Common carp tissue reactions to surgically implanted radio tags with external antennas

Examination of common carp Cyprinus carpio , carrying implanted telemetry transmitters, showed that wound healing was incomplete after 4 months and completed after 1 year. Granulation tissue, which encapsulated the transmitter, adhered to the intestine and liver. The liver was infiltrated by the granulation tissue after 1 year. Epidermal tissue grew down the channel for the antenna, establishing a potential pathway for pathogens. No signs of infections, however, were found.     

Learning from unsuccessful radio tagging of common carp in a Namibian reservoir

Radio tagged common carp Cyprinus carpio in a Namibian reservoir, where this species has been introduced, experienced 100% mortality or tag loss (  n  = 13) with surgically implanted transmitters and 100% survival (  n  = 5) with externally attached transmitters at water temperatures of 24–25° C. It seems that the negative effects of the tagging were greater for surgical implantation than for external attachment. The body shape of common carp make them well suited for external tagging, and this tagging method is recommended for common carp studies at high water temperatures or under other stressful conditions.     

Modified method for external attachment of transmitters to birds using two subcutaneous anchors

ABSTRACT Of the transmitter attachment techniques for birds, the subcutaneous anchor provides a secure attachment that yields relatively few secondary effects. However, the use of subcutaneous anchors has been limited by transmitter size and retention time. Using a modified method of attachment that utilized two subcutaneous anchors, we deployed 69 GPS transmitters, plus 13 VHF transmitters that were similar in size and weight to GPS models, on Pacific Black Brant (Branta bernicla nigricans). Prior to our study, only harnesses were used for attaching GPS transmitters on birds, mainly because GPS transmitters are too large for other external attachment techniques and implantation in the body cavity attenuates the GPS signal. Thus, to increase the size capacity of anchor attachment and to avoid the well‐documented negative effects of harnesses on behavior and survival, we added a second anchor at the transmitter's posterior end. The double‐anchor attachment technique was quickly and easily accomplished in the field, requiring bird handling times of <10 min. Incidental recoveries of tagged Brant indicate a high degree of transmitter retention. Five recaptured birds (4–6 weeks after deployment) and eight killed by hunters (3–6 mo after deployment) retained their GPS transmitters. For studies involving the use of relatively large transmitters, the double‐anchor method appears to provide a viable alternative for external attachment.     

借助斑马鱼EST数据库从鲤鱼微卫星序列中寻找蛋白编码基因(英文）

应用 in silico的方法,利用Blastn 和Blastx 搜索引擎,将鲤鱼微卫星序列与GenBank数据库进行同源序列比对。利用Blastn,将侧翼序列长度>50 bp的875个鲤鱼微卫星序列与斑马鱼的EST数据库首先进行比对,结果找到了121个同源序列。随后采用Blastx搜索蛋白质数据库,有94个微卫星位点存在同源蛋白。除了33个假定和3个未知蛋白外,剩余的58个微卫星位点被成功地进行了功能注释,而且其中的7个位点已经定位在了鲤鱼连锁图谱上。另外,通过PCR-SSCP的方法,将两个与鲤鱼微卫星侧翼序列相匹配的斑马鱼EST序列开发成鲤鱼的STS标记,并将其中的一个标记HLJZe33定位到鲤鱼连锁图谱上。以上研究结果表明,通过比较基因组研究,模式生物斑马鱼的很多遗传和基因组资源都可以被利用到鲤鱼的基因组研究中。     

借助斑马鱼EST数据库从鲤鱼微卫星序列中寻找蛋白编码基因

应用in sifico的方法,利用Blastu和Blastx搜索引擎,将鲤鱼微卫星序列与GenBank数据库进行同源序列比对.利用Blastn,将侧翼序列长度＞50bp的875个鲤鱼微卫星序列与斑马鱼的EST数据库首先进行比对,结果找到了121个同源序列.随后采用Blastx搜索蛋白质数据库,有94个微卫星位点存在同源蛋白.除了33个假定和3个未知蛋白外,剩余的58个微卫星位点被成功地进行了功能注释,而且其中的7个位点已经定位在了鲤鱼连锁图谱上.另外,通过PCR-SSCP的方法,将两个与鲤鱼微卫星侧翼序列相匹配的斑马鱼EST序列开发成鲤鱼的STS标记,并将其中的一个标记HLJZe33定位到鲤鱼连锁图谱上.以上研究结果表明,通过比较基因组研究,模式生物斑马鱼的很多遗传和基因组资源都可以被利用到鲤鱼的基因组研究中.     

