-old lambs was carried out. Progress of the infection was monitored during 3 weeks by daily rectal temperature and blood samples. Eight lambs showed hyperthermia, presence of E. phagocytophila in blood, and lower growth rate while the other two never showed the agent in their blood or any clinical signs. This experimental infection, essentially designed to provide infected samples, also showed that flocks from areas previously thought to be free of E. phagocytophila were actually infected. PCR on blood samples allowed detection of the infection earlier than in case of microscopic examination. This was related to an increased sensitivity (87-91%). PCR yielded positive results on samples with as few as 1.08 infected neutrophils/µl (about 0.1%).
Wildlife can act as reservoir of different tick-borne pathogens of veterinary and zoonotic import... more Wildlife can act as reservoir of different tick-borne pathogens of veterinary and zoonotic importance. To investigate the role of wild ruminants as reservoir of piroplasm infection, 28 red deer, 69 roe deer and 38 chamois from Northern Spain were examined by reverse line blot (RLB) hybridization. The survey detected a prevalence of 85 . 7% in red deer, 62 . 3% in roe deer and 28 . 9% in chamois. Four different piroplasms were identified : Theileria sp. OT3 (previously described in sheep) as the most prevalent (85 . 7% in red deer, 46 . 4% in roe deer and 26 . 3% in chamois) ; Theileria sp. 3185/02 (previously described in a red deer in Central Spain) more abundant in red deer (53 . 6%) than in roe deer (10 . 1%) but absent from chamois; Babesia divergens detected in 6 roe deer; Theileria ovis present in 1 chamois. Mixed infections (Theileria sp. OT3 and Theileria sp. 3185/02) were only found in red and roe deer. Sequencing analysis of the 18S rRNA gene confirmed the RLB results and showed 99 . 7% identity between Theileria sp. 3185/02 and T. capreoli, suggesting that they are the same species. Tick distribution and contact of wild ruminants with domestic animals are discussed in terms of piroplasm infection. The results suggest that a considerable number of wildlife ruminants are asymptomatic carriers that may serve as reservoirs of the infection posing a serious concern in terms of piroplasmosis control.
Ixodid tick abundance was investigated in the Basque region in Spain in two 1-year longitudinal s... more Ixodid tick abundance was investigated in the Basque region in Spain in two 1-year longitudinal studies, in 1992-1993 and 2003-2004. Forty zones were visited monthly and 162 672 ticks (87% larvae, 12% nymphs and 1% adults) were collected by blanket dragging. Eleven tick species belonging to the genera Ixodes, Haemaphysalis, Rhipicephalus and Dermacentor were identified including Haemaphysalis concinna Koch, which had not previously been reported in Spain. Tick species abundance differed between zones, studies and seasons. In 1992-1993, Haemaphysalis punctata Canestrini & Fanzago was the predominant species and distinct spring-summer and autumn-early winter peaks of activity were observed. In 2003-2004, Ixodes ricinus (Linneaus) was the most common species and was active throughout the winter. Larval and nymph seasonal activity patterns coincided in both 1993 and 2003 and this could facilitate co-feeding transmission of pathogens. Higher tick abundance was associated with increased livestock abundance in 1992-1993 and milder winter temperatures in 2003-2004. Tick collection rates in areas with moderate and high tick density were positively associated with the interaction between ambient temperature at sampling and rainfall 7 days prior to sampling. Collection rates were also significantly higher at medium rather than higher altitude, in forested areas than in open grasslands and lower in recreational areas frequented by people and with wet vegetation at sampling.
