Response of Elk to Changes in Plant Production and Nutrition Following Prescribed Burning

Abstract: Researchers have ascribed use of areas by grazers after burning to changes in plant community structure, community composition, nutritional quality, and seasonal availability. Researchers can better evaluate these alternatives if they monitor changes in plant communities following burning concurrently with changes in animal use. We examined responses of elk (Cervus elaphus) to prescribed burning of areas dominated by sagebrush (Artemisia spp.) in south-central Montana, USA, within which we monitored changes in plant production, nutritional quality, and community composition and diversity from 1989 to 1999. Elk increased use of burned sites 1–2 years after burning, then reduced use to levels associated with preburn conditions over the next 3–10 years. Burning transformed low-diversity, sagebrush-dominated communities into relatively high-diversity, grass- and forb-dominated communities that persisted for 10 years, but forage biomass and protein content declined on burned sites after initial short-term increases. Changes in elk use closely tracked changes in production and nutritional quality of plants. Therefore, we concluded that increases in quantity and quality of forage were the primary cause for increased use of burned sites by elk. Managers may observe only short-term responses from elk following burning but can expect longer-term increases in plant diversity and persistence of grass—forb communities on burned sites for >10 years that may be important to elk or other grazing ungulates.     

Patch selection by red deer in relation to energy and protein intake: a re-evaluation of Langvatn and Hanley's (1993) results

Langvatn and Hanley (1993) recently reported that patch use by red deer (Cervus elaphus) was more strongly correlated with short term rates of intake of digestible protein than dry matter. Such short term measures overlook effects of gut filling, which may constrain intake by ruminants over longer time scales (i.e., daily rates of gain). We reanalyzed Langvatn and Hanley's data using an energy intake model incorporating such a processing constraint, to determine whether their conclusions are robust. We found that the use of patches by red deer was just as strongly correlated with an estimate of the daily rate of intake of digestible energy as one of digestible protein during four out of seven trials, but slightly lower in three out of seven trials. In all cases, daily intake of digestible energy was a much better predictor of patch preference by red deer than was the intake of dry matter. Our reanalysis suggests that the daily intake of energy was highly correlated with that of protein in these trials, as may often be the case for herbivores feeding on graminoids. Hence the observed pattern of patch use by red deer could simultaneously enhance rates of both protein and energy intake.     

Recovery after Experimental Cutting and Burning in Three Shrub Communities with Different Dominant Species

The aim of this study is to compare the recovery dynamics in three shrub communities subjected to experimental burning and cutting, and situated on an altitudinal gradient. Climatic features are different in each area, but all had the common characteristic of very homogeneous vegetation cover before the disturbances, with only one shrub species clearly dominant, a different taxon in each area, and with different regeneration strategies. The first area was a heathland dominated by Calluna vulgaris, situated at an altitude of 1600 m, with a continental climate (mean annual precipitation 1320 mm). The second area was a heathland dominated by Erica australis, located at an altitude of 1000 m (mean annual precipitation 840 mm). The third area was a Cistus ladanifer shrubland, located at 900 m altitude, with a Mediterranean climate similar to that of the previous area, but with lower mean annual precipitation (470 mm). Erica australis recovers by vegetative resprouting, but Cistus ladanifer is an obligate seeder, as is Calluna vulgaris in these areas. Each experimental disturbance was carried out over 100 m² in each area. Post-fire recovery is faster in Cistus ladanifer: 2 years after burning there was 40% cover vs. less than 20% in the other two species. However, recovery after cutting was similar for Cistus ladanifer and Erica australis. Calluna vulgaris recovers very slowly, with cover values below 20% even 10 years after both disturbances. Cover of dominant shrub species is negatively correlated with cover of herbaceous species. So different recovery of dominant species lead a different community dynamic in each area.     

