Effects of flipper bands and injected transponders on the survival of adult Little Penguins Eudyptula minor

Tagging is essential for many types of ecological and behavioural studies, and it is generally assumed that it does not affect the fitness of the individuals being examined. However, the tagging of birds has been shown to have negative effects on some aspects of their lives. Here we investigate the influence of tagging on apparent survival. We examined the effects of flipper bands and injected transponders on the apparent survival of adult Little Penguins by comparing the survival probabilities of 2483 Little Penguins marked at Phillip Island, Australia, between 1995 and 2001 in one of three ways: with bands, with transponders or with both. The design of the study and our method of analysis allowed us to estimate tag loss and ensured that tag loss did not bias the survival estimates. Birds marked with flipper bands had lower survival probabilities than those marked with transponders (with apparent survival probabilities in the first year after tagging of 75% for banded birds and 80% for birds fitted with transponders, and in subsequent years of 87% for banded birds and 91% for birds fitted with transponders). We estimated both band and transponder loss probabilities for the first time, and found that transponder loss probabilities were substantially higher than band loss probabilities, particularly in the first year after marking when the tag loss probability was 5% for transponders and 0.7% for bands. Survival probabilities were lower in the first year after marking than in subsequent years for all birds. Studies of penguins that have used flipper bands to identify individuals may have underestimated annual adult survival probabilities, as banded penguins were likely to have lower than average survival probabilities than those of unbanded birds. The higher annual survival probabilities of individuals marked with transponders indicate that this should be the preferred marking technique for Little Penguins. However, future studies will, like ours, need to consider the higher rates of transponder loss when estimating survival, possibly by double‐tagging some birds.     

PIT-tagging Italian spined loach (Cobitis bilineata): Methodology,survival and behavioural effects

The Italian spined loach (Cobitis bilineata) is an elongated, small-sized (<12 cm) spined loach native to northern Italy, Slovenia and Croatia. As for loaches in general, little is known about the individual movements of this loach in nature. Passive integrated transponders (PIT-tags) are small (typically 7–32 mm), relatively cheap and allow tracking of individual fish movements and behaviour. A fundamental assumption in animal telemetry is that the performance of a tagged animal does not deviate substantially from its natural performance. Although PIT-tagged fish often display high survival and tag retention, the effect varies between species and contexts, and few studies have looked at behavioural effects of PIT-tagging. Here we demonstrate a PIT-tagging methodology for spined loaches, and compare survival, activity and provoked escape response (maximum swimming speed) between tagged and control fish. We also track tag retention in the tagged fish. Italian spined loaches tagged with 12 mm PIT-tags displayed high tag retention and no extra mortality, and no effects of tagging on activity or maximum swimming speed were observed. The tag-to-fish weight and length ratios in our study ranged from 2% to 5% and from 10% to 16%, respectively, and we conclude that PIT-tagging, within these ratios, appears suitable for Italian spined loach.     

Migration success and migration time of gastrically radio-tagged v. PIT-tagged adult Atlantic salmon

Adult Atlantic salmon (55–116 cm total length) with PIT-tags ( n = 318) and both PIT-tags and gastric radio-tags ( n = 127) in a regulated river showed similar upstream migration success, yet radio-tagged fish migrated faster than fish with only PIT-tags. Regurgitation rate of radio-tags was 9%.     

Information transfer about roosts in female Bechstein's bats: an experimental field study

Information transfer among group members is believed to play an important part in the evolution of coloniality in both birds and bats. Although information transfer has received much scientific interest, field studies using experiments to test the underlying hypotheses are rare. We used a field experiment to test if communally breeding female Bechstein's bats (Myotis bechsteinii) exchange information regarding novel roosts. We supplied a wild colony, comprising 17 adult females of known relatedness, with pairs of suitable and unsuitable roosts and monitored the arrival of individuals marked with transponders (PIT-tags) over 2 years. As expected with information transfer, significantly more naive females were recruited towards suitable than towards unsuitable roosts. We conclude that information transfer about roosts has two functions: (i) it generates communal knowledge of a large set of roosts; and (ii) it aids avoidance of colony fission during roost switching. Both functions seem important in Bechstein's bats, in which colonies depend on many day roosts and where colony members live together for many years.     

