Ana Cláudia Norte

The design of artificial nestboxes for the study of secondary hole-nesting birds: a review of methodological inconsistencies and potential biases

Abstract. The widespread use of artificial nestboxes has led to significant advances in our knowledge of the ecology, behaviour and physiology of cavity nesting birds, especially small passerines. Nestboxes have made it easier to perform routine monitoring and experimental manipulation of eggs or nestlings, and also repeatedly to capture, identify and manipulate the parents. However, when comparing results across study sites the use of nestboxes may also introduce a potentially significant confounding variable in the form of differences in nestbox design amongst studies, such as their physical dimensions, placement height, and the way in which they are constructed and maintained. However, the use of nestboxes may also introduce an unconsidered and potentially significant confounding variable due to differences in nestbox design amongst studies, such as their physical dimensions, placement height, and the way in which they are constructed and maintained. Here we review to what extent the characteristics of artificial nestboxes (e.g. size, shape, construction material, colour) are documented in the ‘methods’ sections of publications involving hole-nesting passerine birds using natural or excavated cavities or artificial nestboxes for reproduction and roosting. Despite explicit previous recommendations that authors describe in detail the characteristics of the nestboxes used, we found that the description of nestbox characteristics in most recent publications remains poor and insufficient. We therefore list the types of descriptive data that should be included in the methods sections of relevant manuscripts and justify this by discussing how variation in nestbox characteristics can affect or confound conclusions from nestbox studies. We also propose several recommendations to improve the reliability and usefulness of research based on long-term studies of any secondary hole-nesting species using artificial nestboxes for breeding or roosting.

Brominated flame retardants and organochlorines in the European environment using great tit eggs as a biomonitoring tool

Large-scale studies are essential to assess the emission patterns and spatial distribution of organohalogenated pollutants (OHPs) in the environment. Bird eggs have several advantages compared to other environmental media which have previously been used to map the distribution of OHPs. In this study, large-scale geographical variation in the occurrence of OHPs, such as polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and organochlorine pesticides (OCPs), was investigated throughout Europe using eggs of a terrestrial residential passerine species, the great tit (Parus major). Great tit eggs from 22 sampling sites, involving urban, rural and remote areas, in 14 European countries were collected and analysed (5-8 eggs per sampling site). The environmentally most important congeners/compounds of the analysed pollutants were detectable in all sampling locations. For PCBs, PBDEs and OCPs, no clear geographical contamination pattern was found. Sum PCB levels ranged from 143 ng/g lipid weight (lw) to 3660 ng/g lw. As expected, PCB concentrations were significantly higher in the sampled urban compared to the remote locations. However, the urban locations did not show significantly higher concentrations compared to the rural locations. Sum PBDEs ranged from 4.0 ng/g lw to 136 ng/g lw. PBDEs were significantly higher in the urbanized sampling locations compared to the other locations. The significant, positive correlation between PCB and PBDE concentrations suggests similar spatial exposure and/or mechanisms of accumulation. Significantly higher levels of OCPs (sum OCPs ranging from 191 ng/g lw to 7830 ng/g lw) were detected in rural sampling locations. Contamination profiles of PCBs, PBDEs and OCPs differed also among the sampling locations, which may be due to local usage and contamination sources. The higher variance among sampling locations for the PCBs and OCPs, suggests that local contamination sources are more important for the PCBs and OCPs compared to the PBDEs. To our knowledge, this is the first study in which bird eggs were used as a monitoring tool for OHPs on such a large geographical scale.

Variation of adult Great Tit Parus major body condition and blood parameters in relation to sex, age, year and season

In recent years, an increasing number of studies have focused on the ranges of variation of health related biochemical and haematological parameters in wildlife, but information is still scarce for enzymatic activities which can be extremely important in detecting potential responses to environmental change. In a Great Tit (Parus major) population, we describe the variation in relation to age, sex, season and year of: (1) morphological: body condition index, fat and muscle, (2) haematological: hematocrit, haemoglobin, white blood cell count and heterophil/lymphocyte ratio, and (3) biochemical parameters: plasma protein and activities of plasma cholinesterases (acetylcholinesterase and butyrylcholinesterase) activity and red blood cell glutathione peroxidase. Sex had significant effects on all morphological parameters except fat. Age significantly affected cholinesterase activities, H/L ratio and haemoglobin, and there was a significant interaction between sex and age affecting hematocrit. There were significant interactions of year and season affecting almost all parameters studied—body condition index, fat, protein, acetyl and butyrylcholinesterase activities, glutathione peroxidase activity and haemoglobin. This study indicates that these parameters are largely influenced by year and seasonal effects, besides the individual’s intrinsic variation. Therefore, when evaluating experimental or environmental change effects, appropriate controls should be used.

The use of blue tit eggs as a biomonitoring tool for organohalogenated pollutants in the European environment

In the present study, large scale geographical variation in the occurrence of organohalogenated pollutants (OHPs) was investigated throughout Europe using eggs of a terrestrial resident passerine species, the blue tit (Cyanistes caeruleus). Blue tit eggs from 10 sampling locations, involving suburban, rural and remote areas, in 7 European countries were collected and analysed. Sum polychlorinated biphenyl (PCB) levels ranged from 150ng/g lipid weight (lw) to 2003ng/g lw. Sum polybrominated diphenyl ethers (PBDEs) ranged from 3.95ng/g lw to 114ng/g lw. As expected, PCB and PBDE concentrations were significantly higher in the sampled suburban locations compared to the rural and remote locations. Sum organochlorine pesticides (OCPs) ranged from 122ng/g lw to 775ng/g lw. OCP concentrations were, against the expectations, found to be lower in the rural sampling locations compared to the other locations. Contamination profiles of PCBs, PBDEs and OCPs differed also among the sampling locations, which may be due to local contamination sources. Finally, we compared the results of this study with previously reported OHP concentrations in the eggs of a closely related species, the great tit (Parus major), from the same sampling locations in Europe. We found no differences in concentrations between the species. In addition, we found a significant, positive correlation between the sum PCB concentrations in blue tit eggs and great tit eggs, suggesting similar exposure pathways, mechanisms of accumulation and maternal transfer of PCBs. In conclusion, our results suggest the usefulness of eggs from passerine birds as a biomonitoring tool for OHPs on a large geographical scale.

