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Interspecific hybridization is a route for transgenes from genetically modified (GM) animals to invade wild populations, yet the ecological effects and potential risks that may emerge from such hybridization are unknown. Through experimental crosses, we demonstrate transmission of a growth hormone transgene via hybridization between a candidate for commercial aquaculture production, GM Atlantic salmon (Salmo salar) and closely related wild brown trout (Salmo trutta). Transgenic hybrids were viable and grew more rapidly than transgenic salmon and other non-transgenic crosses in hatchery-like conditions. In stream mesocosms designed to more closely emulate natural conditions, transgenic hybrids appeared to express competitive dominance and suppressed the growth of transgenic and non-transgenic (wild-type) salmon by 82 and 54 per cent, respectively. To the best of our knowledge, this is the first demonstration of environmental impacts of hybridization between a GM animal and a closely related species. These results provide empirical evidence of the first steps towards introgression of foreign transgenes into the genomes of new species and contribute to the growing evidence that transgenic animals have complex and context-specific interactions with wild populations. We suggest that interspecific hybridization be explicitly considered when assessing the environmental consequences should transgenic animals escape to nature.
1. Introduction
The production of genetically modified (GM) plants and animals has prompted considerable debate surrounding the environmental and ecological consequences should they escape [1,2]. The majority of research has assessed the performance of such organisms and their potential interactions with conspecifics and other species under experimental conditions [3–6]. In both plants and animals, the potential ecological risks are modulated by the reproductive capabilities of the transgenic organism, the fitness effects of the transgene and the potential for interbreeding with wild conspecifics. Parallel with these risks is that of introgression into gene pools of closely related species, especially between species that hybridize naturally [7]. Natural hybridization rates of wild, non-transgenic organisms are facilitated by reductions in reproductive barriers associated with mating behaviour; this has been observed in the translocations and invasions of species [8,9], including following release or escape of strains that have been exposed to domestication selection [10–12]. Successful artificial transgenic hybridization between two species of loach (genus Misgurnus) has been reported, yet these species are not known to hybridize naturally [13]. In contrast to plants [14], the potential ecological consequences of interspecific hybridization involving transgenic animals are unknown.
Atlantic salmon (Salmo salar), a candidate for growth-enhancing transgenic biotechnology for commercial production, hybridizes naturally with closely related brown trout (Salmo trutta). Natural levels of hybridization rarely exceed 1 per cent [15], yet translocations, as well as escapes and introductions of domesticated salmon [16–18], can increase rates to as much as 41 per cent [17]. Hybridization between Atlantic salmon and brown trout is asymmetric, where the maternal origin of the hybrids varies among regions. The maternal species is predominately salmon within the natural geographical range of sympatry, while it is trout in areas where the species have come into secondary contact ([19] and references therein). Despite such regional differences, mortality prior to exogenous feeding is higher among hybrid offspring of trout mothers relative to hybrids with salmon mothers [19,20]. Given these biological patterns, we hypothesized that the transgene may further affect the outcome of hybridization, especially in terms of hybrid viability and hybrid growth rates.
Here we test the potential for transgene exchange through interspecific hybridization between transgenic Atlantic salmon and brown trout and whether this would generate novel ecological interactions. To do so, we created a series of experimental crosses and reared the offspring in both hatchery-like conditions and contained stream mesocosms designed to more closely emulate nature. These crosses and subsequent rearing experiments were designed to test: (i) whether the transgene can be transmitted into hybrid offspring via artificial mating; (ii) the survival of hybrid offspring; (iii) the phenotypic expression of the transgene in hybrids, particularly in relation to that observed in transgenic salmon; and (iv) the effect of direction of hybridization (i.e. whether the mother was a salmon or a trout) on phenotypic expression of the transgene. We used salmon hemizygous for the transgene, which enabled us to compare full sibling transgenic and non-transgenic hybrids (i.e. largely control for parental and other non-random genetic effects). Subsequently, to investigate potential ecological consequences of such hybridization, we undertook a replicated experiment to (v) quantify the effects of the presence of transgenic hybrids (from both maternal origins) on the growth of non-transgenic (henceforth referred to as wild-type) and transgenic salmon in stream mesocosms under food-limited conditions.
2. Material and methods
(a) Experimental fishes
Experimental crosses (see the electronic supplementary material, table S1) were produced using milt and ova from wild brown trout, wild Atlantic salmon and growth hormone (GH) transgenic Atlantic salmon. Wild salmon were captured from the Exploits River, Newfoundland, Canada, and trout from the Rennie's River, Newfoundland. Gametes from GH transgenic salmon [21] were provided by AquaBounty Farms (PEI, Canada). The gene construct used was a chimeric GH construct (opAFP-GHc2; EO-1α line) that has been shown to greatly enhance growth [21], reduce reproductive performance among precocious mature males [22] and increase risk-taking foraging behaviours [23,24] in transgenic individuals. However, such phenotypic changes are known to be heavily influenced by gene–environment interactions [25,26]. The wild populations used in our experimental crosses were numerically large with no indications of inbreeding depression or low heterozygosity, and unaffected by the potentially confounding influences of domestication selection for growth [27]. The transgenic salmon were hemizygous, with a dominant transgene on a single chromosome that is inherited in classic Mendelian proportions [28]. Thus, families with a transgenic parent are expected to consist of ca. 50 per cent transgenic and 50 per cent wild-type offspring.
Logistical constraints on the availability and quality of transgenic sperm and ova limited the number of crosses created. Initial plans to conduct a full-factorial half-sibling breeding design were not practical because of trauma to many gametes during shipment. Instead, seven experimental groups of fishes were available for comparisons: (i) six families of brown trout; (ii) seven families of Atlantic salmon (including wild-type and transgenic individuals); (iii) four families of hybrids from brown trout mothers (including wild-type and transgenic individuals, henceforth referred to as BT hybrids); and (iv) two families of hybrids from Atlantic salmon mothers (including wild-type and transgenic individuals, henceforth referred to as AS hybrids). To fully utilize the available gametes, four half-sibling families (two of the AS hybrid families were half-siblings with two separate salmon families) were created where gametes from two transgenic females were split to create two crosses each. Similarly, one brown trout male was also used in two separate crosses, bringing half-sibling families to six. (See the electronic supplementary material, table S1 for more information.)
(b) Transmission and phenotypic expression in hatchery-like conditions
To test for GH transgene transmission into salmon-trout hybrid offspring and to compare survival and phenotypic expression among experimental groups, 60 haphazardly selected individuals from each family were reared from the start of exogenous feeding (late March 2009) for ca. 100 days. Each family was reared in separate 25 l circular tanks, one tank per family. Fishes were fed a commercial dry feed (Corey Feed Mills, Fredericton, NB, Canada) continuously during daylight hours with automatic feeders (Lifegard Aquatics, Cerritos, CA, USA). Feeding was at satiation levels, with the weight of feed equivalent to ca. 4–6% of total biomass within a tank, which was adjusted periodically (ca. monthly) to account for changes in biomass resulting from growth and mortality. Photoperiod (controlled by automatic light timers) and water temperatures represented ambient conditions throughout the experiment. Mortalities (n = 216) were removed daily and every second mortality was screened for the presence of the transgene (see below).
At the start of the experiment, we quantified body mass (0.01 g) and length (0.01 mm) from 30 random individuals per family (except in one BT hybrid and Atlantic salmon family where n = 16 and n = 17, respectively) and genotyped 363 individuals that had a transgenic parent. After 100 days of rearing, we re-sampled individuals for length and weight and genotyped ca. 50 per cent of the surviving individuals (n = 360) following completion of the stream mesocosm experiment.
(c) Stream mesocosm environment
To test the competitive effects of hybrids on Atlantic salmon, we compared the growth rates of juvenile salmon in the presence (sympatry) or absence of hybrids (allopatry) in stream mesocosms (2.61 × 0.25 m; see Moreau et al. [29] for detailed description) over a period of ca. 30 days starting in mid-July. These mesocosms, contained within the laboratory, were designed to more closely approximate natural conditions. In contrast to inanimate dry feed, fishes were fed 20.0 g of live Artemia nauplii per day (dry cyst weight, roughly 300 000 hatched individuals, shared among the 12 mesocosms) from drip feeders, spread over two feedings. Previous experiments using these mesocosms with the same food delivery system and where food levels were quantified suggest that food levels in the present study were probably representative of a food-limited environment of a natural stream [29]. Substrate of variable sizes and colours were added and unidirectional current forced individuals to hold position in the current as they would do in nature. Moreover, we observed characteristic territorial behaviour common in wild individuals [30], whereas density in the hatchery tanks precluded territoriality. Thus, we consider the mesocosms to reflect 'semi-natural' conditions compared to the hatchery-like environment.
The mesocosm experiment started immediately following the 100 days in hatchery-like conditions. The sympatric treatments consisted of salmon (n = 12) with either BT hybrids or AS hybrids (n = 12). Both sympatric treatments and the allopatric treatment (n = 24 salmon) were replicated four times. Fishes were selected haphazardly for inclusion in the mesocosm experiment, anaesthetized and measured for initial fork length (mm; via digital photography using a Nikon D300 SLR with a 60 mm macro lens and ImageJ software) and initial weight (0.001 g). They were then individually marked with a passive integrated transponder tag (8 mm Biomark, Boise, ID, USA) and assigned to the experimental mesocosms after a 24 h period of recovery. We controlled for possible family effects by distributing families evenly among mesocosms. Within families, individuals were assigned to experimental streams without knowledge of their genotype, and screening for the presence of the transgene was performed following the experiment. Mortalities were recorded (2.1% of salmon, spread among streams), removed and replaced with a live individual from the same family to maintain densities. Growth rates of replacement fishes were excluded from analyses.
All fish sampling was performed under mild anaesthesia with MS-222 (Western Chemical, Inc., Ferndale, WA, USA). At the termination of the experiment, all fishes were euthanized with an overdose of MS-222.
(d) Genetic screening
We determined individual genotypes as transgenic or wild-type using the polymerase chain reaction (PCR) protocol modified from Deitch et al. [31]. Briefly, DNA was extracted from air-dried fin tissue (stored in 95 per cent ethanol at −20°C) with 50–200 µl of 10 mM NaOH. Samples were placed in a boiling water bath for 10 min, placed on ice for 2 min and then centrifuged at 10 000 rpm for 2 min at 4°C. PCRs were performed using recombinant Taq DNA polymerase (Fermentas International, Inc., Burlington, Ontario, Canada) and primers 2653F (5′-GCTCTTCAACATCGCGGTCA-3′) and 2654R (5′-ATATGGAGCAGCTTCAGGAC). These reactions (50 μl) contained 2 µl DNA, Taq DNA polymerase (2.5 U), the manufacturer's Taq Buffer with KCl (1× final concentration), 1.5 mM MgCl2, 0.2 mM dNTPs and 0.2 μM each of forward and of reverse primer. Thermocycling conditions consisted of 35 iterations of 94°C for 1 min, 60°C for 1 min and 72°C for 1 min. PCRs (25 μl) were analysed using 2 per cent agarose gel electrophoresis with a visual comparison of bands using a DNA size marker. The presence of a 207 bp band was taken as diagnostic of the transgenic genotype [31].
(e) Data analyses
(i) Transmission and phenotypic expression
We used counts of mortalities as the response variable and experimental cross as a fixed categorical factor in a generalized linear model (GLM) with Poisson error structure to test for differences in mortality among crosses. Significance was assessed with an F-test to account for overdispersion. To test whether mortality occurred randomly with respect to genotype, we compared the probability of a dead individual being either transgenic or wild-type against a binomial distribution (0.5 probability of success/failure) using binomial tests.
We used a general linear mixed model (GLMM) to test for differences in growth among experimental cross and genotype. Models were formulated with experimental cross as a fixed categorical factor and genotype (transgenic or wild-type) nested in a random family factor. As individuals were too small to efficiently mark at the start of the experiment, we based analyses on family averages. Tukey post-hoc tests on experimental cross and genotype were conducted using the 'multcomp' package in R [32].
Growth rates were standardized (Ω) to account for differences in initial size among groups (see the electronic supplementary material, table S2) following Ostrovsky [33]: where M2 and M1 are final and initial size, respectively, time is the growth interval and τ is the species-specific allometric coefficient for the relationship between growth rate and size. We set τ = 0.310 and 0.308 for salmon and trout, respectively, based on experimentally determined values from the literature ([34], reviewed by [30]). Failing to account for size differences altered interpretation of growth among groups (see the electronic supplementary material, table S3).
(ii) Stream mesocosm environment
Similar to above, we used a GLMM to test for differences in juvenile salmon growth among treatments. Treatment (three levels: allopatric treatment [salmon only control], and two types of sympatric treatments [BT hybrids and AS hybrids]) and genotype (two levels: transgenic or wild-type) were included as fixed factors, with mesocosm as a random effect nested within treatment to account for potential tank effects and non-independence of growth rates. Interaction terms between treatment and genotype were tested and removed when non-significant; other factors were interpreted individually. Tukey's HSD tests were used to interpret growth differences among treatments.
