scholarly journals The effect of captivity on the skin microbial symbionts in three Atelopus species from the lowlands of Colombia and Ecuador

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e3594 ◽  
Author(s):  
Sandra V. Flechas ◽  
Ailin Blasco-Zúñiga ◽  
Andrés Merino-Viteri ◽  
Valeria Ramírez-Castañeda ◽  
Miryan Rivera ◽  
...  

Many amphibian species are at risk of extinction in their natural habitats due to the presence of the fungal pathogen Batrachochytrium dendrobatidis (Bd). For the most highly endangered species, captive assurance colonies have been established as an emergency measure to avoid extinction. Experimental research has suggested that symbiotic microorganisms in the skin of amphibians play a key role against Bd. While previous studies have addressed the effects of captivity on the cutaneous bacterial community, it remains poorly studied whether and how captive conditions affect the proportion of beneficial bacteria or their anti-Bd performance on amphibian hosts. In this study we sampled three amphibian species of the highly threatened genus, Atelopus, that remain in the wild but are also part of ex situ breeding programs in Colombia and Ecuador. Our goals were to (1) estimate the diversity of culturable bacterial assemblages in these three species of Atelopus, (2) describe the effect of captivity on the composition of skin microbiota, and (3) examine how captivity affects the bacterial ability to inhibit Bd growth. Using challenge assays we tested each bacterial isolate against Bd, and through sequencing of the 16S rRNA gene, we identified species from thirteen genera of bacteria that inhibited Bd growth. Surprisingly, we did not detect a reduction in skin bacteria diversity in captive frogs. Moreover, we found that frogs in captivity still harbor bacteria with anti-Bd activity. Although the scope of our study is limited to a few species and to the culturable portion of the bacterial community, our results indicate that captive programs do not necessarily change bacterial communities of the toad skins in a way that impedes the control of Bd in case of an eventual reintroduction.

2017 ◽  
Vol 83 (9) ◽  
Author(s):  
Carly R. Muletz-Wolz ◽  
Graziella V. DiRenzo ◽  
Stephanie A. Yarwood ◽  
Evan H. Campbell Grant ◽  
Robert C. Fleischer ◽  
...  

ABSTRACT Diverse bacteria inhabit amphibian skin; some of those bacteria inhibit growth of the fungal pathogen Batrachochytrium dendrobatidis. Yet there has been no systematic survey of anti-B. dendrobatidis bacteria across localities, species, and elevations. This is important given geographic and taxonomic variations in amphibian susceptibility to B. dendrobatidis. Our collection sites were at locations within the Appalachian Mountains where previous sampling had indicated low B. dendrobatidis prevalence. We determined the numbers and identities of anti-B. dendrobatidis bacteria on 61 Plethodon salamanders (37 P. cinereus, 15 P. glutinosus, 9 P. cylindraceus) via culturing methods and 16S rRNA gene sequencing. We sampled co-occurring species at three localities and sampled P. cinereus along an elevational gradient (700 to 1,000 meters above sea level [masl]) at one locality. We identified 50 anti-B. dendrobatidis bacterial operational taxonomic units (OTUs) and found that the degree of B. dendrobatidis inhibition was not correlated with relatedness. Five anti-B. dendrobatidis bacterial strains occurred on multiple amphibian species at multiple localities, but none were shared among all species and localities. The prevalence of anti-B. dendrobatidis bacteria was higher at Shenandoah National Park (NP), VA, with 96% (25/26) of salamanders hosting at least one anti-B. dendrobatidis bacterial species compared to 50% (7/14) at Catoctin Mountain Park (MP), MD, and 38% (8/21) at Mt. Rogers National Recreation Area (NRA), VA. At the individual level, salamanders at Shenandoah NP had more anti-B. dendrobatidis bacteria per individual (μ = 3.3) than those at Catoctin MP (μ = 0.8) and at Mt. Rogers NRA (μ = 0.4). All salamanders tested negative for B. dendrobatidis. Anti-B. dendrobatidis bacterial species are diverse in central Appalachian Plethodon salamanders, and their distribution varied geographically. The antifungal bacterial species that we identified may play a protective role for these salamanders. IMPORTANCE Amphibians harbor skin bacteria that can kill an amphibian fungal pathogen, Batrachochytrium dendrobatidis. Some amphibians die from B. dendrobatidis infection, whereas others do not. The bacteria that can kill B. dendrobatidis, called anti-B. dendrobatidis bacteria, are thought to influence the B. dendrobatidis infection outcome for the amphibian. Yet how anti-B. dendrobatidis bacterial species vary among amphibian species and populations is unknown. We determined the distribution of anti-B. dendrobatidis bacterial species among three salamander species (n = 61) sampled at three localities. We identified 50 unique anti-B. dendrobatidis bacterial species and found that all of the tested salamanders were negative for B. dendrobatidis. Five anti-B. dendrobatidis bacterial species were commonly detected, suggesting a stable, functional association with these salamanders. The number of anti-B. dendrobatidis bacteria per individual varied among localities but not among co-occurring salamander species, demonstrating that environment is more influential than host factors in structuring the anti-B. dendrobatidis bacterial community. These anti-B. dendrobatidis bacteria may serve a protective function for their salamander hosts.