Borrelia burgdorferi was found widespread in ixodid ticks from the Basque Country (Spain) during ... more Borrelia burgdorferi was found widespread in ixodid ticks from the Basque Country (Spain) during a two-step study. In the Þrst part, a total of 7,835 ixodids of eight different species was collected from vegetation, classiÞed, and processed using polymerase chain reaction (PCR) for detection of B. burgdorferi ospA DNA. B. burgdorferi DNA was detected in Յ12.5% of adults and Ն0.6% of Ixodes ricinus (L., 1758) nymphs (mean 1.5 and 0.05%, respectively), and in Յ14.3% of adult Hemaphysalis punctata (Canestrini & Fanzago, 1877) analyzed (mean 1.2%). The second part of the study was undertaken 2 yr later to characterize B. burgdorferi distribution by focusing on the areas where I. ricinus was the predominant species. Ten areas were selected from which 1,535 nymphs and adults of I. ricinus were collected and processed by PCR and culture techniques. Infected ticks were found in all zones. B. burgdorferi DNA was detected in a mean of 9.3 and 1.5% of adults and nymphs, respectively. Nine isolates of B. burgdorferi were obtained, belonging to four different genospecies (B. burgdorferi sensu stricto, B. garinii, B. valaisiana, and B. lusitaniae). The results indicate that some areas of Spain have a potential risk for Lyme disease agent exposure and that B. burgdorferi appears to have an increasing occurrence in ticks in the Basque Country.
A new, efficient molecular method for detection of Bartonella, based on the 16S-23S rRNA intergen... more A new, efficient molecular method for detection of Bartonella, based on the 16S-23S rRNA intergenic spacer and 16S rRNA amplification by multiplex PCR combined with reverse line blotting, was designed. This assay could simultaneously detect 20 different known species and other Bartonella species not described previously.
The helminthological study of 26 Eurasian badgers ( Meles meles L.) in the Basque Country (northe... more The helminthological study of 26 Eurasian badgers ( Meles meles L.) in the Basque Country (northern Spain) revealed the presence of two trematode ( Euryhelmis squamula and Brachylaima sp.), three cestode ( Atriotaenia incisa, Mesocestoides sp. and Taenia sp.) and five nematode ( Aonchotheca putorii, Physaloptera sp., Molineus patens, Uncinaria criniformis and Strongyloides sp.) species. All 15 individuals analysed for Trichinella sp. were negative. The coprological analysis (flotation and migration) revealed the excretion of strongylid, capillariid and trematode eggs, and Crenosoma sp. and Angiostrongylus sp. larvae. No sex- or age-related differences were found in the parasite burden or egg and larvae excretion. A. incisa and M. patens abundances were positively correlated, both species being negatively correlated with badger weight. The presence of adults of U. criniformis and strongylid eggs in faeces was not independent. Badgers excreting Angiostrongylus larvae were in poorer condition than those not excreting.
The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several... more The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several areas where Lyme disease is endemic in northern Spain. A low rate of infestation by Ixodes ricinus nymphs was found in the small mammal populations studied that correlated with the near-absence of B. burgdorferi sensu lato in 184 animals tested and with the lack of
Fifteen Borrelia burgdorferi sensu lato isolates from questing ticks and skin biopsy specimens fr... more Fifteen Borrelia burgdorferi sensu lato isolates from questing ticks and skin biopsy specimens from erythema migrans patients in three different areas of Spain were characterized. Four different genospecies were found (nine Borrelia garinii, including the two human isolates, three B. burgdorferi sensu stricto, two B. valaisiana, and one B. lusitaniae), showing a diverse spectrum of B. burgdorferi sensu lato species. B. garinii isolates were highly variable in terms of pulsed-field gel electrophoresis pattern and OspA serotype, with four of the seven serotypes described. One of the human isolates was OspA serotype 5, the same found in four of seven tick isolates. The second human isolate was OspA serotype 3, which was not present in ticks from the same area. Seven B. garinii isolates were able to disseminate through the skin of C3H/HeN mice and to cause severe inflammation of joints. One of the two B. valaisiana isolates also caused disease in mice. Only one B. burgdorferi sensu stri...
