Photo‐identification as a tool to study small‐spotted catshark Scyliorhinus canicula

Photo‐identification (photo‐ID) was tested as a means to identify individual small‐spotted catsharks Scyliorhinus canicula. The spotting pattern of the caudal region of S. canicula was used for the tests and revealed that photo‐ID is an efficient method to identify individuals. Photo‐ID is logistically simple, making it a potential alternative to traditional tagging to provide information on the distribution patterns and population dynamics of S. canicula and related species.     

The use of a non‐invasive tool for capture–recapture studies on a seahorse Hippocampus guttulatus population

In this study, the spot pattern in Hippocampus guttulatus was analysed using a computer programme algorithm that allowed individual comparison. This methodology was first tested in a controlled environment using 51 adult and 55 juvenile H. guttulatus. Positive matches were obtained in 86·3 and 83·6% of the adults and juveniles, respectively. In a second experiment, monthly surveys were carried out in five selected locations in the Ria Formosa Lagoon, south Portugal, over the course of a year and a total of 980 photographs were analysed. Photographed H. guttulatus were re‐sighted one to nine times during the course of the survey period with an overall re‐sight record of over 30%. Photo‐identification was therefore shown to be a useful tool for non‐invasive mark–recapture studies that can be successfully used to survey the population abundance of H. guttulatus aged 6 months or older in consecutive years. This could be of great value when considering the assessment of H. guttulatus populations and understanding changes over time.     

Microdot technology for individual marking of small arthropods

• 1 Individual mark–release–recapture is an important method for gathering data on insect movement, although it is limited by the constraints of tagging small insects with individual information.
• 2 Microdots, originally developed for covert security applications, are small polymer discs (diameter 0.5 mm) bearing up to 26 characters of information and have the potential as an alternative to the larger bee tags. In the present study, we test microdots for the individual marking of a 9‐mm parasitoid wasp.
• 3 We individually marked 505 wasps. The recapture rate was 24% of individuals over 189 recapture events, for which 84% retained legible microdot labels. Movement was in the range 0–161 m with a mean displacement 21.2 ± 2.7 m. A captive survival experiment showed no difference in lifespan between marked and unmarked wasps.
• 4 The present study shows that microdots can provide an effective, durable, low‐cost method for individually tagging small insects. The technique offers new opportunities by greatly expanding the capability for individually marking small insects, shifting the minimum size below that of bee tags, which is the only other manufactured option for individualized miniature marking.

     

In situ tagging technique for fishes provides insight into growth and movement of invasive lionfish

Information on fish movement and growth is primarily obtained through the marking and tracking of individuals with external tags, which are usually affixed to anesthetized individuals at the surface. However, the quantity and quality of data obtained by this method is often limited by small sample sizes owing to the time associated with the tagging process, high rates of tagging‐related mortality, and displacement of tagged individuals from the initial capture location. To address these issues, we describe a technique for applying external streamer and dart tags in situ, which uses SCUBA divers to capture and tag individual fish on the sea floor without the use of anesthetic. We demonstrate this method for Indo‐Pacific lionfish (Pterois volitans/P. miles), species which are particularly vulnerable to barotrauma when transported to and handled at the surface. To test our method, we tagged 161 individuals inhabiting 26 coral reef locations in the Bahamas over a period of 3 years. Our method resulted in no instances of barotrauma, reduced handling and recovery time, and minimal post‐tagging release displacement compared with conventional ex situ tag application. Opportunistic resighting and recapture of tagged individuals reveals that lionfish exhibit highly variable site fidelity, movement patterns, and growth rates on invaded coral reef habitats. In total, 24% of lionfish were resighted between 29 and 188 days after tagging. Of these, 90% were located at the site of capture, while the remaining individuals were resighted between 200 m and 1.1 km from initial site of capture over 29 days later. In situ growth rates ranged between 0.1 and 0.6 mm/day. While individuals tagged with streamer tags posted slower growth rates with increasing size, as expected, there was no relationship between growth rate and fish size for individuals marked with dart tags, potentially because of large effects of tag presence on the activities of small bodied lionfish (i.e., <150 mm), where the tag was up to 7.6% of the lionfish's mass. Our study offers a novel in situ tagging technique that can be used to provide critical information on fish site fidelity, movement patterns, and growth in cases where ex situ tagging is not feasible.     

