Overcoming threats to montane amphibian: Approach to ecosystem conservation

Frogs and their pathogenic diseases

Amphibians are known as an ideal model for indicating environmental status because they
are vulnerable to the potential threats of global climate change and habitat destruction. They are
also susceptible to the emergent fungal pathogen called Batrachochytrium dendrobatidis (Bd), that
causes the chythridiomycosis disease which have been associated with the declines of their
population worldwide. Studies are conducting investigations with their survival in relation with
the fungal disease by modeling individual variation in susceptibility. This is not the only disease
that montane amphibians suffer from but also in recent studies the interaction of Bd and other
pathogenic factors can influence their development. Davidson et al., [5] investigated the potential
threat of Bd interaction to carbaryl pesticides and found out that it can inhibit their growth by a
half and it can suppress their production of skin peptides which increases the frog’s susceptibility
to catching the disease. Other diseases may contribute to the continuous decline of frog population
including the Saprolegnia fungal infections [2], iridovirus and bacterial infections [3], and
trematode infections [12]. Adults and sub-adults are highly infected by the disease compared to
their other life stages [17] but the mortality of juveniles is greatly increased when they are exposed
(see figure 1) [6].

Figure 1. Plots showing the estimated effects (solid lines) on Bd infection intensity of (A) Rana sierrae life stage, and
(B) day of the year. Plots are arranged in order of the strength of each predictor variable’s effect, and confidence
intervals (95%) are shown as dashed lines. Hatch marks above the x-axis in (B) indicate the observed values of the
predictor variable. In (B), x-axis values correspond to the following dates: 160 ¼ 10 June, 200 ¼ 20 July, and 240 ¼
29 August [6].

Amphibian skin microbes as defense for pathogenic diseases

Amphibians group are experiencing significant population reductions and extinctions as a
result of chytridiomycosis, making management and conservation research a top concern for
amphibian conservation biologists. Based on the threat of the fungal Bd to montane amphibians,
the risk of spreading it to the whole population is high. Several research has been done to devise
prevention methods to fight the spread and impacts of Bd in nature. One study prevented the
impacts of Bd by using bioaugmentation of the microbial community found in the frog’s skins by
introducing the Janthinobacterium lividum bacteria [10]. All the frogs that were exposed to the
bacteria+Bd treatments all survived and those exposed to the disease are mostly dead and one
individual showed a symptoms of advance-stage chytridiomycosis (see figure 2). Survival of frogs
was strongly associated with presence of violacein, an anti-Bd metabolite produced by J. lividum
[10]. However, violacein were only present in the frogs that bathe in J. lividum solution suggesting
that it acts in a way that it selectively influenced the growth of skin bacterial strains that produce
antibiotic metabolites when the amphibian is exposed to skin pathogens.

Figure 2. The effect of treatment group on survival [10].

The introduction of anti-Bd bacteria to the skin of the frog may be employed as a
conservation technique in the future.

Antifungal drugs development

Successful efforts on clinical trials were also done to safely treat adult infected frogs with
the antifungal drugs and intraconazole baths [2]. Although these treatments were very effective in
adults, the reported juveniles had been diagnosed with depigmentation [7] because early
developmental stages are highly sensitive to toxins. Controlled trials using PCR testing were used
to test different life stages on different species and showed that treatments using fluconazole
appeared to be benign for tadpoles at higher doses [16].

Other factors on decline of frog population

The impact of Bd on host populations may be connected to the density of amphibian hosts
and the variety of the host community aside from environmental factors. With higher host density,
there is an expected increased rate of transmission across individuals. Host variety can impact
pathogen prevalence by modifying host density, changing its behavior, and allowing weakly
reservoir-competent hosts to increase the power of infection [13]. While other alternative methods
can change the decline by increasing the population size of frogs to have a better chance of
surviving than the small size populations. The variation of individuals in a large population has a
higher chance to resist the impacts and threats of Bd disease from spreading and may improve their
survivability [14].
Results from resurveys of researchers showed that elevation has also a link to the spread
of Bd and mostly infect montane amphibians [8] and found that in warmer areas Bd is not common
[5]. As the season warms, adults may have a greater ability to resist since their immune capacity
is temperature sensitive [18] thus, leading to reduced prevalence later in the summer. Warmer
temperatures have been linked to be an effective preventive measure against the spread of
chytridiomycosis in some species.
Montane forests restoration efforts in Costa Rica
Deforestation and conversion of pasture lands are the primary cause of tropical montane
forest degradation in Costa Rica. An estimated 46% is covered with pasture lands [20] which were
subsequently used for agriculture and cattle grazing and the disturbances increased the presence
of non-native species in the area.
Holl et. al [11], investigated the factors that limit the recovery of the abandoned pasture in
Costa Rica and compared their results to overcome tropical montane forest degradation. Seed
dispersal, germination, and high predation are the most common limiting factors that may
contribute to the successful recovery (see figure 3).