Revue scientifique et technique (International Office of Epizootics), 2004
The authors investigated the prevalence of Salmonella spp. in 205 wild birds and mammals belongin... more The authors investigated the prevalence of Salmonella spp. in 205 wild birds and mammals belonging to 45 species during the years 2001 and 2002 in the Basque Country (Spain). Salmonella was isolated from 16 (7.8%) animals. The prevalence was 8.5% (7/82) in birds, and 7.2% (9/123) in mammals. Nine serotypes, all of them belonging to the species Salmonella enterica, were identified: two isolates of Typhimurium (from 1/3 griffon vultures [Gyps fulvus], and 1/5 sparrowhawks [Accipiter nisus]); one of 6,14:z4, z23: (subsp. houtenae, 1/1 common kestrel [Falco tinnunculus]); one of Muenchen (1/1 captive Harris's hawk [Parabuteo unicinctus]); two of Enteritidis (1/5 tawny owls [Strix aluco], and 1/14 foxes [Vulpes vulpes]); one of Give, Newport and Umbilo and one untyped islolate (4/22 badgers [Meles meles]); two of Worthington and one of 38:IV:z35 (subsp. arizonae, 3/40 wild boars [Sus scrofa]); and three other untyped isolates (1/1 northern fulmar [Fulmarus glacialis], 1/11 buzzards [...
Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish ... more Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n = 53), 29% of foxes (n = 48), 29% of stone martens (n = 21) and in 25-50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.
An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic... more An experimental infection with highly pathogenic avian influenza virus (HPAIV) and low pathogenic avian influenza virus (LPAIV) was carried out in red-legged partridges (Alectoris rufa) in order to study clinical signs, gross and microscopic lesions, and viral distribution in tissues and viral shedding. Birds were infected with a HPAIV subtype H7N1 (A/Chicken/Italy/5093/1999) and a LPAIV subtype H7N9 (A/Anas crecca/Spain/1460/2008). Uninoculated birds were included as contacts in both groups. In HPAIV infected birds, the first clinical signs were observed at 3 dpi, and mortality started at 4 dpi, reaching 100% at 8 dpi. The presence of viral antigen in tissues and viral shedding were confirmed by immunohistochemistry and quantitative real time RT-PCR (qRRT-PCR), respectively, in all birds infected with HPAIV. However, neither clinical signs nor histopathological findings were observed in LPAIV infected partridges. In addition, only short-term viral shedding together with seroconversion was detected in some LPAIV inoculated animals. The present study demonstrates that the red-legged partridge is highly susceptible to the H7N1 HPAIV strain, causing severe disease, mortality and abundant viral shedding and thus contributing to the spread of a potential local outbreak of this virus. In contrast, our results concerning H7N9 LPAIV suggest that the red-legged partridge is not a reservoir species for this virus.
We describe the distribution of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red... more We describe the distribution of tuberculosis-like lesions (TBL) in wild boar (Sus scrofa) and red deer (Cervus elaphus) in Spain. Animals with TBL were confirmed in 84.21% of mixed populations (n = 57) of red deer and wild boar and in 75% of populations of wild boar alone (n = 8) in central and southern Spain (core area). The prevalence of TBL declined towards the periphery of this region. In the core area, the prevalence ranged up to 100% in local populations of wild boar (mean estate prevalence 42.51%) and up to 50% in red deer (mean estate prevalence 13.70%). We carried out exploratory statistical analyses to describe the epidemiology of TBL in both species throughout the core area. Prevalence of TBL increased with age in both species. Wild boar and red deer mean TBL prevalence at the estate level were positively associated, and lesion scores were consistently higher in wild boars than in red deer. The wild boar prevalence of TBL in wild boar did not differ between populations that were or were not cohabiting with red deer. Amongst the wild boars with TBL, 61.19% presented generalized lesions, and the proportion of generalized cases was similar between sex and age classes. In red deer, 57.14% of TBL-positive individuals presented generalized lesions, and the percentage of generalized cases increased with age class, but did not differ between the sexes. These results highlight the potential importance of wild boar and red deer in the maintenance of tuberculosis in south central Spain.