Analysis of variance (ANOVA) was used to investigate if juvenile growth rate in the mesocosms differed between transgenic and non-transgenic salmon and hybrids. Specifically, we assessed standardized growth as a function of genotype, cross (salmon or hybrid) and their interaction set as fixed factors. All analyses were performed in R [35].
3. Results
(a) Transmission, viability and growth enhancement
PCR analysis confirmed successful transmission of the transgene into hybrid offspring of Atlantic salmon and brown trout, and was reflected in body size differences among groups after ca. 100 days of feeding (figure 1). Of the 363 individuals screened at the start of the experiment, 43 per cent of individuals tested positive for the transgene. Among the families of hybrids with salmon mothers (AS hybrids; n = 60 screened) and hybrids with trout mothers (BT hybrids; n = 106 screened), 37 and 42 per cent of individuals, respectively, were transgenic. Only the AS hybrid group differed significantly from expectations of 50 per cent transgene transmission (, p = 0.03).
In laboratory conditions, rates of mortality differed significantly among experimental groups. BT hybrids died at higher rates (45%) than salmon, trout and AS hybrids (14, 11 and 6%, respectively; electronic supplementary material, figure S1; Poisson GLMM, p < 0.001), while mortality rates of the latter three did not differ from each other (p > 0.1). The direction of hybridization appeared to influence transgenic viability; 57 per cent of AS hybrid mortalities (n = 7; binomial test, p = 1) were transgenic, whereas only 27 per cent of BT hybrid mortalities were transgenic (n = 55; binomial test, p < 0.001). Lower mortality of transgenic compared to wild-type siblings was detected in every family of BT hybrids (i.e. 69, 82, 77 and 68% of mortalities were wild-type individuals in four families). In summary, the patterns of mortality within BT hybrid crosses indicate that transgenic BT hybrids died at statistically lower rates than their wild-type siblings, although the mortality rate of transgenic BT hybrids (24%) was still greater than that experienced by transgenic AS hybrids (7%).
Growth rates differed among experimental crosses and genotypes after ca. 100 days of rearing (figure 2). For brevity, we report growth in body mass because analyses based on length metrics yielded similar interpretations (see the electronic supplementary material, tables S2 and S3). One family of BT hybrids had insufficiently few surviving individuals (n = 12) and were excluded from further analyses. GLMMs revealed significant differences in growth among crosses (p < 0.001) and between transgenics and wild-types (p < 0.001). Among wild-type individuals, growth of trout (1.7%) and AS hybrids (1.6%) were significantly faster than that of BT hybrids (1.4%) and salmon (1.5%; p < 0.01). The transgene resulted in faster growth relative to wild-type counterparts in all cases (figure 2 and see the electronic supplementary material, table S3). Transgenic AS (2.1%) and BT (2.1%) hybrids grew significantly faster than transgenic salmon (1.9%, p < 0.01). Differences in growth were not consistent with density-dependent effects, as no relationship between mortality rates in tanks and growth rates was detected (ordinary least squares, p = 0.87, r2 = 0).
(b) Stream mesocosm environment
Hybrids suppressed the growth (% change in mass day−1) of both transgenic and wild-type juvenile salmon in stream mesocosms (figure 3; electronic supplementary material, table S4). Results for growth based on changes in length (mm day–1) and condition factor (103 × weight length−3) were congruent (see the electronic supplementary material, table S5). An experimental error in identifying one family resulted in all replicates of the AS hybrid sympatric treatment containing eight hybrids and 16 salmon, rather than the intended 12 of each. Despite this, growth of both transgenic and wild-type juvenile salmon was significantly lower in sympatric treatments compared to allopatric treatments (GLMM, p < 0.001; figure 3). In sympatric treatments, wild-type salmon growth rate in mass was reduced by 54 per cent whereas transgenic juvenile salmon growth was reduced by 82 per cent compared to allopatric treatments. Similar growth rates between the sympatric treatments indicated that hybrid maternal origin did not affect the growth of juvenile salmon, despite a difference in the proportion of hybrids present between treatments (GLMM, p = 0.87; figure 3). Growth rates of juvenile transgenic and wild-type salmon differed in both the allopatric and sympatric treatments (figure 3). Transgenic salmon had lower growth rates than wild-type salmon in all treatments (GLMM, p < 0.001).
Transgenic AS and BT hybrids grew 86 and 87 per cent faster than transgenic salmon in the same treatments, while wild-type salmon and wild-type hybrids grew at comparable rates (figure 4a,b). Specifically, we observed a marginally non-significant genotype×cross interaction in the AS hybrid sympatric treatment (ANOVA, p = 0.051; figure 4a) and a significant interaction in the BT hybrid sympatric treatment (ANOVA, p = 0.023; figure 4b).
4. Discussion
To the best of our knowledge, this is the first study demonstrating transmission and ecological consequences from interspecific hybridization between a GM animal and a naturally hybridizing species. These data confirm successful transmission of a GH transgene into interspecific hybrid offspring, marking the successful first steps towards potential introgression of the transgene into the gene pool of another species. Transgenic F1 hybrids expressed the enhanced growth phenotype; however, the direction of hybridization affected transgenic hybrid viability. The effect of the transgene on juvenile growth was environmentally dependent, with transgenic individuals growing faster than their wild-type counterparts in hatchery-like conditions, but slower in stream mesocosms. Juvenile transgenic hybrids suppressed the growth of both transgenic and wild-type juvenile salmon in stream mesocosms. Taken together, these findings suggest that complex competitive interactions associated with transgenesis and hybridization could have substantial ecological consequences for wild Atlantic salmon should they ever come into contact in nature.
Considerable differences in offspring survival were observed between genotypes (transgenic or wild-type) and among species groups (Atlantic salmon, brown trout, AS hybrids or BT hybrids) during early rearing in hatchery conditions. BT hybrids, whether transgenic or not, had higher mortality than all other groups, as has been observed previously [19,36]. Surprisingly, the transgenic BT hybrids had consistently lower mortality than their wild-type siblings, a pattern not seen in the AS hybrids. The underlying mechanism for this apparent interaction between transgenesis and hybrid maternal origin remains unclear. One possible explanation would be that transgenic BT hybrids died at higher rates during the egg and larval stages (prior to the start of the experiment); however, this seems unlikely as the number of transgenic BT hybrids at the start of the experiment was not significantly different than 50 per cent expectations. By contrast, AS hybrids differed significantly from 50 per cent expectations at the start of the experiment, although transgenic and non-transgenic siblings died at equal probabilities.
Results indicated that transgensis could combine with hybridization to result in rapid growth, but heterosis, or hybrid vigour, was not apparent in juveniles. Transgenic AS hybrids and transgenic BT hybrids expressed the enhanced growth phenotype and grew faster than other groups in hatchery conditions, including transgenic salmon. However, the brown trout groups grew faster than wild-type hybrid groups, a pattern that is inconsistent with heterosis at this life stage. Wild-type BT hybrids and wild-type salmon juvenile growth rates were not significantly different, but transgenic BT hybrids grew significantly faster than transgenic salmon. Taken together, these data suggest that growth may be influenced by complex interactions between transgenesis and hybridization.
To the best of our knowledge, this is the first study reporting a competitive advantage of transgenic interspecific hybrids over a parent species. Although hybridization with non-native species is often an important conservation concern [37,38], studies that quantify the ecological effects of non-transgenic hybrids on sympatric parent species are rare (but see [39,40]). Juvenile brown trout have demonstrated competitive dominance over juvenile Atlantic salmon, and can reduce salmon growth rates in stream mesocosms ([41] and references therein). If hybrids are of intermediate dominance, this may have contributed to decreased salmon growth rates.
Although both hybrid genotypes were present in the mesocosms, we suggest that the observed decrease in salmon growth was predominately owing to transgenic rather than wild-type hybrids. Owing to chance and differential survival during the first 100 days of rearing, transgenic hybrids were on average more common in mesocosms than wild-type hybrids (on average, 64% of the hybrids in sympatric treatments were transgenic). We consider a greater influence of transgenic hybrids to be likely as transgenic salmon have higher foraging motivation than non-transgenics [23,24], which can affect access to food, growth and survival for non-transgenic salmon [5,42]. Increased foraging motivation is also probably manifested in transgenic hybrids, compounding any competitive advantage hybrids may have over salmon. Correspondingly, we observed higher growth of transgenic hybrids over transgenic salmon in the same treatments.
In contrast to the hatchery-like conditions, the stream mesocosms were food limited, which may explain some of the differences observed between salmon genotypes in response to hybrids. Transgenic salmon grew less and incurred a greater reduction in mass growth (82%) than wild-type salmon (54%) in the presence of hybrids. This is consistent with previous studies on GH transgenic coho salmon [5,24] and GH-implanted Atlantic salmon [43], where these fish perform poorly relative to their wild-type counterparts in natural or semi-natural conditions (but see [29]). Although growth in general was low, it was within the range observed elsewhere [30]. In this study, the decreased growth of transgenic relative to wild-type juveniles was probably owing to their higher metabolic demands [44] in combination with limited food availability and warm water temperatures (mean 16.9 ± 2.5°C during the experiment). Regardless of the mechanism, our observations of reduced growth of transgenic salmon in stream mesocosms but increased growth in hatchery-like conditions contribute to evidence suggesting a complex, dynamic interplay between transgenic growth potential and the environment [25,26].
Several caveats are worthy of discussion. First, the number of hybrid families for the two cross directions used in analyses was small, owing to logistical constraints on the availability of transgenic gametes. Family and cross effects were difficult to completely disentangle in the hatchery-like environment; we observed less variation in growth within families of brown trout and BT hybrids than among crosses, but greater variation within families of AS hybrids and Atlantic salmon. However, our results from the mesocosms are independent of potential parental influence as families were equally distributed among treatments. Thus, we interpret the differences in growth among crosses and the ecological consequences of sympatric rearing of transgenic hybrids on salmon to not be primarily the result of family effects. Second, only the juvenile stage was examined and it is possible that competitive relationships or the impact of the transgene in the reproductive life stage in hybrids would have yielded different interpretations of their performance and viability. Even so, the juvenile life stage is known to be a critical period influencing the dynamics of salmon populations [30]. Third, we focused on the ecological interactions of transgenic hybrids on only one parent species, Atlantic salmon. A lack of hybrids precluded mesocosm experiments with the trout parental species; however, exhaustive risk assessments concerning transgenic interspecific hybridization ought to explore interactions with both parent species and other species as well [2].
(a) Implications and conclusions
We report that a foreign transgene that conveys remarkable growth advantage was successfully transmitted from GM Atlantic salmon and expressed in F1 interspecific salmon × brown trout hybrids. It remains unclear, however, whether the transgene could have successfully invaded the brown trout genome through viable backcrosses to the trout parental species. Despite an observed successful first step, several lines of evidence from the literature combine to suggest that introgression of the transgene into the brown trout genome via backcrossing is unlikely. First, the reproductive abilities of transgenic salmonids are reduced compared to non-transgenic fishes [22,45,46]. Second, low natural rates of hybridization [15] and poor hybrid survival [19,36] further reduce the likelihood of transgene introgression into the brown trout genome. On the other hand, poor reproductive capacity of captive salmonids improves following wild exposure [45,47–49], and the reproductive behaviour of transgenic salmonids in truly wild environments is unknown. Additionally, increased rates of salmon–trout hybridization have occurred in areas with high rates of escaped farmed or stocked heterospecific individuals ([12,15–18], but see [50]) and this has contributed to the introgression of trout genes into salmon populations and vice versa [12]. Third, only experimental backcrosses of hybrids to the salmon parental species have produced viable offspring [51,52], although evidence from wild systems suggests that hybrid backcrosses to trout can be viable [12]. Fourth, transgene introgression into the trout genome requires that a functionally intact transgene be included in the seemingly limited genetic material transferred into backcrossed offspring [12].
Despite the apparent low probability for genetic introgression into the brown trout genome, the ecological consequences of decreased salmon growth in the presence of transgenic hybrids indicate that hybridization is relevant to risk assessments. Although transgenic hybrids would probably be rarer in the wild than in our experiment, our results indicate that transgenic hybrids have a competitive advantage over salmon in at least some semi-natural conditions. Still, it is entirely unclear whether this would be observed in truly wild environments. If this advantage is maintained in the wild, transgenic hybrids could detrimentally affect wild salmon populations. Ultimately, we suggest that hybridization of transgenic fishes with closely related species represents potential ecological risks for wild populations and a possible route for introgression of a transgene, however low the likelihood, into a new species in nature.
Fish care followed the Canadian Council on Animal Care guidelines and was approved by Memorial University's Institutional Animal Care Committee (AUP 09-03-IF).
We thank C. Conway, A. Fitzpatrick, and D. Ings for assistance in data collection and fish rearing. We thank the employees of AquaBounty Technologies for kindly providing the transgenic salmon gametes for our crosses. J. Hall of the CREAIT Network of Memorial University of Newfoundland ran the PCR screening. This work was funded by NSERC Discovery grant to I.A.F., K.B.O. was supported by two NSERC USRAs and funding from Memorial University's MUCEP program. Support to P.A.H.W. was provided by the Institute of Biodiversity & Ecosystem Sustainability and the Atlantic Salmon Federation.
- Received April 24, 2013.
- Accepted May 7, 2013.
- © 2013 The Author(s) Published by the Royal Society. All rights reserved.