Sucking and allosucking duration in farmed red deer (Cervus elaphus)

Sucking duration in ungulates does not only mean milk transfer, but is also associated with maternal care in general. It seems to be a reflection of offspring demand rather than solely milk transfer rate. Thus, the objective of this study was to discriminate between sucking and allosucking (i.e. sucking non-maternal hind) behaviour in red deer according to the sucking duration.We hypothesized that: (1) calves should suck longer from their mothers than allosuck from non-maternal hinds; (2) sucking duration of calves frequently nursed by a particular non-maternal hind should be longer than that of calves occasionally allonursed; (3) sucking duration should be longer for bouts including one calf than two or more calves sucking simultaneously; (4) male calves should suck and allosuck longer than female calves; and (5) primiparous hinds should nurse and allonurse longer than multiparous hinds. We observed sucking behaviour of 25 hinds and their 38 calves (from birth until the youngest calf reached one month of age) in two seasons. We recorded 1730 sucking bouts, of which 11.62% in the first season and 4.37% in the second season were non-filial. The duration of filial sucking was significantly longer than non-filial sucking. A large individual variance in the incidence of non-filial sucking in both the calves and hinds was found. Therefore, the non-filial hind–calf pairs were categorized in two clusters according to the frequency of nursing non-filial calves for one hind in relationship to all nursing events for this hind by a cluster analysis (PROC CLUSTER, SAS). We used a general linear mixed model, GLMM (PROC MIXED, SAS) to test the influence of hind relationship to the nursed calf (filial, frequently allosucking non-filial, or occasionally allosucking non-filial pair). Sucking duration of occasionally allosucking non-filial calves was only marginally different from that of filial calves. There was no difference between the two groups of non-filial calves. Multiple sucking bouts were shorter than those with one calf. Male calves sucked longer than female calves; however, the greatest difference was recorded between frequently allosucking non-filial pairs of both sexes. Frequently allosucking non-filial males sucked the longest and differently from occasionally allosucking non-filial males. Frequently allosucking non-filial females sucked the shortest and differently from filial calves of both sexes. It is more likely that allosucking seems to be more important for male rather than female calves.Therefore, it is concluded that allosucking calves differ in their sucking behaviour and two types of allosuckers (frequent and occasional) should be taken into account when analyzing allosuckling behaviour.     

Genetic diversity, gene flow and drift in Bavarian red deer populations (Cervus elaphus)

The red deer (Cervus elaphus) populationof Bavaria in Southern Germany was severelyreduced during the 19th century due toover-hunting. The species has since recoveredwithin designated areas. Subsequent habitatfragmentation presumably has changed thegenetic structure of Bavarian red deer.In order to assess the genetic diversity, weanalysed samples obtained from nine differentBavarian and two adjacent (Thueringen andCzech-Republic) red deer populations,genotyping 19 microsatellite loci. Our analysesrevealed moderate and significant differencesin diversity. Referring to assignment tests,the genetic differentiation of Bavarian reddeer was sufficient to assign an individual'sorigin to the correct population at an averageof 91.6%. The correlation of genetic andgeographic distance matrices revealed noevidence for isolation by distance. Thecoalescent model analysis suggests that thegenetic structure used to be characterized by adrift–gene flow equilibrium and is nowinfluenced by drift and disruption of the geneflow. Only three of the examined populationsshowed a probability of less than 10% that twogenes within these populations share a commonancestor (F _IS-value). Twopopulations had high F _IS values,indicating the influence of drift.Additionally, the intrapopulation indicesrevealed a low variability in thesepopulations. The estimated effective populationsizes (N _e) were generally in thesame range as the actual population sizes. Theinbreeding rates, based on the estimated N _e, and the inbreeding coefficientssuggested that the Bavarian red deerpopulations are in a stable state.     

Changes in the floristic composition of a Quercus coccifera L. garrigue in relation to different fire regimes

Plant Ecology - To analyse the impact of fire on plants, an experiment has been set up in a Quercus coccifera L. garrigue near Montpellier. The objectives of the study were to follow the changes of...     