Population dynamics in a long-lived seabird: I. Impact of breeding activity on survival and breeding probability in unbanded king penguins

Understanding the trade-off between current reproductive effort, future survival and future breeding attempts is crucial for demographic analyses and life history studies. We investigated this trade-off in a population of king penguins (Aptenodytes patagonicus) marked individually with transponders using multistate capture-recapture models. This colonial seabird species has a low annual proportion of non-breeders (13%), despite a breeding cycle which lasts over 1 year. To draw inferences about the consequences of non-breeding, we tested for an effect of reproductive activity on survival and on the probability of subsequent breeding. We found that birds non-breeding in year t show the same survival rate as breeders (two-states analysis: breeding and non-breeding). However, breeders had a lower probability of breeding again the following year. This negative phenotypic correlation suggests the existence of reproductive costs affecting future breeding probability, but it might also be strengthened by late arrival for courtship in year t. A three-state analysis including breeding success revealed that failed breeders in year t have a lower probability to reproduce successfully in year t + 1 than non-breeders in year t, providing some evidence for the existence of reproductive costs. Moreover, successful breeders showed higher survival probability. This positive phenotypic correlation between current reproduction and subsequent survival supports the hypothesis of an heterogeneity in individual quality. Males breeding in year t had a lower probability to breed again in year t + 1 than females, suggesting higher reproductive costs for this sex. Such additional costs might be due to higher male parental investment in the final phase of chick-rearing, which also delays the arrival of males in year t + 1, and decreases their breeding probability. Our study is the first to explore the breeding biology and the demography of penguins without the disturbance of flipper-bands.     

Potential consequences of angling on nest-site fidelity in largemouth bass

Breeding site fidelity has evolved in many vertebrate taxa, suggesting both that site selection has an important influence on fitness potential and that the decision to reuse a nesting site is related to the individual’s prior nesting success at that location. For a species that provides parental care, such as the Largemouth Bass Micropterus salmoides, catch-and-release angling impacts individual nesting success and fitness through physiological disturbance and by removing the nest-guarding male from its brood, thereby allowing temporary access to eggs and hatchlings by brood predators. To assess the impact of catch-and-release angling on nest site fidelity, we compared the consequences of angling on individually marked (i.e., with passive integrated transponders) nest-guarding male Largemouth Bass in Ontario. An extremely high degree of nest site fidelity in year two was observed for males that were angled only once during year one (87% within 10 m of the previous year’s nest), 96.7% of which remained on the nest and completed parental care activities. There was significantly lower fidelity in year two, however, for males that were angled multiple times during year one (27% within 10m of the previous year’s nest), only 5.6% of which remained on the nest and completed parental care activities. This observed difference suggests that angling nesting bass may cause them to avoid previously used nest sites and instead search for alternative sites during future reproductive seasons. This human-induced impact on nest site choice may impact the future reproductive success of those Largemouth Bass.     

A NEW METHOD FOR INDIVIDUALLY MARKING SLUGS (ARION LUSITANICUS (MABILLE)) BY MAGNETIC TRANSPONDERS

A new method for marking anonid slugs is described. By the useof magnetic transponders they can be marked individually withoutsignificantly altering their vitality High numbers of differentcodes, long duration, and easy, safe and reliable readabilityare the main advantages of this method. (Received 25 January 1996; accepted 26 April 1996)     