Telemetry tag effects on juvenile lingcod Ophiodon elongatus movement: a laboratory and field study

This study tested the behavioural effects of tagging subyearling and yearling lingcod Ophiodon elongatus with acoustic telemetry tags in laboratory tanks and in the natural environment (Puget Sound, WA). In the laboratory, tagged individuals showed less movement and feeding behaviour soon after tagging than untagged controls. The effect dissipated after c. 1 week, presumably as the tagged O. elongatus recovered from surgery or adjusted to the presence of the tags. This dissipation enabled a field study that compared early‐tagged individuals with a long recovery period after tagging to recently‐tagged individuals with a short recovery period after tagging. Consistent with findings from the laboratory experiment, recently tagged individuals showed less movement away from three release sites in Puget Sound than early‐tagged individuals. Together, the laboratory and field results provide evidence of temporary tag effects on actual movement in the natural environment and provide a method for testing tag effects in the field. This study suggests that subyearling and yearling O. elongatus should be held for a recovery period before release. If holding after tagging is not an option, then movement data collected during the first week should be interpreted cautiously.     

Reconstructing the lionfish invasion: insights into Greater Caribbean biogeography

Aim Lionfish (Pterois volitans and P. miles) are popular ornamental fishes native to the Indo‐Pacific that were introduced into Florida waters and are rapidly spreading and establishing throughout the Western Atlantic (WA). Although unfortunate, this invasion provides an excellent system in which to test hypotheses on conservation biology and marine biogeography. The goals of this study are: (1) to document the geographical extent of P. volitans and P. miles; (2) to determine whether the progression of the lionfish invasion is the result of expansion following the initial introduction event or the consequence of multiple introductions at various WA locations; and (3) to analyse the chronology of the invasion in conjunction with the genetic data in order to provide real‐time assessments of hypotheses of marine biogeography. Location The Greater Caribbean, including the US east coast, Bermuda, the Bahamas and the Caribbean Sea. Methods Mitochondrial control region sequences were obtained from lionfish individuals collected from Bermuda and three Caribbean locations and analysed in conjunction with previously published data from five native and two non‐native locations (US east coast and the Bahamas; a total of six WA locations). Genetic variation within and among groups was quantified, and population structure inferred via spatial analyses of molecular variance, pairwise Φ_ST, exact tests, Mantel tests and haplotype networks. Results Mitochondrial DNA screening of WA lionfish shows that while P. miles is restricted to the northernmost locations (Bermuda and the US east coast), P. volitans is ubiquitous and much more abundant. Invasive populations of P. miles and P. volitans have significantly lower levels of genetic diversity relative to their native counterparts, confirming that their introduction resulted in a strong founder effect. Despite the relative genetic homogeneity across the six WA locations, population structure analyses of P. volitans indicate significant differentiation between the northern (US east coast, the Bahamas and Bermuda) and the Caribbean populations. Main conclusions Our findings suggest that the ubiquity of WA lionfish is the result of dispersal from a single source of introduction in Florida and not of multiple independent introductions across the range. In addition, the progression of the lionfish invasion (as documented from sightings), integrated with the genetic evidence, provides support for five of six major scenarios of connectivity and phylogeographical breaks previously inferred for Caribbean organisms.     

Individually mark–mass release–resight study elucidates effects of patch characteristics and distance on host patch location by an insect herbivore

1. How organisms locate their hosts is of fundamental importance in a variety of basic and applied ecological fields, including population dynamics, invasive species management and biological control. However, tracking movement of small organisms, such as insects, poses significant logistical challenges. 2. Mass‐release and individual–mark–recapture techniques were combined in an individually mark–mass release–resight (IMMRR) approach to track the movement of over 2000 adult insects in an economically important plant–herbivore system. Despite its widespread use for the biological control of the invasive thistle Carduus nutans, the host‐finding behaviour of the thistle head weevil Rhinocyllus conicus has not previously been studied. Insects were released at different distances from a mosaic of artificially created host patches with different areas and number of plants to assess the ecological determinants of patch finding. 3. The study was able to characterize the within‐season dispersal abilities and between‐patch movement patterns of R. conicus. Weevils found host plant patches over 900 m away. Large patches, with tall plants, situated close to the nearest release point had the highest first R. conicus resights. Patch area and plant density had no effect on the number of weevils resighted per plant; however, R. conicus individuals were more likely to disperse out of small patches and into large patches. 4. By understanding how R. conicus locates host patches of C. nutans, management activities for the control of this invasive thistle can be better informed. A deeper mechanistic understanding of host location will also improve prediction of coupled plant–herbivore spatial dynamics in general.     

