Effects of stress exposure in captivity on physiology and infection in avian hosts: no evidence of increased Borrelia burgdorferi s.l. infectivity to vector ticks.

Exposure to environmental stressors, an increasingly recurring event in natural communities due to anthropogenic-induced environmental change, profoundly impacts disease emergence and spread. One mechanism through which this occurs is through stress-induced immunosuppression increasing disease susceptibility, prevalence, intensity and reactivation in hosts. We experimentally evaluated how exposure to stressors affected both the physiology of avian hosts and the prevalence of the zoonotic bacteria Borrelia burgdorferi sensu lato (s.l.), in two model species-the blackbird Turdus merula and the robin Erithacus rubecula captured in the wild, using xenodiagnoses and analysis of skin biopsies and blood. Although exposure to stressors in captivity induced physiological stress in birds (increased the number of circulating heterophils), there was no evidence of increased infectivity to xenodiagnostic ticks. However, Borrelia detection in the blood for both experimental groups of blackbirds was higher by the end of the captivity period. The infectivity and efficiency of transmission were higher for blackbirds than robins. When comparing different methodologies to determine infection status, xenodiagnosis was a more sensitive method than skin biopsies and blood samples, which could be attributed to mild levels of infection in these avian hosts and/or dynamics and timing of Borrelia infection relapses and redistribution in tissues.

Francisella species in ticks and animals, Iberian Peninsula.

The presence of Francisella species in 2134 ticks, 93 lagomorphs and 280 small mammals from the Iberian Peninsula was studied. Overall, 19 ticks and 6 lagomorphs were positive for Francisella tularensis subsp. holarctica, suggesting, as described for other regions, that lagomorphs may have an important role in the maintenance of F. tularensis in nature. Of the 6 positive lagomorphs, 4 were identified as the European rabbit, Oryctogalus cuniculus. Additionally, 353 ticks and 3 small mammals were PCR positive for Francisella-like endosymbionts (FLEs) and one small mammal was also positive for Francisella hispaniensis-like DNA sequences. Among FLE positive specimens, a variety of sequence types were detected: ticks were associated with 5 lpnA sequence types, with only one type identified per tick, in contrast to 2 lpnA sequence types detected in a single wood mouse (Apodemus sylvaticus). To our knowledge, this is the first report of FLEs in free-living small mammals as well as the first detection of F. hispaniensis-like sequences in a natural setting.

Patterns of tick infestation and their Borrelia burgdorferi s.l. infection in wild birds in Portugal.

Wild birds may act as reservoirs for zoonotic pathogens and may be mechanical carriers of pathogen infected vector ticks through long distances during migration. The aim of this study was to assess tick infestation patterns in birds in Portugal and the prevalence of tick infection by Borrelia burgdorferi s.l. using PCR techniques. Seven tick species were collected from birds including Haemaphysalis punctata, Hyalomma spp., Ixodes acuminatus, Ixodes arboricola, Ixodes frontalis, Ixodes ricinus and Ixodes ventalloi. We found that I. frontalis and Hyalomma spp. were the most common ticks infesting birds of several species and that they were widespread in Portugal. Turdus merula was the bird species that presented the highest diversity of infesting ticks and had one of the highest infestation intensities. B. burgdorferi s.l. was detected in 7.3% (37/505) of Ixodidae ticks derived from birds. The most common genospecies was Borrelia turdi (6.9%), detected in ticks collected from Parus major, T. merula and Turdus philomelos, but Borrelia valaisiana (0.2%) and one Borrelia sp. (0.2%) similar to Borrelia bissettii (96% of similarity of the flaB gene in Blastn) were also detected. This study contributed to a better knowledge of the Ixodidae tick fauna parasitizing birds in Western Europe and to the assessment of the prevalence of B. burgdorferi s.l. associated with birds and their ticks.

Tularaemia: a challenging zoonosis.

In recent years, several emerging zoonotic vector-borne infections with potential impact on human health have been identified in Europe, including tularaemia, caused by Francisella tularensis. This remarkable pathogen, one of the most virulent microorganisms currently known, has been detected in increasingly new settings and in a wide range of wild species, including lagomorphs, rodents, carnivores, fish and invertebrate arthropods. Also, a renewed concern has arisen with regard to F. tularensis: its potential use by bioterrorists. Based on the information published concerning the latest outbreaks, the aim of this paper is to review the main features of the agent, its biology, immunology and epidemiology. Moreover, special focus will be given to zoonotic aspects of the disease, as tularaemia outbreaks in human populations have been frequently associated with disease in animals.

Do ticks and Borrelia burgdorferi s.l. constitute a burden to birds?

Ticks consume resources from their hosts shaping their life-history traits and are vectors of many zoonotic pathogens. Several studies have focused on the health effects of blood-sucking ectoparasites on avian hosts, but there is limited information on the effects of ticks on adult and sub-adult birds, which may actively avoid ticks and are likely to present low infestation intensities. We evaluated the effects of the presence of feeding ticks and intensity of infestation on health variables of avian hosts. We also evaluated whether these variables were affected by tick infection by Borrelia burgdorferi sensu lato (s.l.) and by the presence of Borrelia infection on the birds' skin. Presence of parasite association among ticks, haemosporidea and Borrelia within the bird-host was also tested. We found that infestation by ticks significantly increased heterophyl/lymphocyte ratio in Turdus merula suggesting increased stress. This was especially evident at high infestation intensities when a significant decrease in body mass and body condition (body mass corrected for size) was also observed. Erithacus rubecula infested with more than 10 larvae tended to have lower haematocrit and blood haemoglobin. Plasma globulin concentration in T. merula tended to be affected by the presence of attached ticks and their infection with Borrelia, but this depended on the age of the bird. No association was detected among ticks, haemosporidea and Borrelia infection. We showed that ticks have detrimental effects on their avian hosts even under natural infestation conditions and that confirmed Borrelia reservoir hosts may also present symptoms of infection, though these may be subtle.

