Bundles of Spider Silk,Braided into Sutures,Resist Basic Cyclic Tests: Potential Use for Flexor Tendon Repair

Repair success for injuries to the flexor tendon in the hand is often limited by the in vivo behaviour of the suture used for repair. Common problems associated with the choice of suture material include increased risk of infection, foreign body reactions, and inappropriate mechanical responses, particularly decreases in mechanical properties over time. Improved suture materials are therefore needed. As high-performance materials with excellent tensile strength, spider silk fibres are an extremely promising candidate for use in surgical sutures. However, the mechanical behaviour of sutures comprised of individual silk fibres braided together has not been thoroughly investigated. In the present study, we characterise the maximum tensile strength, stress, strain, elastic modulus, and fatigue response of silk sutures produced using different braiding methods to investigate the influence of braiding on the tensile properties of the sutures. The mechanical properties of conventional surgical sutures are also characterised to assess whether silk offers any advantages over conventional suture materials. The results demonstrate that braiding single spider silk fibres together produces strong sutures with excellent fatigue behaviour; the braided silk sutures exhibited tensile strengths comparable to those of conventional sutures and no loss of strength over 1000 fatigue cycles. In addition, the braiding technique had a significant influence on the tensile properties of the braided silk sutures. These results suggest that braided spider silk could be suitable for use as sutures in flexor tendon repair, providing similar tensile behaviour and improved fatigue properties compared with conventional suture materials.     

Failure of low-velocity swimming to enhance recovery from exhaustive exercise in largemouth bass (Micropterus salmoides)

This study was intended to discover whether forcing largemouth bass (Micropterus salmoides) to swim at 0.5 body lengths/second following exercise would expedite recovery relative to fish recovered in static water. Exercise resulted in a suite of physiological disturbances for largemouth bass that included a depletion of anaerobic energy stores, an accumulation of lactate, and increased cardiac output. At 1 h following exercise, exhaustively exercised largemouth bass forced to swim exhibited expedited recovery relative to fish in static water, evidenced by lower concentrations of lactate in white muscle, elevated concentrations of phosphocreatine in white muscle, and reduced concentrations of glucose in plasma. By 4 h postexercise, largemouth bass forced to swim during recovery exhibited signs of physiological disturbance that were absent in fish recovered in static water. These signs of disturbance included a loss of osmotically active particles from plasma, elevated lactate in plasma, reductions of phospocreatine in white muscle, and increased cardiac output. These results are discussed in relation to the body of work with salmonid fishes showing physiological benefits to recovering fish in flowing water.     

Experimental trophic ecology of juvenile largemouth bass,Micropterus salmoides,and blue tilapia,Oreochromis aureus

Synopsis The potential for feeding competition between largemouth bass, Micropterus salmoides, and blue tilapia, Oreochromis areus, in Lake Fairfield, Texas was evaluated experimentally. Largemouth bass and blue tilapia were grown in cages alone and in combination with each other. The fish were allowed to feed on the natural food within the lake. Largemouth bass grown in combination with blue tilapia were significantly shorter and weighed less than largemouth bass grown alone. Blue tilapia grown in combination with largemouth bass were statistically significantly longer and heavier than blue tilapia grown alone. Largemouth bass grown alone had diets (volume and number of food items) significantly different than the largemouth bass grown with the blue tilapia. Largemouth bass fed primarily on chironomid larvae and pupae, and odonates, whereas blue tilapia consumed vegetable matter, detritus, and chironomid larvae. Length and weight differences between large-mouth bass grown alone and in combination with blue tilapia, in conjunction with the largemouth bass diet shift, support the theory that these two species compete for food resources.     

Antipredator Behavior of American Bullfrogs (Lithobates catesbeianus) in a Novel Environment