Drought effects on fine-root and ectomycorrhizal-root biomass in managed Pinus oaxacana Mirov stands in Oaxaca, Mexico

The effects of a severe drought on fine-root and ectomycorrhizal biomass were investigated in a forest ecosystem dominated by Pinus oaxacana located in Oaxaca, Mexico. Root cores were collected during both the wet and dry seasons of 1998 and 1999 from three sites subjected to different forest management treatments in 1990 and assessed for total fine-root biomass and ectomycorrhizal-root biomass. Additionally, a bioassay experiment with P. oaxacana seedlings was conducted to assess the ectomycorrhizal inoculum potential of the soil for each of the three stands. Results indicated that biomasses of both fine roots and ectomycorrhizal roots were reduced by almost 60% in the drought year compared to the nondrought year. There were no significant differences in ectomycorrhizal and fine-root biomass between the wet and dry seasons. Further, the proportion of total root biomass consisting of ectomycorrhizal roots did not vary between years or seasons. These results suggest that both total fine-root biomass and ectomycorrhizal-root biomass are strongly affected by severe drought in these high-elevation tropical pine forests, and that these responses outweigh seasonal effects. Forest management practices in these tropical pine forests should consider the effects of drought on the capacity of P. oaxacana to maintain sufficient levels of ectomycorrhizae especially when there is a potential for synergistic interactions between multiple disturbances that may lead to more severe stress in the host plant and subsequent reductions in ectomycorrhizal colonization.     

Production and nutritional quality of the rotifer Brachionus plicatilis fed marine Chlorella sp. at different cell densities

The effect of different cell densities of marine Chlorella sp. on the growth rate, doubling time and production of the rotifer Brachionus plicatilis was investigated. A significant increase in rotifer production was achieved at a density of 50 × 10⁶ Chlorella cells ml^–1. The nutritional quality of rotifers grown at different concentrations of Chlorella is discussed.     

Importance of a pilot study for non-invasive genetic sampling: genotyping errors and population size estimation in red deer

Population size information is critical for managing endangered or harvested populations. Population size can now be estimated from non-invasive genetic sampling. However, pitfalls remain such as genotyping errors (allele dropout and false alleles at microsatellite loci). To evaluate the feasibility of non-invasive sampling (e.g., for population size estimation), a pilot study is required. Here, we present a pilot study consisting of (i) a genetic step to test loci amplification and to estimate allele frequencies and genotyping error rates when using faecal DNA, and (ii) a simulation step to quantify and minimise the effects of errors on estimates of population size. The pilot study was conducted on a population of red deer in a fenced natural area of 5440 ha, in France. Twelve microsatellite loci were tested for amplification and genotyping errors. The genotyping error rates for microsatellite loci were 0–0.83 (mean=0.2) for allele dropout rates and 0–0.14 (mean=0.02) for false allele rates, comparable to rates encountered in other non-invasive studies. Simulation results suggest we must conduct 6 PCR amplifications per sample (per locus) to achieve approximately 97% correct genotypes. The 3% error rate appears to have little influence on the accuracy and precision of population size estimation. This paper illustrates the importance of conducting a pilot study (including genotyping and simulations) when using non-invasive sampling to study threatened or managed populations.     

Individual variation in migratory and exploratory movements and habitat use by adult red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis> L.) in a mountainous temperate forest

Using data from a global positioning system (GPS), seven adult red deer (Cervus elaphus L.) were tracked in the Parc National des Cévennes, southern France, between November 1998 and December 2000 to assess the factors affecting large-range movement patterns and habitat use. The home range varied from a single compact area for females to three distinct seasonal ranges for males, which used alternative migratory strategies (i.e. non-, downward- and upward-migrants). The migrants used mainly southerly and easterly aspects, and wintered in areas having steeper slopes than were used during summer or the rut season. For males, the time of rut migration was mid-September and they finally entered wintering ranges from mid-December to the beginning of January. Exploratory behaviour (i.e. individuals found outside the limits of their familiar area but returning to it a few days later) occurred in both sexes and for all individuals monitored during at least a 6-month period. Velocity and efficiency of exploratory movements were higher than usual movements. During these exploratory movements, hinds may have used different landscape attributes (elevation, slope, canopy cover) while stags did not. These results provide new empirical information that could be used for building and applying broad-scale spatial and landscape use models in ecological research.     