Low Rates of PIT-Tag Loss in an Insectivorous Bat Species

The rate of loss of tags used to mark individuals is an important consideration in wildlife research and monitoring. Passive integrated transponder (PIT) tags (or microchips) generally have high retention rates; however, tag loss rates for small mammals such as insectivorous bats are poorly understood. We double-marked a population of Gould's wattled bats (Chalinolobus gouldii) with forearm bands and PIT tags (with the injection site sealed with surgical adhesive) in January and February 2020 to determine rates of subsequent tag loss over the short- (1–2 months) and medium- (13–14 months) term. Loss of PIT tags occurred in 4 (2.7%) of 146 recaptured individuals, all within 2 months of microchipping. We also recorded 1 occurrence of band loss 11 months after banding. Our study supports assertions that PIT-tag retention rates in small mammals are high, and suggests that rates of tag loss in small bat species are low when surgical adhesive is applied. Quantifying the rate of tag loss enables this variable to be incorporated into mark-recapture models. © 2021 The Wildlife Society.     

Dissipative particle dynamics simulations for biological tissues: rheology and competition

In this work, we model biological tissues using a simple, mechanistic simulation based on dissipative particle dynamics. We investigate the continuum behavior of the simulated tissue and determine its dependence on the properties of the individual cell. Cells in our simulation adhere to each other, expand in volume, divide after reaching a specific size checkpoint and undergo apoptosis at a constant rate, leading to a steady-state homeostatic pressure in the tissue. We measure the dependence of the homeostatic state on the microscopic parameters of our model and show that homeostatic pressure, rather than the unconfined rate of cell division, determines the outcome of tissue competitions. Simulated cell aggregates are cohesive and round up due to the effect of tissue surface tension, which we measure for different tissues. Furthermore, mixtures of different cells unmix according to their adhesive properties. Using a variety of shear and creep simulations, we study tissue rheology by measuring yield stresses, shear viscosities, complex viscosities as well as the loss tangents as a function of model parameters. We find that cell division and apoptosis lead to a vanishing yield stress and fluid-like tissues. The effects of different adhesion strengths and levels of noise on the rheology of the tissue are also measured. In addition, we find that the level of cell division and apoptosis drives the diffusion of cells in the tissue. Finally, we present a method for measuring the compressibility of the tissue and its response to external stress via cell division and apoptosis.     

Protein analysis through Western blot of cells excised individually from human brain and muscle tissue

Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually.     

Efficacy and accuracy of portable PIT-antennae when locating fish in ice-covered streams

Active tracking of passive integrated transponder (PIT)-tags using portable antennae is becoming an increasingly common technique in fish habitat studies in shallow rivers. We carried out “blind testing” to test the efficacy (% tags found) and accuracy (distance between predicted and true tag location) of a portable antenna system (Texas Instruments) in winter conditions using 23-mm PIT-tags. Up to 90 cm reading range was achieved and signals penetrated ice, rock, wood and water. In the “blind test” trials, a majority of the hidden tags (N = 12–30) were found indicating high tracking efficacy. PIT-tags that were oriented with their cylindrical axis parallel to the plane of the antenna coil inductor loop resulted in a bimodal detection field that had low detection range in the centre of the loop. The utilization of this bimodal detection field proved to be a very accurate method for identifying tag position (mean ± SE distance from predicted to true tag location 10.9 ± 1.4 cm) and thus well suited for microhabitat and activity studies in winter conditions. Aggregations of tags (multiple tags within 1 m²) and obstacles for the antenna maneuvering (e.g., boulders, logjams) reduced the pinpointing accuracy (mean ± SE 13.3 ± 1.8 cm), but the reduction in accuracy was statistically non-significant between the single and aggregated tags.     

Evaluation of marking and tagging methods for genetic studies in carp

A variety of marking and tagging methods were tested on common carp (Cyprinus carpio L.) with the aim of identifying suitable methods for genetic studies in this and other species of carp. Elastomer and Alcian blue dye marking; Cold and Silver nitrate branding; Floy, Fingerling, Carlin disc and visible implant tags; and fin clipping were all tested on a range of sizes of common carp (from mean weights of 10–25 g up to 600–800 g). The branding and tagging methods tested did not give satisfactory retention rates. A combination of elastomer marking and fin clipping was then tested as a method for strain identification in a growth comparison trial on catla (Catla catla Hamilton) and found to be satisfactory for this purpose. Passive integrated transponders (PIT) tags were used to individually identify catla of wild or hatchery origin being grown for use as broodstock. These had almost 100% (98.8%) retention rates, but are expensive compared to most other tagging methods.     