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By Frank Tobe, Editor and Publisher, The Robot Report
Business Insider wrote recently of a list of seven stock picks discovered in a Credit Suisse report on long term trends. One of those trends was increasing global automation (particularly in China) and the seven picks relate to companies poised to benefit from those needs.
The article and list prompted a series of quick personal opinions concerning almost all of their selections which I thought might be interesting to followers of the robotics industry.
Below are the seven Credit Suisse stock picks with a brief description of what the companies do, my best understanding of Credit Suisse's reason for selecting the stock for the list, and my opinions as to the particular company.
1. Siemens AG (NYSE:SI)
CS Reason: low PE
|Source: Siemens Annual Report
FT Comment: Because Siemens is into so many different activities, they are a good conglomerate but not necessarily a good bet for automation and robotics. Their stock moves to a different drummer then a company that is devoted only to robotics such as, for example, FANUC. It may be a good conservative bet for a long-term hold... if you want to invest in conglomerates. Other conglomerates with some involvement in robotics include Textron (NYSE:TST) and Teledyne (NYSE:TDY) for example.2. Rockwell Automation (NYSE:ROK)
CS Reason: possible acquisition target
|Source: Rockwell Automation Annual Report
FT Comment: Up until recently the computing power necessary to compute the complex math of robotic movement and control the multiple motors that drive a robot were offloaded into separate boxes for that purpose. Rockwell has been a leader in this field for a long time. But with the cost and size of CPUs dropping and the need for mobility in all the newer-tech robots, Rockwell may face an uncertain future and a diminishing share of the robotics ancillary products and control systems marketplace.3. FANUC Corp. (TYO:6954)
CS Reason: Has 20% of global robot market
FT Comment: FANUC is predominantly a robotics company and hasn't suffered too much from the tsunami disaster. They have been ramping up because there is great demand for their robots and systems in China as well as the rest of the world. Unless there emerges a Chinese competitor in the near term -- such as Foxconn might become -- FANUC has the potential for success over the next many years. Other robot manufacturers similar to FANUC include ABB (NYSE:ABB), KUKA (ETR:KU2), Yaskawa Electric (TYO:6506) and Adept (NASDAQ:ADEP).4. Delta Electronics (TPE:2308)
CS Reason: Competitive in Chinese market with high operating profits
FT Comment: Taiwan-based Delta Electronics' motors are a staple in China and barring anything unforeseen, they are a good bet to continue that trend. However, industrial automation products are a small portion of their gross revenue, are considered a commodity product, and consequently may not be an important component of the value of the company. It may be an interesting company for other purposes, but not automation.5. Keyence (TYO:6861)
CS Reason: Will benefit from aging societies investment in automation
|Source: Keyence Annual Report
FT Comment: Microsoft and PrimeSense blindsided many of the laser-scanner companies with their new-tech low-cost scanners and eloquent software. The marketplace now expects that low cost but, at present, cannot meet its needs for scanning with Kinect-like devices. [This is changing however with better and better software.] That, combined with a slow recovery from the twin economic and tsunami crises have hurt Keyence, a Japan-based company. Even though they are cash rich and profitable at present, they haven't fully recovered from the economic crisis and their annual reports don't seem to indicate research efforts that will enable lower cost technologies in the future. Currently there is sharp pressure on pricing and Keyence, Optech, V-Gen, Velodyne and other LIDAR and laser-scanning manufacturers are beginning to feel it. A few of these companies are embracing Kinect-like devices to supplement their product lines but I see the automation marketplace finding newer technologies at much lower cost from newly emerging companies to fill this very necessary vision/mobility task. Notable alternative vendors providing vision systems for robotics and automation include Cognex (NASDAQ:CGNX) and Raytheon (NYSE:RTN). The biggest and most known provider of laser scanners is SICK AG, a privately held company.6. Mitsubishi Electric (TYO:6503)
CS Reason: Market leader in computers that control automated production machines
|Source: Mitsubishi Electric Annual Report
FT Comment: Mitsubishi has been a market leader in industrial robotics and the computers that control them. And they have a big and well-respected sales presence in China. They do not have much of a presence in the emerging service robotics sector however, and this suggests that they will do well as an industrial robotics provider but may not capture market share in either the emerging SME or service robotics markets.7. TECO (TPE:1504)
CS Reason: Expanding 10% market share of motors and servos
|Teco AC motor drives
FT Comment: Robots and factory automation systems require all sorts of equipment and systems to do their tasks. Electrical motors and their control systems are integral to that process. But there are many providers and TECO is just one -- Yaskawa Electric and ABB are names more familiar to the robotics world that provide similar servos and electrical equipment and systems. To some extent, this is a commodity business with ever-lower prices and global competition; not a particularly appealing market for the long term.
* * *
What I See:
|Frank Tobe
The Robot Report
Industrial robotics as we know it -- caged arms welding, painting and manipulating long streams of mass-produced automotive and electrical goods -- will continue to grow at a healthy pace as more of the world stays competitive by automating their factories. The major players in this field are robot manufacturers KUKA, ABB, Yaskawa Motoman, Mitsubishi, FANUC and providers of ancillary systems and equipment like Rockwell Automation, Delta Electronics, Keyence, Cognex and others. Many research firms have projected steady growth for this sector at 3-5% CAGR for the rest of this decade.
Other types of factory automation may not be as visible as a robotic arm; instead it might be a process control or material handling system that is robotic in nature. This robotic-like form of automation and industrial robotics are both moving toward produce-on-demand systems which require great mechanical flexibility and complex software and systems. This is a moving target which progresses as technology does -- literally changing day by day -- and utilizes advanced sensors, fast computer processing, massive data manipulation and complex algorithms.
It's not just visible robots but whole robotic-like automation systems that are enabling companies to compete with low-cost labor from off shore. This is true even in countries like China which is beginning to upgrade their product quality and worker efficiency while coping with higher wages -- and they are doing this through automating as much as they can.
Far more companies are beginning to automate to stay competitive than just big factories. Small and medium-sized enterprises (SMEs) are propelling the service robotics segment of the robotics industry forward to new capabilities and expectations. SMEs need robots that are plug and play, easily trainable, have vision capabilities, are super flexible and safe to work alongside -- just like their iPads and apps from the App Store. Thus manufacturers of stand-alone control systems and companies that provide systems integration may not be as needed in the SME marketplace as they were (and for the time being are) for big factory robotics. New co-robot manufacturers like Rethink Robotics, Redwood Robotics and Universal Robots (Denmark), and software developers like Universal Robotics (US) and their Neocortex control system for 3D sensing of random objects and then integrating that data into user systems (e.g., into Yaskawa's Motoman 2-armed robots) -- all privately held -- may become disruptive and take away market share from the bigger companies.
Up until recently, neither vision-enabled systems nor mobility in manufacturing has been economically feasible, but that is changing. Mobile robots today move things from here to there and can be summoned and instructed by iPads. Buyers expect that mobile robotic devices be self-contained and encapsulate their various systems within the device; again, no need for stand-alone control systems or systems integration except, perhaps, in the Cloud. And vision-enabled robotic systems are proliferating and making it possible to get more from existing robots.
Case in point is Tesla Motors which uses multiple tools for their robots to allow them to do a quick tool changes and perform multiple functions, e.g., welding, riveting, bonding and installing a component, yet the core robots are old-style fixed-to-the-floor KUKA, Fanuc and Fronius robots and mobile SmartCarts from Motion Controls.
Source: Tesla Motors
Almost every non-industrial robotic system (which, for lack of a better term is presently called Service Robotics) involved in making, moving or handing products, has the expectation of mobility, safety, vision, trainability without programming, and great flexibility. That is where the industry seems to be heading and the industrial players are slowly transitioning to be able to provide for that need. The companies mentioned above are beginning that transition -- but not very fast because they have a healthy and profitable growing industrial market already. Perhaps newer more adventurous companies will move more quickly to capture this market segment.There are publicly-traded exciting companies out there that will likely benefit from the trend toward automation. 3D printer companies like Stratasys (NASDAQ:SSYS) and 3D Systems (NYSE:DDD) will play a key role in this emerging market sector but privately held companies like MakerBot may register to go public and give them some competition. Kiva Systems and their innovative warehousing system was recently acquired by Amazon (the acquisition occurred in lieu of Kiva going public). This reminded me how important Amazon (NASDAQ:AMZN) is in providing material handing in addition to being an online sales force. To quote Bruce Welty in the Huffington Post:
During the California Gold Rush, there were many fortunes made and lost but one of the more enduring successes was Levi-Strauss, a clothier to all miners. Levi's never had to worry about which miners were going to discover gold; they all needed pants. In the New American Economy, these trends all converge around advanced fulfillment capabilities. Amazon has invested in its fulfillment capability for just that reason. The "Levi-Strauss-like" opportunity of today is in Fulfillment or, more specifically, Robotic Fulfillment.
Finally, Credit Suisse didn't mention other areas for automation and there are many. One example is hospital automation -- a big market for pill dispensing robots, mobile tugs to deliver linens, meals, trash and medicines, autonomous cleaners of all types and remote presence robots. Three public companies of note are Swisslog (SW:SLOG), Adept (NASDAQ:ADEP) and iRobot (NASDAQ:IRBT) and two at-present privately-held companies are InTouch Health and Aethon.