Invasive species capable of recognizing potential predators may have increased establishment rates in novel environments. Individuals may retain historical predator recognition and invoke innate responses in the presence of taxonomically or ecologically similar predators, generalize antipredator responses, or learn to avoid risky species in novel environments. Invasive amphibians in aquatic environments often use chemical cues to assess predation risk and learn to avoid novel predators via direct experience and/or associated chemical cues. Ontogeny may also influence recognition; experience with predators may need to occur at certain developmental stages for individuals to respond correctly. We tested predator recognition in invasive American bullfrog ( Lithobates catesbeianus) tadpoles that varied in experience with fish predators at the population and individual scale. We found that bullfrog tadpoles responded to a historical predator, largemouth bass ( Micropterus salmoides), only if the population was locally sympatric with largemouth bass. Individuals from a population that did not co‐occur with largemouth bass did not increase refuge use in response to either largemouth bass chemical cues alone or chemical cues with diet cues (largemouth bass fed bullfrog tadpoles). To test whether this behavioral response was generalized across fish predators, we exposed tadpoles to rainbow trout ( Oncorhynchus mykiss) and found that tadpoles could not recognize this novel predator regardless of co‐occurrence with other fish species. These results suggest that environment may be more important for predator recognition than evolutionary history for this invasive species, and individuals do not retain predator recognition or generalize across fish predators.     

Influence of water temperature and net tending frequency on the condition of fish bycatch in a small-scale inland commercial fyke net fishery

To date, most studies of commercial fisheries bycatch have focused on mortality at time of capture as an endpoint. However, sub-lethal indicators of organismal condition have the potential to reveal mechanisms associated with mortality (both at time of capture and post-release) and opportunities for improving fish welfare. In this study, we simulated commercial fishing efforts in inland lakes with fyke nets during a typical fishing season (early April to late June) in southeastern Ontario, Canada, where bycatch of non-target fish species had previously been documented. Using non-target gamefish (i.e., largemouth bass [Micropterus salmoides, Lacépède], northern pike [Esox Lucius, L.]), as well as a target species (i.e., bluegill [Lepomis macrochirus, Rafinesque]), we examined the sub-lethal consequences of capture (e.g., blood physiology, reflex impairment, and injury) and compared the effects of being retained in the net for two different durations (i.e., two or six days) over a range of water temperatures (i.e., 3–28 °C). Sub-lethal physiological disturbances (i.e., blood glucose and lactate) in largemouth bass and bluegill tended to be greater at higher water temperatures. However, fish retained for six days generally did not exhibit greater stress than those retained for two days, with the exception of plasma glucose in largemouth bass. Reflex impairment was similar among temperature and retention periods. Fish retained in nets experienced a range of injuries (including fin frays, scale loss, and mouth damage) that had the potential to facilitate the development of opportunistic pathogenic infections. Greater incidences of injury on fish bycatch tended to be associated with higher temperatures and longer retention. To reduce physiological disturbances and injury that could lead to delayed mortality, we suggest that regulations for inland commercial fishers require them to check their nets more frequently as water temperatures increase. We suggest that future studies of bycatch incorporate sub-lethal endpoints given that they serve as an objective measure of fish welfare and can provide quantitative mechanistic information to support management actions.     

The Impact of Surgical Experience on Major Intraoperative Aneurysm Rupture and Their Consequences on Outcome: A Multivariate Analysis of 538 Microsurgical Clipping Cases

The incidence and associated mortality of major intraoperative rupture (MIOR) in intracranial aneurysm surgery is diverse. One possible reason is that many studies failed to consider and properly adjust the factor of surgical experience in the context. We conducted this study to clarify the role of surgical experience on MIOR and associated outcome. 538 consecutive intracranial aneurysm surgeries performed on 501 patients were enrolled in this study. Various potential predictors of MIOR were evaluated with stratified analysis and multivariate logistic regression. The impact of surgical experience and MIOR on outcome was further studied in a logistic regression model with adjustment of each other. The outcome was evaluated using the Glasgow Outcome Scale one year after the surgery. Surgical experience and preoperative Glasgow Coma Scale (GCS) were identified as independent predictors of MIOR. Experienced neurovascular surgeons encountered fewer cases of MIOR compared to novice neurosurgeons (MIOR, 18/225, 8.0% vs. 50/313, 16.0%, P = 0.009). Inexperience and MIOR were both associated with a worse outcome. Compared to experienced neurovascular surgeons, inexperienced neurosurgeons had a 1.90-fold risk of poor outcome. On the other hand, MIOR resulted in a 3.21-fold risk of unfavorable outcome compared to those without it. Those MIOR cases managed by experienced neurovascular surgeons had a better prognosis compared with those managed by inexperienced neurosurgeons (poor outcome, 4/18, 22% vs. 30/50, 60%, P = 0.013).     