Scaling marine fish movement behavior from individuals to populations

Understanding how, where, and when animals move is a central problem in marine ecology and conservation. Key to improving our knowledge about what drives animal movement is the rising deployment of telemetry devices on a range of free‐roaming species. An increasingly popular way of gaining meaningful inference from an animal's recorded movements is the application of hidden Markov models (HMMs), which allow for the identification of latent behavioral states in the movement paths of individuals. However, the use of HMMs to explore the population‐level consequences of movement is often limited by model complexity and insufficient sample sizes. Here, we introduce an alternative approach to current practices and provide evidence of how the inclusion of prior information in model structure can simplify the application of HMMs to multiple animal movement paths with two clear benefits: (a) consistent state allocation and (b) increases in effective sample size. To demonstrate the utility of our approach, we apply HMMs and adapted HMMs to over 100 multivariate movement paths consisting of conditionally dependent daily horizontal and vertical movements in two species of demersal fish: Atlantic cod (Gadus morhua; n = 46) and European plaice (Pleuronectes platessa; n = 61). We identify latent states corresponding to two main underlying behaviors: resident and migrating. As our analysis considers a relatively large sample size and states are allocated consistently, we use collective model output to investigate state‐dependent spatiotemporal trends at the individual and population levels. In particular, we show how both species shift their movement behaviors on a seasonal basis and demonstrate population space use patterns that are consistent with previous individual‐level studies. Tagging studies are increasingly being used to inform stock assessment models, spatial management strategies, and monitoring of marine fish populations. Our approach provides a promising way of adding value to tagging studies because inferences about movement behavior can be gained from a larger proportion of datasets, making tagging studies more relevant to management and more cost‐effective.     

Biotin tagging coupled with amino acid‐coded mass tagging for efficient and precise screening of interaction proteome in mammalian cells

In mammalian cells, when tandem affinity purification approach is employed, the existence of untagged endogenous target protein and repetitive washing steps together result in overall low yield of purified/stable complexes and the loss of weakly and transiently interacting partners of biological significance. To avoid the trade‐offs involving in methodological sensitivity, precision, and throughput, here we introduce an integrated method, biotin tagging coupled with amino acid‐coded mass tagging, for highly sensitive and accurate screening of mammalian protein–protein interactions. Without the need of establishing a stable cell line, using a short peptide tag which could be specifically biotinylated in vivo, the biotin‐tagged target/bait protein was then isolated along with its associates efficiently by streptavidin magnetic microbeads in a single step. In a pulled‐down complex amino acid‐coded mass tagging serves as “in‐spectra” quantitative markers to distinguish those bait‐specific interactors from non‐specific background proteins under stringent criteria. Applying this biotin tagging coupled with amino acid‐coded mass tagging approach, we first biotin‐tagged in vivo a multi‐functional protein family member, 14‐3‐3ε, which was expressed at close to endogenous level. Starting with approximately 20 millions of 293T cells which were significantly less than what needed for a tandem affinity purification run, 266 specific interactors of 14‐3‐3ε were identified in high confidence.     