A flat-bed passive integrated transponder antenna array for monitoring behaviour of Atlantic salmon parr and other fish

A system for monitoring automatically the movements of individually identified small fish in natural habitats is described. The system incorporates novel flat-bed antennae to detect the passage of miniature passive integrated transponders.     

Litter loss triggers estrus in a nonsocial seasonal breeder

Sexually selected infanticide (SSI) is often presumed to be rare among seasonal breeders, because it would require a near immediate return to estrus after the loss of an entire litter during the mating season. We evaluated changes in reproductive strategies and the reproductive fate of females that experienced litter loss during the mating season in a seasonal breeder with strong evidence for SSI, the brown bear. First, we used a long‐term demographic dataset (1986–2011) to document that a large majority of females (>91%) that lose their entire litter during the mating season in fact do enter estrus, mate, and give birth during the subsequent birthing season. Second, we used high‐resolution movement data (2005–2011) to evaluate how females changed reproductive strategies after losing their entire litter during the mating season. We hypothesized that females would shift from the sedentary lifestyle typical for females with cubs‐of‐the‐year to a roam‐to‐mate behavior typical for receptive females in no more than a few (~3) days after litter loss. We found that females with cubs‐of‐the‐year moved at about 1/3 of the rate and in a less bimodal diurnal pattern than receptive females during the mating season. The probability of litter loss was positively related with movement rate, suggesting that being elusive and sedentary is a strategy to enhance cub survival rather than a relic of cub mobility itself. The movement patterns of receptive females and females after litter loss were indistinguishable within 1–2 days after the litter loss, and we illustrate that SSI can significantly reduce the female interbirth interval (50–85%). Our results suggest that SSI can also be advantageous for males in seasonally breeding mammals. We propose that infanticide as a male reproductive strategy is more prevalent among mammals with reproductive seasonality than observed or reported.     

Micro-capillary tube in situ hybridisation: a novel method for processing small individual samples

We have developed a strategy to individually analyse large numbers of small tissue samples by RNA in situ hybridisation. Samples of approximately 0.4 mm x 0.5 mm are processed in rectangular capillary tubes fitted with nylon mesh and glass beads using standard protocols. Eighteen samples can be assayed simultaneously without loss, and background is low. Specifically, mouse Sox2 RNA expression is examined in the chorion of extraembryonic tissue of 7.5 days post-coitum embryos. This technique works equally well for double RNA labelling and could potentially be used for antibody staining of proteins.     

Film tomography as a tool for three-dimensional image construction and gene expression studies

In order to observe three-dimensional (3D) expression patterns of genes in whole animals, whole organs, or whole tissues, in situ hybridization (ISH) of many sections must be carried out and then used to construct a 3D image. For this purpose, we have developed an automatic microtome to prepare tissue sections with an adhesive film. We used commercially available film suitable for sectioning and ISH. We constructed a microtome and, after adherence of the film to a paraffin-embedded tissue block, cut the block with a blade to prepare sections on film. Then, the sections-on-film were automatically set in a plastic frame that was the same size as a conventional glass slide. With this automatic microtome, tissue sections can be made for ISH or immunohistochemistry in addition to conventional hematoxylin and eosin staining without specific training. We demonstrate that we can construct 3D images of gene expression patterns obtained by ISH on sections prepared with this automatic microtome. We have designated this method as 'Film Tomography (FITO)'.     