大兴安岭南段马鹿生境适宜性分析与生态廊道构建

构建生态廊道在缓解生境破碎化对生物多样性的影响、维持濒危物种的遗传多样性、维护自然生态系统结构完整与功能稳定方面具有重要作用。以内蒙古大兴安岭南段分布的马鹿(Cervus elaphus)种群为研究对象,利用MaxEnt模型对其生境适宜性进行分析,并利用最小累积阻力模型构建潜在生态扩散廊道,探讨大兴安岭南段区域隔离马鹿种群的栖息地连通方案。结果显示,马鹿栖息地呈破碎化状态,种群有明显的隔离分布趋势,现有适宜栖息地具有海拔较低(800—1200 m)、坡度较缓(<15°)、靠近水源、植被类型多为靠近山林的灌丛或草地等特点。所构建12条生态廊道具有经过河流浅水节段、远离村落等特点,便于落实栖息地生态恢复管理措施。研究从区域尺度综合分析了大兴安岭南段马鹿栖息地现状及连通性,有助于优化适宜栖息地格局,促进马鹿扩散和栖息地连通,为该物种隔离种群及其栖息地保护规划提供现实指导和基础资料。     

Use of mires and food habits of sika deer in the Oze Area,central Japan

Sika deer Cervus nippon entered the Oze Area, a snowy area in central Japan, in the 1990s. The use of mires and the food habits of this species were studied in 1999 and 2000. Deer used the mires immediately after snow melted and did not use the mires in mid summer, although they did use them lightly in autumn. The summer food habits differed from those of the deer population living in Nikko in the lower area: non-Sasa graminoids and other monocotyledonous leaves occupied more (approximately 30%) of the feces in the former population, whereas Sasa nipponica, a dwarf bamboo, was the dominant food (60%) of the latter population. Seasonal changes in fecal composition in the Oze deer were that mire graminoids occupied a considerable portion of their food in spring (11.6%) and summer (17.0%), whereas the amount of dwarf bamboo increased in fall (26.0%). Despite the small size of the mires, Oze deer appear to prefer mires to forests. Nitrogen concentrations of the mire plants did not differ from those of the forest plants; however, foraging efficiency would be greater in the mires and this may explain the preference observed. Despite the small total biomass, leaf biomass of the mire was equivalent to that of the adjacent forest floor. In addition, biomass was concentrated near the ground, or the biomass concentration was greater than that of the forest floor.     

Status of the regal fritillary (<Emphasis Type="Italic">Speyeria idalia</Emphasis>) and effects of fire management on its abundance in northeastern Kansas,USA

The Regal Fritillary (Speyeria idalia), which once occupied prairies and meadows in North America from the upper Great Plains to the Atlantic coast, has disappeared in recent decades from nearly the entirety of the eastern half of its range and has declined westward. In the Great Plains, where the species is limited to native prairie remnants, several large populations are thought to exist, but patterns of occurrence and abundance in the region have not been described in detail. We surveyed prairies within a three county area of northeastern Kansas using distance-sampling along line transects and found Regal Fritillaries present at 70 of 87 sites. Population density varied considerably among sites but was generally much higher at those that had not been burned in the past year. Despite the loss of >99% of its original prairie landcover and the small sizes of remnants we estimate that our study area supports a globally significant population of ∼12,000 adult individuals. Given the rapidity of decline of Regal Fritillary populations elsewhere, this study establishes important population benchmarks and a practical protocol for future monitoring efforts.     