Predation of Japanese dace, Tribolodon hakonensis, by largemouth bass, Micropterus salmoides, in experimental aquaria

 To test the size range of prey fish that largemouth bass, Micropterus salmoides, can successfully consume, live Japanese dace, Tribolodon hakonensis, were given as prey fish to individual largemouth bass in aquaria. The ratio of maximum standard length (SL) of the Japanese dace consumed by largemouth bass was 46–69% of bass SL. The maximum length of Japanese dace consumed did not differ significantly between largemouth bass and smallmouth bass (M. dolomieu) previously studied, although largemouth bass have relatively larger mouth sizes than smallmouth bass. Largemouth bass occasionally injured and killed Japanese dace larger than the limit that could be consumed.     

Annual cycles of growth and reproduction in hatchery–reared Florida largemouth bass, Micropterus salmoides floridanus, raised on forage or pelleted diets

Annual cycles of growth and reproduction of hatchery–reared Florida largemouth bass, Micropterus salmoides floridanus , were investigated. Animals were raised on either forage (goldfish, Carassius auratus ) or a pelleted salmon feed. Male and female year–class 1 largemouth bass were sampled throughout one complete yearly cycle (January–December). A biphasic growth cycle was observed in both forage–fed and pellet–fed fish. No increase in body length or weight was observed until approximately midway through the spawning period (May), after which fish grew at a consistent rate for the remainder of the study. The reproductive cycle of forage–fed fish was characterized by a rapid increase in gonadosomatic index (GS1) between January and April, followed by a prolonged spawning period (April–July) during which GSI progressively declined. Fully regressed gonads were observed in September and October, and a resumption of gonadal recrudescence was observed between October and December. Visceral adipose deposits (expressed as mesenteric fat index; MFI) were resorbed during gonadal growth and the initial stages of the spawning period, and restored during the post–spawning phase. Fish raised on pelleted feed had growth and reproductive cycles that parallelled those of forage–fed fish, but several significant differences were observed between the two diet groups. During the growth phase of the cycle, pellet–fed largemouth bass grew significantly faster than forage–fed largemouth bass, and had significantly larger MFIs than forage–fed largemouth bass at all times of the year. Pellet–fed fish also had significantly larger GSIs than forage–fed fish. These data indicate that diet composition may be an important determinant of growth and reproductive function in this species.     

A comparison of prey capture kinematics in hatchery and wild Micropterus salmoides floridanus: effects of ontogeny and experience

Analysis of high‐speed videography demonstrated that juvenile wild Florida largemouth bass Micropterus salmoides floridanus captured live prey with very rapid movements and large excursions. Hatchery fish of the same age, raised on pelleted feed, however, used slower kinematics with smaller excursions, yielding strikes with a higher degree of 'suction'. Capture events of hatchery Florida largemouth bass fed live prey for the first time were characterized by movements that had smaller excursion measurements than wild fish and resulted in a decreased level of capture success. After five exposures to elusive mosquitofish Gambusia holbrooki , hatchery Florida largemouth bass adapted their behaviour to capture prey at the kinematic level of wild fish.     

Suture materials and other factors associated with tissue reactivity, infection, and wound dehiscence among plastic surgery outpatients

The most common complications in plastic surgery are tissue reactivity, infections, and wound dehiscence. In the literature, there are only a few studies with sample sizes large enough and methods of statistical analysis appropriate for evaluating the role of suture materials in inducing such complications. In the 1000 plastic surgery outpatients in this study, the association of different suture materials, individual patient characteristics, surgeon skill, and wound site and length with postoperative wound complications (i.e., tissue reactivity, infection rate, and wound dehiscence) were investigated. No substantial differences were found between the different suture materials and suturing techniques. A moderate increase in the risk of tissue reactivity for silk and polyglactin 910 and a protective effect of thinner internal sutures were observed. In multivariate analysis, such differences were not statistically significant. Male sex [odds ratio (OR), 1.7; 95 percent confidence interval (CI), 1.06 to 2.72] and older age (OR, 2.34; 95 percent CI, 1.36 to 4.05) were found to be the most important risk factors for tissue reactivity and infection rate (male sex: OR, 5.1; 95 percent CI, 1.7 to 15.9; older age: OR, 5.6; 95 percent CI, 1.9 to 16), whereas younger age was associated with an increased risk of dehiscence (OR, 3.06; 95 percent CI, 1.41 to 6.65). Wounds on the lower limbs showed a lower risk of tissue reactivity and wounds on the back a higher risk of dehiscence. Wound length was associated with the risk of tissue reactivity in one-layer sutures (OR, 2.92; 95 percent CI, 1.51 to 5.65). An increased risk of both tissue reactivity (OR, 1.53; 95 percent CI, 1.03 to 2.27) and dehiscence (OR, 2.44; 95 percent CI, 1.1 to 5.43) was observed for operations performed by less-experienced surgeons. Rather than factors related to suture materials and different surgical techniques, and with the exception of surgeon experience, general characteristics of the patients (i.e., sex and age) and of the wounds (i.e., length and site) seemed to be primarily responsible for local wound complications.     

