Moulting Performances Evaluation of Female Orange Mud Crab, Scylla olivacea (Herbst, 1796) In-Captivity: Effects of Water Salinity and Limb Autotomy.

Female Scylla olivacea has become more popular in Malaysia as emerging species mainly for soft-shell crabs and crab fattening (to increase weight, size and ovary maturation so that they can be sold at a higher price). To harvest crabs in soft-shell conditions and fattening, both conditions depend mostly on moulting events. To accelerate the moulting process, the manipulation of water parameter (salinity) and autotomy of the limb is commonly used. In this study, the evaluation of the moulting performances of full limb autotomy (the removal of all the appendages except for the swimming legs) and non-ablated (control) using immature S. olivacea cultured in three different salinity treatments (10 ppt, 20 ppt and 30 ppt) were performed. Results indicate there were significant differences between mud crab's culture duration, BW increments, growth performances and feeding efficiency with salinity. However, CW increments and survival indicate no significant effect with salinity. Meanwhile, limb autotomy proved to affect the culture duration, BW increments, survival and feeding efficiency of S. olivacea. The study concludes that both salinity and limb autotomy play significant roles in moulting performances of S. olivacea, with 20 ppt being the best salinity to stimulate S. olivacea moulting and development compared with the other two treatments (10 ppt and 30 ppt). Limb autotomy also indicates promising results as this technique proved to accelerate the moulting duration of S. olivacea with a 100% moulting percentage within 30 days. Therefore, the outcome would certainly benefit in the aquaculture production of this species of commercial importance mainly on soft-shell crabs production and also emerge as crabs fattening technique.

Discovery of a neuropeptide that acts as an autotomy-promoting factor.

One of the most remarkable adaptations to survive attacks from predators is to detach an appendage-a process known as autotomy. This occurs in a variety of animals, including lizards (tail), crabs (legs), and starfish (arms). There has been extensive investigation of the evolution, ecology, and biomechanical impact of autotomy,1,2,3 but little is known about neural mechanisms controlling autotomy in animals. However, evidence for the existence of a peptide that acts as an autotomy-promoting factor in starfish has been reported.4 While investigating in vivo effects of a sulfakinin/cholecystokinin-type neuropeptide (ArSK/CCK1) in the starfish Asterias rubens,5,6 we observed that this peptide triggered arm autotomy in some animals. Furthermore, when injection of ArSK/CCK1 was combined with mechanical clamping of an arm, autotomy of the clamped arm occurred in 85% of animals tested, with 46% also autotomizing one or more other arms. In contrast, no autotomy was observed in clamped animals that were injected with water (control). To examine the physiological relevance of these findings, we analyzed expression of ArSK/CCK1 in the autotomy plane, a specialized region at the base of the arms in A. rubens.7,8 In accordance with its in vivo effects, nerve fibers expressing ArSK/CCK1 were revealed in the tourniquet muscle, a band of muscle that mediates constriction of the arm during and after autotomy. We conclude that ArSK/CCK1 acts as an autotomy-promoting factor in starfish and as such it is the first neuropeptide to be identified as a regulator of autotomy in animals.

Basement Membranes, Brittlestar Tendons, and Their Mechanical Adaptability.

Basement membranes (BMs) are thin layers of extracellular matrix that separate epithelia, endothelia, muscle cells, and nerve cells from adjacent interstitial connective tissue. BMs are ubiquitous in almost all multicellular animals, and their composition is highly conserved across the Metazoa. There is increasing interest in the mechanical functioning of BMs, including the involvement of altered BM stiffness in development and pathology, particularly cancer metastasis, which can be facilitated by BM destabilization. Such BM weakening has been assumed to occur primarily through enzymatic degradation by matrix metalloproteinases. However, emerging evidence indicates that non-enzymatic mechanisms may also contribute. In brittlestars (Echinodermata, Ophiuroidea), the tendons linking the musculature to the endoskeleton consist of extensions of muscle cell BMs. During the process of brittlestar autotomy, in which arms are detached for the purpose of self-defense, muscles break away from the endoskeleton as a consequence of the rapid destabilization and rupture of their BM-derived tendons. This contribution provides a broad overview of current knowledge of the structural organization and biomechanics of non-echinoderm BMs, compares this with the equivalent information on brittlestar tendons, and discusses the possible relationship between the weakening phenomena exhibited by BMs and brittlestar tendons, and the potential translational value of the latter as a model system of BM destabilization.

