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1.
Common carp (Cyprinus carpio, [Linnaeus, 1758]) have long been established in the United States and in some cases their populations can be very dense, leading biologists to expend significant effort towards reducing numbers of common carp in some populations. Estimating abundance of common carp prior to removal efforts would be useful in evaluating success of these removal efforts, but marking large numbers of fish can be expensive. Therefore, a low-cost tagging option is needed. In this study, we used tank and field experiments to determine the retention and longevity of hole-punch marks in the opercula of common carp. For the tank experiment, fish were double marked with a size-3 self-piercing tag and an operculum hole-punch mark (using a paper hole-punch tool with a hole diameter of 6.4 mm) on opposite sides of the fish. Over the entirety of the 180–day tank experiment, retention of the self-piercing tags and hole-punch marks was 100% and no marking mortality was observed. For the field experiment, 883 common carp were tagged at random in two groups, a double-marked group (n = 416, both self-piercing tags and hole-punch) and a single-mark group (n = 467, self-piercing tag). Fish were sampled monthly for 398 days. Because the length distribution sampled was bimodal, we evaluated tag retention of fish <330 mm TL (small fish, n = 273) and > 331 mm TL (large fish, n = 143), separately. Hole-punch mark retention was high for both size classes throughout most of the field experiment. For large fish, retention of hole-punch marks was 100% for the entire 398-day experiment. For small fish, retention of hole-punch marks was 100% through 184-day and remained above 93% through 328-days, but declined to 0% by day 398. Our results suggest that the operculum hole-punch mark is a valuable low-cost, long-term technique for tagging common carp.  相似文献   

2.
Growth, and potential marking‐related mortality of small European eel Anguilla anguilla (L.) after marking with visible implant elastomer tags (VIE) and coded wire tags (CWT) were investigated over a 6‐month period in a laboratory experiment. In addition, mark retention after marking was examined for another 16.5 months. Neither marking method had a significant effect on growth (P > 0.05) or mortality (P > 0.05). After 32 days detection of the VIE marks implanted on the ventral surface and along the base of the ventral tail fin margin was 98 and 100%, respectively, but decreased to 9 and 66% after 512 days. Retention of the CWT implanted in the dorsal musculature of A. anguilla was 99% after 32 days and did not change by day 512. It was therefore concluded that marking‐induced mortality was nil for both marking types over the 6‐month period. Generally, both methods are found to be suitable for marking young A. Anguilla. VIE tags, however, should be used for short‐time mark‐recapture experiments only, and should be injected at the base of the ventral fin margin. The use of CWTs seems to be the more suitable marking method for long‐term mark‐recapture experiments. Nevertheless, field tests are necessary to evaluate these marking methods.  相似文献   

3.
The lack of a reliable and innocuous mark-recapture method has limited studies that would provide essential information for the management of commercial sea cucumbers. Tagging sea cucumbers is notoriously difficult because of their plastic nature and autolysis capacities. The markers that have so far been tested, mainly on or through the body wall, were either lost rapidly or had major drawbacks (e.g. suitable only for batch identification, requiring complex analysis, causing infections, necrosis, behavioural changes and mortality). The present study explored the efficacy of passive integrated transponder (PIT) tags for individually marking sea cucumbers by assessing retention rates and long-term side effects of tags inserted in previously unstudied tissues/organs. Individuals of the species Cucumaria frondosa were tagged in the body wall, aquapharyngeal bulb and at the base of the oral tentacles. They were monitored closely for evidence of stress, infection, change in feeding and spawning behaviour and tag retention rate. Implanting the tag in an oral tentacle to reach the hydrovascular system of the aquapharyngeal bulb achieved the best retention rates in full-size individuals: from a maximum of 92% after 30 days to 68% at the end of the experimental period (300 days). Efficacy was lower in smaller individuals (84% after 30 d and 42% after 300 d). Following a slight increase in cloacal movements for 15 h post tagging, no side effect was noted in sea cucumbers tagged in the aquapharyngeal bulb via the tentacles. Feeding and spawning behaviours were not affected and no signs of infections or abnormal cell development in the vicinity of the tags were observed. This study indicates that marking sea cucumbers with 8.2 mm long PIT tags implanted via the oral tentacle is an effective technique, yielding relatively high retention rates over long periods without any detectable physiological or behavioural effects.  相似文献   