Figure 3. Factors limiting pasture recovery. Processes that influence seedling establishment are in boxes with solid
lines. Factors affecting the magnitude of these processes are in boxes with dashed lines [11].

Artificial bird perching

Artificially placed perching structures attracts bird to the area which increases seed
dispersal capabilities that facilitates recovery, especially in disturbed forest [15]. Perches is
particularly effective in the rain forest where vegetations are adapted to developed mutualistic
relationship with animals. However, seed dispersal is low at open pasture areas [11] and it does
not facilitate other factors in recovery obstacles. By incorporating perches and efforts through
clipping and using herbicides may improve the rate of recovery [9].

Planting native species

Native tree species play a critical role in recovery of disturbed areas by increasing seed
dispersal through species recruitment [15], altering microclimatic conditions, and improving soil
nutrient composition [1]. Some tree species developed at faster rate when grown in native habitat
and in abandoned pasture areas [11], which may increase the success of forest recovery. It can also
serve as habitat for birds and Samuels [19], recorded 27 species were perching into remnant trees
in pasture lands. In addition, it increases the animal-dispersed seeds in pasture lands with remnant
trees compared to open pastures (see table 1).
Table 1. Number of animal- and wind-dispersed seeds falling in open pasture, below remnant pasture trees, and in
forest. Seeds were collected between 1 May 1995 and 30 April 1996. Values are means 6 1 SE for n 5 6 sites, which
are averages of three seed traps at each site [11].

Studies showed that leaf litters fall under the trees enhanced the growth of seedlings and
the animal dispersal improved similarly to forest habitats. Mycorrhizal fungi from trees help in the
growth of seedlings and the amount of sunlight under trees is favorable for optimal development
thus, these mechanisms increase the seedling growth [11].
Restoration strategies is crucial in understanding the forest ecology to do more experiments
to enhance the methods and acknowledge the factors that contribute to recovery. Restoring a large
area with native species can significantly improved the distribution of species and provide habitats
for them.

Conclusion
The high level of sensitivity to slight changes in climate and disturbances in montane forest
made the species that lives in it also susceptible to diseases. Although, some adult species can
tolerate the Bd infection, the likelihood of increase mortality rate among juveniles still persists.
Bioaugmentation of bacteria into the frog’s skin can only prevent it from spreading and treatment
of infected individuals may allow reintroduction and interact with native species. Whether to adopt
bioaugmentation technique to mend the spread of chytridiomycosis in nature presents a
challenging ecological question.
Antifungal drugs and continuous research efforts can mitigate the spread of Bd disease on
adults but juveniles are intolerant to toxins. By providing adequate evidence through monitoring
before trials on threatened species would be ideal to ensure the safety and feasible results.
Treatments that have been shown to be beneficial in captive populations can subsequently be tested
in the wild amphibians. This approach is expensive and evidence for safety and effectiveness of
trials were varied among species. Therefore, it needs intensive research and monitoring to optimize
fully the usage of antifungal solutions. It is important to understand the risk in environmental
problems interacting with Bd to predict an immediate and proper action in conservation of native
species in montane environments.
Habitat degradation is widespread worldwide and human activities posed a high threat to
destruction. The presence of limiting factors contributes to the slow progress of recovery in pasture
lands. Bird perching and planting native tree species showed an effective approach in restoring
small-scale areas of montane forest by comparing restoration strategies used in forest pasture
lands. Understanding these factors is crucial in creating an effective restoration program in the
future and determine which is the best strategies can be useful.
References:

[1] Belsky, A. J., R. G. Admunson, J. M. Duxbury, S. J. Riha, A. R. Ali, and S. M. Mwonga.

1989. The effects of trees on their physical, chemical, and biological environments in a
semi-arid savanna in Kenya. Journal of Applied Ecology 26:1005–1024.