Effects of Acetic Acid and Morphine in Shore Crabs, Carcinus maenas: Implications for the Possibility of Pain in Decapods.

Noxious chemicals, coupled with morphine treatment, are often used in studies on pain in vertebrates. Here we show that injection of morphine caused several behavioural changes in the crab, Carcinus maenas, including reduced pressing against the sides of the enclosure and more rubbing and picking at the mouth parts and, at least for a short time, more defensive displays. Subsequent injection of acetic acid into one rear leg caused rubbing of the injected leg and the injected leg was held vertically off the ground. These activities directed at or involving the specific leg are consistent with previous observations of directed behaviour following noxious stimuli and are consistent with the idea that decapods experience pain. Further, acetic acid but not injection of water induced autotomy of the injected leg in these animals. Because autotomy is temporally associated with directed behaviour, it is possible that the autotomy is a pain-related response. Acetic acid is clearly a noxious substance when applied to decapods. However, morphine had no effect on the activities associated with acetic acid injection and thus there is no evidence for an analgesic effect. Further, the injection of acetic acid did not interfere with behavioural effects of morphine. The activities directed towards the site of injection are like those observed with injection, or with external application, of various noxious substances and the present study adds to a growing body of knowledge about possible pain in decapods.

Shake it off: exploring drivers and outcomes of autotomy in marine invertebrates.

Autotomy refers to self-amputation where the loss of a limb or organ is generally said to be (1) in response to stressful external stimuli; (2) voluntary and nervously mediated; (3) supported by adaptive features that increase efficiency and simultaneously mediate the cost; and (4) morphologically delineated by a predictable breakage plane. It is estimated that this phenomenon has evolved independently nine different times across the animal kingdom, appearing in many different taxa, including vertebrate and invertebrate as well as aquatic and terrestrial animals. Marine invertebrates use this behaviour in a diversity of manners that have yet to be globally reviewed and critically examined. Here, published data from marine invertebrate taxa were used to explore instances of injury as an evolutionary driver of autotomy. Findings suggest that phyla (e.g. Echinodermata and Arthropoda) possibly experiencing high rates of injury (tissue damage or loss) are more likely to be able to perform autotomy. Additionally, this review looks at various morphological, physiological and environmental conditions that have either driven the evolution or maintained the behaviour of autotomy in marine invertebrates. Finally, the use of autotomic abilities in the development of more sustainable and less ecologically invasive fisheries is explored.

Self-Amputating and Interfusing Machines.

Biological organisms exhibit phenomenal adaptation through morphology-shifting mechanisms including self-amputation, regeneration, and collective behavior. For example, reptiles, crustaceans, and insects amputate their own appendages in response to threats. Temporary fusion between individuals enables collective behaviors, such as in ants that temporarily fuse to build bridges. The concept of morphological editing often involves the addition and subtraction of mass and can be linked to modular robotics, wherein synthetic body morphology may be revised by rearranging parts. This work describes a reversible cohesive interface made of thermoplastic elastomer that allows for strong attachment and easy detachment of distributed soft robot modules without direct human handling. The reversible joint boasts a modulus similar to materials commonly used in soft robotics, and can thus be distributed throughout soft robot bodies without introducing mechanical incongruities. To demonstrate utility, the reversible joint is implemented in two embodiments: a soft quadruped robot that self-amputates a limb when stuck, and a cluster of three soft-crawling robots that fuse to cross a land gap. This work points toward future robots capable of radical shape-shifting via changes in mass through autotomy and interfusion, as well as highlights the crucial role that interfacial stiffness change plays in autotomizable biological and artificial systems.