4.
Tagging or marking of fish is instrumental to fisheries biologists and managers seeking to distinguish groups of fish, track movement or migration patterns, and monitor population characteristics. However, tag loss can inhibit the ability of biologists and managers to reach these objectives. The ability of Lake Sturgeon to live for long periods of time and reach large sizes, in combination with their dynamic spawning activity, requires tags to be retained under a variety of environmental and physically demanding conditions. This study evaluated tag retention of loop, monel, and passive integrated transponder (PIT) tags on wild, free‐ranging Lake Sturgeon in Lake Huron and the St. Clair – Detroit River System. Lake Sturgeon in this study were double‐tagged with both a PIT tag under one of the three anterior‐most dorsal scutes and an external tag (loop or monel) at the base of the dorsal fin. Fish were at large for up to 16 years. Overall, tag loss for PIT tags was 1% followed by monel tags at 12% and loop tags at 36%. Tag loss for loop tags was higher when the initial length of Lake Sturgeon tagged was smaller. Tag loss for monel tags increased with time at large but was not related to length at initial tagging. Monel tags left behind abrasion marks when attached to smaller Lake Sturgeon. PIT tag retention was higher than reported in previous studies that tagged other sturgeon species in different body locations. Monel tag retention was higher than other external tag types evaluated in previous studies while loop tags had similar retention rates to external tag types. Most previous studies on tag retention of sturgeon species were of shorter duration and conducted in laboratory settings, therefore loop tags may have performed more favorably during studies under short term laboratory settings. Results of this study suggest that PIT tags inserted below dorsal scutes represent a viable option for long‐term tracking of Lake Sturgeon. Monel tags attached at the base of the dorsal fin also seem to be a viable option relative to other external tag types, but should be limited to larger sturgeon as they can leave behind abrasion marks.  相似文献   

5.
The present study deals with the impact of juvenile stocking sizes of the major carp on production in a minor reservoir, Bibinagar, Nalgonda, India. Reservoir surface area equaled over 23.8 ha and water from the reservoir was used for irrigation and fisheries. Four experiments were conducted for 4 years from 2000–2001 to 2003–2004. The experiments were planned in such a way that every year juvenile stocking size was held constant and subsequent fish production analyzed. During the first year’s experiment (2000–2001) a stocking size of 25–30 mm (fry) was maintained. Similarly 50–55 (advanced fry), 75–80 mm (fingerling) and 100–105 mm (advanced fingerling) were stocked during 2001–2002, 2002–2003 and 2003–2004 respectively. Uniform yearly stocking densities (2000/ha) were established in each experiment in the month of July and fish were harvested in June of the subsequent year. Major carp production was enhanced with larger stocking size. Yearly productions equaled 144.00, 231.48, 632.91 and 1005.03 kg/ha/year with the above stocking sizes. Variation in stocking sizes had significant (P<0.05) effects on fish production. The number of fish per kg production decreased gradually with increases in stocking size with an average value of 3.97. Reservoir production of catla was the most abundant species, followed by rohu, common carp, mrigal and grass carp. These results show that stocking size has a great impact on fish production and the stocking of advanced fingerlings will provide maximum carp production in minor reservoirs in India.  相似文献   

