Dynamics of Horizontal Gene Transfer and the Ecological Risk Assessment of Genetically Engineered Organisms

The extensive published discussion of potential ecological impacts of introduced genetic sequences and genetically engineered organisms has lacked a quantified delineation of the critical questions for the estimation of risk. Ultimately, the ecological risk assessment of introduced gene sequences is the application of evolution, population genetics, and ecology to risk estimation and decision making. This paper provides a framework for the estimation of risk due to introduced sequences in bacteria, and the principles should also hold for many diploid species. Horizontal genetic exchange poses new challenges for ecological risk assessment. Plasmid transfer can occur without any impacts, although the sequence can become ubiquitous in the population. Conversely, the introduction of a plasmid can change the dynamics of the host population, potentially altering the population minimum and maximum characteristics of its dynamics. Because of genetic exchange, new genetic information is unlikely to be constrained among one type of prokaryote. An example of the use of the model is given using genetic exchange data from a series of published soil microcosm experiments. The model demonstrates the increase in plasmid frequency when using experimentally derived conjugation frequencies. Application of these results to ongoing discussion of the risks of genetically engineered organisms is presented. Particular attention is paid to the transfer of genetic material and the resultant changes in host population dynamics.     

Risk assessment of the crayfish pet trade based on data from the Czech Republic

The pet trade in freshwater crustaceans, including crayfish, has grown rapidly in recent decades and become an important pathway for introducing new non-indigenous species into Europe. This paper provides the first overview of non-indigenous crayfish species (NICS) traded as ornamental and their potential impact in the Czech Republic, which is the second leading importer into Europe. The paper presents a full list of traded crayfish species, their market availability, and trade names or misnomers used in the country. In total, 27 crayfish species from all three families are advertised and marketed, of which Astacus astacus is the only indigenous species. Only three NICS were recognized as very common on the market. The invasiveness and risk associated with ornamental crayfish trade were assessed using the Freshwater Invertebrate Invasiveness Scoring Kit. Five NICS were classified into the high-risk category, the highest score being for Procambarus fallax f. virginalis. The invasiveness of crayfish indigenous to North America is significantly greater than that of crayfish from the rest of the world, and therefore regulation in this regard is recommended.     

From Regional to National Clouds: TV Coverage in the Czech Republic

Media, and particularly TV media, have a great impact on the general public. In recent years, spatial patterns of information and the relevance of intangible geographies have become increasingly important. Gatekeeping plays a critical role in the selection of information that is transformed into media. Therefore, gatekeeping, through national media, also co-forms the generation of mental maps. In this paper, correspondence analysis (a statistical method) combined with cloud lines (a new visual analytics technique) is used to analyze how individual major regional events in one of the post-communist countries, the Czech Republic, penetrate into the media on a national scale. Although national news should minimize distortions about regions, this assumption has not been verified by our research. Impressions presented by the media of selected regions that were markedly influenced by one or several events in those regions demonstrate that gatekeepers, especially news reporters, functioned as a filter by selecting only a few specific, and in many cases, unusual events for dissemination.     

Mexican diabroticite beetles: I. Laboratory test on host breadth of Acalymma and Diabrotica spp.