Robustness to leg loss in Opiliones: A review and framework considerations for future research.

Animals have evolved behavioral and morphological traits that allow them to respond to environmental challenges. However, these traits may have long-term consequences that could impact an animal's performance, fitness, and welfare. Several species in a group of the arachnid order of Opiliones release their legs voluntarily to escape predators. These animals use their legs for locomotion, sensation, and reproduction. Here, we first compile data across species in the suborder Eupnoi, showing that more than half of individuals are found missing legs. Then, we review recent work on the ultimate and proximate implications of leg loss in Opiliones. Field and laboratory experiments showed that leg loss (a) did not affect their survival or mating success and (b) compromised the kinematics and energetics of locomotion, but individuals recovered velocity and acceleration quickly. These findings demonstrate that these animals display robustness, i.e., the ability to withstand and overcome the potential consequences of bodily damage. This may explain why leg loss is so common and prevalent in Opiliones. Additionally, we encourage researchers to consider expanding their hypotheses beyond traditional adaptationist and ableist lenses and incorporate a comprehensive examination of animal welfare when studying animals' responses to bodily damage. Finally, we highlight avenues for future research in Opiliones, namely assessing how individuals move in three-dimensional environments, the neural plasticity aiding recovery post-leg loss, applications for bio-inspired design, and evidence-based animal welfare measures.

Report of abnormal tail regeneration of Eremias yarkandensis (Sauria: Lacertidae) and its locomotor performance.

Caudal autotomy is a phenomenon observed in many reptile taxa, and tail loss is a pivotal functional trait for reptiles, with potentially negative implications for organism fitness due to its influence on locomotion. Some lizard species can regenerate a lost tail, which sometimes can lead to the development of more than one tail (i.e., abnormal tail regeneration) in the process. However, little is currently known about the impact of abnormal tail regeneration on locomotor performance. In this study, we document abnormal tail regeneration in Eremias yarkandensis, a reptile species native to northwestern China. Additionally, we investigated the sprint speed and endurance performance of these lizards. This study provides the first report on abnormal tail regeneration and its locomotor performance on a Chinese reptile. We suggest that the abnormal regeneration of tails may contribute to the accumulation of food reserves in the species. In light of our findings, we propose that herpetologists continue to share their sporadic observations and assess the locomotor performance of species experiencing abnormal tail regeneration, further expanding our understanding of this intriguing phenomenon.

Adaptation of Glucose Metabolism to Limb Autotomy and Regeneration in the Chinese Mitten Crab.

Limb autotomy and regeneration represent distinctive responses of crustaceans to environmental stress. Glucose metabolism plays a pivotal role in energy generation for tissue development and regeneration across various species. However, the relationship between glucose metabolism and tissue regeneration in crustaceans remains elusive. Therefore, this study is aimed at analyzing the alterations of glucose metabolic profile during limb autotomy and regeneration in Eriocheir sinensis, while also evaluating the effects of carbohydrate supplementation on limb regeneration. The results demonstrated that limb autotomy triggered a metabolic profile adaption at the early stage of regeneration. Hemolymph glucose levels were elevated, and multiple glucose catabolic pathways were enhanced in the hepatopancreas. Additionally, glucose and ATP levels in the regenerative limb were upregulated, along with increased expression of glucose transporters. Furthermore, the gene expression and activity of enzymes involved in gluconeogenesis were repressed in the hepatopancreas. These findings indicate that limb regeneration triggers metabolic profile adaptations to meet the elevated energy requirements. Moreover, the study observed that supplementation with corn starch enhanced limb regeneration capacity by promoting wound healing and blastema growth. Interestingly, dietary carbohydrate addition influenced limb regeneration by stimulating gluconeogenesis rather than glycolysis in the regenerative limb. Thus, these results underscore the adaptation of glucose metabolism during limb autotomy and regeneration, highlighting its essential role in the limb regeneration process of E. sinensis.