A conceptual ecological model to aid restoration of Cootes Paradise Marsh,a degraded coastal wetland of Lake Ontario,Canada

An ecological model is derived from recent studies, based on 60 years of empirical observations and experimental data, that conceptualizes how Cootes Paradise Marsh was transformed from a lush emergent marsh with considerable ecological diversity in all trophic levels, to one that is currently turbid, devoid of vegetation, and dominated by a few exotic plant and fish species. This conceptual model contains 17 key components that interact and contribute to the overall unhealthy state of the marsh. The most influential component is high water level which caused the initial loss of emergent vegetation in the 1940s and 1950s. In the absence of plants to attenuate sediment and assimilate nutrients, the marsh became turbid and windswept, and this led to the disappearance of submergent vegetation over the next two decades. Currently, high water turbidity is being maintained by wind re-suspension, high sediment loading from the watershed during the summer, high algal biomass resulting from excessive nutrient loads from sewage effluent and surface runoff, and the feeding and spawning activities of a very large population of common carp ( Cyprinus carpio). Due to vegetation loss, the substrate has become mostly loose sediment that is no longer suitable for the diverse assemblage of aquatic insect larvae that lived on the plants and detrital material in the 1940s. Benthic grazers have been kept in low abundances due to predation by benthivorous carp; consequently, epiphytic algae have proliferated and further contribute to light limitation of macrophytes. High nutrient loadings contribute to high diurnal fluxes in dissolved oxygen levels that tend to select against less tolerant organisms such as insect larvae (other than chironomids) and piscivores (northern pike and largemouth bass). Without piscivores in the marsh, the planktivores have become dominant and have virtually eliminated all of the large herbivorous zooplankton (e.g., Daphnia), except for a few pockets in the marsh inlets close to residual macrophyte beds. Because of the dominance of small-bodied inefficient grazers (rotifers and small cladocerans), algal biomass is high, and the community has a large proportion of heterotrophic forms that tolerate low light environments. This ecological model suggests that the current turbid un-vegetated state of Cootes Paradise may be very stable. It will persist as long as water levels remain unfavorable for natural re-colonization by the emergent flora, and/or water turbidities remain sufficiently high to suppress the growth of submergent vegetation. Using this conceptual model, I developed a model of how Cootes Paradise Marsh may have functioned as a healthy marsh prior to the 1940s, and use these models as a basis to explore a number of restoration and management options and discuss their implications on the aquatic foodweb.     

A novel method to prepare size-regulated spheroids composed of human dermal fibroblasts

A simple method to prepare size-regulated spheroids has been successfully developed by combining a temperature responsive polymer, poly-N-isopropyl-acrylamide (PNIPAAm), conjugated with collagen and ultraviolet (UV) irradiation with photomasks. The coating layer composed of PNIPAAm conjugated with collagen functions as a cell substratum at 37 degrees C, then when lowering the temperature of culture medium the cells attached to it detach as a self-supporting sheet. This is because PNIPAAm dissolves into the culture medium below the lower critical solution temperature LCST; about 30 degrees C, but it is insoluble above the LCST. The detached cell sheet forms a multicellular spheroid. On the other hand, UV effectively immobilized collagen in the coating layer because UV generates crosslinkages in collagen molecules. Crosslinkages were quantitatively introduced by controlling the energy of UV-irradiation thus the ability of human dermal fibroblasts to attach to and detach from the surface was tightly controlled. When the collagen content in the coating layer was 9 mug/cm(2) (collagen ratio, 4.5%), UV-irradiation energy of 2000 J/m(2) was suitable to obtain 100% of the attachability and detachability. However, the cells did not attach to the nonirradiated surface at this collagen content because insufficient collagen was immobilized. Using photomakes to apply UV-irradiation, it was possible to obtain cell-adhesive areas(irradiated areas) and nonadhesive areas (nonirradiated areas) on the same surface. Consequently, spheroids of any size and in any number from one dish were prepared. The viability of cells in spheroids 350 mum in diameter was maintained at a high level for 28 days; however, viability of spheroids 800 mum in diameter rapidly decreased for 2 days. The size was very important to maintain the viability. This novel method is useful to develop size-regulated spheroids for different applications; for example, in toxicology tests. (c) 1994 John Wiley & Sons, Inc.     