Choice tests were conducted to determine relative degree of specialization of feeding behavior of 11 Mexican diabroticite species in the genera Acalymma and Diabrotica (Chrysomelidae: Luperini). Adult beetles were offered a choice between cotyledons of a non-bitter (not containing cucurbitacin) cucurbit (C. pepo L. var. Crookneck), corn (Zea mays L.), and beans (Phaseolus vulgaris L.). In a second assay a bitter (containing cucurbitacin) cucurbit (C. pepo L. var. Ambassador) was added to the array of plants offered. Neonates of two species of Acalymma and one species of Diabrotica were offered a choice between roots of a non-bitter and a bitter cucurbit, and between a bitter cucurbit and corn.Adult insects showed distinct preferences in the first assay. All Acalymma spp. tested accepted only the non-bitter cucurbit as host, whereas Diabrotica spp. preferred either the cucurbit or the noncucurbit hosts. When the bitter cucurbit was offered together with the other three hosts, all species changed their host choice and significantly preferred the bitter cucurbit. Neonates of all three species tested significantly preferred the bitter cucurbit roots over the non-bitter roots, and the corn roots over the bitter cucurbit.The observation that all Mexican diabroticite species tested left suitable hosts when bitter cucurbits were offered in a choice situation supported the hypothesis that the association between diabroticites and Cucurbitaceae is mediated by plant chemical compounds. For both, the Acalymma spp., which were found to be cucurbit specialists, as well as for the polyphagous Diabrotica spp., cucurbitacin B acted as a strong feeding arrestant which implies that the chemical mediation of this interaction might be an evolutionary conservative trait within the tribe.     

美国主要转基因作物发展现状及启示

以美国农业部国家农业统计局公布的相关数据为基础,首先简要介绍美国主要农作物种植变化情况,然后对2000~2011年美国主要转基因作物种植面积、性状等进行分析,得出了美国主要转基因作物种植的发展趋势。最后简要介绍我国相应转基因作物的发展情况,并据此提出我国发展转基因作物的几点建议。     

Health Risk Assessment of Lead in the Republic of Korea

The purpose of this study was to estimate the daily exposure to lead due to food ingestion, air inhalation, and soil ingestion in the Republic of Korea's general population, and to evaluate the level of risk associated with the current lead exposure level using the proportional daily dose (3–4 μg/kg body weight/day) corresponding to the Provisional Tolerable Weekly Intake (PTWI) suggested by the Joint FAO/WHO Expert Committee on Food Additives as the toxicological tolerance level. The estimation of the daily exposure to lead via three pathways including food, soil ingestion and air inhalation was conducted as a chronic exposure assessment. For the lead exposure assessment through dietary intake, 1,389 lead residue data for 45 commodities investigated by the Korea Food and Drug Administration during the period 1995–2000 were utilized (KFDA 1996, 1997, 1998). Six hundred seventy-two air monitoring data from 7 major cities during the period 1993–2000 and 4,500 soil residue data at 1,500 sites during the period 1999–2001 were considered for the lead exposure assessment involving air inhalation and soil ingestion, respectively. The total daily exposure to lead was estimated by combining dietary intake, inhaled amount and soil intake corresponding to the typical activity of the general population, which was treated as a group of adults with a body weight of 60 kg. For risk characterization, the daily exposure to lead was compared with the toxicological tolerance level. The level of risk due to lead exposure was calculated using the hazard ratio (HR). The dietary intake of lead was 9.71 × 10_?4 mg/kg/day and the total daily exposure level, including air inhalation and soil ingestion, was 9.97 × 10_?4 mg/kg/day. The exposure contributions of foods, air and soil induced from the percentage of each media to the total daily exposure were 97.4%, 2.1% and 0.5%, respectively. Of the different commodity groups, the highest contribution to the total exposure came from grain, which represented 47.7% of the total. Additional exposure to lead occurs in certain population groups due to the use of tobacco, alcoholic beverages, and the intake of other foods, all factors not considered in this study. Through the comparison of the daily exposure to lead with the tolerance level based on the PTWI, the hazard ratio was estimated as being 0.25–0.33. This value implies that no increase in blood lead level is to be expected in the general population at the current lead exposure levels.     

Ex-Ante Economic Impact Assessment of Genetically Modified Banana Resistant to Xanthomonas Wilt in the Great Lakes Region of Africa

Background

Credible empirical evidence is scanty on the social implications of genetically modified (GM) crops in Africa, especially on vegetatively propagated crops. Little is known about the future success of introducing GM technologies into staple crops such as bananas, which are widely produced and consumed in the Great Lakes Region of Africa (GLA). GM banana has a potential to control the destructive banana Xanthomonas wilt disease.