At present few studies have been carried out on the distribution and incidence of Coxiella burnet... more At present few studies have been carried out on the distribution and incidence of Coxiella burnetii infection in wildlife. Therefore, the aim of this study was to investigate the distribution of C. burnetii in the main wild species in the Basque Country (Northern Spain), such as carnivores, cervids, wild boar, lagomorphs and several species of birds. Tissues from a total of 601 animals and 340 adult ticks collected from them were analyzed by PCR. DNA of C. burnetii was detected in 5.1% of roe deer (Capreolus capreolus), 4.3% of wild boar (Sus scrofa), 9.1% of European hare (Lepus europaeus), and among wild birds, in 11% of vultures (Gyps fulvus) and 14% of black kites (Milvus migrans). These results showed that C. burnetii circulates in wildlife in Spain participating in the cycle of Q fever in nature. All of the adult ticks analyzed were negative for C. burnetii, suggesting that ticks do not play an important role in the transmission of C. burnetii in this area.
Information on lesion distribution and characteristics is essential to determine the significance... more Information on lesion distribution and characteristics is essential to determine the significance of a species as a reservoir host for tuberculosis (TB). Herein, we describe the extension and distribution of lesions in 127 Mycobacterium tuberculosis Complex culture positive European wild boars (Sus scrofa), and use this information to discuss the role of this wildlife species in TB epidemiology in Mediterranean Spain. Macroscopic TB-compatible lesions were detected in 105 of 127 wild boars (82.68%). Only microscopic lesions were found in 11 wild boars (8.66%). Lesions were not evident in 11 wild boars (8.66%). A total of 49 wild boars had lesions confined to one anatomical region (42.2%, localized TB), while 67 animals had lesions in more than one anatomical region (57.8%, generalized TB). Head lymph nodes (LNs), particularly the mandibular LNs, were most frequently affected (107/116, 92.24%), and 43 wild boar had only mandibular LN lesions. Histopathology evidenced TB lesions in 38.1% of the lungs, 23% of the livers and 13% of the spleens examined. Mammary gland lesions were observed in three cases. When TB lesions were localized, granulomas characterized by a mixed inflammatory cell population were more predominant, whereas strongly necrotic-calcified granulomas were more prevalent in generalized cases of TB infection. Large lesions in more than one anatomical region were more frequent among juveniles. The histopathological characteristics of the tuberculous reaction and the associated tissue damage in various organs, together with the gross pathology, indicate that at least those wild boar with large lesions and generalized infections have the potential to excrete mycobacteria by several routes. This finding, in the context of unusually high densities of wild boar and fencing and feeding, reinforces the suggestion that wild boar can act as a true TB reservoir under the particular circumstances of Mediterranean Spain. Further studies on the routes of excretion as well as the effect of altering management methods would be of interest to confirm the role of wild boar in TB epidemiology in Spain.
This study was aimed at investigating risk factors associated with prevalence of tuberculosis (TB... more This study was aimed at investigating risk factors associated with prevalence of tuberculosis (TB)-like lesions and associated mycobacteria in Ethiopian cattle slaughtered. The study was carried out during 2006-2007 in five selected municipal and export abattoirs. Methods of investigation involved detailed necropsy examination of carcasses and isolation of mycobacteria from pathologic tissue specimens. Factors of animal and environment origin were recorded and examined as explanatory variables in relation to the presence or absence of TB-like lesions and the presence of viable mycobacteria. Multivariable logistic regression analysis was used to identify risk factors associated with prevalence of TB-like lesions and mycobacteria. Out of 3,322 carcasses inspected, 10.2% (95% confidence interval (CI) [9.2-11.3]) and 3.2% (95% CI [2.6-3.8]) were positive, respectively, based on necropsy and bacteriologic examinations. The highest and lowest lesion prevalence was recorded in Adama (24.7%) and Yabello (4.2%), respectively. Multivariable logistic regression analysis identified age, breed, abattoir location, geographic origin and management system as being risk factors for prevalence of TB-like lesions and occurrence of viable mycobacteria in Ethiopian cattle. The study demonstrated mycobacterial infection as important infectious disease of Ethiopian cattle. The reported confirmed cases of the disease in different management and geographic settings in Ethiopia disproved the earlier held opinion of its occurrence as a low sporadic profile. In view of a dietary proclivity of Ethiopian communities (milk and meat are predominantly consumed raw) and lifestyle (close contact of people with animals), the risk of bovine tuberculosis as a public health threat is eminent.