Distribution and diet of Suwannee bass and largemouth bass in the lower Santa Fe River,Florida

Synopsis Suwannee bass,Micropterus notius, and largemouth bass,Micropterus salmoides, were collected by electrofishing in six habitats in the lower Santa Fe River, Florida during May 1981–March 1982. Both bass species were collected concomitantly in all habitats and habitat segregation was not evident. Crayfish (Procambarus spp.) were the primary food of Suwannee bass. Fish were the primary food of largemouth bass, but crayfish were common in the diet of largemouth bass ≥300 mm long. Suwannee bass have a greater throat width and consumed longer and wider forage than did largemouth bass of equal length. Available evidence suggests that Suwannee bass exhibit a positive selection for crayfish and a diverse forage resource, including abundant crayfish, is necessary for a Suwannee bass to coexist with a largemouth bass. This is Journal Series Number 6034 of the Florida Agricultural Experiment Station.     

Projected shifts in fish species dominance in Wisconsin lakes under climate change

Temperate lakes may contain both coolwater fish species such as walleye (Sander vitreus) and warmwater fish species such as largemouth bass (Micropterus salmoides). Recent declining walleye and increasing largemouth bass populations have raised questions regarding the future trajectories and management actions for these species. We developed a thermodynamic model of water temperatures driven by downscaled climate data and lake‐specific characteristics to estimate daily water temperature profiles for 2148 lakes in Wisconsin, US, under contemporary (1989–2014) and future (2040–2064 and 2065–2089) conditions. We correlated contemporary walleye recruitment and largemouth bass relative abundance to modeled water temperature, lake morphometry, and lake productivity, and projected lake‐specific changes in each species under future climate conditions. Walleye recruitment success was negatively related and largemouth bass abundance was positively related to water temperature degree days. Both species exhibited a threshold response at the same degree day value, albeit in opposite directions. Degree days were predicted to increase in the future, although the magnitude of increase varied among lakes, time periods, and global circulation models (GCMs). Under future conditions, we predicted a loss of walleye recruitment in 33–75% of lakes where recruitment is currently supported and a 27–60% increase in the number of lakes suitable for high largemouth bass abundance. The percentage of lakes capable of supporting abundant largemouth bass but failed walleye recruitment was predicted to increase from 58% in contemporary conditions to 86% by mid‐century and to 91% of lakes by late century, based on median projections across GCMs. Conversely, the percentage of lakes with successful walleye recruitment and low largemouth bass abundance was predicted to decline from 9% of lakes in contemporary conditions to only 1% of lakes in both future periods. Importantly, we identify up to 85 resilient lakes predicted to continue to support natural walleye recruitment. Management resources could target preserving these resilient walleye populations.     

Examination of the effects of largemouth bass (Micropterus salmoides) and bluegill (Lepomis macrochirus) on the ecosystem attributes of lake Kawahara-oike,Nagasaki, Japan