基于红外相机技术的贺兰山同域分布阿拉善马鹿和岩羊活动规律研究

为了探究同域分布的阿拉善马鹿(Cervur alashanicus)与岩羊(Pseudois nayaur)的日活动节律,及两者在时间生态位上的分化情况。于2014.10—2015.10,在贺兰山利用红外相机技术,对两者进行野外监测。相关监测数据,在R软件中利用"overlap"统计包,采取核密度估计(Kernel density estimation)、重叠指数(Coefficient of overlap):■进行分析。结果表明,马鹿日活动节律模式属于晨昏活动类型(日活动高峰期:5:00—10:00、16:00—21:00),存在季节性变化(冷暖季日活动节律重叠指数■=0.77),由暖季到冷季晨昏活动高峰期均向中午移动(暖季:4:00—8:00、16:00—22:00,冷季:6:00—11:00、15:00—20:00),昼间活动强度大于暖季,而夜间低于暖季。岩羊日活动节律为主要在昼间活动的模式,其活动高峰期在8:00—10:00、13:00—15:00两个时间段。由暖季到冷季存在季节性变化特征(■=0.74),昼间活动高峰期向后推迟(暖季7:00—9:00、12:00—14:00,冷季9:00—11:00、13:00—17:00),夜间活动降低。两者在日活动节律模式上的相似度较高(■=0.67),存在冷季(■=0.66)高于暖季(■=0.61)的季节变化。在日活动节律模式分离特征为:马鹿为晨昏活动模式,岩羊为昼间活动模式;前者昼间活动强度低于后者,而夜间强于后者;两者活动在高峰期上大体错开。本研究不仅提供了马鹿与岩羊的日活动节律模式,而且还分析了两者在时间生态位上的分化方式和共存机制,并为两者行为生态学研究的深入提供必要的科学依据。     

Multiple herbivores and coevolutionary interactions in an Ipomopsis hybrid swarm

Studies focusing on pairwise interactions between plants and herbivores may not give an accurate picture of the overall selective effect of herbivory, given that plants are often eaten by a diverse array of herbivore species. The outcome of such interactions may be further complicated by the effects of plant hybridization. Hybridization can lead to changes in morphological, phenological and chemical traits that could in turn alter plant–herbivore interactions. Here we present results from manipulative field experiments investigating the interactive effects of multiple herbivores and plant hybridization on the reproductive success of Ipomopsis aggregata formosissima X I. tenuituba. Results showed that ungulate herbivores alone had a net positive effect on plant relative fitness, increasing seed production approximately 2-fold. Caterpillars had no effect on plant relative fitness when acting alone, with caterpillar-attacked plants producing the same number of flowers, fruits and seeds as the uneaten controls. Caterpillars, however, significantly reduced flower production of ungulate browsed plants. Flower production in these plants, however, was still significantly greater (approximately 1.7-fold greater) than uneaten controls, likely leading to an increase in reproductive success through the paternal component of fitness given that fruit and seed production was not significantly different from that of herbivore-free controls. Although results suggest that herbivore imposed selection is pairwise, ungulates likely have a large influence on the abundance of, and hence the amount of damage caused by, caterpillar herbivores. Thus, because of the ecological interactions between ungulates and caterpillars, selection on Ipomopsis may be diffuse rather than pairwise, assuming such interactions translate into differential effects on plant fitness as herbivore densities vary. Plant hybridization had no significant effect on patterns of ungulate or caterpillar herbivory; i.e., no significant interactions were detected between herbivory and plant hybridization for any of the fitness traits measured in this study nor did plant hybridization have any significant effect on host preference. These results may be due to patterns of introgression or the lack of species-specific differences between I. aggregate formosissima and I. tenuituba. Plant hybridization per se resulted in lowered reproductive success of white colored morphs due in part to the effects of pollination. Although it appears that there would be strong directional selection favoring darker flower colors due to the lower reproductive success of the white colored morphs in the short run, the natural distribution of hybrids suggest that over the long run selection either tends to average out or there are no fitness differences among morphs in most years due to the additive fitness effects of hawkmoth and hummingbird pollinators.     