6.
Population augmentation and propagation of pallid sturgeon (Scaphirhynchus albus) have been a focus of scientists since the mid‐1990s. Various tag types have been used to gain a better understanding of population characteristics, stocking success, and movement patterns. Passive integrated transponder (PIT) tags have been consistently used since the inception of recovery efforts to mark age‐1 and older pallid sturgeon. This tag has been successfully used with other sturgeon species, and tagging protocols for pallid sturgeon were developed in response to those successes. Tag retention rates in pallid sturgeon have been reported as variable or unknown and there has been no formal study to determine PIT tag retention in pallid sturgeon, particularly with age‐1 individuals that are propagated in hatcheries to be stocked into the wild. The objective was to evaluate retention rates of PIT tags inserted into the operculum and along the base of the dorsal fin of age‐1 pallid sturgeon (total fork length [FL] range = 214–358 mm). Tagged sturgeon (n = 80) were held in a large holding tank and inspected for tag loss twice a month for a total of 189 days. Final tag retention was 83% for tags inserted into the operculum and 85% for tags inserted near the dorsal fin. Tags shed from the operculum were from smaller fish (mean FL = 265 mm) and losses occurred during the first 60 days of the experiment. Tags shed from the dorsal site were predominantly from larger individuals (mean FL = 305 mm) and losses were continual throughout the experiment. Future research should determine size‐specific tag retention rates so that hatcheries can maximize retention in either tag placement location.  相似文献   

7.
Innovative tools that benefit conservation are critical as freshwater fishes are lost at unprecedented rates. Mark-recapture methods can characterize population demographics and life-history traits of diverse fishes, but suitable techniques for tagging for individual recognition of small-bodied fishes are rare. Passive integrated transponder (PIT) tag technology may facilitate the tagging of small fishes and early life stages of larger species. However, relatively little research has evaluated the suitability of these small (8.4 × 1.4 mm) tags for many groups of small fishes. Tag loss (retention and survival) and growth of individuals implanted with PIT tags relative to control and sham treatments were compared for eight fishes of differing morphologies. Additionally, the utility of cyanoacrylate to improve tag retention was tested on a subset of taxa. Fish of each species were equally divided and randomly assigned to one of three treatment groups (handled [control], surgical incision [sham], or surgical incision and PIT tag implantation [PIT]). During the 42-d study period, mortalities and expelled tags were counted daily and growth was measured weekly. Researchers can generally expect little tag loss and uncompromised growth rates for a variety of small-bodied fishes; however, initial fish length was related to the magnitude of physiological effects for some taxa (i.e., blacknose dace Rhinichthys atratulus, johnny darter Etheostoma nigrum, juvenile white sucker Catostomus commersonii). Relatively poor survival (<80%) was observed for two benthic species: johnny darter and blacknose dace, often when incision wounds became inflamed before healing. Prevalent tag loss for johnny darter, tadpole madtom Noturus gyrinus, and a dorsal-laterally compressed cyprinid can be reduced by closing the wounds with cyanoacrylate, but with substantially increased mortality rates. This research demonstrates the broad applicability of PIT tagging technology for ecological studies of small-bodied fishes and alleviates many concerns when surgically implanting tags into several fishes.  相似文献   

8.
ABSTRACT Individual color‐marking is an essential tool for studying the behavior of free‐living birds. Hummingbirds represent a particular challenge for traditional avian color‐marking techniques because of their small size and short tarsi. Although several techniques have been successfully used, the retention time of color‐markers and their safety and ease of construction could be improved. I developed two new color‐marking techniques for marking Little Hermits (Phaethornis longuemareus): (1) a plastic back tag constructed by fusing colored beads, and (2) a leg tag attached to a metal band fitted around the tarsus. Both tag designs were visible in field conditions, and neither appeared to adversely affect behavior. Both back and leg tags had high retention rates within seasons, but back tags had a poor retention rate between years. Although these marking techniques were designed for use on one species of hummingbird, they would likely also be useful with other species of hummingbirds.  相似文献   

9.
The efficacy and sub‐lethal consequences of single and double tagging European catfish Silurus glanis with Petersen disc and passive integrated transponder (PIT) tags were examined in short (laboratory) and longer‐term (field) experiments. Tag retention in the laboratory was 100%, with normal behaviour (i.e. feeding) in all fish returning within 36 h. In the field, 65 of 120 tagged S. glanis were recaptured from five small study ponds, with 85% retaining their PIT tags, though recapture rates and tagging efficacy were highly variable amongst locations. This is consistent with literature for other fishes, suggesting that tagging efficiency is variable across species and largely context dependent (fish length, tagging location, habitat).  相似文献   