Objective

To gain a better understanding of future adoption and consumption of GM banana in the GLA countries which are yet to permit the production of GM crops; specifically, to evaluate the potential economic impacts of GM cultivars resistant to banana Xanthomonas wilt disease.

Data Sources

The paper uses data collected from farmers, traders, agricultural extension agents and key informants in the GLA.

Analysis

We analyze the perceptions of the respondents about the adoption and consumption of GM crop. Economic surplus model is used to determine future economic benefits and costs of producing GM banana.

Results

On the release of GM banana for commercialization, the expected initial adoption rate ranges from 21 to 70%, while the ceiling adoption rate is up to 100%. Investment in the development of GM banana is economically viable. However, aggregate benefits vary substantially across the target countries ranging from US$ 20 million to 953 million, highest in countries where disease incidence and production losses are high, ranging from 51 to 83% of production.

Conclusion

The findings support investment in the development of GM banana resistant to Xanthomonas wilt disease. The main beneficiaries of this technology development are farmers and consumers, although the latter benefit more than the former from reduced prices. Designing a participatory breeding program involving farmers and consumers signifies the successful adoption and consumption of GM banana in the target countries.     

Large‐scale mating disruption of Ephestia spp. and Plodia interpunctella in Czech Republic,Greece and Italy

Field trials were carried out to evaluate the use of the pheromone (9Z,12E)‐tetradecadienyl acetate (TDA/ZETA) for mating disruption (MD) of Pyralidae moths associated with stored products, in most cases the Raisin moth, Ephestia cautella (Walker), Mediterranean flour moth, Ephestia kuehniella Zeller and Indianmeal moth, Plodia interpunctella (Hübner). The experiments were conducted in the Czech Republic, Greece and Italy during 2007 and 2008 in storage facilities that varied in their size and type, and included flour mills, retail stores, storage rooms with currants and raw grain stores. After a summer pre‐treatment monitoring period to assess moth population in, dispensers containing TDA were placed in the fall. Adjacent facilities without dispensers were used as control units. Pheromone‐baited traps were used to monitor the population fluctuation of the pyralid moths during the entire experimental periods. The presence of MD dispensers notably reduced the number of adults found in the traps in comparison with control rooms. Monitoring of female oviposition, measured as number of hatched larvae in cups containing food, indicated that there was a reduction in the number of larvae in the areas with MD dispensers. The results of the present work indicate that the use of mating disruption is feasible against pyralid moths in storage facilities, and should be further evaluated as a component of an integrated pest management based control strategy.     

Flight activity of Diabrotica spp. at the borders of cornfields and its relation to ovarian stage in D. barberi

Flight activity of corn rootworm beetles, the northern (NCR), Diabrotica barberi (Coleoptera: Chrysomelidae), and western (WCR), D. virgifera virgifera, was monitored using malaise traps placed at the edges of two small cornfields. Populations of beetles in the fields peaked while corn was flowering, but capture in malaise traps remained low until silks had dried. Capture of NCR and (to a lesser extent) WCR then increased and remained high through the season. Malaise traps were constructed to permit separate collections of beetles that entered on each of two opposite sides. Female NCR that were captured in sides that faced toward corn tended to contain fewer mature eggs than those in sides facing away from corn. These findings support the hypothesis that female NCR emigrate from cornfields to feed when fresh silk and corn pollen (favored foods) become unavailable, but that egg-laden NCR actively seek corn when searching for oviposition sites.

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Résumé Les activités de vol de D. barberi (NCR) et D. virgifera virgifera (WCR) ont été contrôlées par l'utilisation de pièges malais placés en lisière de deux petits champs de maïs. Les populations de ces coléoptères sont maximales quand le maïs fleurit, mais leurs captures sont restées faibles jusqu'à ce que les barbes du maïs aient été sèches. Les captures de NCR et, dans une plus faible mesure, celles de WCR ont alors augmenté et sont restées importantes pendant toute la saison. Les pièges malais ont été conçus pour que les adultes pénétrant par les côtés opposés soient récoltés séparément. Les femelles de NCR capturées face au champ de maïs avaient tendance à contenir moins d'ovocytes mûrs que celles capturées du côté opposé. Ces observations appuient l'hypothèse que les femelles NCR émigrent des champs de maïs pour s'alimenter, quand les barbes et le pollen de maïs, aliments préférés, deviennent indisponibles, mais que les femelles NCR pleines d'ovocytes mûrs recherchent le maïs quand elles sont en quête de lieux de ponte.