text: These strings are all about spin. They bite, and they impart spin. I use these on the Prince Triple Threat Warrior Mid Plus racket (spin friendly) at 57-58 lbs. I generate nasty slices and and kicks and get compliments from 4.0 and some 4.5 players. Sometimes the spin I generate leaves me with my mouth open in disbelief. Power is adequate, although with this racket, I need to generate my own power. I have a very quick swing which also helps with the spin. If you are all about spin, these are a great value for a spin friendly string. They last about 6-8 in singles play with a lot of base line rallies and hard serves, and that probably fluctuates 2-3 hours depending on the players style. BTW, if you are thinking about getting a Focus Hex, 18 guage, I'd suggest going with this one instead. I am a 4.0.


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In his latest book Niche, James Harkin, an expert on international social and economic change, analyzes the case histories of different brands that stand apart from the rest for their marketing aimed at well defined niches, on a global scale. These include Apple, The Economist, HBO and Moleskine.
In the chapter on Moleskine, the British journalist recounts how the beloved notebook of Bruce Chatwin has returned to stores and to the hands of contemporary artists and thinkers, finding its place between the old custom of note-taking and sketching and the evolution of the digital world with its blogs, virtual files and the Web.
James Harkin is director of Flockwatching and contributes regularly to the Financial Times and the Guardian. Niche is Harkin's third book, following his successes with Cyburbia and Big Ideas.