Production of Perennial Vegetation in an Oasis-desert Transition Zone in NW China - Allometric Estimation,and Assessment of Flooding and Use Effects

River oases at the southern fringe of the Taklamakan desert in NW China are surrounded by belts of spontaneous vegetation that protect the oases from sand drift. As an important source of forage, fuel and construction wood, this foreland vegetation is also a component part of the agricultural system of the oases but has been, and still is, destroyed through overuse. Within a broader study that aimed to provide a basis for a sustainable management of this foreland vegetation, biomass and production were studied in four vegetation types dominated either by Alhagi sparsifolia, Calligonum caput-medusae, Populus euphratica, or Tamarix ramosissima that were thought to occur under different regimes of natural flooding in the foreland of Qira (Cele) oasis, Xinjiang, NW China. Shoot biomass components were closely correlated to basal area (Calligonum, Populus, Tamarix) or shrub volume and projection area (Alhagi), enabling non-destructive estimation of stand biomass from shoot diameters or shrub dimensions with sufficient precision using allometric regression equations. Relationships between shoot basal area and biomass of the woody species (Calligonum, Populus and Tamarix) agreed with predictions by a theoretical model of plant vascular systems, suggesting that they are determined by hydraulic and mechanical requirements for shoot architecture. Average aboveground biomass densities of typical stands in late summer were 2.97 Mg/ha in Alhagi, 3.6 Mg/ha in a row plantation and 10.9 Mg/ha in homogenous stands of Calligonum, 22–29 Mg/ha in 22 year-old Populus forests and 1.9–3.1 Mg/ha in Tamarix-dominated vegetation. Annual aboveground production including wood and assimilation organs ranged from 2.11 to 11.3 Mg/ha in plantations of Calligonum, 3.17 to 6.12 Mg/ha in Populus, and 1.55 to 1.74 Mg/ha (based on total ground area) or 3.10 to 7.15 Mg/ha (in homogenous stands) in Tamarix. Production of Alhagi is equal to peak biomass. A thinning treatment simulating use by the local population enhanced productivity of Calligonum, Populus and Tamarix. A complete harvest of Alhagi in late August decreased production in the following year. An artificial flood irrigation treatment did not sufficiently increase soil water content except in the uppermost layer and had no clear beneficial effect on growth of the four species and even a negative effect on Alhagi, which was due to increased competition from annual species. As biomass and production with or without artificial irrigation were much higher than values expected for rain-fed desert vegetation at a mean annual precipitation of 35 mm, it is concluded that the existence of all vegetation types studied is probably based on permanent access to groundwater and that natural floods or precipitation do not contribute to their water supply. The effects of agricultural groundwater use in the oasis on groundwater in the foreland of the oasis need further study. Sustainable use of this productive vegetation is possible but requires proper management.     

Changes in Diet and Food Consumption of Largemouth Bass Following Large-scale Hydrilla Reduction in Lake Seminole, Georgia