The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whethe... more The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whether sympatric carnivores are reservoirs of pathogens posing a disease risk for the lynx, evidence of exposure to 17 viral, bacterial and protozoan agents was investigated in 176 carnivores comprising 26 free-living lynx, 53 domestic cats, 28 dogs, 33 red foxes (Vulpes vulpes), 24 Egyptian mongooses (Herpestes ichneumon), 10 common genets (Genetta genetta) and 2 Eurasian badgers (Meles meles) in the areas inhabited by the last two populations of Iberian lynx, both in Andalusia (South-Western Spain).
Methods available for the diagnosis of Ehrlichia phagocytophila infection are limited and of low ... more Methods available for the diagnosis of Ehrlichia phagocytophila infection are limited and of low sensitivity. Since polymerase chain reaction (PCR) is a highly sensitive and specific method for the detection of infectious agents in animals, it was considered useful to develop a PCR method specific for this agent. An experimental protocol in which blood from 10 different flocks was inoculated into 10 naive 1 month-old lambs was carried out. Progress of the infection was monitored during 3 weeks by daily rectal temperature and blood samples. Eight lambs showed hyperthermia, presence of E. phagocytophila in blood, and lower growth rate while the other two never showed the agent in their blood or any clinical signs. This experimental infection, essentially designed to provide infected samples, also showed that flocks from areas previously thought to be free of E. phagocytophila were actually infected. PCR on blood samples allowed detection of the infection earlier than in case of microscopic examination. This was related to an increased sensitivity (87-91%). PCR yielded positive results on samples with as few as 1.08 infected neutrophils/µl (about 0.1%).
Wildlife can act as reservoir of different tick-borne pathogens of veterinary and zoonotic import... more Wildlife can act as reservoir of different tick-borne pathogens of veterinary and zoonotic importance. To investigate the role of wild ruminants as reservoir of piroplasm infection, 28 red deer, 69 roe deer and 38 chamois from Northern Spain were examined by reverse line blot (RLB) hybridization. The survey detected a prevalence of 85 . 7% in red deer, 62 . 3% in roe deer and 28 . 9% in chamois. Four different piroplasms were identified : Theileria sp. OT3 (previously described in sheep) as the most prevalent (85 . 7% in red deer, 46 . 4% in roe deer and 26 . 3% in chamois) ; Theileria sp. 3185/02 (previously described in a red deer in Central Spain) more abundant in red deer (53 . 6%) than in roe deer (10 . 1%) but absent from chamois; Babesia divergens detected in 6 roe deer; Theileria ovis present in 1 chamois. Mixed infections (Theileria sp. OT3 and Theileria sp. 3185/02) were only found in red and roe deer. Sequencing analysis of the 18S rRNA gene confirmed the RLB results and showed 99 . 7% identity between Theileria sp. 3185/02 and T. capreoli, suggesting that they are the same species. Tick distribution and contact of wild ruminants with domestic animals are discussed in terms of piroplasm infection. The results suggest that a considerable number of wildlife ruminants are asymptomatic carriers that may serve as reservoirs of the infection posing a serious concern in terms of piroplasmosis control.