The introduction of largemouth bass (Micropterus salmoides) and bluegill sunfish (Lepomis macrochirus) into the freshwater ecosystems of Japan has resulted in the suppression and/or replacement of native species, generating considerable concerns among resource managers. The impacts of largemouth bass and bluegill on native fauna have been examined in aquaria and isolated farm ponds, but there is limited work examining the likelihood to fundamentally modifying Japan's lakes. The objective of the present study is to examine the direct and synergistic ecological effects of largemouth bass and bluegill on the biotic communities of Lake Kawahara-oike, Nagasaki, Japan, using an ecosystem (Ecopath) modeling approach. Specifically, we examine whether the two fish species have played a critical role in shaping the trophodynamics of the lake. We attempt to shed light on the trophic interactions between largemouth bass and bluegill and subsequently evaluate to what extent these interactions facilitate their establishment at the expense of native species. We also examine how these changes propagate through the Lake Kawahara-oike food web. Our study suggests that the introduction of bluegill has induced a range of changes at multiple trophic levels. The present analysis also provides evidence that largemouth bass was unable to exert significant top-down control on the growth rates of the bluegill population. Largemouth bass and bluegill appear to prevail over the native fish species populations and can apparently coexist in large numbers in invaded lakes. Future management strategies controlling invasive species are urgently required, if the integrity of native Japanese fish communities is to be protected.     

Effects of pressure changes induced by commercial navigation traffic on mortality of fish early life stages

Mortality of fish early life stages was measured in a pressure vessel to simulate vertical displacement within the water column. Mortality was measured for three pressure regimes for four fish species: larval bigmouth buffalo Ictiobus cyprinellus, larval blue catfish Ictalurus furcatus, juvenile bluegill Lepomis macrochirus, and juvenile largemouth bass Micropterus salmoides. The maximum pressure‐change tested, 344.8 kPa, equivalent to a 35.2 m displacement of fish within the water column, did not cause significant mortality of larvae or juveniles. Since 32.5 m exceeds depths in most inland navigation channels and possibly the depth to which rapid propeller induced water mixing occurs, the range of pressure changes that could be experienced by early life stages during towboat mixing of the water column will not result in significant mortality.     

The environmental basis of diet variation in pumpkinseed sunfish, Lepomis gibbosus, and largemouth bass, Micropterus salmoides, along an Iberian river basin

The diet composition of largemouth bass, Micropterus salmoides, and pumpkinseed sunfish, Lepomis gibbosus, and its relation to environmental factors was studied in intermittent watercourses of the lower Guadiana basin (southern Iberia) during the particularly dry summer of 1994. Overall, both species took food items similar to those found in their North American and acclimatised ranges, with pumpkinseed consuming invertebrates (chiefly Chironomidae) and bass preying on invertebrates (chiefly Micronecta meridionalis) and fish (chiefly L. gibbosus and Gambusia holbrooki). Both bass and pumpkinseed were opportunistic feeders, feeding on the most frequent and abundant prey. However, Diptera larvae were apparently preferred by pumpkinseed and avoided by bass, while the contrary occurred with respect to Heteroptera. The two main fish prey of bass were eaten on the basis of random encounter. The relative abundance of macro-prey (i.e. fish and Atyephira desmarestii) were the principal environmental variables constraining bass dietary variation along the basin. On the contrary, pumpkinseed diet variation was mainly related to pumpkinseed size and the presence of piscivorous bass nearby, although habitat size and cyprinid abundance were also influential. This revised version was published online in July 2006 with corrections to the Cover Date.     

Pathogen Host Switching in Commercial Trade with Management Recommendations

Global wildlife trade exacerbates the spread of nonindigenous species. Pathogens also move with hosts through trade and often are released into naïve populations with unpredictable outcomes. Amphibians are moved commercially for pets, food, bait, and biomedicine, and are an excellent model for studying how wildlife trade relates to pathogen pollution. Ranaviruses are amphibian pathogens associated with annual population die-offs; multiple strains of tiger salamander ranaviruses move through the bait trade in the western United States. Ranaviruses infect amphibians, reptiles, and fish and are of additional concern because they can switch hosts. Tiger salamanders are used as live bait for freshwater fishing and are a potential source for ranaviruses switching hosts from amphibians to fish. We experimentally injected largemouth bass with a bait trade tiger salamander ranavirus. Largemouth bass became infected but exhibited no signs of disease or mortality. Amphibian bait ranaviruses have the potential to switch hosts to infect fish, but fish may act as dead-end hosts or nonsymptomatic carriers, potentially spreading infection as a result of trade.     