Largemouth bass Micropterus salmoides Lacepède growth (in length) increased an average of 14% and bioenergetics modeling predicted a 38% increase in total annual food consumption following a large-scale reduction of hydrilla Hydrilla verticillata L.f. Royle in Spring Creek, a 2,343-ha embayment of Lake Seminole, Georgia. Coverage of submersed aquatic vegetation (SAV) declined from 76% to 22% in 1 year due to a drip-delivery fluridone treatment. In contrast, largemouth bass growth only increased an average of 4% and bioenegetics modeling predicted a 13% increase in total food consumption over the same time period in the Chattahoochee River embyament, where SAV coverage naturally declined from 26% to 15%. Diets were collected from a total of 4,409 largemouth bass over a 2.5-year period in the two embayments; the primary diet item (by weight) for largemouth bass in both embayments was sunfish (mostly Lepomis spp.). Diets before and after SAV reduction were generally similar for fish greater than stock-size (≥203 mm) in the Spring Creek arm; however, fewer invertebrates were consumed after SAV reduction. Low diet similarity was observed in smaller fish, caused by a decline in consumption of grass shrimp and sunfishes and an increase in use of damselflies, shiners Notropis spp., and topminnows Fundulus spp. after SAV reduction. Diets were similar between the same time periods for all sizes of fish in the Chattahoochee River arm. These results agreed with many laboratory results describing the effects of aquatic plant density on largemouth bass food consumption and growth, and demonstrated that increased predation efficiency resulting from decreased plant abundance was likely a stronger factor determining growth rates than any potential diet shift that may occur as a result in vegetation decline.     

Use of plant extracts and biocontrol agents for the management of brown spot disease in rice

Fifty plant extracts, four oil cakes and eight antagonistic organisms were tested against Bipolaris oryzae (Cochliobolus miyabeanus), the causal agent of brown spot disease of rice. In vitro studies indicated that two leaf extracts, Nerium oleander and Pithecolobium dulce exerted the higher percent inhibition to mycelial growth (77.4, 75.1%) and spore germination (80.3, 80.0%) of B. oryzae. Among the four oil cake extracts tested in vitro against B. oryzae, neem cake extract showed the maximum inhibition percent to mycelial growth (80.18%) and spore germination (81.13%) of the pathogen followed by mahua cake extract, castor and gingelly cake extract. Trichoderma viride (Tv2) was significantly effective in inhibiting the mycelial growth (62.92%) and spore germination (77.03%) of the pathogen followed by Trichoderma harzianum (Th5) and Trichoderma reesei (Tr3). The promising leaf extracts, oil cake extracts and antagonistic microorganisms were further evaluated for their efficacies in disease management under glasshouse and field conditions. In glasshouse studies, post-infectional spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) was significantly effective in reducing the incidence of brown spot of rice by 66, 52 and 45 percent respectively. Two rounds of spraying of rice plants with neem cake extract, N. oleander leaf extract and T. viride (Tv2) in the field at initial appearance of disease and 15 days later reduced the incidence of brown spot (70, 53 and 48% disease reduction respectively) and increased the yield by 23, 18 and 15 percent respectively.     

Production of the main surface antigen of Toxoplasma gondii in tobacco leaves and analysis of its antigenicity and immunogenicity

We adapted a previously described Agrobacterium-mediated transient expression system to test the expression level of three constructs carrying the surface antigen 1 (SAG1) of Toxoplasma gondii. Two constructs were based in a Potato virus X (PVX) amplicon. In one of them, the PVX movement protein genes were replaced by the sag1 gene. In the other, the sag1 gene was placed under the control of an additional coat protein subgenomic promoter. In the third construct, the sag1 gene was fused to an apoplastic peptide signal under the CaMV 35S promoter. Western blot analysis of leaf extracts infiltrated with each construct revealed a protein of 35 kDa. SAG1 accumulation in leaves ranged from 0.1 to 0.06% of total soluble protein (equivalent to 10 μg and 6 μg of SAG1 per gram of fresh leaf tissue, respectively). Three of five human seropositive samples reacted with tobacco-expressed SAG1 in Western blot analysis. The C3H mice were immunized with SAG-expressing leaf extracts and perorally challenged with a nonlethal dose of the T. gondii Me49 strain. Mice vaccinated with SAG1 showed significantly lower brain cyst burdens compared to those from the control group. Immunization with SAG1-expressing leaves elicited a specific humoral response with predominant participation of type IgG2a. In conclusion, a functional SAG1 version could be transiently expressed in tobacco leaves.