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HBO has renewed Veep and Silicon Valley for a fourth and second season, respectively, the channel announced today. Great news for Zach Woods, who is on both of those shows! And on The Good Wife sometimes, making him Sunday's most popular white guy. HBO already renewed Game of Thrones for two more seasons, plus Girls, Looking, and Getting On will all be back; mainstays True Blood and Boardwalk Empire are going into their final seasons later this year, as is The Newsroom. Fear not, though. As the HBO bible tells us, to everything there is a season; a time to renew, a time to cancel; a time to order 12 episodes of a comedy, a time to order a scanty 8; a time to order a one-off finale special for Hello Ladies, a time to let Larry David make his own decisions about whether Curb Your Enthusiasm will ever return.
HBO Renews Veep, Silicon Valley
Photo: Paul Schiraldi/HBO


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Jefferson City, Missouri
Jeremiah Wilson "Jay" Nixon (born February 13, 1956) is the 55th and current Governor of the U.S. state of Missouri. A member of the Democratic Party, he previously served as Missouri's Attorney General before his election in 2008. Nixon holds both an undergraduate degree and a law degree from the University of Missouri in Columbia.
He made his first bid for statewide office in 1988, an unsuccessful effort to oust incumbent U.S. Senator John Danforth. Nixon was defeated by Danforth, winning 32% of the vote compared to Danforth's 68%. Nixon was first elected as Missouri's Attorney General on November 3, 1992, on a platform of fighting crime, cleaning up government corruption, and protecting consumers and the environment. Nixon followed William L. Webster as Attorney General. The 1992 race for Attorney General between Nixon and former State House Minority Leader David Steelman was especially heated; however Nixon beat Steelman 51% to 45%, with an unusually high 4% of the votes going to the Libertarian Party candidate.


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Taking psilocybe mushrooms and MDMA (ecstacy) together. Actually regarded quite highly within the drug subculture. The effects rarely last longer than 8 hours. The effects of the MDMA lend a very upbeat feeling to the user, usually resulting in a positive mushroom trip. Because MDMA is a fairly "reliable" good high, but mushrooms aren't (there can be bad trips), the MDMA high helps to stabablize the mushroom trip.
"What are you doing for the rave on Thursday?"
"I'm going hippy flipping."
"I'm going hippy flipping."
by WILDmushroomSHROOMER October 31, 2006
the experience of taking mushrooms and ecstacy.
by Anonymous August 15, 2003
Taking extasy and mushrooms in a combination with each other. The tripp can last until the person goes to sleep(up to several days). Highly discouraged by most veteran drug users.
I've got a couple of tabs of x and a few shroom caps, do you want to go hippy flipping.
by Chaos(not-the-same-as-entropy) October 27, 2005
Taking mushrooms and acid together, as these are the drugs the most connected to hippies. This is different than candyflipping which is lsd and mdma.
"Yo im about to hippy flip on 3 hits and an eigth of shrooms i'm ready to go into space man."
"DUde you're gonna be fucked, last time i was hippyflipping everything exploded into infinity"
"DUde you're gonna be fucked, last time i was hippyflipping everything exploded into infinity"
by dudeonacid May 26, 2012


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Love nachos but hate the grease they leave behind on your hands and the way you feel stuffed in a bad way after you inhale a batch while out with friends or sitting on your couch in front of a hockey game or movie? Here are some great tips on making nachos in a healthier – yet still extremely tasty – way.
First of all ditch those high sodium, high fat chips you've been using and choose a low sodium whole wheat or multigrain variety. Or you can use baked tortilla chips – make your own or pick up a bag at your grocery store. Next, the meat – you can swap out the beef for turkey but if you have to have the beef – make it lean. Once you have browned the meat – drain off all excess fat and add a few sprinkles of seasoning – do not use one of those pre mixed taco seasoning bags! They are loaded with sodium and a whole bunch of other unpronounceable ingredients. Instead choose a blend of chili powder, garlic, turmeric, oregano, cilantro, or parsley. All of these spices have great health benefits.
As for the rest of the toppings – use veggies! Olive slices, tomato slices, shredded lettuce, onions, peppers – whatever you like. Corn and black beans are great on nachos as well and black beans are a very good source of fiber. And yes, you can still have the cheese – just don't load it on. You've got enough other yummy stuff on there that you don't need a ton.
As for dips – guacamole is an OK choice. It is fairly high in fat but it's not bad fat so you can have a bit. Sour cream is not your best option so swap that out and use hummus! Hummus is quite healthy and will add a lot of flavor. And do not forget the salsa! Salsa is an extremely healthy choice and is also a nacho staple.
So enjoy the upcoming hockey playoffs with a plate of nachos. Invite your friends over – they will think they are so yummy and won't believe they are actually healthy!


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Welcome to The Press Advertising Department:
Your key to the Eastern Maumee Bay Market.
No other newspaper delivers the Eastern Maumee Bay Communities like The Press. Our weekly circulation of more than 34,642 connects you to 23 communities in Lucas, Wood, Ottawa and Sandusky counties. Total Market Coverage, carrier delivered.
We offer two zoned editions to target your customers with display advertising and more than 70 carrier routes to pinpoint neighborhoods with flyers. (For details click on Circulation on menu bar)
Metro Edition:
Weekly news coverage of East Toledo, Oregon, Northwood and Jerusalem Township.
Suburban Edition
Weekly news coverage of communities east of Toledo including Genoa, Elmore, Woodville, Walbridge, Millbury, Oak Harbor, Pemberville and Gibsonburg.
Maumee Bay Press
A tourist publication delivered throughout the state of Ohio and at more than 100 stops along Western Lake Erie and Maumee Bay State Park. With a total print run of 50,000, there are four seasonal issues featuring local events and locator map. You can read our last issue of Maumee Bay by Clicking Here!
With 20,000 printed, our Maumee Bay Statewide Edition is distributed at Travel Information Center, Turnpike Welcome Centers, AAA Offices and at Maumee Bay State Park and Lodge. Save big when you sign up for all 4!!
Target Marketing
Our Extra Sections (Entertainment, Family, Education and Outdoors) allows you to place your ads next to related news. In addition, we offer numerous promotions throughout the year.
Updated Circulation Figures are on the "Circulation Information" page.
|Weekly Highlights: Hit your target audience with The Press Monthly Extra Sections!
|Spotlight Pages: In an effort to better serve both our readers and advertisers, The Press runs special Features that focus on specific topics or events: In an effort to better serve both our readers and advertisers, The Press runs special Features that focus on specific topics or events.
|Annual Events
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• Genoa Street Fair
|Special Sections/Pubs
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|Inserts: Our services can include custom design or just delivery to one or more of our 70 targeted routes. Click here, for more delivery area details.
|Maumee Bay Visitor's Guide: A visitors' guide to Maumee Bay & Western Lake Erie. 46,000 printed & distributed!