The effects of surgically implanted acoustic transmitters on laboratory growth, survival and tag retention in hatchery yearling Chinook salmon

Telemetry has proven an effective means for studying the movement of fishes, however, biases associated with tagged animals requires careful scrutiny if accurate conclusions are to be made from field studies. The objective of this study was to evaluate growth, survival, and tag retention in hatchery yearling Chinook salmon (Oncorhynchus tshawytscha) juveniles with intracoelomic surgically implanted acoustic transmitters representing 2.6 to 5.6% of body weight. The first trial consisted of three treatments; passive integrated transponder (PIT) tag-only (25 fish), acoustic-tag+PIT-tag (25 fish), and sham-surgery+PIT-tag (25 fish). There were no significant differences in relative growth rate (% change in weight day^?1) among treatments over the 221 day trial. Survival in the acoustic-tag treatment (80%) was not significantly different from the PIT-tag-only and sham treatments (92 and 88% respectively). The second trial consisted of three treatments; PIT-tag-only (22 fish), acoustic-tag+PIT-tag with absorbable sutures (12 fish) and acoustic-tag+PIT-tag with non-absorbable sutures (12 fish). There were no significant differences in relative growth rate among treatments over the 160 day trial. Survival in the second trial was 100%. Fish with absorbable sutures healed sooner and with less inflammation compared to fish with non-absorbable sutures. Tag retention was 100% in both trials. The results of this study suggest that acoustic transmitters of less than 5.6% body weight can be effectively used in 1-year old Chinook salmon.     

The diet of largemouth bass, Micropterus salmoides, in Lake Naivasha, Kenya

Lake Naivasha is a freshwater lake situated in the eastern rift valley of Kenya. Only five species of fish are present, all of which have been introduced. Of these, Oreochromis leucostictus, Tilapia zillii and Micropterus salmoides (largemouth black bass) support an important gillnet fishery with bass also being taken for sport. Until bass reached 260 mm f.l. they depended upon invertebrate food organisms. Thereafter crayfish, fish and frogs became increasingly important the larger the size of the bass. The most important invertebrate prey species was the water boatman, Micronecta scutellaris , followed by chironomid and culicid pupae. Zooplank-ton was consumed but only in large quantity by fish smaller than 80 mm. For bass over 260 mm the crayfish, Procambarus darkii , was the principal food. The largemouth bass in Lake Naivasha are generalized macro-predators, feeding principally on free-living animals of a kind most likely to be found in the littoral zones.     

The influence of environmental temperature and oxygen concentration on the recovery of largemouth bass from exercise: implications for live–release angling tournaments

The impact of variation in water temperature and dissolved oxygen on recovery of largemouth bass Micropterus salmoides from exercise was examined. For this, largemouth bass were first exercised and recovered for either 1, 2 or 4 h at ambient water temperatures (25° C) in fully oxygenated water. Results showed that exercise forced fish to utilize anaerobic metabolism to meet energy demands, and resulted in reductions in anaerobic energy stores adenosine triphosphate (ATP), Phosphocreatine (PCr) and glycogen. Exercise also resulted in a seven‐fold increase in lactate within white muscle. After 2 h of recovery in oxygenated water at acclimation temperature, physiological recovery from exercise was under way, and by 4 h most variables examined had returned to control levels. Next, largemouth bass were exercised at ambient temperatures and recovered for 2 h in environments with either elevated temperature (32° C), reduced temperature (14 and 20° C), hypoxia or hyperoxia. Both elevated and reduced temperature impaired recovery of tissue lactate and tissue ATP relative to fish recovered in water at acclimation temperature, while hyperoxic water impaired recovery of tissue ATP. Moderately hypoxic waters impaired the recovery of plasma glucose, plasma lactate and tissue PCr relative to fish recovered in fully oxygenated water. Results from this study are discussed in the context of critical oxygen and temperature guidelines for largemouth bass. In addition, several recommendations are made concerning remedial treatments used in livewells (tanks) during angling tournaments when fish are recovering from exercise associated with angling.     

The influence of fish size size on the avoidance of hypoxia and oxygen selection by largemouth bass

When an oxygen gradient ranging from c . 10 to 95% air saturation was formed in a 5 m chamber, largemouth bass Micropterus salmoides avoided water in which dissolved oxygen values were <27% air saturation. There was a significant ( P <0·05) correlation between fish mass and the level of dissolved aquatic oxygen that was selected. Small fish (23–500 g) utilized waters of lower oxygen levels than did the larger fish (1000–3000 g). The results of this study suggest that largemouth bass are able to sense and avoid hypoxic water, and select aquatic oxygen tensions that maintain their metabolic scope for growth and activity.