Introduction
One benefit of invasive plant treatment for a land manager is the opportunity for native plants to grow in the treatment site (Boyce Reference Boyce2015). Of special concern are endangered species that may be particularly vulnerable to the effects of the invasive plants (Albrecht and McCue Reference Albrecht and McCue2009). For example, invasive shrubs may be threatening the survival of populations of rare herbaceous perennial plants found in the cedar glades of the Tennessee central basin. The cedar glades are named for the eastern redcedar (Juniperus virginiana L.; which is not a true cedar) found in these forest openings, which also harbor a suite of grasses and herbaceous perennials adapted to full sunlight, spring flooding, summer drought, and shallow soils with extensive exposed limestone bedrock (Baskin and Baskin Reference Baskin and Baskin2004). One of these herbaceous perennials is wild dill [Perideridia americana (Nutt. ex DC.) Rchb.; also called eastern Eulophus and eastern yampah], a geophyte that is state listed as an endangered species in Tennessee (Crabtree Reference Crabtree2016). The species is also state listed as endangered in Indiana (Indiana Department of Natural Resources 2016), as threatened in Kentucky (Kentucky State Nature Preserves Commission 2011), and as extirpated in Ohio (Ohio Division of Wildlife 2015) but is not listed at the federal level (USDA 2018). In Tennessee, P. americana is restricted to the cedar glades in the state’s central basin where Baskin and Baskin (Reference Baskin and Baskin1993) studied the species and found that P. americana emerges in March and shoot senescence occurs in June. The nonnative shrub Amur honeysuckle [Lonicera maackii (Rupr.) Herder] has invaded Tennessee central basin cedar glades (Cofer et al. Reference Cofer, Walck and Hidayati2008) and shades out early vernal native species such as P. americana, because it grows taller than the herbaceous perennials and its leaf out occurs before the native shrub species leaf out and at roughly the same time P. americana emerges (Chen and Matter Reference Chen and Matter2017).
Research on L. maackii treatment has investigated the response of perennial herbaceous species following the treatment of L. maackii plants. Miller and Gorchov (Reference Miller and Gorchov2004) found greater growth and reproduction for the spring-flowering perennial narrowleaf wild leek [Allium burdickii (Hanes) A.G. Jones] following L. maackii treatment. Christopher et al. (Reference Christopher, Matter and Cameron2014), Peebles-Spencer et al. (Reference Peebles-Spencer, Gorchov and Crist2017), Shields et al. (Reference Shields, Saunders, Gibson, Zollner, Dunning and Jenkins2015), and Ward et al. (Reference Ward, Williams and Linske2017) demonstrated that treatment of L. maackii was related to an increase in spring herbaceous perennials.
The overpopulation of white-tailed deer (Odocoileus virginianus Zimmermann) in the eastern United States has resulted in the overbrowsing of herbaceous and woody plants (Loeb et al. Reference Loeb, Germeraad, Griffin and Ward2011). This effect gives rise to the possibility of O. virginianus causing a reduced recovery of herbaceous perennials after L. maackii treatments. Christopher et al. (Reference Christopher, Matter and Cameron2014) found overabundant O. virginianus population caused a reduction in the population of spring perennial species following L. maackii treatment. Ward et al. (Reference Ward, Williams and Linske2017) showed the response of short perennial species to L. maackii treatment peaked after 3 yr, and O. virginianus browsing reduced the population of short perennial species. In contrast, Peebles-Spencer et al. (Reference Peebles-Spencer, Gorchov and Crist2017) reported that O. virginianus did not have a significant effect on spring perennial species after L. maackii treatment.
Lonicera maackii treatment and endangered species management, including P. americana, are two Radnor Lake State Natural Area, Nashville, TN (RLSNA), land-management goals. However, O. virginianus population size control is not a land-management goal (Barnes Reference Barnes2016). Also, long-term L. maackii treatment efforts have been successful at RLSNA (Loeb et al. Reference Loeb, Germeraad, Treece, Wakefield and Ward2010). This research focuses on the response of the endangered plant P. americana to L. maackii treatments in five cedar glades located in RLSNA. The first adaptive management research question (Loeb Reference Loeb2011) is: Will the treatment of L. maackii be followed by an increase in the P. americana plant population when comparing pretreatment and posttreatment P. americana population sizes? Odocoileus virginianus have overbrowsed tree seedlings and saplings in RLSNA (Loeb et al. Reference Loeb, Germeraad, Griffin and Ward2011, Reference Loeb, King and Helton2015) but no evidence of O. virginianus browsing of P. americana was observed before L. maackii treatment. The second adaptive management research question is: Does the post–L. maackii treatment P. americana plant population size change from P. americana leaf out to flowering?