2015 ◽  
Vol 81 (19) ◽  
pp. 6589-6600 ◽  
Author(s):  
Jenifer B. Walke ◽  
Matthew H. Becker ◽  
Myra C. Hughey ◽  
Meredith C. Swartwout ◽  
Roderick V. Jensen ◽  
...  

ABSTRACTCurrently, it is estimated that only 0.001% to 15% of bacteria in any given system can be cultured by use of commonly used techniques and media, yet culturing is critically important for investigations of bacterial function. Despite this situation, few studies have attempted to link culture-dependent and culture-independent data for a single system to better understand which members of the microbial community are readily cultured. In amphibians, some cutaneous bacterial symbionts can inhibit establishment and growth of the fungal pathogenBatrachochytrium dendrobatidis, and thus there is great interest in using these symbionts as probiotics for the conservation of amphibians threatened byB. dendrobatidis. The present study examined the portion of the culture-independent bacterial community (based on Illumina amplicon sequencing of the 16S rRNA gene) that was cultured with R2A low-nutrient agar and whether the cultured bacteria represented rare or dominant members of the community in the following four amphibian species: bullfrogs (Lithobates catesbeianus), eastern newts (Notophthalmus viridescens), spring peepers (Pseudacris crucifer), and American toads (Anaxyrus americanus). To determine which percentage of the community was cultured, we clustered Illumina sequences at 97% similarity, using the culture sequences as a reference database. For each amphibian species, we cultured, on average, 0.59% to 1.12% of each individual's bacterial community. However, the average percentage of bacteria that were culturable for each amphibian species was higher, with averages ranging from 2.81% to 7.47%. Furthermore, most of the dominant operational taxonomic units (OTUs), families, and phyla were represented in our cultures. These results open up new research avenues for understanding the functional roles of these dominant bacteria in host health.


2018 ◽  
Vol 373 (1740) ◽  
pp. 20160508 ◽  
Author(s):  
Sarah Benson-Amram ◽  
Geoff Gilfillan ◽  
Karen McComb

Playback experiments have proved to be a useful tool to investigate the extent to which wild animals understand numerical concepts and the factors that play into their decisions to respond to different numbers of vocalizing conspecifics. In particular, playback experiments have broadened our understanding of the cognitive abilities of historically understudied species that are challenging to test in the traditional laboratory, such as members of the Order Carnivora. Additionally, playback experiments allow us to assess the importance of numerical information versus other ecologically important variables when animals are making adaptive decisions in their natural habitats. Here, we begin by reviewing what we know about quantity discrimination in carnivores from studies conducted in captivity. We then review a series of playback experiments conducted with wild social carnivores, including African lions, spotted hyenas and wolves, which demonstrate that these animals can assess the number of conspecifics calling and respond based on numerical advantage. We discuss how the wild studies complement those conducted in captivity and allow us to gain insights into why wild animals may not always respond based solely on differences in quantity. We then consider the key roles that individual discrimination and cross-modal recognition play in the ability of animals to assess the number of conspecifics vocalizing nearby. Finally, we explore new directions for future research in this area, highlighting in particular the need for further work on the cognitive basis of numerical assessment skills and experimental paradigms that can be effective in both captive and wild settings. This article is part of a discussion meeting issue ‘The origins of numerical abilities’.