[2] Berger, L.; Speare, R.; Pessier, A.; Voyles, J.; Skerratt, L.F. (2010). Treatment of
chytridiomycosis requires urgent clinical trials. Diseases Aquatic Organisms. 92: 165–174.

[3] Blaustein, A.R.; Hokit, D.G.; O’Hara, R.K.; Holt, R.A. 1994b. Pathogenic fungus
contributes to amphibian losses in the Pacific Northwest. BiologicalConservation. 67: 251–
254.

[4] Carey, C.; Cohen, N.; Rollins-Smith, L. 1999. Amphibian declines: an immunological
perspective. Developmental and Comparative Immunology 23: 459–472.

[5] Davidson C.; Benard, M.F.; Shaffer, H.B.; Parker, J.M.; O’Leary, C.; Conlon, J.M.;
Rollins-Smith, L.A. (2007). Effects of chytrid and carbaryl exposure on survival, growth
and skin peptide defenses in foothill yellow-legged frogs. Environmental Science and
Technology. 41: 1771–1776.

[6] Garcia, T.S.; Romansic, J.M.; Blaustein, A.R. (2006). Survival of three species of anuran
metamorphs exposed to UV-B radiation and the pathogenic fungus Batrachochytrium
dendrobatidis. Diseases of Aquatic Organisms. 72: 163–169.

[7] Garner TWJ, Garcia G, Carroll B, Fisher MC (2009) Using itraconazole to clear
Batrachochytrium dendrobatidis infection, and subsequent depigmentation of Alytes
muletensis tadpoles. Dis Aquat Org 83:257–260.

[8] Fisher, M.C., Garner, T.W.J., Walker, S.F., (2009). Global emergence of Batrachochytrium
dendrobatidis and amphibian chytridiomycosis in space, time, and host. Annual Review of
Microbiology 63, 291–310.

[9] Ferguson, B. G. 1995. Overcoming barriers to forest regeneration in a degraded tropical

pasture: an evaluation of restoration techniques. M.S. thesis. University of Michigan, Ann
Arbor.

[10] Harris R.; Brucker, R.; Minbiole, K.; Walke, J.; Becker, M.; Schwantes, C. et al. (2009).
Skin microbes on frogs prevent morbidity and mortality caused by a lethal skin fungus.
ISME Journal. 3(7): 818–824.

[11] Holl, K., Loik, M., Lin, E., Samuels, I. (2000). Tropical Montane Forest Restoration in
Costa Rica: Overcoming Barriers to Dispersal and Establishment. Restoration Ecology
Vol. 8 No. 4, pp. 339–349. doi.org/10.1046/j.1526-100x.2000.80049.x
[12] Johnson, P.T.; Lunde, K.B.; Ritchie, E.G.; Launer, A.E. 1999. The effect of trematode
infection on amphibian limb development and survivorship. Science.284: 802–804.

[13] Keesing, F., Holt, R.D., Ostfeld, R.S., 2006. Effects of species diversity on disease risk.
Ecology Letters 9, 485–498.

[14] Knapp, R.A.; Briggs, C.J.; Smith, T.C.; Maurer, J.R. (2011). Nowhere to hide: impact of a
temperature-sensitive amphibian pathogen along an elevation gradient in the temperate
zone. Ecosphere. 2: 1–26.

[15] McClanahan, T. R., and R. W. Wolfe. (1993). Accelerating forest succession in a

fragmented landscape: the role of birds and perches. Conservation Biology 7:279–288

[16] McInnes K (1999) An investigation into the treatment of chytrid fungus in the larval stages
of Australian amphibians. Honours thesis, Deakin University, Melbourne.

[17] Piovia-Scott, J.; Pope, K.L.; Lawler, S.P.; Cole, E.M.; Foley, J.E. (2011). Factors related
to the distribution and prevalence of the fungal pathogen Batrachochytrium dentrobatidis
in Rana cascadae and other amphibians in the Klamath Mountains. Biological
Conservation. 144: 2913–2921.

[18] Raffel, T.R., Rohr, J.R., Kiesecker, J.M., Hudson, P.J., (2006). Negative effects of
changing temperature on amphibian immunity under field conditions. Functional Ecology
20, 819–828.

[19] Samuels, I. A. 1998. The role of riparian corridors and isolated pasture trees in facilitating
bird movement in tropical pasture. B.S. thesis. University of California, Santa Cruz.

[20] World Resources Institute (WRI). (1998). World Resources 1998–1999. Oxford University
Press, Oxford.