Materials and Methods
Site Description and Perideridia americana Biology
RLSNA is located on the southern boundary of Nashville, TN (Loeb et al. Reference Loeb, Germeraad, Treece, Wakefield and Ward2010). Cedar glades in RLSNA are located on the Fernvale and Arnheim Limestone Formations, which are below the Fort Payne Chert and above the Chattanooga Shale (Wilson and Miller Reference Wilson and Miller1972). Only small parts of the Fernvale and Arnheim Formations are covered with cedar glades. As well, only seven cedar glades in RLSNA contain P. americana growing on some segment of the cedar glade. Two of the seven cedar glades containing P. americana have fewer than 10 P. americana plants, and these two sites were not selected for treatment. The five research sites are remote locations within the nearly 540-ha area of RLSNA. No formal trails lead to the five cedar glades. The public is not permitted to go off the formal trails in RLSNA. The names and elevations of the centers of the five research sites in RLSNA are: Cheek, 313 m; East Hall Farm, 319 m; Harris 2, 300 m; Hideaway, 313 m; and Norfleet, 297 m. Longitudes and latitudes for the research sites are not provided to protect the P. americana plants from being harvested. However, the authors welcome requests for a guided visit to a research site.
The only disturbance and historical information concerning P. americana sites in RLSNA was found in the Tennessee Division of Natural Areas Biotics Database. There were four records dated May 17, 1979. Three of the sites contained 2 or fewer plants; however, one site had about 1,000 P. americana plants (P Somers and L Smith, personal communication). A search for the four sites in 2013 revealed P. americana was not found at two sites. In contrast, Cheek is the place noted as having approximately 1,000 plants, and Hideaway is a location that had 2 plants in 1979.
Perideridia americana is a perennial herb with tuberous storage roots in a cluster of two or more radiating from the stem and slender delicate leaves that dry completely after flowering but before fruiting (Chuang Reference Chuang1970). In RLSNA, P. americana leaf out starts in early March and flowers appear by the beginning of May. Greenhouse research on P. americana indicates: 51% seed germination in the first year; less than 1% seed germination after the first year; only cotyledons develop in the first growing season; and flowering does not occur until the third growing season, with the vast majority of plants flowering in the fifth year (Baskin and Baskin Reference Baskin and Baskin1993). Bidwell and Bidwell (Reference Bidwell and Bidwell2018) suggest P. americana is susceptible to O. virginianus browsing.
Plant Survey and Statistical Analysis
The narrow Fernvale and Arhheim Limestone Formations and small extent of P. americana in the cedar glades permitted the entire population of P. americana at pretreatment to be counted in a grid of 60 adjacent 2 m by 4 m plots laid out in six rows of 10 plots at each cedar glade, except for the Hideaway site. The Hideaway site is on the ridgeline, which left a triangle of limestone 4-m wide at the base and 8-m long to the tip of the triangle at the lower elevation boundary of the plots, and this area was not included in the research. The highest-elevation boundary of the plots was placed at the contact of the Fort Payne Chert with the Fernvale and Arhheim Limestone Formations, and the lowest-elevation row of plots encompassed the boundary of the Fernvale and Arnheim Formations and the Chattanooga Shale.
Hideaway and East Hall Farm were the first two sites discovered to contain P. americana in 2013. The decision to begin to conduct the research was made in the winter of 2014. Grid placement was done after leaf out in the Hideaway and East Hall Farm sites during the second week of March 2014. The grid of plots was placed at Cheek following flowering in the first week of May 2014. As additional sites were discovered, the grids were placed at Harris 2 after leaf out in the second week of March 2015 and at Norfleet following flowering in the first week of May 2015. The posttreatment counts were done in the second week of March 2018 for leaf out and the first week of May 2018 for flowering.