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Here's the latest article from the 12 Step Recovery site at BellaOnline.com.
During my growth through the 12 step program I realized how much of my nature was not only as an addict but also as a serious co-dependent.
Please visit 12steprecovery.bellaonline.com for even more great content about 12 Step Recovery. To participate in free, fun online discussions, this site has a community forum all about 12 Step Recovery located here -
I hope to hear from you sometime soon, either in the forum or in response to this email message. I thrive on your feedback!
Have fun passing this message along to family and friends, because we all love free knowledge!
Lara F., 12 Step Recovery Editor
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For the past three years, the Facebook Platform has been built on top of a session-based authentication system that many developers found complex. In order to make any API calls, you have to understand the details of signature algorithms. It's a common source of problems for new developers using the Facebook Platform. We have been searching for a way to make it simpler.
The OAuth community has faced similar issues. If a new developer just wants to start using the Twitter API, suddenly they have to understand things like HMAC-SHA256 and how to sign their base string. That is just overkill for a simple web API.
Because of this complexity, I am really excited about the next version of OAuth. OAuth 2.0 largely solves the performance and usability issues from OAuth 1.0a. It relies on SSL instead of signatures as the default way to interact with applications, which means you can play with it in your browser. While there are drawbacks to exclusive reliance on SSL, it is so simple to get started that it can't be beat for most developers. OAuth 2.0 also splits out flows for different security and performance contexts (i.e., desktop apps vs. web apps vs. mobile apps). That means that when a developer starts coding, they can see exactly what they need and start running.
How did we get here? Last fall, a small group wrote a proposal called OAuth WRAP, which introduced the core innovations of what would become OAuth 2.0 – using SSL and multiple flows. After Bret Taylor implemented OAuth WRAP in Friendfeed, we realized that it lived up to the promise. In the time since, we have been working with Yahoo, Twitter, Google, Microsoft, and many community members within the IETF to produce a new draft.
At f8, Facebook shipped a new Graph API, which relies entirely on OAuth 2.0. In this post, I'll go into detail about exactly what we shipped and our plans for the future.
The access token is king
The OAuth 2.0 spec is divided into two sections:
* First, you get an access token
* Second, you use the token to access protected resources.
Using a token is the easy part, so we'll start there.
To use an access token, just append it to the end of a protected resource with the
oauth_token parameter. (To alleviate some developer confusion, Facebook also accepts
access_token as the parameter name.)
All calls are required to go over HTTPS.
The new Graph API demonstrates how easy it is to get started. You can fetch data about any object on Facebook by hitting
https://graph.facebook.com/. For instance, this is my profile without an access token – all you can see are a few public fields:
{ "id": "2901279", "name": "Luke Shepard", "first_name": "Luke", "last_name": "Shepard", "link": "http://www.facebook.com/luke.shepard" }
Now, if you append an access token (notice the protocol switches to https), you can see much more information about me, such as my employer:
{ "id": "2901279", "name": "Luke Shepard", "first_name": "Luke", "last_name": "Shepard", "link": "http://www.facebook.com/luke.shepard", "work": [ { "employer": { "id": 109719699048814, "name": "Facebook" }, "location": { "id": 104022926303756, "name": "Palo Alto, California" }, "position": { "id": 109981385691387, "name": "Software Development Engineer" }, "start_date": "2007-05" }, ....
The new Graph API only accepts OAuth tokens. All existing API calls are backwards compatible with our existing auth system, but they do accept an OAuth token as well. So the Graph API is a carrot to encourage OAuth adoption.
Getting an Access Token
It used to be that people just accessed services from their desktop or laptop computers, but in the past few years, that has changed. Now, people use laptops, all sorts of web browsers, mobile phones, devices connected to TVs, etc. One of the most common criticisms of OAuth 1.0a is that there is only one way to do things – the new version of OAuth 2.0 addresses that by allowing for multiple types of flows to get a token.
In this initial launch, Facebook supports three ways of getting a token.
Web server flow
The web server flow is intended for use by server-side developers that don't really like JavaScript. The whole flow works by redirecting the user to the authorization server (Facebook) and back to your site. It is baked into the Facebook docs as the default auth flow (if you don't pass "type" param, then this flow is assumed).
You must pre-register a "Connect URL" with the domain and path of their site. When the user tries to authorize, Facebook checks the "redirect_uri" to make sure it begins with the URL registered for the given "client_id".
You can also pass an optional "display" parameter to customize, primarily for mobile devices. Values accepted are "page" (default), "popup", "wap", and "touch" (the last two for mobile sites).
User-agent flow
While the code for web applications typically lives on the web server, far away from the user, sometimes it lives on the user's machine. The main examples are desktop apps (Tweetdeck) and JavaScript-based applications (StreamDiff). Because the code actually runs on the client device, it can't really rely on embedded secret keys for security – in JavaScript, anyone can look at the source and trivially extract the secret. So, we need something else.
The user agent flow is created for applications that cannot embed a secret key. The access token is just returned directly in the redirect response instead of requiring an extra server call. Security is handled in two ways:
* Facebook makes sure that the access token is not sent to a random webserver by validating the redirect_uri matches a pre-registered URL.
* The access token never goes across the wire in the clear. Even if redirect_uri is an HTTP url, the token itself is returned after the fragment (#) and so the browser will never send it to the server.
Facebook encourages the User Agent flow for use in Desktop applications. We plan to incorporate it into the the JavaScript client library
Client Credentials flow
This is perhaps the simplest flow – just exchange your client_id and secret for an access token, no user involved. We support this for accessing application-only resources. In particular, it's required to use our new subscriptions API, modify developer settings, etc.
Session Exchange flow
Finally, for backwards compatibility, there is an endpoint where developers can exchange existing session keys for access tokens. We considered using the "Assertion Flow" for this, but we didn't because a) it uses a bunch of parameters and is optimized for SAML tokens, and b) this is just a migration strategy and not really core to the spec.
Open Areas
The OAuth 2.0 spec is in a draft state, which means it's still subject to change – especially around the edges. Since it's much easier to add a feature later than to remove or change one, we have tried to implement the stable core while postponing controversial or unstable features. In particular, the following areas are really interesting:
Identity. As David Recordon posted, we would love to "get OAuth 2.0 to the point – fairly quickly – where we can start to architect the next version of OpenID on top of it." Since Facebook provides both identity as well as authorization, it's critical that we get this piece right in order to have a complete solution.
Signatures. Most of the benefit of OAuth 2.0 is its lack of signatures – however, there are some use cases where it is difficult or nonperformant to make SSL requests, and in those situations we want to use signatures. Signatures are also required for identity – the server needs to be able to verify that a given message came from Facebook without making another HTTP request. We will eventually add support for signatures.
Immediate mode. In order to do single sign in with JavaScript, we need the ability to check if the user is currently logged in with an iframe. Today, Facebook uses an endpoint called login_status.php to handle this, but we plan to use the immediate mode in the future.
Device flow. I am intrigued by the Netflix-style device flow, and I think it's important for someone to implement it and try it out in the wild.
Refresh tokens. Most of the access tokens issued by Facebook are short-lived – they last for an hour or at most a day. In the OAuth 2.0 spec, the official way to handle long lived tokens is by issuing a "refresh token", which is then exchanged repeatedly for new, short term access tokens. In the Facebook API, the developer just asks for the "offline_access" extended permission, and then their access token just lasts forever (or until the user revokes it). We may look into using refresh tokens in the future.
Error formats. The error messages are currently JSON encoded, but they should be form-encoded – we are planning to fix that.
Client state parameter. The "state" parameter was removed and then added back in, but we don't support it. I believe that client state should be tracked in the redirect_uri, as it offers more protection.
Display parameter. We use a "display" parameter to support mobile flows, even though the spec does not officially include it (yet). We hope to see it included in a future draft.
What comes next?
The OAuth 2.0 draft is still a work in progress. As we work through the open issues, the spec will evolve. I'm looking forward to other companies shipping their own endpoints, and eventually building OAuth 2.0 into the fundamental fabric of how the Internet works. Please leave a comment or join the mailing list to get involved!


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Jose Landaverde was inspired to cook by memories of making pupusas with his late father, who was killed in El Salvador. Jorge Perez's interest in food was cultivated by his grandfather, a caterer who introduced him to exotic spices on a trip to Thailand. And Lucile Flores was practically raised in kitchens, especially at the Jack in the Box restaurant where both of her parents work.
Drawn to food by powerful family ties, the Los Angeles Unified student chefs took their culinary interest a grand step further Thursday as they vied to win the local round of a national high school healthful cooking competition. The school district entered the Cooking up Change contest for the first time this year as part of a broader move to give students a larger voice in its healthful school food initiative.
L.A. Unified dramatically altered its menu a few years ago, throwing out chicken nuggets and nachos, for instance, in favor of more whole grains, fresh produce and less sodium and fat. But students rejected many of the new dishes, prompting school officials to more actively involve them through surveys, taste tests, work with culinary teams and, now, a cooking contest.
"What we're trying to do is get kids to eat more healthfully and nutritionally but also have them learn how it impacts their health," said David Binkle, the district's food services director. "It all starts with kids helping us do that."