A non‐invasive method to assess the impact of electronic tag insertion on stress levels in fishes

The present study investigated, for the first time, a non‐invasive technique for monitoring cortisol levels in fishes that had been implanted with electronic transmitters. This technique involved the measurement of cortisol released into the water by individual fish. Carp Cyprinus carpio and roach Rutilus rutilus that had been tagged with miniature dummy acoustic transmitters responded to the surgical implantation of the tags with an immediate (1 to 4 h) increase in cortisol concentrations in the water. These amounts of cortisol were higher than in 'handled only' and 'handled and anaesthetized' groups. Estimated cortisol release rates for both species correlated well with the direct measurements in the water. In both species, water cortisol concentration and cortisol release rates returned to pre‐stress levels within 12 h of tag‐insertion, and remained low for the remainder of the experiment. The calculation of cortisol release rates highlighted a difference of a factor of about four between peak release rates in carp and roach. There was no evident long‐term cortisol stress response to the presence of a tag in the body cavity in either species.     

A tool to facilitate implantation of electrodes for electromyographic telemetry experiments

Conventional techniques for implanting electrodes into axial swimming musculature of fish are reviewed. A new device is described that reduces time for electrode implantation, ensures constancy in electrode orientation, implantation depth, and separation distance. This device is inexpensive, simple to build, and easy to use.     

A wireless communication system for determining site fidelity of juvenile dusky groupers Epinephelus marginatus (Lowe, 1834) using coded acoustic transmitters

The value of marine reserves to protect threatened and vulnerable species, such as the dusky grouper (Epinephelus marginatus), is often uncertain because of a lack of knowledge about their ecology and habitat utilisation. To address this issue, a Wireless Communication System for location-based positioning of fish tagged with coded acoustic transmitters was installed to determine site fidelity of juvenile dusky groupers (mean total length = 30.4 cm, range: 20.5–40.0 cm; mean total weight = 470 g, range: 113–950 g) within a marine reserve (Island of Ustica, Sicily, Italy). The system was based on five wireless hydrophones distributed approximately 200 m apart along the coastline of the reserve, each monitoring acoustic signals from transmitters within a reception cell. The hydrophones retransmitted detected signals as a radio signal to a land-based receiver and data logger. Unique identifiers for each hydrophone permitted the identification of the detection cell containing the tagged fish. The wireless hydrophones had partly overlapping detection ranges, dividing the marine reserve into more than five different detection cells. The average detection range was 232 m, but coverage varied from site to site and with time, depending on many factors, such as ambient noise, wave action and signal strength of the transmitters. Despite some limitations, the Wireless Hydrophone System (WHS) combined with manual tracking provided a valuable tool for investigating site fidelity in the dusky grouper. Preliminary home ranges for the fish were estimated by the kernel method (probability level of 95%), based on a limited set of manual tracking data, to be a median 5312 m² (range 1673–18626 m²).     

A new method to model change in cutaneous blood flow due to mechanical skin irritation part II: parameter identification procedure

Mechanical skin irritation, for example a light scratch with a needle, induces histamine and neuropeptide release on the line of stroke and in the surrounding tissue. Both histamine and neuropeptides are vasodilators. They cause vasodilation by changing the contraction state of the vascular smooth muscles and hence vessel compliance. Smooth muscle contraction state is very difficult to measure in vivo. For that reason we propose in this article an identification procedure to establish an irritation law. The law gives change in vessel compliance as a function of space, time and the intensity of the stroke. We have showed that vessel compliance increases immediately after the stroke not only on the line of stroke, but also in the surrounding tissue. Then, after a short delay, vessel compliance starts decreasing in the surrounding tissue, whereas vessel compliance on the line of stroke keeps increasing. Hence, blood is transported from the surrounding tissue to the line of stroke. In this way, higher blood volume on the line of stroke can be obtained than by only changing vessel compliance locally.     