Prior research demonstrated that repeated L. maackii treatment resulted in lower counts of L. maackii plants than treatment in only 1 yr at RLSNA (Loeb et al. Reference Loeb, Germeraad, Treece, Wakefield and Ward2010). However, the prior research did not encompass the cedar glades. No research is available concerning relationships between L. maackii and P. americana. Greenhouse research showed that P. americana first flowers 3 to 4 yr after seed germination (Baskin and Baskin Reference Baskin and Baskin1993); therefore, posttreatment counts were done at least 3 yr after treatment. The adaptive management research questions called for counting plants, which was performed by counting the individual root crowns to determine individual L. maackii and P. americana plants. The L. maackii and P. americana plants were counted pretreatment, which was immediately after grid placement and before the first treatment of L. maackii plants at each site. The treatment of L. maackii in the sites ceased after 2016. To match the timing of the pretreatment counts, the posttreatment counts for L. maackii and P. americana were in the second week of March 2018 for East Hall Farm, Harris 2, and Hideaway and in the first week of May 2018 for Cheek and Norfleet. Perideridia americana was also counted in the second week of March and the first week of May 2018 at all five sites for a comparison of posttreatment leaf out and flowering.
Log natural–transformed counts were used to provide normally distributed values for the statistical tests. Mixed-model analysis (P-value of 0.05 was the significance level) was done to first determine whether the five sites were similar and thus could be treated as representing the entire RLSNA. Mixed-model analysis was also used to determine whether the sites with 3 yr since treatment were different from the sites with 4 yr since treatment. The mixed model was: Perideridia americana = Treatment (fixed factor) + Lonicera maackii (fixed covariate) + site (fixed factor) + years (fixed factor) + plot (random factor). For each of the five sites as different research locations, t-tests (P-value of 0.05 was the significance level) were used to assess whether there were significant differences between pretreatment and posttreatment for L. maackii and P. americana and leaf out and flowering for P. americana in 2018. Statistics were calculated using SPSS Amos v. 25 (IBM, 1 New Orchard Road, Armonk, New York 10504-1722).
Lonicera maackii Treatment
With extraordinary attention given to not disturb the P. americana plants, L. maackii plants were pulled from the ground, except for the few shrubs > 1-m tall that were treated following the standard technique at RLSNA for larger L. maackii plants (Loeb et al. Reference Loeb, Germeraad, Treece, Wakefield and Ward2010). The stems of shrubs >1-m tall were cut with a lopper at the time the smaller plants were pulled. The day of cutting off the stems, a backpack sprayer (Solo, 5100 Chestnut Avenue, Newport News, VA 23605) was used to apply glyphosate (Rodeo®, Dow AgroSciences, 9330 Zionsville Road, Indianapolis, IN 46268) at a concentration of 20 ml L−1 to the L. maackii stumps when there was more than 4 h of direct sunlight remaining, no prediction of rain for 3 d, and the temperature exceeded 4 C. The treatment of L. maackii plants <1-m tall described above occurred once annually from the first treatment through 2016, but the treatment of plants >1-m tall was only needed and performed during the first treatment. Treatment was ceased in 2017 to enable posttreatment comparison. Finally, the adaptive management research question does not ask whether the treatment process had an impact on the P. americana plants despite the best efforts to not disturb P. americana plants. Instead, the adaptive management research question inquires whether the P. americana population increases after the L. maackii treatment was performed.
Results and Discussion
The mixed-model analysis found that only the East Hall Farm and Harris 2 sites were similar. The comparison of number of years of treatment, that is 3 yr versus 4 yr, also showed no significant difference. Based on these results, the following analysis will focus on the individual sites and not consider the five sites as representing the entire RLSNA. In addition, the number of years since treatment will not be used to group the five separate sites. The statistical analysis indicates the results from the five sites cannot be generalized. Therefore, comparisons with flowering perennials similar to P. americana and cedar glade–like environments are limited.
The means and standard deviations for: pretreatment and posttreatment counts at the five sites are presented for L. maackii in Table 1 and P. americana in Table 2; and the posttreatment comparison of P. americana counts at leaf out and flowering are presented in Table 3. The t-test comparison of means for pretreatment and posttreatment L. maackii counts indicate all five sites had significant (P-value < 0.001) decreases in L. maackii (Table 1). The largest decrease in the means for L. maackii plants occurred for the East Hall Farm site at 2.25 plants m−2 which had the largest pretreatment mean at 2.88 plants m−2. The smallest decrease in the means was 0.09 plants m−2 which occurred for the Norfleet site that also had the smallest pretreatment mean of 0.12 plants m−2. The pretreatment means for Cheek, Harris 2 and Hideaway were between 1.65 and 1.94 plants m−2 and the decreases ranged from 1.31 to 1.77 plants m−2. The L. maackii treatment land management goal was achieved because there were significantly fewer L. maackii shrubs at posttreatment in all five cedar glades in RLSNA.