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State Sen. Lee Bright, R-Roebuck, has filed legislation to allow South Carolina schools to offer a class in firearms marksmanship as an elective.
State Sen. Lee Bright, R-Roebuck, has filed legislation to allow South Carolina schools to offer a class in firearms marksmanship as an elective.
According to the legislation, the course would be called the South Carolina Gun Safety Program. It would teach firearm operations, history of the constitutional right to keep and bear arms and "the role of firearms in preserving peace and freedom." Students would be taught to fire a gun at moving and stationary targets and would successfully complete the course by safely discharging a firearm.
"I asked around to some of my colleagues in the legislature if schools could offer a course like this and it wasn't concisely expressed whether they could. I figured I'd sponsor this bill and give schools that option," Bright said.
The bill states schools or school districts offering the course should arrange for adequate time on a shooting range and does not repeal prohibition of firearms on school campuses.
Gun control and particularly guns in schools has been a central issue of national debate since the tragic death of 26 people, including 20 students, in a mass shooting at Sandy Hook Elementary School in Newtown, Conn.
"We need to move the debate," Bright said. "The guns are objects; they don't create these crimes."
Bright said he thinks society should have more guns and students should be educated in how to use them safely.
Some area school administrators questioned the need for such a course and whether schools were the proper conduit for a shooting class.
"I believe there are commercial agencies that are better suited for this," Spartanburg District 5 Superintendent Scott Turner wrote in an email. "We have more than enough curriculum and not enough time to cover it now. I am not in favor of adding more."
With all the academic, and increasingly non-academic, standards schools are required to meet, Spartanburg District 4 Superintendent Rallie Liston said he couldn't see his district taking on teaching something readily available in the private sector.
"We've got enough on our hands already," he said. "… It's not that it isn't something good to learn, but so is playing the piano, so are certain kinds of dance."
The bill instructs the South Carolina Law Enforcement Division to design the course in consultation with the South Carolina Department of Natural Resources, the South Carolina Department of Public Safety and private firearms organizations. It would be taught by a certified firearms instructor.
The Department of Natural Resources already partners with several public schools to offer a hunter safety course that awards students credit along with fulfilling a requirement for a hunting license. The course typically does not offer shooting practice and does not focus on the historical areas addressed in the bill, said DNR spokesman Greg Lucas.
DNR also partners with the Youth Shooting Foundation to create extracurricular shooting clubs at some high schools.
Individual elective classes do not require an act of the legislature and are usually created by the schools or the state Department of Education, said Jay Ragley, director of legislative affairs for the South Carolina Department of Education.
"It's not common that we see bills that create courses, but it does happen," he said.
Most of those bills don't pass. The last time a similar bill was passed was in 2006 when the legislature voted to allow academic credit for Bible study done on release time. Last year, the state education department lobbied against a bill that would have required CPR certification for every graduating senior and the measure did not pass. Since Bright's bill is optional and not a mandate and has no budgetary obligations, it likely will not meet with the same objections, Ragley said.
The need to appropriate money effectively also weighed on the minds of local superintendents as they considered the bill.
"We have many more high tech courses needed to ready our students for our global economy rather than teaching them to fire a gun," Turner wrote.
In some rural areas, including Woodruff, a shooting class would be redundant to many students.
"Elective courses are based on student needs," Liston said. "Most of our students already know how to shoot. Most of them are pretty good at it."


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U.S. Equal Employment
Opportunity Commission
Indianapolis Bar Fired Bartender/Server Because She Was Pregnant, Federal Agency Charged
INDIANAPOLIS – WBS Broad Ripple, Inc., doing business as the Wild Beaver Saloon in Indianapolis, unlawfully fired a female bartender/server because of her pregnancy, the U.S. Equal Employment Opportunity Commission (EEOC) charged in a lawsuit it filed March 16, 2011.
The suit, EEOC v. WBS Broad Ripple, Inc., d/b/a Wild Beaver Saloon, Civil Action No. 1:11-cv-00373-TWP-DML, filed in U.S. District Court for the Southern District of Indiana, Indianapolis Division, charged that the company violated Title VII of the Civil Rights Act of 1964, which prohibits employment discrimination based on sex, including pregnancy.
"Employees who become pregnant should not lose their jobs because of their condition," said Laurie A. Young, regional attorney of the Indianapolis District Office. "The Commission will vigorously prosecute employers who engage in pregnancy discrimination or other forms of sex discrimination."
The EEOC is seeking compensatory and punitive damages against the company, as well as other relief, including a permanent injunction to prevent the company from engaging in any employment practice that discriminates on the bases of sex.
EEOC enforces federal laws prohibiting employment discrimination. Further information about the EEOC is available on its web site at www.eeoc.gov.


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What textbook is this?

 The Orphika I am reading from is in a textbook with fragments, and other -fairly easy- Greek texts.

Many random facts are lodged in my brain. I'm not sure how this one got there.

 how did you know about "Consaidin"? (amazed!)


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American Traders Canoe & Furniture Collection
No single product better represents the true American spirit than the classic wooden canoe. They are a direct link to the adventures and legends of outdoor life. They capture and reflect nature's wonders and help you enjoy the simple pleasures of life as you take in the beauty of the world around you. With over 35 years of canoe building experience American Traders is the largest builder of classic cedar canoes in the world. Our commitment to the craft and the careful attention to detail in each canoe really defines the exceptional quality of our boats.
With most of us living insanely busy and stressful lives it's important to find time to enjoy the simple pleasures of life'rican Traders' canoes are the perfect escape, what could be better than spending a few hours connecting with the outside, exercising to keep fit and unwinding on the water. As beautiful as our canoes are, they are not museum pieces. Yes, they are nice to look at but all are intended for use in the water. And now the same classic designs availble in lightweight E-Fusion Kevlar or carbon and wood trim, built with the same high quality and attention to detail.


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The 301.8C mini hydraulic excavator features an 18.1 hp engine and increased hydraulic flow and pressure. The two-speed excavator's variable displacement hydraulic pump prevents stalling, and the independent swing boom enables the machine to work next to a wall or obstruction. Its cab has a retractable front window, a suspension seat, console-mounted implement controls, and adjustable wrist supports. The mini excavator has a 200° bucket-rotation, a 12 ft 6 in. maximum reach, and a 7 ft 7 in. maximum digging depth.
For more information, visit www.cat.com.
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Brief SummaryRead full entry
BiologyAs its name implies, the fishing cat predominantly preys on fish (6). Largely active at night, fishing cats are good swimmers and have been observed diving for fish, as well as scooping them out of the water with their paws (8). These cats will also prey on frogs, crustaceans, snakes, birds, calves, goats, and dogs, and will scavenge on carcasses of larger animals (5). Although capable of breeding all year round (6), birth peaks have been noted in March and May in north-eastern India (8). One to four kittens are born after a gestation period of 63 days (7). Young suckle until they are six months old (5) and reach independence at ten months (8). In captivity, males have been recorded to aid in the rearing of young (5). Fishing cats live an average of 12 years (8), but have been known to live more than 15 years in captivity (5).


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Sesame Street: Once Upon a Monster
Gaming platform: Kinect for Xbox 360
ESRB Rating: E for Everyone
Appropriate for ages: 4+
Price: $43.91 on amazon.com
If you've ever played a Kinect game on the Xbox 360, you know that it's an experience like no other. Unencumbered by controllers, kids and adults simply need to use their body movements to play the game and interact with the characters on screen – and what better characters to interact with than friends from Sesame Street.
Sesame Street: Once Upon a Monster is a fantastic game for kids ages 4+ that takes them on an adventure inside of a storybook. With Elmo and Cookie Monster leading the way, kids (and their parents) can engage in many different games and activities within the many chapters of the game. To keep young kids' attention, the game moves pretty quickly through activities like tapping a drum, choosing colors and dancing and promotes important lessons like friendship and responsibility.
I like that this game keeps kids active and also promotes activity. If there are activities that are a bit more complex for young children, parents can easily hop in and out to help since the game easily accommodates multi-player gaming. The one caveat of this game is that kids need to unlock levels to continue to progress through its entirety. But once that is done, players are free to jump around from chapter to chapter, focusing on the games or starring characters that have become their favorites.
Take a peek at the game:
If you have a video game that you'd like considered for Screen Play, please contact firstname.lastname@example.com.