Walking leg regeneration in the sea spider Nymphon brevirostre Hodge, 1863 (Pycnogonida).

Regeneration is widespread across all animal taxa, but patterns of its distribution and key factors determining regeneration capabilities stay enigmatic. A comparative approach could shed light on the problem, but its efficacy is limited by the fact that data is only available on a few species from derived taxa. Pycnogonida are nested basally within the Chelicerata. They can shed and replace their walking legs and have a high regeneration capacity. In this work, we carried careful observation on leg appendotomy and regeneration processes in a sea spider under laboratory settings. The limb structure and in vivo observation reveal autotomy as the most likely appendotomy mechanism. High regeneration capabilities were ascertained: an anatomically normal but small leg appeared in a single molting cycle and the full functionality regained in 2-3 cycles. Wound closure after appendotomy in N. brevirostre primarily relies on hemolymph coagulation, which apparently differs from both xiphosurans and crustaceans. Regeneration is provided by proliferation in the leg cutpiece. Regenerative morphogenesis resembles the normal ontogenetic morphogenesis of a walking leg, but accelerated. Unlike in most arthropods, in N. brevirostre, regeneration does not necessarily correspond to the molting cycle, inferring a plesiomorphic state.

MicroRNA sequencing analysis reveals injury-induced immune responses of Scylla paramamosain against cheliped autotomy.

During pond culture or intensive culture system of crabs (mainly Eriocheir sinensis, Portunus trituberculatus and Scylla paramamosain), high-density farming has typically contributed to a higher limb autotomy level in juvenile animals, especially in S. paramamosain which has a high level of cannibalism. Due to the high limb autotomy level, the survival and growth rates in S. paramamosain farming are restricted, which limit the growth of the mud crab farming industry. MicroRNAs (miRNAs) are small noncoding RNAs that regulate a series of biological processes including innate immune responses by post-transcriptional suppression of their target genes. MiRNAs are believed to be crucial for innate immune process of host wound healing. Many miRNAs have been verified to be required in host immune responses to repair wound and to defense pathogen after tissue damage. However, to our best knowledge, the miRNAs functions of crustacean innate immune reactions against injury induced by limb autotomy have not been studied yet. Here in this study, for the first time, miRNAs involved in the S. paramamosain immune reactions against injury induced by cheliped autotomy were obtained by high-throughput sequencing. A total of 575 miRNAs (518 known miRNAs and 57 novel predicted miRNAs) were obtained, of which 141 differentially expressed microRNAs (93 up-regulated microRNAs and 48 down-regulated microRNAs) were revealed to be modified against cheliped autotomy, and the qPCR results of randomly selected miRNAs confirmed the expression patterns in the miRNAs sequencing data. Numerous immune-related target genes associated with innate immune system were mediated by miRNAs to induce host humoral immune and cellular immune defense to minimize acute physical damage. Furthermore, the genes expression in hemolymph coagulation and melanization pathways, as well as Toll and Imd signaling pathways were mediated by miRNAs to activate host immune responses including melanization and antimicrobial peptides for rapid wound healing and killing invaded pathogens. These results will help to understand injury-induced immune responses in crabs and to develop an effective control strategy of autotomy rate in crabs farming.

The tailless gecko gets the worm: prey type alters the effects of caudal autotomy on prey capture and subjugation kinematics.