The effect of close approaches for tagging activities by small research vessels on the behavior of humpback whales (Megaptera novaeangliae)

Small research vessels are often used as platforms for tagging activities to collect behavioral data on cetaceans and they have the potential to disturb that group or individual. If this disturbance is ignored, results and conclusions produced by that study could be inaccurate. Here land‐based behavioral data of migrating humpback whales (Megaptera novaeangliae) (n = 29) were used to determine the effect of close approaches for tagging by research vessels on their diving, movement and surface behaviors. Groups of whales were tagged, using digital recording tags, by small research vessels, as part of a behavioral response study. In groups that were approached for tagging, temporary changes in movement behaviors during close approaches were found, with subsequent recovery to “pre‐approach” levels. In female‐calf groups more long‐term changes in travel speed were found. Results suggest that, although close approaches for tagging by small vessels may cause behavioral changes in humpback whales, this change may be small and temporary. However, in female‐calf groups, the behavioral change may be greater and longer lasting. This study shows that when using small vessels for behavioral research, disturbance, and recovery should be measured to ensure integrity of data used for other analyses.     

Validation of back‐calculated body lengths and timing of growth mark deposition in Hawaiian green sea turtles

Somatic growth rate data for wild sea turtles can provide insight into life‐stage durations, time to maturation, and total lifespan. When appropriately validated, the technique of skeletochronology allows prior growth rates of sea turtles to be calculated with considerably less time and labor than required by mark‐–recapture studies. We applied skeletochronology to 10 dead, stranded green turtles Chelonia mydas that had previously been measured, tagged, and injected with OTC (oxytetracycline) during mark–recapture studies in Hawaii for validating skeletochronological analysis. We tested the validity of back‐calculating carapace lengths (CLs) from diameters of LAGs (lines of arrested growth), which mark the outer boundaries of individual skeletal growth increments. This validation was achieved by comparing CLs estimated from measurements of the LAG proposed to have been deposited closest to the time of tagging to actual CLs measured at the time of tagging. Measureable OTC‐mark diameters in five turtles also allowed us to investigate the time of year when LAGs are deposited. We found no significant difference between CLs measured at tagging and those estimated through skeletochronology, which supports calculation of somatic growth rates by taking the difference between CLs estimated from successive LAG diameters in humerus bones for this species. Back‐calculated CLs associated with the OTC mark and growth mark deposited closest to tagging indicated that annual LAGs are deposited in the spring. The results of this validation study increase confidence in utilization of skeletochronology to rapidly obtain accurate age and growth data for green turtles.     

Inferring parturition and neonate survival from movement patterns of female ungulates: a case study using woodland caribou

Analyses of animal movement data have primarily focused on understanding patterns of space use and the behavioural processes driving them. Here, we analyzed animal movement data to infer components of individual fitness, specifically parturition and neonate survival. We predicted that parturition and neonate loss events could be identified by sudden and marked changes in female movement patterns. Using GPS radio‐telemetry data from female woodland caribou (Rangifer tarandus caribou), we developed and tested two novel movement‐based methods for inferring parturition and neonate survival. The first method estimated movement thresholds indicative of parturition and neonate loss from population‐level data then applied these thresholds in a moving‐window analysis on individual time‐series data. The second method used an individual‐based approach that discriminated among three a priori models representing the movement patterns of non‐parturient females, females with surviving offspring, and females losing offspring. The models assumed that step lengths (the distance between successive GPS locations) were exponentially distributed and that abrupt changes in the scale parameter of the exponential distribution were indicative of parturition and offspring loss. Both methods predicted parturition with near certainty (>97% accuracy) and produced appropriate predictions of parturition dates. Prediction of neonate survival was affected by data quality for both methods; however, when using high quality data (i.e., with few missing GPS locations), the individual‐based method performed better, predicting neonate survival status with an accuracy rate of 87%. Understanding ungulate population dynamics often requires estimates of parturition and neonate survival rates. With GPS radio‐collars increasingly being used in research and management of ungulates, our movement‐based methods represent a viable approach for estimating rates of both parameters.     

Tobacco mutants with reduced microtubule dynamics are less susceptible to TMV

A panel of seven SR1 tobacco mutants (ATER1 to ATER7) derived via T‐DNA activation tagging and screening for resistance to a microtubule assembly inhibitor, ethyl phenyl carbamate, were used to study the role of microtubules during infection and spread of tobacco mosaic virus (TMV). In one of these lines, ATER2, α‐tubulin is shifted from the tyrosinylated into the detyrosinated form, and the microtubule plus‐end marker GFP–EB1 moves significantly slower when expressed in the background of the ATER2 mutant as compared with the SR1 wild type. The efficiency of cell‐to‐cell movement of TMV encoding GFP‐tagged movement protein (MP‐GFP) is reduced in ATER2 accompanied by a reduced association of MP‐GFP with plasmodesmata. This mutant is also more tolerant to viral infection as compared with the SR1 wild type, implying that reduced microtubule dynamics confer a comparative advantage in face of TMV infection.     