     

Notes on the genusOenothera subsect.Oenothera (Onagraceae) in the Czech Republic

Up-to-date results of taxonomic and phytogeographic studies ofOenothera subsect.Oenothera in the eastern part of the Czech Republic (Moravia and Silesia) are presented. A new hybridogenous species,Oe. moravica Jehlík etRostański (=Oe. fallax × Oe. victorini), is described from S Moravia. Several taxa and hybrids are reported for the first time for the Czech Republic:Oe. victorini Gates etCatcheside (=Oe. rostanskii Jehlík),Oe. × wienii Renner exRostański,Oe. acutifolia × Oe. victorini. Other taxa are recorded from the eastern part of the Czech Republic for the first time:Oe. acutifolia Rostański,Oe. hoelscheri Renner exRostański,Oe. parviflora L.,Oe. subterminalis Gates. The taxonomy ofOe. pycnocarpa Atkinson etBartlett is discussed. A survey of species and hybrids ofOenothera subsect.Oenothera in the Czech Republic is added.     

Molecular and microscopical evidence of Ehrlichia spp. and Borrelia burgdorferi sensu lato in patients,animals and ticks in the Czech Republic

We report moderately severe cases of human ehrlichiosis and a lethal one caused by human granulocytic Ehrlichia, the HGE agent, closely related to Ehrlichia phagocytophila and Ehrlichia equi. Their vector is the Ixodes ricinus tick, which also transmits Borrelia burgorferi sensu lato in central, west and east regions of the Czech Republic. The diagnosis was established by PCR with sequence analysis of the genes encoding 16S rRNA of Ehrlichia and with reverse hybridization by using enzyme linked immunosorbent assay with different covalently coupled probes to the activated plate.Ten out of 47 patients and 10 huntsmen were PCR positive and 7 of them seroconverted to the HGE. Coinfection of Ehrlichia phagocytophila with Borrelia burgdorferi sensu lato was detected in 3 patients. Ehrlichia spp., the HGE agent, was isolated and propagated only from one patient in the HL-60 promyelocytic cell line. The maintenance of Ehrlichia in culture and in patients was assayed also by immunocytological staining and electron microscopy. Sequence or hybridization analysis of PCR results in different wild mammals and birds showed significant sources of Ehrlichia fagocytophila in nature. Three variants of E. phagocytophila in wild roe deer and boars, as well as for the first time in birds, have been described. Cultures from the blood of horses, and from the spleen and kidney specimens of roes and boars, PCR positive for Ehrlichia spp., displayed a disappearing level of the pathogen or contamination with other bacteria.     

Unusual finding of Trichobilharzia sp. in Motacilla alba in the Czech Republic

Adult male worms of Trichobilharzia sp. recovered from a pied wagtail (Motacilla alba) in the Czech Republic were found to belong to a new species of the genus. The finding of Trichobilharzia sp. in a passeriform bird in Europe represents an important discovery, as only anseriform birds have thus far been reported as final hosts of the European Trichobilharzia species.     