10.
Amphibians have been declining worldwide and the comprehension of the threats that they face could be improved by using mark–recapture models to estimate vital rates of natural populations. Recently, the consequences of marking amphibians have been under discussion and the effects of toe clipping on survival are debatable, although it is still the most common technique for individually identifying amphibians. The passive integrated transponder (PIT tag) is an alternative technique, but comparisons among marking techniques in free‐ranging populations are still lacking. We compared these two marking techniques using mark–recapture models to estimate apparent survival and recapture probability of a neotropical population of the blacksmith tree frog, Hypsiboas faber. We tested the effects of marking technique and number of toe pads removed while controlling for sex. Survival was similar among groups, although slightly decreased from individuals with one toe pad removed, to individuals with two and three toe pads removed, and finally to PIT‐tagged individuals. No sex differences were detected. Recapture probability slightly increased with the number of toe pads removed and was the lowest for PIT‐tagged individuals. Sex was an important predictor for recapture probability, with males being nearly five times more likely to be recaptured. Potential negative effects of both techniques may include reduced locomotion and high stress levels. We recommend the use of covariates in models to better understand the effects of marking techniques on frogs. Accounting for the effect of the technique on the results should be considered, because most techniques may reduce survival. Based on our results, but also on logistical and cost issues associated with PIT tagging, we suggest the use of toe clipping with anurans like the blacksmith tree frog.  相似文献   

11.
To test the reliability of PIT tags and VIE marks as new marking techniques for the bullhead Cottus gobio , different tagging treatments were assayed. The relatively high recapture rates suggest the applicability of both marking techniques for this small benthic fish species.  相似文献   

12.
Two new shrimp-associated gobies,Amblyeleotris yanoi sp. nov. andA. masuii sp. nov. are described on the basis of specimens from Iriomote-jima Island and Okinawa-jima Islands, Okinawa Prefecture, Japan.A. yanoi is distinguished from other members of the genus by the combination of the following characters: 14 anal fin soft rays, 19 pectoral fin rays, 97–103 longitudinal scales, a candle flame-shaped marking on the caudal fin, a very low membrane connecting the pelvic fins and absence of a ventral frenum.A. masuii differs from all other congeners by having 92–97 longitudinal scales, the length of the interpelvic connecting membrane relative to the longest pelvic fin ray (0.43–0.66), black blotches on the sides of the chin, and blue spots on the opercle and preopercle.  相似文献   

13.
In laboratory fish research, the zebrafish Danio rerio (Cyprinidae) represents the equivalent of the mouse in mammalian research. This species has become a major model for studies in developmental and behavioural genetics, neurophysiology, biomedicine, ecotoxicology, and behavioural and evolutionary ecology. To meet the need for accurate and reproducible data in both fundamental and applied sciences, it is of primary importance to be able to tag and/or recognize individual zebrafish. However, classic methods used in fish ecology and aquaculture are generally difficult to apply to such small fish. Recently, various new tagging methods have been developed. This paper presents a first review of current identification and marking methods applied to zebrafish, from external observation methods (such as skin pattern recognition, fin clipping, scale regeneration, colour and transgenic methods) to the most advanced technological developments in electronic (low- and high- radio-frequencies PIT tags, microchip) and image analysis methods (video tracking). This review aims to help researchers and zebrafish facility managers select the identification method (ID) best adapted to their needs. The main characteristics of each ID method are examined (including detection range, durability, speed and repetitiveness, ID code combination, size dependence and ethical considerations), and their pros and cons are summarized in a decision table to help select the most appropriate option for a research or management program. Finally, contextual applications of these ID methods and future developments are discussed.  相似文献   