Prey capture and subjugation are complex behaviors affected by many factors including physiological and behavioral traits of both the predator and the prey. The western banded gecko (Coleonyx variegatus) is a small generalist predator that consumes both evasive prey items, such as spiders, wasps, and orthopterans, and non-evasive prey items, including larvae, pupae, and isopterans. When consuming certain prey (e.g., scorpions), banded geckos will capture and then rapidly oscillate, or shake, their head and anterior part of their body. Banded geckos also have large, active tails that can account for over 20% of their body weight and can be voluntarily severed through the process of caudal autotomy. However, how autotomy influences prey capture behavior in geckos is poorly understood. Using high-speed 3D videography, we studied the effects of both prey type (mealworms and crickets) and tail autotomy on prey capture and subjugation performance in banded geckos. Performance metrics included maximum velocity and distance of prey capture, as well as velocity and frequency of post-capture shaking. Maximum velocity and distance of prey capture were lower for mealworms than crickets regardless of tail state. However, after autotomy, maximum velocity increased for strikes on mealworms but significantly decreased for crickets. After capture, geckos always shook mealworms, but never crickets. The frequency of shaking mealworms decreased after autotomy and additional qualitative differences were observed. Our results highlight the complex and interactive effects of prey type and caudal autotomy on prey capture biomechanics.

Snow flies self-amputate freezing limbs to sustain behavior at sub-zero temperatures.

Temperature profoundly impacts all living creatures. In spite of the thermodynamic constraints on biology, some animals have evolved to live and move in extremely cold environments. Here, we investigate behavioral mechanisms of cold tolerance in the snow fly (Chionea spp.), a flightless crane fly that is active throughout the winter in boreal and alpine environments of the northern hemisphere. Using thermal imaging, we show that adult snow flies maintain the ability to walk down to an average body temperature of -7°C. At this supercooling limit, ice crystallization occurs within the snow fly's hemolymph and rapidly spreads throughout the body, resulting in death. However, we discovered that snow flies frequently survive freezing by rapidly amputating legs before ice crystallization can spread to their vital organs. Self-amputation of freezing limbs is a last-ditch tactic to prolong survival in frigid conditions that few animals can endure. Understanding the extreme physiology and behavior of snow insects holds particular significance at this moment when their alpine habitats are rapidly changing due to anthropogenic climate change. VIDEO ABSTRACT.

Hedgehog signaling is essential in the regulation of limb regeneration in the Chinese mitten crab, Eriocheir sinensis.

Tissue autotomy is a unique adaptive response to environmental stress, followed by regeneration process compensating for the loss of body parts. The crustaceans present remarkable activity of appendage autotomy and regeneration, however, the molecular mechanism is still unclear. In this study, the Eriocheir sinensis Hedgehog (EsHH) and Smoothened (EsSMO) were identified in the regenerative limbs, and the function of Hedgehog signaling pathway on limb regeneration was evaluated. At the blastema growth stage of limb regeneration, the expression of EsHH and EsSMO was up-regulated in response to limb autotomy stress, and down-regulated at blastema differentiation stage. To clarify the effect of Hedgehog pathway during limb regeneration, the regenerative efficiency was evaluated with Smoothened inhibitor cyclopamine or RNAi (ds-HH) injection. We observed that the regenerative efficiency was significantly repressed with blockage of Hedgehog pathway at both the basal growth stage and the proecdysial growth stage, which was indicated by the delay of wound healing and blastema growth, as well as a decrease in the size of newly formed limbs. In addition, gene expression and BrdU incorporation assay showed that the proliferation and myogenic differentiation of blastema cells were suppressed with either cyclopamine or ds-HH injection. Thus, these results suggest that Hedgehog signaling pathway is essential for the establishment of limb regeneration in E. sinensis through promoting the proliferation and myogenic differentiation of blastema cells.

Behavioral toxicity, histopathological alterations and oxidative stress in Tubifex tubifex exposed to aromatic carboxylic acids- acetic acid and benzoic acid: A comparative time-dependent toxicity assessment.