Table 1 Means and standard deviations for Lonicera maackii plant counts in 60 plots at pretreatment and posttreatment in the Cheek, East Hall Farm, Harris 2, Hideaway, and Norfleet cedar glades located in Radnor Lake State Natural Area, Nashville, TN. a
a Comparison of pretreatment versus posttreatment used a t-test of the log natural–transformed plant counts for the 60 plots at pretreatment and posttreatment. The dates of pretreatment and posttreatment were: Cheek pretreatment, May 2014, and posttreatment, May 2018; East Hall Farm pretreatment, March 2014, and posttreatment, March 2018; Harris 2 pretreatment, May 2014, and posttreatment, May 2018; Hideaway pretreatment, March 2014, and posttreatment, March 2018; Norfleet pretreatment, May 2015, and posttreatment, May 2018. Lonicera maackii treatment ceased in 2016 to provide a year for recovery before posttreatment counts.
b An asterisk (*) indicates a significant t-test comparison at 0.05 level.
Table 2 Means and standard deviations for Perideridia americana plant counts in 60 plots at pretreatment and posttreatment in the Cheek, East Hall Farm, Harris 2, Hideaway, and Norfleet cedar glades located in Radnor Lake State Natural Area, Nashville, TN. a
a Comparison of pretreatment versus posttreatment used a t-test of the log natural–transformed plant counts for the 60 plots at pretreatment and posttreatment. The dates of pretreatment and posttreatment were: Cheek pretreatment, May 2014, and posttreatment, May 2018; East Hall Farm pretreatment, March 2014, and posttreatment, March 2018; Harris 2 pretreatment, May 2014, and posttreatment, May 2018; Hideaway pretreatment, March 2014, and posttreatment, March 2018; Norfleet pretreatment, May 2015, and posttreatment, May 2018.
b An asterisk (*) indicates a significant t-test comparison at 0.05 level.
Table 3 Means and standard deviations for Perideridia americana plant counts in 60 plots at posttreatment leaf out in March 2018 and flowering in May 2018 for the Cheek, East Hall Farm, Harris 2, Hideaway, and Norfleet cedar glades located in Radnor Lake State Natural Area, Nashville, TN. a
a Comparison of leaf out versus flowering used a t-test of the log natural–transformed plant counts for the 60 plots at leaf out in March 2018 and flowering in May 2018.
b An asterisk (*) indicates a significant t-test comparison at 0.05 level.
The t-test comparisons of means for pretreatment and posttreatment P. americana counts showed significant increases in Cheek, Harris 2, Hideaway, and Norfleet, but East Hall Farm had an increase that was not significant (Table 2). East Hall Farm had the smallest mean for P. americana at pretreatment at 0.07 plants m−2 and the smallest mean at posttreatment at 0.12 plants m−2 among the five cedar glades. Considering the change from pretreatment to posttreatment, the largest increase in P. americana plants occurred in Cheek at 1.74 plants m−2. Cheek had the second-largest pretreatment population at 2.39 plants m−2. Cheek also had the second-largest posttreatment population of P. americana at 4.13 plants m−2. The largest pretreatment P. americana population was in Hideaway at 5.87 plants m−2. Hideaway had the largest posttreatment population at 7.43 plants m−2. The greatest percentage increase for P. americana (353%) occurred at Norfleet, but this increase represents only 0.76 plants m−2.
In regard to the first research question, the treatment resulted in a significant decrease for L. maackii, and an increase of P. americana at all five sites in RLSNA, but the P. americana increase was not significant at East Hall Farm. These outcomes at the five sites are in agreement with the research on L. maackii treatment effectiveness and the response of flowering perennials post–L. maackii treatment conducted by Christopher et al. (Reference Christopher, Matter and Cameron2014), Miller and Gorchov (Reference Miller and Gorchov2004), Peebles-Spencer et al. (Reference Peebles-Spencer, Gorchov and Crist2017), Shields et al. (Reference Shields, Saunders, Gibson, Zollner, Dunning and Jenkins2015), and Ward et al. (Reference Ward, Williams and Linske2017).