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Within minutes of the opening arguments before the Florida Supreme Court Friday, two justices lashed into lawyers for opponents of the legislatively-drawn map when they concluded that the court rejected more than eight flawed districts when it threw out the Senate map March 9.
"Speaking for the person that wrote it, it was pretty clear that there were certain districts that were specifically invalidated,'' said Justice Barbara Pariente, the author of the 5-2 opinion that forced the state Legislature to return in an extraordinary session to revise its Senate map.
She suggested that it would be unfair to the Senate if the court were to reopen districts where challenges previously were rejected "or the burden of the challenges were unmet."
Justice Peggy Quince also raised doubts about the arguments raised by the Florida Democratic Party and the coalition of voter groups that want the court to reject the entire map. "Why would want we want to go back and look at others? We looked at the whole thing,'' she said. "It just doesn't resonate."
But Paul Smith, lawyer for the Florida chapter of the League of Women Voters, the National Council of La Raza and Common Cause, said that the court must make sure that the new map adheres to the Fair District standards approved by voters and the new map clearly does not do that.
Pariente conceded that that argument "is, in my view, your strongest challenge'' but suggested it could also be "reasonable legislative discretion."
Smith countered that there were "plenty of competitive districts they could have drawn" that didn't divide Daytona Beach but "they just didn't'want to do it." He said they "gratuitously split a county" in "a very clear effort to make sure they cancel out Democratic votes in that region of the state."
The arguments are part of the required court review of the Legislature's redistricting maps. This is the lawmakers' second attempt to get its Senate map approved by the state's high court. The court rejected their first attempt by a 5-2 vote as violating the state Constitution because of eight flawed districts, a numbering system that protected incumbents and a plan "rife with indicators of improper intent.''
The court unanimously approved the House's redistricting map, but legislators were forced to come back for a 15-day extra session to redraw its Senate boundaries.
The court has until May 14 to review the Legislature's second attempt at drawing its redistricting maps and if the court rejects it this time, the justices must propose a fix themselves.
The Republican-led Legislature, faced with its last strike, has hired former Florida Supreme Court Justice Raoul Cantero, now a lawyer with the Miami law firm of Case and White, to argue its case. He will be paid $695 an hour. Cantero commended his former colleagues but urged them to give the Legislature deference in the once-a-decade process.
"You can always make a better plan,'' Cantero told the justices, "… but that's not the purpose of this court. The purpose of this court is to determine if our plan is legitimate."
Pariente conducted most of the questioning, just as she did during the hearing into the first round of maps. This time, she made it clear that if the court were to take control of the once-a-decade mapmaking process, it would revise the map selectively.
"If we decided that one district was invalid, nobody would suggest that the whole map should be redrawn," Pariente said.
She zeroed in on the proposed District 8, drawn by Republican senators to split Volusia County, a Democrat-dominated county, into three districts while leaving the adjacent Republican-dominated counties intact.
Democrats claim it was designed to dilute the Democratic vote, protect incumbent Sen. John Thrasher, R-St. Augustine in the adjacent District 6, and elect a Republican in the newly opened District 8 seat. The NAACP says it also isolates black voters in Daytona Beach.
"That is a concern of mine,'' Pariente said, adding that she thinks the configuration of these districts is the Democrats' "strongest argument."
Smith, the lawyer for the coaltion, underscored that point and said the newly drawn black-majority district centered in Duval County, District 9, packs Democrats in such a way as to make every other surrounding district "Republican-leaning or very likely Republican held."
But Cantero countered, saying the Senate put an end to the sprawling district it previously proposed and created a black majority district within the confines of Duval County because the court wanted it that way.
"I've never seen a case in my life that somebody gets reversed because this is what a court told us to do,'' he said.
Unlike the first round, when the NAACP commented on the first map but did not ask the court to reject it, the group this time vigorously urged the court to reject the Senate map.
Allison Riggs of the Southern Coalition for Social Justice argued that the new Senate map dilutes minority voting strength in District 9 in Duval County. Also, in District 31, a sprawling district in Broward and Palm Beach counties now held by Democrat Chris Smith, the number of black voters who are citizens is estimated at 43 percent.
The result is that a black candidate could lose to a white candidate because of the state's history of "racially polarized voting,'' in which white Democrats won't vote for a black Democrat, Riggs told the court. The Senate maps don't provide enough information about that practice, she said, and absent that, risk violating the Constitution. She disagreed, however, that the NAACP's claims are too late.
"I think this court owes more in fairness to minority voters in the state than it does to the Senate,'' she said after the hearing. "If someone is going to have to be confronted with unfairness, it should not be black voters in Northeast Florida."


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2019 Jeep Compass
Latitude 4dr SUV (2.4L 4cyl 6A)
$470/mo est.
Price Analysis
63 Days on Lot
Vehicle Details
Ext. Color:
Granite Crystal Metallic Clear Coat
Int. Color:
Black vinyl/cloth
Drive Type:
front wheel drive
All Features and Specs
UConnect w/Bluetooth wireless phone connectivity
Digital Audio Input
AM/FM/Satellite-prep Radio
Total Number of Speakers: 6
Braking Assist
ABS and Driveline Traction Control
Cornering Lights
Stability controll with anti-roll
Privacy glass: Deep
Silver w/painted accents aluminum rims
Wheel Diameter: 17
Wheel Width: 7
Driver knee airbags
Front fog/driving lights
Leather/metal-look steering wheel trim
Leather/metal-look shift knob trim
Metal-look dash trim
Metal-look center console trim
Roof rails
Video Monitor Location: Front
Trip computer
External temperature display
Auxilliary engine cooler
Engine block heater
Manufacturer's 0-60mph acceleration time (seconds): 7.6 s
Power remote driver mirror adjustment
Heated driver mirror
Heated passenger mirror
Power remote passenger mirror adjustment
Dual illuminated vanity mirrors
Daytime running lights
Audio controls on steering wheel
Front reading lights
Cloth/vinyl seat upholstery
Bucket front seats
Rear bench
Fold forward seatback rear seats
Rear seats center armrest
Tilt and telescopic steering wheel
Electric power steering
Suspension class: Regular
Interior air filtration
Automatic front air conditioning
Dual front air conditioning zones
Tire Pressure Monitoring System: Tire specific
Remote activated exterior entry lights
Cargo area light
Max cargo capacity: 60 cu.ft.
Vehicle Emissions: LEV3-SULEV30
Fuel Type: Regular unleaded
Fuel Capacity: 13.5 gal.
Instrumentation: Low fuel level
Clock: In-radio display
Headlights off auto delay
Dusk sensing headlights
Driver Seat Head Restraint Whiplash Protection
Head Restraint Whiplash Protection with Passenger Seat
Coil front spring
Regular front stabilizer bar
Independent front suspension classification
Strut front suspension
Four-wheel Independent Suspension
Coil rear spring
Rear Stabilizer Bar: Regular
Independent rear suspension
Strut rear suspension
Front and rear suspension stabilizer bars
Variable intermittent front wipers
Black grille w/chrome accents
Center Console: Full with covered storage
Overhead console: Mini
AC power outlet: 1
Curb weight: 3,184 lbs.
Gross vehicle weight: 4,400 lbs.
Overall Length: 173.0"
Overall Width: 73.8"
Overall height: 64.8"
Wheelbase: 103.8"
Front Head Room: 39.2"
Rear Head Room: 38.5"
Front Leg Room: 41.8"
Rear Leg Room: 38.3"
Front Shoulder Room: 56.7"
Rear Shoulder Room: 55.1"
Front Hip Room: 54.1"
Rear Hip Room: 49.2"
Two 12V DC power outlets
Transmission hill holder
Seatbelt pretensioners: Front
Rear center seatbelt: 3-point belt
Door reinforcement: Side-impact door beam
Engine immobilizer
Cargo tie downs
Floor mats: Carpet front and rear
Cupholders: Front and rear
Door pockets: Driver, passenger and rear
Seatback storage: 2
Tires: Width: 225 mm
Tires: Profile: 60
Tires: Speed Rating: T
Diameter of tires: 17.0"
Type of tires: AS
Tires: Prefix: P
Left rear passenger door type: Conventional
Rear door type: Liftgate
Right rear passenger door type: Conventional
Body-colored bumpers
Integrated roof antenna
4 Door
Driver airbag
Manual child safety locks
Rear View Camera
Keyless ignition w/ push button start & keyless door entry
2 USB ports
Halogen aero-composite headlights
Ambient lighting
Dealer Description
What a fantastic deal! This Jeep won't be on the lot long! You'll appreciate its safety and technology features! Top features include front fog lights, a trip computer, an outside temperature display, and remote keyless entry. Under the hood you'll find a 4 cylinder engine with more than 170 horsepower, and for added security, dynamic Stability Control supplements the drivetrain. Our experienced sales staff is eager to share its knowledge and enthusiasm with you. We'd be happy to answer any questions that you may have. Call now to schedule a test drive.
Dealer Information
Contact Information
Ed Tomko Chrysler Dodge Jeep Ram
33725 Walker Rd, Avon Lake, OH 44012
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MSRP is the Manufacturer's Suggested Retail Price for this vehicle as provided to Edmunds by the dealer.
Price excludes all federal, state and local taxes, and all title, license and registration fees. Dealer is under no obligation to include or offer any additional equipment, products or services.
This vehicle is offered subject to prior sale; accordingly, there is no guarantee that this vehicle will not be sold to another buyer. Offer expires on the close of business on the date set forth above.
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