A 90-Day Feeding Study in Rats to Assess the Safety of Genetically Engineered Pork

Our laboratory recently produced genetically engineered (GE) Meishan pigs containing a ZFN-edited myostatin loss-of-function mutant. These GE pigs develop and grow as normal as wild type pigs but produce pork with greater lean yield and lower fat mass. To assess any potential subchronic toxicity risks of this GE pork, a 90-day feeding study was conducted in Sprague-Dawley rats. Rats were randomly divided into five groups, and fed for 90 days with basic diet and basic diets formulated with low dose and high dose pork prepared from wild type pigs and GE pigs, respectively. Animal behaviors and clinical signs were monitored twice daily, and body weight and food consumption were measured and recorded weekly. At days 45 and 90, blood tests (lipid panel, electrolytes, parameters related to liver and kidney functions, and complete blood counts) were performed. Additionally, gross pathology and histopathological analyses were performed for major organs in each group. Data analysis shows that there were no significant differences in growth rate, food consumption, and blood test parameters between rat groups fed with GE pork and wild type pork. Although differences in some liver function parameters (such as aspartate aminotransferase, total proteins, albumin, and alkaline phosphatase) and white blood cell counts (such as lymphocyte percentage and monocyte percentage) were observed between rats fed with high dose GE pork and basic diet, all test results in rats fed with GE pork are in the normal range. Additionally, there are no apparent lesions noted in all organs isolated from rats in all five feeding groups on days 45 and 90. Overall, our results clearly indicate that food consumption of GE pork produced by ZFN-edited myostatin loss-of-function mutant pigs did not have any long-term adverse effects on the health status in rats.     

A Forward-Design Approach to Increase the Production of Poly-3-Hydroxybutyrate in Genetically Engineered Escherichia coli

Biopolymers, such as poly-3-hydroxybutyrate (P(3HB)) are produced as a carbon store in an array of organisms and exhibit characteristics which are similar to oil-derived plastics, yet have the added advantages of biodegradability and biocompatibility. Despite these advantages, P(3HB) production is currently more expensive than the production of oil-derived plastics, and therefore, more efficient P(3HB) production processes would be desirable. In this study, we describe the model-guided design and experimental validation of several engineered P(3HB) producing operons. In particular, we describe the characterization of a hybrid phaCAB operon that consists of a dual promoter (native and J23104) and RBS (native and B0034) design. P(3HB) production at 24 h was around six-fold higher in hybrid phaCAB engineered Escherichia coli in comparison to E. coli engineered with the native phaCAB operon from Ralstonia eutropha H16. Additionally, we describe the utilization of non-recyclable waste as a low-cost carbon source for the production of P(3HB).     

Tracking Host Sources of Cryptosporidium spp. in Raw Water for Improved Health Risk Assessment

Recent molecular evidence suggests that different species and/or genotypes of Cryptosporidium display strong host specificity, altering our perceptions regarding the zoonotic potential of this parasite. Molecular forensic profiling of the small-subunit rRNA gene from oocysts enumerated on microscope slides by U.S. Environmental Protection Agency method 1623 was used to identify the range and prevalence of Cryptosporidium species and genotypes in the South Nation watershed in Ontario, Canada. Fourteen sites within the watershed were monitored weekly for 10 weeks to assess the occurrence, molecular composition, and host sources of Cryptosporidium parasites impacting water within the region. Cryptosporidium andersoni, Cryptosporidium muskrat genotype II, Cryptosporidium cervine genotype, C. baileyi, C. parvum, Cryptosporidium muskrat genotype I, the Cryptosporidium fox genotype, genotype W1, and genotype W12 were detected in the watershed. The molecular composition of the Cryptosporidium parasites, supported by general land use analysis, indicated that mature cattle were likely the main source of contamination of the watershed. Deer, muskrats, voles, birds, and other wildlife species, in addition to sewage (human or agricultural) may also potentially impact water quality within the study area. Source water protection studies that use land use analysis with molecular genotyping of Cryptosporidium parasites may provide a more robust source-tracking tool to characterize fecal impacts in a watershed. Moreover, the information is vital for assessing environmental and human health risks posed by water contaminated with zoonotic and/or anthroponotic forms of Cryptosporidium.     