The second research question was answered with the P. americana at all five sites showing decreases from leaf out to flowering during the posttreatment period, but the only significant decreases were at Cheek and Hideaway (Table 3). Hideaway had the largest mean for posttreatment leaf out at 7.43 plants m−2. The largest change in the means from leaf out to flowering was a decrease of 4.48 plants m−2 that occurred at Hideaway. The mean at leaf out for Cheek was close to Hideaway, at 6.18 plants m−2, but the decrease at Cheek of 2.05 plants m−2 was less than half of what occurred at Hideaway. The range of differences for the means at leaf out and flowering for East Hall Farm, Harris 2, and Norfleet was 0.11 to 0.53 plants m−2.
The P. americana losses from leaf out to flowering and lack of significance for the increase from pretreatment to posttreatment at the East Hall Farm site can be explained by the conclusion of Baskin and Baskin (Reference Baskin and Baskin1993) that P. americana is susceptible to disturbance. One apparent cause of disturbance is O. virginianus, which have overbrowsed tree seedlings and saplings in RLSNA (Loeb et al. Reference Loeb, Germeraad, Griffin and Ward2011). Although P. americana floral head browsing was found at Hideaway in 2018, O. virginianus trampling of the delicate P. americana plants appears to have caused the losses evident in comparing the leaf out to flowering results as well as the nonsignificant P. americana population increases at East Hall Farm. The research on the effects of O. virginianus following L. maackii treatment of Christopher et al. (Reference Christopher, Matter and Cameron2014) and Ward et al. (Reference Ward, Williams and Linske2017) is in agreement with decreases at the five sites in RLSNA. However, the research of Peebles-Spencer et al. (Reference Peebles-Spencer, Gorchov and Crist2017) did not reveal losses in perennial herbs related to O. virginianus. Reducing the impact of O. virginianus by fencing the portions of the cedar glades containing P. americana was not done, because the fencing would be a location signal for P. americana poachers. The alternative of hunting is not allowed in RLSNA. Installing fencing and reducing the O. virginianus population also would not be well accepted by the public (Loeb et al. Reference Loeb, Germeraad, Griffin and Ward2011).
Identifying a comparable species to P. americana that was examined in prior research on the response of perennial species following L. maackii treatment (Christopher et al. Reference Christopher, Matter and Cameron2014; Cipollini et al. Reference Cipollini, Ames and Cipollini2009; Miller and Gorchov Reference Miller and Gorchov2004; Peebles-Spencer et al. Reference Peebles-Spencer, Gorchov and Crist2017; Shields et al. Reference Shields, Saunders, Gibson, Zollner, Dunning and Jenkins2015; Swab et al. Reference Swab, Zhang and Mitsch2007; Ward et al. Reference Ward, Williams and Linske2017) is a challenge. One candidate from a different taxonomic family is Canadian wildginger (Asarum canadense L.), which does share characteristics with P. americana of being a spring perennial with a tuberous root system. Asarum canadense is not listed as endangered at the federal level or by any state, but it is listed as threatened in Maine (Maine Natural Areas Program 2015). Jull (Reference Jull2001) indicates O. virginianus do not favor A. canadense. The research of Cipollini et al. (Reference Cipollini, Ames and Cipollini2009) reveals few effects from L. maackii treatment or O. virginianus damage on transplanted A. canadense. The study of Swab et al. (Reference Swab, Zhang and Mitsch2007) showed higher abundance and frequency of A. canadense plants in sites with no L. maackii clearance than cleared sites. The research on L. maackii treatment and A. canadense shows the opposite response to the increases for P. americana after L. maackii treatment in RLSNA. Likewise, P. americana experienced losses from leaf out to flowering related to O. virginianus trampling, but O. virginianus had little effect on A. canadense. Although land managers may expect an increase in a native species following L. maackii treatment, the A. canadense results demonstrate that this expectation may not be met.