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.

Variation in clutch size in relation to nest size in birds

Nests are structures built to support and protect eggs and/or offspring from predators, parasites, and adverse weather conditions. Nests are mainly constructed prior to egg laying, meaning that parent birds must make decisions about nest site choice and nest building behavior before the start of egg-laying. Parent birds should be selected to choose nest sites and to build optimally sized nests, yet our current understanding of clutch size-nest size relationships is limited to small-scale studies performed over short time periods. Here, we quantified the relationship between clutch size and nest size, using an exhaustive database of 116 slope estimates based on 17,472 nests of 21 species of hole and non-hole-nesting birds. There was a significant, positive relationship between clutch size and the base area of the nest box or the nest, and this relationship did not differ significantly between open nesting and hole-nesting species. The slope of the relationship showed significant intraspecific and interspecific heterogeneity among four species of secondary hole-nesting species, but also among all 116 slope estimates. The estimated relationship between clutch size and nest box base area in study sites with more than a single size of nest box was not significantly different from the relationship using studies with only a single size of nest box. The slope of the relationship between clutch size and nest base area in different species of birds was significantly negatively related to minimum base area, and less so to maximum base area in a given study. These findings are consistent with the hypothesis that bird species have a general reaction norm reflecting the relationship between nest size and clutch size. Further, they suggest that scientists may influence the clutch size decisions of hole-nesting birds through the provisioning of nest boxes of varying sizes.

Repeatability and Method-Dependent Variation of Blood Parameters in Wild-Caught Great Tits Parus major

When interpreting responses to experimental manipulations or particular environmental cues, it is necessary to have previous knowledge about the natural variation of the response traits. We studied how nine blood parameters, including four enzymatic activities, varied with time in wild Great Tits by assessing their repeatabilities over periods of 45 days, in the same season (Spring or Autumn/Winter), in the same year and over a four years period. The accuracy of the measurements of these blood parameters was also assessed. Measurement error reflected essentially sample and time of storage rather than assay effects. Hematocrit and haemoglobin had low repeatabilities within Spring, ranging from 0.26 to 0.31; Heterophil/Lymphocyte ratio (H/L), white blood cell count (WBC), total plasma cholinesterase and red blood cell glutathione peroxidase (GSH-Px) activities had moderate to high repeatabilities over periods of 45 days (repeatabilities ranged from 0.47 to 0.81 for GSH-Px and H/L, respectively), but also during longer periods such as dur- ing Spring (total plasma cholinesterase activity) and Autumn/Winter (WBC, H/L and GSH-Px). Of the blood parameters measured, total plasma cholinesterase, glutathione peroxidase and the haematological parameters WBC and H/L seem relatively constant and therefore reliable indicators of Great Tits physiological condition within, at least, a 45 days time frame.

Clutch-size variation in Western Palaearctic secondary hole-nesting passerine birds in relation to nest box design

Secondary hole-nesting birds that do not construct nest holes themselves and hence regularly breed in nest boxes constitute important model systems for field studies in many biological disciplines with hundreds of scientists and amateurs involved. Those research groups are spread over wide geographic areas that experience considerable variation in environmental conditions, and researchers provide nest boxes of varying designs that may inadvertently introduce spatial and temporal variation in reproductive parameters. We quantified the relationship between mean clutch size and nest box size and material after controlling for a range of environmental variables in four of the most widely used model species in the Western Palaearctic: great tit Parus major, blue tit Cyanistes caeruleus, pied flycatcher Ficedula hypoleuca and collared flycatcher F.albicollis from 365 populations and 79610 clutches. Nest floor area and nest box material varied non-randomly across latitudes and longitudes, showing that scientists did not adopt a random box design. Clutch size increased with nest floor area in great tits, but not in blue tits and flycatchers. Clutch size of blue tits was larger in wooden than in concrete nest boxes. These findings demonstrate that the size of nest boxes and material used to construct nest boxes can differentially affect clutch size in different species. The findings also suggest that the nest box design may affect not only focal species, but also indirectly other species through the effects of nest box design on productivity and therefore potentially population density and hence interspecific competition.

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.

Blackbirds Turdus merula as competent reservoirs for Borrelia turdi and Borrelia valaisiana in Portugal: evidence from a xenodiagnostic experiment

To confirm that thrushes, such as blackbirds Turdus merula, play a role as reservoir for some Borrelia genospecies, we performed a xenodiagnostic experiment with blackbirds captured in a mixed wood located in Western Portugal where Borrelia turdi, an uncommon genospecies in Europe, was the most prevalent genospecies associated with birds. Two out of five birds harboured B. turdi infected Ixodes frontalis at the time of capture. Four out of five birds transmitted spirochaetes to Ixodes ricinus xenodiagnostic ticks: two birds transmitted Borrelia valaisiana to 25.7% and 10.5% of ticks, and two transmitted B. turdi to 6.4% and 5.4% of ticks. Our results showed that blackbirds transmit B. valaisiana and B. turdi to I. ricinus feeding larvae, acting as reservoir hosts for these genospecies in nature.