This study evaluated Acetic acid (AA) and Benzoic acid's (BA) acute and sublethal toxicity by observing mortality, behavioral responses, and changes in the levels of oxidative stress enzymes in Tubifex tubifex. Exposure-induced changes in antioxidant activity (Catalase, Superoxide dismutase), oxidative stress (Malondialdehyde concentrations), and histopathological alterations in the tubificid worms were also noted across exposure intervals. The 96 h LC50 values of AA and BA to T. tubifex were 74.99 and 37.15 mg/l, respectively. Severity in behavioral alterations (including increased mucus production, wrinkling, and reduction in clumping) and autotomy showed concentration-dependent trends for both toxicants. Although histopathological effects also showed marked degeneration in the alimentary and integumentary systems in highest exposure groups (worms exposed to 14.99 mg/l for AA and 7.42 mg/l for BA) for both toxicants. Antioxidant enzymes (catalase and superoxide dismutase) also showed a marked increase of up to 8-fold and 10-fold for the highest exposure group of AA and BA respectively. While species sensitivity distribution analysis revealed T. tubifex as most sensitive to AA and BA compared to other freshwater vertebrates and invertebrates, General Unified Threshold model of Survival (GUTS) predicted individual tolerance effects (GUTS-IT), with slower potential for toxicodynamic recovery, as a more likely pathway for population mortality. Study findings demonstrate BA with greater potential for ecological effects compared to AA within 24 h of exposure. Furthermore, ecological risks to critical detritus feeders like T. tubifex may have severe implications for ecosystem services and nutrient availability within freshwater habitats.

How does the timing of weapon loss influence reproductive traits and trade-offs in the insect Narnia femorata?

A longstanding goal of evolutionary biology is to understand among-individual variation in resource allocation decisions and the timing of these decisions. Recent studies have shown that investment in elaborate and costly weapons can result in trade-offs with investment in testes. In this study, we ask the following questions: At what point plasticity in resource allocation to these different structures ceases during development, if at all? Furthermore, can individuals tailor their reproductive behavior to accompany structural changes? We experimentally addressed these questions in the insect Narnia femorata, quantifying resource reallocation across development for the first time, using a phenotypic engineering approach. To investigate whether allocation plasticity diminishes throughout ontogeny, we induced weapon loss at a range of different developmental stages and examined subsequent testes mass and reproductive behavior. We found that relative testes mass increased as weapon investment decreased, implying a direct trade-off between testes and weapon investment. However, autotomy postadulthood ceased to induce larger testes mass. Intriguingly, losing a weapon while young was associated with extended adult mating duration, potentially enabling compensation for reduced fighting ability. Our results highlight the importance of examining the ontogeny of trade-offs between reproductive traits and the flexibility of the relationship between reproductive morphology and behavior.

Unwrapping broken tails: Biological and environmental correlates of predation pressure in limbless reptiles.

Studying species interactions in nature often requires elaborated logistics and intense fieldwork. The difficulties in such task might hinder our ability to answer questions on how biotic interactions change with the environment. Fortunately, a workaround to this problem lies within scientific collections. For some animals, the inspection of preserved specimens can reveal the scars of past antagonistic encounters, such as predation attempts. A common defensive behaviour that leaves scars on animals is autotomy, the loss of a body appendage to escape predation. By knowing the collection site of preserved specimens, it is possible to assess the influence of organismal biology and the surrounding environment in the occurrence of autotomy. We gathered data on tail loss for 8189 preserved specimens of 33 snake and 11 amphisbaenian species to investigate biological and environmental correlates of autotomy in reptiles. We applied generalized linear mixed effect models to evaluate whether body size, sex, life-stage, habitat use, activity pattern, biome, tropicality, temperature and precipitation affect the probability of tail loss in limbless reptiles. We observed autotomy in 23.6% of examined specimens, with 18.7% of amphisbaenian and 33.4% of snake specimens showing tail loss. The probability of tail loss did not differ between snakes and amphisbaenians, but it was higher among large-sized specimens, particularly in adults and females. Chance of tail loss was higher for diurnal and arboreal species, and among specimens collected in warmer regions, but it was unaffected by biome, precipitation, and tropicality. Autotomy in limbless reptiles was affected by size-dependent factors that interplay with ontogeny and sexual dimorphism, although size-independent effects of life-stage and sex also shaped behavioural responses to predators. The increase in probability of tail loss with verticality and diurnality suggests a risk-balance mechanism between species habitat use and activity pattern. Although autotomy is more likely in warmer regions, it seems unrelated to seasonal differences in snakes and amphisbaenians activity. Our findings reveal several processes related to predator-prey interactions involving limbless reptiles, demonstrating the importance of scientific collections to unveil ecological mechanisms at different spatio-temporal scales.