This research on treatment of the invasive shrub L. maackii in RLSNA cedar glades containing the endangered perennial P. americana was the first step in an iterative land-management methods assessment process (Loeb Reference Loeb2011). Intensive, annual L. maackii clearing is no longer needed, because the invasive treatment resulted in a significant reduction in L. maackii and was followed by an expansion of the P. americana population. Therefore, in the future, periodic treatment of L. maackii plants with glyphosate will occur before P. americana leaf out or after P. americana senescence. An alternative recommended by Cipollini et al. (Reference Cipollini, Ames and Cipollini2009) would be to perform basal treatment on L. maackii and leave dead stems standing to reduce O. virginianus damage. An implication of this treatment method is the L. maackii plants would not be treated for many years to permit the shrubs to grow to sufficient size to be an impediment to O. virginianus transit. Regardless of which L. maackii treatment method is selected in the future, the populations of L. maackii and P. americana will be monitored to determine whether the P. americana population shows a decline in more than 1 yr. If a 2-yr decline occurs, without an obvious explanation such as prolonged drought or extended cold in the spring, then L. maackii treatment will be implemented. If additional invasive species appear in the future, as sometimes occurs in treatment areas (Loeb Reference Loeb2011), then the new invasive species will be treated as well.
Land managers can benefit from a full explanation of the research basis for the adaptive management decisions made regarding treatment of the invasive species L. maackii for the endangered species P. americana. The research method implemented to measure the pretreatment versus posttreatment population changes for the endangered and invasive species was based on long-term research on L. maackii treatment at RLSNA (Loeb et al. Reference Loeb, Germeraad, Treece, Wakefield and Ward2010) and research information concerning the ecological life cycle of P. americana (Baskin and Baskin Reference Baskin and Baskin1993). The key biological characteristic that led to the placement of 2 m by 4 m permanent plots is L. maackii shrubs will become reestablished in a treatment site despite repeated treatments. The reproductive biology of P. americana led to waiting at least 3 yr for the pretreatment versus posttreatment comparison, because P. americana requires at least 3 yr after seed germination to display flowers. The treatment method of L. maackii removal by pulling out the plants required giving extraordinary attention to not disturbing the individual P. americana plants. This focus on protecting the endangered species also was thoroughly adhered to during subsequent invasive treatments to eliminate new or reestablished L. maackii plants. The invasive species treatment was ended 1 yr before the posttreatment measurement to enable a pretreatment versus posttreatment comparison for the invasive species. A second benefit of permanent plots is that periodic monitoring of the endangered species as well as the invasive species can continue to be done to inform management decisions in the future, such as changing the invasive species treatment method.
Based on observations that the P. americana populations did not show evidence of browsing, we did not expect detrimental effects from O. virginianus, even though we were aware that the endangered species P. americana was sensitive to disturbance. We adapted the research methodology to include the comparison of plant populations at leaf out and at flowering in order to provide data on the damage caused by O. virginianus disturbance. Looking to the future, we are considering additional research on an invasive treatment change. This invasive management research would compare the recommendation of Cipollini et al. (Reference Cipollini, Ames and Cipollini2009) to perform basal treatment and leave the stems in the plot to reduce O. virginianus damage with our method of pulling out the plants from the plots. Implicit in testing the management change is monitoring the P. americana and L. maackii in the plots to determine whether the P. americana population declines as the L. maackii plants grow to sufficient size to serve as impediments to the O. virginianus.
Acknowledgments
We appreciate permission to conduct the research, provision of records, and assistance with field work provided by Roger McCoy, Director, Division of Natural Areas, Tennessee Department of Environment and Conservation. Thanks are extended to RLSNA Rangers Brandon Jarrett and Paul Schauer as well as former rangers Dustin Crowell, Jesse Germeraad, and Samuel King for assistance with fieldwork. We appreciate the plot grid preparation work performed by the AmeriCorps team members: Douglas Campbell, Matthew Eaton, Ashley Foust, Kailyn Gramly, Maya Ortiz, Elizabeth Richmond, William Titus, and Tony Williams. We value the assistance with data entry provided by Kiersten Stauffer and Taylor Walborn. We thank Kevin Quinlan and Christian Schmid of the Statistical Consulting Center, Department of Statistics, Pennsylvania State University, for their advice on and assistance with statistical analysis. The support for this research provided by the Friends of Radnor Lake, DuBois Educational Foundation, and Robert and Joyce Umbaugh Faculty Research Fund is deeply appreciated. No conflicts of interest have been declared.