14.
Interleukin-10 (IL-10) is a pleiotropic immune-regulatory cytokine that is expressed in various species of fish and higher vertebrates, and is activated during infection. In spite of its important role, IL-10 has not been well characterized either functionally or structurally in fish. To analyze its properties and function, we constructed a 3D model of IL-10 in the Indian major carp, the catla (Catla catla), which is a highly preferred fish species and the most commercially important one in the Indian subcontinent. The catla IL-10 model was constructed by comparative modeling using human IL-10 (2ILK) as the template, and a 5 ns molecular dynamics (MD) simulation was carried out to characterize its structural and dynamical features, which was validated by the SAVES, WHAT IF and MolProbity servers. Analysis using the VAST server revealed a comparatively low level of homology between catla and human IL-10 amino acids at the N-terminal (22.7%) compared to the C-terminal (38.29%). Six conserved domains (A–F) were predicted in catla that threaded well with human IL-10, but their putative interaction sites varied significantly. The amino acid residues in helices A and F differed in length between catla and human IL-10, which may lead to the differences in the IL-10/IL-10R complexes of these two species. The existence of two highly conserved amino acid residues (Cys5 and Cys10) in fish IL-10 but not in higher vertebrate (including human) IL-10 was analyzed in this 3D model. CastP, cons-PPISP and InterProSurf server identified several binding pockets with various probe radii, but Cys5 and Cys10 did not form any significant bonds relating to structural stabilization or protein–protein interactions.  相似文献   

15.
Recent advances in micro-electronics make the study of the migration of even small marine animals (>12 cm) over many 1000s of kilometres a serious possibility. Important assumptions in long-term studies are that rates of tag loss caused by mortality or tag shedding are low, and that the tagging procedure does not have an unacceptable negative effect on the animal. This paper reports results from a study to examine the retention of relatively large (24 × 8 mm) surgically-implanted dummy acoustic tags over a 7-month period in steelhead pre-smolts (O. mykiss), and the effects of implantation on growth and survival. Although there was some influence on growth to week 12, survival was high for animals > 13 cm FL. In the following 16-week period, growth of surgically implanted pre-smolts was the same as the control population and there was little tag loss from mortality or shedding. Currently available acoustic tags can be implanted in salmonid fish ≥12 cm FL, although combined losses from mortality and tag shedding were 33–40% for animals in the 12 and 13 cm FL size classes. By 14 cm FL, combined rates of tag loss (mortality plus tag shedding) for surgically implanted tags dropped to <15% and growth following surgery was close to that of the controls. Our results suggest that studies of ocean migration and survival over periods of many months are now feasible even for animals as small as salmon smolts. Surgically implanted salmon smolts are therefore good candidates for freshwater and coastal ocean-tracking studies on relatively long time scales (months). On such time scales, even relatively small salmon smolts may move thousands of kilometers in the ocean.  相似文献   

16.
The purpose of this study was to mark endangered sterlet (Acipenser ruthenus) with visible implant elastomer (VIE) in order to assess mortality, compatibility, retention, persistence and histological reactions resulting from this tagging technique. It was hypothesized there would be only minor effects on the fishes' health, and assumed that acute effects would be more pronounced than long‐term effects. On 11 September 2013, 20 specimens were tagged ventrally with visible implant elastomer, 20 received a subcutaneous injection with 0.9% NaCl solution, and another 20 served as untreated control. Mean total length was 28.0 ± 1.8 cm and mean body mass 64.1 ± 12.0 g. The sterlets were kept in four 4,000‐L tanks filled with 2,400‐L water. Acute effects were monitored for 95 days, where fish were held at temperatures between 2.4°C and 15.2°C, reflecting outdoor conditions. Chronic effects were examined 282 days post‐tagging through histological sections of the tagging region in five sterlets. During the first 95 days of observation, tag retention was 100%. No signs of incompatibility were detected. Body mass did not significantly differ between VIE‐tagged fish and controls. At day 282 post‐tagging, however, distinct tissue reactions were visible at the tagging sites of nine fish. Histological examination of five fish revealed a variable degree of infiltration with leukocytes in the areas around the elastomer, which did not necessarily correspond with the externally visible degree of inflammation. After medical treatment, the lesions healed without complications, whereas the retention rate of the VIE tags was 5%. According to the findings, the tag location rather than the tag itself was responsible for the externally visible irritations, indicating that the ventral subcutis of sterlet is not a suitable site, even for small VIE tags in long‐term studies. The results of this study also suggest that VIE marking should in general be critically evaluated before application in field studies.  相似文献   