Thermophilous oak forests in the Czech Republic: Syntaxonomical revision of theQuercetalia pubescenti-petraeae

Using the Braun-Blanquet approach, a syntaxonomical revision of the order ofQuercetalia pubescenti-petraeae, based on the synthesis of 634 relevés, distinguished three alliances of the thermophilous oak forests in the Czech Republic: (1)Quercion pubescenti-petraeae (3 communities:Pruno mahaleb-Quercetum pubescentis, Lathyro versicoloris-Quercetum pubescentis, Corno-Quercetum), which includes peri-alpine and peri-Carpathian oak forests on calcareous bedrock, (2)Aceri tatarici-Quercion (2 communities:Quercetum pubescenti-roboris, Carici fritschii-Quercetum roboris), which comprises Pontic-Pannonian oak forests on loess or sand that occur in southern Moravia only, (3)Quercion petraeae (5 communities:Potentillo albae-Quercetum, Brachypodium pinnatum-Quercus robur community,Sorbo torminalis-Quercetum, Genisto pilosae-Quercetum petraeae, Asplenio cuneifolii-Quercetum petraeae), which includes Central European thermophilous oak forests distributed outside the range ofQuercus pubescens and some other submediterranean species.     

Heracleum mantegazzianum in the Czech Republic: Dynamics of spreading from the historical perspective

The spread ofHeracleum mantegazzianum in the Czech Republic has been reconstructed on the basis of floristic data. The species has spread from several spatially independent localities which have originated from the West Bohemian population introduced in the 19th century. Its expansion first followed the courses of great rivers. An exponential increase in the number of localities reported in particular years and in the cummulative number of localities was found. The species abundance decreases from the region of original introduction, which is located in the westernmost part of the country, to the east. The rate of spread in the last sixty years was the same in the Czech Republic and in Great Britain. The species is, however, more widespread in the North of Europe presumably because of the more favourable climate. It is found most frequently in habitats that encourage the greatest movement of diaspores. Floristic data can be used to infer retrospectively the dynamics of invasion, of a species in a given area provided that there is a sufficient amount of floristic research over time.     

Monitoring of ozone risk for forests in the Czech Republic: preliminary results

Ozone (O3) is supposed to represent a significant risk for the health of forest ecosystems in Central Europe. So far, however, its impact on stands growing under natural conditions has not been clearly proved. A new project of the National Agency for the Research in Agriculture is focused on the O3 effect on selected parameters of forest health. This paper presents the results of the first year of monitoring, 2005. In 2005, high O3 concentrations were measured, mainly in the spring. In the summer, due to wet and cold weather, the O3 load was comparatively low. In the plots investigated, the concentrations of O3 were higher with the altitude. The amount of epicuticular waxes on 1-year-old Norway spruce needles was the only factor showing significant correlation to O3 concentration. Defoliation of the stands depended only on the stand age. The amount of malondialdehyde (MDA), an oxidative stress marker, was related to the altitude, and only for European beech. The results are preliminary, as the summer O3 development was not typical in 2005, and the results may change over the next monitoring periods.     

Impact of mandatory microchipping on traceability of sheltered dogs in the Czech Republic

Records of 4,455 sheltered dogs were analyzed to compare the numbers of traceable (able to be reunited with original caregivers) and untraceable (unable to be reunited with original caregivers) dogs before (Period 1) and after (Period 2) introduction of a mandatory microchipping. Records were also used to compare the numbers of microchipped dogs, to compare dogs’ reunification with caregivers, and to estimate possible financial savings. In Period 1, 51.8% of the dogs were untraceable (median length of stay = 14 days) and 48.2% were traceable (median length of stay = 1 day). Of all traceable dogs, 24.5% were microchipped, and 67.5% of microchipped dogs were traceable. In Period 2, 57.1% of dogs were traceable (median length of stay = 0 days), and 42.9% were untraceable (median length of stay = 12 days). Of all traceable dogs, 61.4% had a microchip, and 80% of microchipped dogs were traceable. In Period 2, authorities saved more than US$43,400 on unreclaimed dog care. Mandatory microchipping of dogs appears to be an effective measure to increase reunification of sheltered dogs, decrease length of stay, and save authority funds.