Biocompatibility of a Novel Cyanoacrylate Based Tissue Adhesive: Cytotoxicity and Biochemical Property Evaluation

Cyanoacrylate (CA) is most widely used as a medical and commercial tissue adhesive because of easier wound closure, good cosmetic results and little discomfort. But, CA-based tissue adhesives have some limitations including the release of cytotoxic chemicals during biodegradation. In previous study, we made prepolymerized allyl 2-CA (PACA) based tissue adhesive, resulting in longer chain structure. In this study, we investigated a biocompatibility of PACA as alternative tissue adhesive for medical application, comparing with that of Dermabond® as commercial tissue adhesive. The biocompatibility of PACA was evaluated for short-term (24 hr) and long-term (3 and 7 days) using conventional cytotoxicity (WST, neutral red, LIVE/DEAD and TUNEL) assays, hematoxylin-eosin (H&E) and Masson trichrome (MT) staining. Besides we examined the biochemical changes in cells and DNA induced by PACA and Dermabond® utilizing Raman spectroscopy which could observe the denaturation and conformational changes in protein, as well as disintegration of the DNA/RNA by cell death. In particular, we analyzed Raman spectrum using the multivariate statistical methods including principal component analysis (PCA) and support vector machine (SVM). As a result, PACA and Dermabond® tissue adhesive treated cells and tissues showed no difference of the cell viability values, histological analysis and Raman spectral intensity. Also, the classification analysis by means of PCA-SVM classifier could not discriminate the difference between the PACA and Dermabond® treated cells and DNA. Therefore we suggest that novel PACA might be useful as potential tissue adhesive with effective biocompatibility.     

Individual genetic tagging for teleosts: an empirical validation and a guideline for ecologists

The efficiency of individual genetic tagging was determined by using passive integrated transponders (PIT) as a comparative conventional tagging method. Fifty-five common dace Leuciscus leuciscus were captured in the wild, PIT tagged and fin clipped (for DNA analysis). Thirty fish were recaptured on three occasions and tissue samples were collected. Using 18 microsatellite loci, 79-94% of the recaptures were correctly assigned. Experience with scoring L. leuciscus microsatellites led to more individuals correctly assigned. Allowing matches that differed by one or two alleles resulted in 100% of all recaptures successfully assigned irrespective of the observer. Reducing the set of loci to five to six loci appropriately selected did not affect the assignment rate, demonstrating that costs can be subsequently reduced. Despite their potential benefits, the application of genetic tags for teleosts has been limited. Here, it was demonstrated that genetic tagging could be applied, and a clear guideline (flowchart) is provided on how this method can be developed for teleosts and other organisms, with subsequent practical applications to ecology, evolutionary biology and conservation management.     

Factors predicting individual variability in diet-induced weight loss in MF1 mice

The effectiveness of caloric restriction (CR) as a treatment for obesity varies considerably between individuals. Reasons for this interindividual variation in weight loss in response to CR may lie in pre-existing individual differences and/or individual differences in compensatory responses. Here we studied the responses of 127 MF1 mice to 30% CR over four weeks, and investigated whether pre-existing differences or compensatory changes in body temperature, resting metabolic rate (RMR) and behavior explained the variation observed in body mass (BM) and fat mass (FM) changes. Mice showed considerable variation in BM loss (36-1%), and in the type of tissue lost (FM or fat free mass, FFM). About 50% of the variation in BM and FM loss could be predicted by pre-existing differences in food intake, RMR, and general activity, where BM loss was greater when food intake was lower and activity and RMR were higher. Compensatory changes in activity and body temperature together explained ~50% of the variation in BM and FM loss in both sexes. In models incorporating baseline variables and compensatory changes, food intake, and activity were the strongest predictors of weight loss in both sexes; i.e., lower baseline food intake and increased changes in activity resulted in greater BM and FM loss. Interestingly, increased baseline activity was a significant predictor of weight loss independent of compensatory changes in activity. Identifying factors involved in individual variability in weight loss may give insights into the mechanisms that underlie this variability, and is important to develop individually tailored weight-management strategies.