Attraction of wild‐like and colony‐reared Bactrocera cucurbitae (Diptera:Tephritidae) to cuelure in the field

The attraction of wild tephritids to semiochemical‐based lures is the ideal basis for trap network design in detection programmes, but in practice, mass‐reared colony insects are usually used to determine trap efficiency. For Bactrocera cucurbitae Coquillett, a lower response by wild males compared with colony‐derived individuals, usually used to estimate attraction parameters, could mean that the sensitivity of detection networks targeting this pest is reduced. We describe the results of mark–release–recapture experiments with wild‐ and colony‐derived B. cucurbitae males in a grid of cuelure‐baited traps within a macadamia nut orchard in Hawaii Island designed to quantify the attraction of cuelure to each fly type. For colony males, we estimate a 65% probability of capture at 27 m, intermediate with previous estimates on the attraction of methyl eugenol to Bactrocera dorsalis Hendel (36 m) and trimedlure to Ceratitis capitata Wiedemann (14 m) at the same site. Results suggest similar response over distance by wild‐derived B. cucurbitae compared with colony in the field, but there may be qualitative differences in response between wild and colony based on very low response of wild males in a standard bioassay of attraction. For both fly types, the estimates of attraction in the smaller of two grid sizes tested were lower than for the larger spacing, suggesting trap competition was a factor at an intertrap distance of 75 m. Dispersal patterns within the grid were generally to the south for the colony‐derived males and more variable for the wild‐derived males. In neither case was the direction of recapture correlated with the prevailing direction of the wind.     

Effects of VIE tagging and partial tissue sampling on the immune response of three‐spined stickleback Gasterosteus aculeatus

A 14 day experiment on effects of visible implant elastomer (VIE) tagging and spine‐clipping of three‐spined stickleback Gasterosteus aculeatus showed significant increases in immune response, particularly in the granulocyte:lymphocyte ratio, in both treatments and the sham control. A minimum two‐week recovery after handling, anaesthesia, tagging and spine‐clipping is recommended to minimize effect of manipulation on the immune system.     

Genetic connectivity of lionfish (Pterois volitans) in marine protected areas of the Gulf of Mexico and Caribbean Sea

Lionfish (Pterois volitans) have rapidly invaded the tropical Atlantic and spread across the wider Caribbean in a relatively short period of time. Because of its high invasion capacity, we used it as a model to identify the connectivity among nine marine protected areas (MPAs) situated in four countries in the Gulf of Mexico and the Caribbean Sea. This study provides evidence of local genetic differentiation of P. volitans in the Gulf of Mexico and the Caribbean Sea. A total of 475 lionfish samples were characterized with 12 microsatellites, with 6–20 alleles per locus. Departures from Hardy–Weinberg equilibrium (HWE) were found in 10 of the 12 loci, all caused by heterozygous excess. Moderate genetic differentiation was observed between Chiriviche, Venezuela and Xcalak, México localities (F_ST = 0.012), and between the Los Roques and the Veracruz (F_ST = 0.074) sites. STRUCTURE analysis found that four genetic entities best fit our data. A unique genetic group in the Gulf of Mexico may imply that the lionfish invasion unfolded both in a counterclockwise manner in the Gulf of Mexico. In spite of the notable dispersion of P. volitans, our results show some genetic structure, as do other noninvasive Caribbean fish species, suggesting that the connectivity in some MPAs analyzed in the Caribbean is limited and caused by only a few source individuals with subsequent genetic drift leading to local genetic differentiation. This indicates that P. volitans dispersion could be caused by mesoscale phenomena, which produce stochastic connectivity pulses. Due to the isolation of some MPAs from others, these findings may hold a promise for local short‐term control of by means of intensive fishing, even in MPAs, and may have regional long‐term effects.     