Birds as reservoirs for Borrelia burgdorferi s.l. in Western Europe: circulation of B. turdi and other genospecies in bird-tick cycles in Portugal.

Birds are important in the ecology of Borrelia burgdorferi sensu lato (s.l.) because they are important hosts for vector tick immature stages and are known reservoirs for some Borrelia genospecies. The aim of our study was to assess the role of common passerine bird species as reservoirs for B. burgdorferi s.l. in Western Europe. We surveyed birds in enzootic areas in Portugal, where no information is available for birds as reservoirs for this aetiologic agent and where B. lusitaniae, for which few reservoirs have been identified, is the dominant genospecies. Twenty-three birds (2.9%), including Turdus merula, T. philomelos, Parus major and Fringilla coelebs harboured infected ticks, but only Turdus sp. harboured infected tick larvae. In one study area, although B. lusitaniae was dominant in questing Ixodes ricinus, no ticks feeding on birds were infected with this genospecies, and B. valaisiana was the dominant genospecies in I. ricinus larvae feeding on birds. In the other area ticks collected from birds were mainly I. frontalis which were infected with B. turdi. Two skin biopsies (4.2%) from two T. merula were positive, one for B. valaisiana and the other for B. turdi. This is the first report for B. turdi in Western Europe.

Role of the lizard Teira dugesii as a potential host for Ixodes ricinus tick-borne pathogens.

PCR screening of ticks and tissue samples collected from 151 Teira dugesii lizards seems to indicate a potential role of this lizard species in the maintenance and transmission cycle of some Ixodes ricinus tick-borne agents, such as Rickettsia monacensis, Rickettsia helvetica, and Borrelia lusitaniae, that are circulating on Madeira Island.

Borrelia hispanica in Ornithodoros erraticus, Portugal.

Tick-borne relapsing fever (TBRF) is a spirochetal infection caused by the genus Borrelia. The disease is distributed in the Old and New World with many different species reported. In Europe, TBRF is caused by B. hispanica transmitted to man by Ornithodoros erraticus, a soft tick usually found in old premises to shelter pig herds. In Portugal, the first human case of TBRF was reported in 1942 but since the beginning of the 1960s, the disease has rarely been described and seems to either have disappeared or have been undiagnosed. Therefore, in 2009 a survey was undertaken to evaluate the presence of the tick in this type of premises and to evaluate its role as a reservoir of Borrelia. The work was carried out where the ticks were previously reported in the Alentejo and Algarve regions. Of 63 pigpens surveyed, O. erraticus was collected from 19% (n = 12) of these pigpens using CO(2) traps. To evaluate potential Borrelia hosts, both pigs (n = 25) and small rodents (n = 10) inhabiting these pigpens were surveyed for Borrelia presence, by whole blood PCR and/or tissue culture, respectively. All results for pigs and rodents were negative for the presence of B. hispanica. PCR assays targeting the 16S rRNA gene and intergenic spacer region of Borrelia were used. Sequence analysis of the positive samples confirmed the presence of B. hispanica in 2.2% (n = 5) of ticks from a pigpen in Alentejo. These results confirm natural, but albeit low, persistence of this agent in Portugal.

[Tularemia]. / Tularémia.

Tularemia is a zoonotic disease caused by Francisella tularensis. The etiological agent is transmitted to man by direct contact with infected animals, air, water or contaminated food, or through hematophagous vectors. In Portugal, in 1998, after an outbreak in Spain, the Direcção Geral de Saúde issued a warning, alerting the clinicians to the possibility of the disease spreading throughout the national territory. In this work, an epidemiological review of tularemia is made in order to contribute to the better knowledge of this disease.

Laboratory diagnosis of Lyme borreliosis at the Portuguese National Institute of Health (1990-2004).

Lyme borreliosis is considered to be an emerging infection in some regions of the world, including Portugal. The first Portuguese human case of Lyme borreliosis was identified in 1989. Since 1999, this disease is considered a notifiable disease (DDO) in Portugal, but only a few cases are reported each year, which does not allow consistent analysis of risk factors and the impact on public health. In this study the authors analyse the data available at the Centre for Vectors and Infectious Diseases Research (CEVDI) laboratory, at the Instituto Nacional de Saude Dr. Ricardo Jorge (National Institute of Health, INSA) during the past 15 years (1990-2004) and evaluate them against the registry of national reported cases (1999-2004). Serological tests were the basis for laboratory diagnosis. Data on year of diagnosis, sex, age, geographical origin and clinical signs are available for 628 well documented Portuguese positive cases. The number of cases per year varied between 2 and 78, with the highest number of cases reported in 1997. Of the positive cases, 53.5% were female and the age group most affected was 35-44 years old. Neuroborreliosis was the most common clinical manifestation (37.3%). Human cases were detected in 17 of the 20 regions of Portugal, and the highest number of laboratory confirmed cases were from the Lisbon district. The comparison of the number of notified cases and the number of positive cases confirmed by our laboratory show that Lyme borreliosis is clearly an underreported disease. Due to the scattered distribution of the positive cases and the low prevalence of the tick species Ixodes ricinus, the most effective prevention measure for Lyme borreliosis in Portugal is education of the risk groups on how to prevent tick bites.