Regenerative peripheral nerve interface prevents neuroma formation after peripheral nerve transection.

Neuroma formation after peripheral nerve transection often leads to severe neuropathic pain. Regenerative peripheral nerve interface has been shown to reduce painful neuroma in the clinic. However, no reports have investigated the underlying mechanisms, and no comparative animal studies on regenerative peripheral nerve interface and other means of neuroma prevention have been conducted to date. In this study, we established a rat model of left sciatic nerve transfection, and subsequently interfered with the model using the regenerative peripheral nerve interface or proximal nerve stump implantation inside a fully innervated muscle. Results showed that, compared with rats subjected to nerve stump implantation inside the muscle, rats subjected to regenerative peripheral nerve interface intervention showed greater inhibition of the proliferation of collagenous fibers and irregular regenerated axons, lower expressions of the fibrosis marker α-smooth muscle actin and the inflammatory marker sigma-1 receptor in the proximal nerve stump, lower autophagy behaviors, lower expressions of c-fos and substance P, higher expression of glial cell line-derived neurotrophic factor in the ipsilateral dorsal root ganglia. These findings suggested that regenerative peripheral nerve interface inhibits peripheral nerve injury-induced neuroma formation and neuropathic pain possibly via the upregulation of the expression of glial cell line-derived neurotrophic factor in the dorsal root ganglia and reducing neuroinflammation in the nerve stump.

Integrative biology of injury in animals.

Mechanical injury is a prevalent challenge in the lives of animals with myriad potential consequences for organisms, including reduced fitness and death. Research on animal injury has focused on many aspects, including the frequency and severity of wounding in wild populations, the short- and long-term consequences of injury at different biological scales, and the variation in the response to injury within or among individuals, species, ontogenies, and environmental contexts. However, relevant research is scattered across diverse biological subdisciplines, and the study of the effects of injury has lacked synthesis and coherence. Furthermore, the depth of knowledge across injury biology is highly uneven in terms of scope and taxonomic coverage: much injury research is biomedical in focus, using mammalian model systems and investigating cellular and molecular processes, while research at organismal and higher scales, research that is explicitly comparative, and research on invertebrate and non-mammalian vertebrate species is less common and often less well integrated into the core body of knowledge about injury. The current state of injury research presents an opportunity to unify conceptually work focusing on a range of relevant questions, to synthesize progress to date, and to identify fruitful avenues for future research. The central aim of this review is to synthesize research concerning the broad range of effects of mechanical injury in animals. We organize reviewed work by four broad and loosely defined levels of biological organization: molecular and cellular effects, physiological and organismal effects, behavioural effects, and ecological and evolutionary effects of injury. Throughout, we highlight the diversity of injury consequences within and among taxonomic groups while emphasizing the gaps in taxonomic coverage, causal understanding, and biological endpoints considered. We additionally discuss the importance of integrating knowledge within and across biological levels, including how initial, localized responses to injury can lead to long-term consequences at the scale of the individual animal and beyond. We also suggest important avenues for future injury biology research, including distinguishing better between related yet distinct injury phenomena, expanding the subjects of injury research to include a greater variety of species, and testing how intrinsic and extrinsic conditions affect the scope and sensitivity of injury responses. It is our hope that this review will not only strengthen understanding of animal injury but will contribute to building a foundation for a more cohesive field of 'injury biology'.