A new method to model change in cutaneous blood flow due to mechanical skin irritation part I: comparison between experimental and numerical data

Mechanical skin irritation creates vasodilation. Vasodilation of vascular networks induces increase in blood volume and blood velocity. Both can be measured by Laser Doppler Velocimetry. We propose in this article a method permitting comparison between experimental and numerical results. Experimental data was obtained by Laser Doppler Velocimetry. Numerical results were obtained by a continuous model of the vascular network. The model consists of three layers. First and last layer are described by anisotropic and heterogeneous porous media. They represent the irrigation and the drainage of the vascular system. The intermediate layer is described by a lumped parameter model that does not permit horizontal fluxes. All vessels are compliant. The permeabilities depend on the volumes of the specific layer. Skin irritation is modeled by a change in compliance of small arterial blood vessels. The comparison between experimental and numerical data is based on the model proposed by Bonner and Nossal [1981. Model for laser Doppler measurements of blood flow. Appl. Opt. 20, 2097-2107]. The model describes the Doppler frequency spectrum S(omega) as a function of the optical phenomena creating the frequency shift. The comparison is based on the model of the first moment M1 approximately integralomegaS(omega)domega. The variables of the first moment can be determined by results of the numerical model. We have shown, that it exists a linear relation between the change in compliance and the following increase in first moment. Using this linear relation experimental and numerical data can be compared.     

A rapid method to screen fungi for resistance to toxic chemicals

Although many species of fungi are able to degrade highly toxic chemicals, only a few species have been evaluated for resistance to toxic effects of these chemicals. In this paper we demonstrate the successful application of a method to rapidly screen several species of fungi for toxicity to chemicals or mixtures of chemicals using pentachlorophenol (PCP) as a model toxic compound. Cellulose antibiotic assay disks were soaked in solutions containing different concentrations of PCP (5, 10, 25, 50, and 80 mg l^–1) and then placed in a triangular pattern outside the growing edge of the mycelia of eighteen species of white rot fungi. The plates were incubated and observed for development of inhibition zones (non-growth areas) around the disks. The short-term (24 h) growth of all eighteen species of fungi was inhibited by 5–10 mg-PCP l^–1, a range similar to that observed using previously reported techniques. Long-term growth studies using this screening method were not useful since PCP diffused from the disk into the agar, decreasing the applied dose.     

A method to construct confidence interval for expected response to multi-trait selection

Summary Confidence intervals are constructed for the expected responses to three types of multi-trait selection. The influence of numbers of replicates and genotypes used in a progeny test experiment on the precision of response of multi-trait selection is discussed based on the structure of the established intervals. Special attention is paid to the characteristics of the intervals constructed for the conventional least square selection indices.     

A method for determining binding kinetics applied to thiamine-binding protein

A rapid and simple method for assaying the binding activity of thiamine-binding protein is described. By this assay method, the binding characteristics of rice bran thiamine-binding protein have been evaluated with [14C]thiamine as ligand. Analysis of these data by Scatchard plot resulted in linear plots giving a dissociation constant (Kd) for thiamine of 0.55 microM and a maximum binding (Bmax) of 14.5 pmol of ligand bound/microgram of protein. Thiamine binding to the binding protein was time dependent and reached equilibrium at approximately 20 min. The Kob was 0.18 min-1 and the k1 was 1.25 X 10(5) min-1 M-1. Reversibility of thiamine binding at equilibrium was completed at 60 min with a k2 value of 0.052 min-1. The Kd calculated from the reverse rate constant was 0.42 microM. These results indicated that this binding assay method was substantially reliable and accurate.     

A computational fluid dynamics (CFD) based method for assessing the hydrodynamic impact of animal borne data loggers on host marine mammals

Animal‐borne data loggers (ABDLs) or “tags” are regularly used to elucidate animal ecology and physiology, but current literature highlights the need to assess associated deleterious impacts including increased resistive force to motion. Previous studies have used computational fluid dynamics (CFD) to estimate this impact, but many suffer limitations (e.g., inaccurate turbulence modeling, neglecting boundary layer transition, neglecting added mass effects, and analyzing the ABDL in isolation from the animal). A novel CFD‐based method is presented in which a “tag impact envelope” is defined utilizing simulations with and without transition modeling to define upper and lower drag limits, respectively, and added mass coefficients are found via simulations with sinusoidally varying inlet velocity, with modified Navier‐Stokes conservation of momentum equations enforcing a shift to the animal's noninertial reference frame. The method generates coefficients for calculating total resistive force for any velocity and acceleration combination, and is validated against theory for a prolate spheroid. An example case shows ABDL drag impact on a harp seal of 11.21%–16.24%, with negligible influence on added mass. By considering the effects of added mass and boundary layer transition, the approach presented is an enhancement to the CFD‐based ABDL impact assessment methods previously applied by researchers.