Food stoichiometry affects the outcome of Daphnia–parasite interaction

Phosphorus (P) is an essential nutrient for growth in consumers. P‐limitation and parasite infection comprise one of the most common stressor pairs consumers confront in nature. We conducted a life‐table study using a Daphnia–microsporidian parasite model, feeding uninfected or infected Daphnia with either P‐sufficient or P‐limited algae, and assessed the impact of the two stressors on life‐history traits of the host. Both infection and P‐limitation negatively affected some life‐history traits tested. However, under P‐limitation, infected animals had higher juvenile growth rate as compared with uninfected animals. All P‐limited individuals died before maturation, regardless of infection. The numbers of spore clusters of the microsporidian parasite did not differ in P‐limited or P‐sufficient hosts. P‐limitation, but not infection, decreased body phosphorus content and ingestion rates of Daphnia tested in separate experiments. As parasite spore production did not suffer even under extreme P‐limitation, our results suggest that parasite was less limited by P than the host. We discuss possible interpretations concerning the stoichiometrical demands of parasite and suggest that our results are explained by parasite‐driven changes in carbon (C) allocation of the hosts. We conclude that the impact of nutrient starvation and parasite infection on consumers depends not only on the stoichiometric demands of host but also those of the parasite.     

Does underwater flash photography affect the behaviour,movement and site persistence of seahorses?

The effect of flash photography on seahorse species has never been tested. An experiment was established to test the effect of flash photography and the handling of Hippocampus whitei, a medium‐sized seahorse species endemic to Australia, on their behavioural responses, movements and site persistence. A total of 24 H. whitei were utilized in the experiment with eight in each of the three treatments (flash photography, handling and control). The effect of underwater flash photography on H. whitei movements was not significant; however, the effect of handling H. whitei to take a photograph had a significant effect on their short‐term behavioural responses to the photographer. Kaplan–Meier log‐rank test revealed that there was no significant difference in site persistence of H. whitei from each of the three treatments and that flash photography had no long‐term effects on their site persistence. It is concluded that the use of flash photography by divers is a safe and viable technique with H. whitei, particularly if photographs can be used for individual identification purposes.     

Invertebrate and Vertebrate Seed Predation in the Amazonian Palm Attalea maripa1

Spatial‐scale‐dependent patterns of Attalea maripa seed predation and removal by the beetle Pachymerus cardo and vertebrates were examined in a palm patch and its adjacent forest in southeastern Amazon. Seed survivorship depended on the distance from the palm patch, but was unrelated to distance from individual fruiting palms.     

Mutual benefit interactions between banded pine weevil Pissodes castaneus and blue‐stain fungus Leptographium serpens in maritime pine

• 1 Ecological interactions between banded pine weevil Pissodes castaneus and blue‐stain fungus Leptographium serpens, when simultaneously sharing the same host plant (maritime pine Pinus pinaster) in winter and spring, were investigated. Temporal components of the interaction were taken into account by either introducing the weevils and the pathogen simultaneously or sequentially, with the weevils being introduced 1 month after the fungal inoculation.
• 2 We measured larval mortality, development time, offspring number, sex ratio and body size of P. castaneus. Phloem phosphorus and nitrogen concentrations were also assessed. Furthermore, we tested whether: (i) emerging offspring transported propagules of the fungus; (ii) artificially‐contaminated weevils may transmit the disease to healthy trees; and (iii) field collected P. castaneus carry the fungus.
• 3 The fungus enhanced weevil colonization and brood production in both seasons. During winter and spring, adults from trees where the pathogen was inoculated prior to weevil introduction emerged earlier than weevils from trees where they had been introduced simultaneously with the fungus. During winter, weevils from pre‐inoculated trees were also larger. Sex ratio and larval mortality were not affected. Leptographium serpens did not affect phloem nitrogen content but phosphorus content was greater in plants inoculated with the pathogen, which may explain the findings on weevil growth.
• 4 Sixty‐five percent of the weevils that emerged from inoculated trees carried spores of L. serpens, although no successful isolation was made from field collected weevils. The fungus was recovered from 25% of the trees infested with artificially‐contaminated weevils.
• 5 These results suggest that P. castaneus benefits from the presence of L. serpens and may contribute to its spread.