Ixodid tick abundance was investigated in the Basque region in Spain in two 1-year longitudinal s... more Ixodid tick abundance was investigated in the Basque region in Spain in two 1-year longitudinal studies, in 1992-1993 and 2003-2004. Forty zones were visited monthly and 162 672 ticks (87% larvae, 12% nymphs and 1% adults) were collected by blanket dragging. Eleven tick species belonging to the genera Ixodes, Haemaphysalis, Rhipicephalus and Dermacentor were identified including Haemaphysalis concinna Koch, which had not previously been reported in Spain. Tick species abundance differed between zones, studies and seasons. In 1992-1993, Haemaphysalis punctata Canestrini & Fanzago was the predominant species and distinct spring-summer and autumn-early winter peaks of activity were observed. In 2003-2004, Ixodes ricinus (Linneaus) was the most common species and was active throughout the winter. Larval and nymph seasonal activity patterns coincided in both 1993 and 2003 and this could facilitate co-feeding transmission of pathogens. Higher tick abundance was associated with increased livestock abundance in 1992-1993 and milder winter temperatures in 2003-2004. Tick collection rates in areas with moderate and high tick density were positively associated with the interaction between ambient temperature at sampling and rainfall 7 days prior to sampling. Collection rates were also significantly higher at medium rather than higher altitude, in forested areas than in open grasslands and lower in recreational areas frequented by people and with wet vegetation at sampling.
Borrelia burgdorferi was found widespread in ixodid ticks from the Basque Country (Spain) during ... more Borrelia burgdorferi was found widespread in ixodid ticks from the Basque Country (Spain) during a two-step study. In the Þrst part, a total of 7,835 ixodids of eight different species was collected from vegetation, classiÞed, and processed using polymerase chain reaction (PCR) for detection of B. burgdorferi ospA DNA. B. burgdorferi DNA was detected in Յ12.5% of adults and Ն0.6% of Ixodes ricinus (L., 1758) nymphs (mean 1.5 and 0.05%, respectively), and in Յ14.3% of adult Hemaphysalis punctata (Canestrini & Fanzago, 1877) analyzed (mean 1.2%). The second part of the study was undertaken 2 yr later to characterize B. burgdorferi distribution by focusing on the areas where I. ricinus was the predominant species. Ten areas were selected from which 1,535 nymphs and adults of I. ricinus were collected and processed by PCR and culture techniques. Infected ticks were found in all zones. B. burgdorferi DNA was detected in a mean of 9.3 and 1.5% of adults and nymphs, respectively. Nine isolates of B. burgdorferi were obtained, belonging to four different genospecies (B. burgdorferi sensu stricto, B. garinii, B. valaisiana, and B. lusitaniae). The results indicate that some areas of Spain have a potential risk for Lyme disease agent exposure and that B. burgdorferi appears to have an increasing occurrence in ticks in the Basque Country.
A new, efficient molecular method for detection of Bartonella, based on the 16S-23S rRNA intergen... more A new, efficient molecular method for detection of Bartonella, based on the 16S-23S rRNA intergenic spacer and 16S rRNA amplification by multiplex PCR combined with reverse line blotting, was designed. This assay could simultaneously detect 20 different known species and other Bartonella species not described previously.
The helminthological study of 26 Eurasian badgers ( Meles meles L.) in the Basque Country (northe... more The helminthological study of 26 Eurasian badgers ( Meles meles L.) in the Basque Country (northern Spain) revealed the presence of two trematode ( Euryhelmis squamula and Brachylaima sp.), three cestode ( Atriotaenia incisa, Mesocestoides sp. and Taenia sp.) and five nematode ( Aonchotheca putorii, Physaloptera sp., Molineus patens, Uncinaria criniformis and Strongyloides sp.) species. All 15 individuals analysed for Trichinella sp. were negative. The coprological analysis (flotation and migration) revealed the excretion of strongylid, capillariid and trematode eggs, and Crenosoma sp. and Angiostrongylus sp. larvae. No sex- or age-related differences were found in the parasite burden or egg and larvae excretion. A. incisa and M. patens abundances were positively correlated, both species being negatively correlated with badger weight. The presence of adults of U. criniformis and strongylid eggs in faeces was not independent. Badgers excreting Angiostrongylus larvae were in poorer condition than those not excreting.
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