17.
The effects of passive integrated transponder (PIT) tagging on cortisol release, standard metabolic rate (SMR) and daily specific growth rate (GS) were evaluated in the Gulf killifish, Fundulus grandis, a small estuarine fish native to the Gulf of Mexico. Cortisol release by individual fish was measured non-invasively prior to PIT tagging, immediately after tagging and once per week for 1 month following tagging. Within the first 2 h of tagging, cortisol release rates were significantly elevated compared with values measured prior to tagging and significantly higher than that of fish handled identically except not implanted with PIT tags. By 1 week after PIT tagging, cortisol release rates returned to control levels. SMR, determined by intermittent-flow respirometry and GS, defined as per cent change in body mass per day, were measured prior to PIT tagging and weekly for 1 month after tagging. Neither SMR nor GS was significantly different in tagged v. untagged fish for the duration of the study. One month after tagging, haematocrit, plasma cortisol, blood glucose and blood lactate did not differ between tagged and untagged individuals. Therefore, after a transient stress response that subsides within 1 week, PIT tagging had no significant effects on these physiological variables in F. grandis, validating its use as a method of marking this and other small fishes.  相似文献   

18.
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.  相似文献   

19.
Uptake and processing of biofilm (BF) and free-cell (FC) vaccines of Aeromonas hydrophila were studied in the Indian major carps catla Catla catla, and rohu Labeo rohita and in the common carp Cyprinus carpio following a single dose oral vaccination of 10(11) CFU g(-1) fish. Fish were sampled at 0.5, 1, 3, 6, 12, 24 h and at 2, 3, 5 and 10 d following vaccination and antigen localization was studied in the gut, kidney and spleen employing monoclonal antibody based immunofluorescence and immunoperoxidase. The general pattern of antigen localization was similar in catla, rohu and common carp. Initially, both the BF and FC antigens were localized in the gut lumen, followed by their uptake by intraepithelial vacuoles and macrophages. Antigen administered orally was also seen in the spleen and kidney. Both BF and FC antigens were detected in the gut lumen of carp within 30 min following oral delivery. However, BF antigen remained in the lumen of the hindgut for 48 h compared to 6 h in the case of FC antigen. In the early stages, BF antigen was localized in the gut epithelial vacuoles while FC antigen was associated with the small macrophages of the hindgut. Antigen localization in spleen and kidney was observed at 3 h and persisted even up to 10 d following oral delivery. In general, there was a distinct difference between BF and FC vaccines in the duration of retention and quantity of uptake in the gut, kidney and spleen.  相似文献   

20.
Management of red snapper stocks is one of the most controversial issues in the Gulf of Mexico fisheries. Fish tagging programs can help to address many management issues if an appropriate tag is selected. In the study three types of tags were tested to determine tag retention and effects on red snapper growth and survival. Hatchery‐produced juveniles (43.7 ± 5.6 g) were either not tagged or given: internal anchor tags anchored in the abdominal cavity; passive integrated transponders (PIT) tags injected into the body cavity; or metal self‐piercing fish tags clamped around the dentary bone. Fish were stocked at 25 m?3 in three replicate circular tanks/tag type, and fed a commercial feed for a 150 days period. Fish survival did not differ by treatment. Average weight gains after 150 days were similar. Tag retention differed by treatment (P = 0.0002), with 100% retention of PIT tags, 98.4 ± 2.8% retention of jaw tags, and 90.4 ± 1.6% retention of internal anchor tags. Loss of internal anchor tags occurred primarily within the first month. Inflammation was frequently observed at the internal anchor tag insertion site. Irritation and cell proliferation was sometimes observed in areas around the jaw tags.  相似文献   

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