2021 ◽  
Vol 4 ◽  
Author(s):  
Omneya Osman ◽  
Johan Andersson ◽  
Tomas Larsson ◽  
Mats Töpel ◽  
Alexander Eiler

National monitoring programs provide the basis for evaluating the integrity of ecosystems, their responses to disturbances, and the success of actions taken to conserve or recover biodiversity. In this study, we successfully established a national program for the invasive chytrid fungus Batrachochytrium dendrobatidis (Bd) based on dual TaqMan assays. Amphibian diversity based on metabarcoding of the mitochondrial 12S rRNA gene was also performed. Assays were optimized for sensitive detection of target species from a wide range of amphibian ponds with variable potential of inhibitions for eDNA based detection. An amphibian mock community of 5 species was used to validate the metabarcoding approach while internal standards were used in the case of the dual TaqMan assays. First sampling of over 170 ponds in Norway resulted in Bd detection in 12 environmental samples and one swab sample taken over multiple years indicating the establishment of Bd in Norway. Five amphibian species Bufo bufo, Lissotriton vulgaris, Triturus cristatus, Rana arvalis and Rana temporaria as predicted from data in long-term citizen science reporting systems were widely detected in the collected eDNA samples. Our large scale-monitoring program indicates a low risk of a Bd outbreak and amphibian decline caused by chytridiomycosis but continued monitoring is recommended in the future. These findings indicate that eDNA is an effective method to detect invasive species, and to monitor endangered amphibian species. Still, several shortcomings (such as PCR inhibitors and sample volume) were identified that need to be addressed to improve eDNA-based monitoring at the national level.


2010 ◽  
Vol 23 (6) ◽  
pp. 811-822 ◽  
Author(s):  
Michelle M. Gehringer ◽  
Jasper J. L. Pengelly ◽  
William S. Cuddy ◽  
Claus Fieker ◽  
Paul I. Forster ◽  
...  

The nitrogen-fixing cyanobacterium Nostoc is a commonly occurring terrestrial and aquatic cyanobacterium often found in symbiosis with a wide range of plant, algal, and fungal species. We investigated the diversity of cyanobacterial species occurring within the coralloid roots of different Macrozamia cycad species at diverse locations throughout Australia. In all, 74 coralloid root samples were processed and 56 endosymbiotic cyanobacteria were cultured. DNA was isolated from unialgal cultures and a segment of the 16S rRNA gene was amplified and sequenced. Microscopic analysis was performed on representative isolates. Twenty-two cyanobacterial species were identified, comprising mostly Nostoc spp. and a Calothrix sp. No correlation was observed between a cycad species and its resident cyanobiont species. The predominant cyanobacterium isolated from 18 root samples occurred over a diverse range of environmental conditions and within 14 different Macrozamia spp. Phylogenetic analysis indicated that endosymbionts were not restricted to previously described terrestrial species. An isolate clustering with Nostoc PCC7120, an aquatic strain, was identified. This is the first comprehensive study to identify the endosymbionts within a cycad genus using samples obtained from their natural habitats. These results indicate that there is negligible host specialization of cyanobacterial endosymbionts within the cycad genus Macrozamia in the wild.


2018 ◽  
Vol 11 ◽  
pp. 00004
Author(s):  
Irina Barsukova ◽  
Alexei Astashenkov

Reproductive biology and biological productivity of Prunella vulgaris L. were studied in natural growth conditions in Khakasiya and under introduction. It was established that in the wild a potential seed productivity of the species varied from 85,4 to 160,3 it./ a shoot, a real one – from 73,6 to 149,2 it.; erem germination under optimal conditions reached 90,5%. Under introduction the indicators of seed productivity increase by 2-5 times, of erem germination – by 1,5 times. Biological productivity of individuals in cultivation is by 5-10 times higher compared to that in natural habitats.


2015 ◽  
Vol 282 (1805) ◽  
pp. 20142881 ◽  
Author(s):  
Matthew H. Becker ◽  
Jenifer B. Walke ◽  
Shawna Cikanek ◽  
Anna E. Savage ◽  
Nichole Mattheus ◽  
...  

Symbiotic microbes can dramatically impact host health and fitness, and recent research in a diversity of systems suggests that different symbiont community structures may result in distinct outcomes for the host. In amphibians, some symbiotic skin bacteria produce metabolites that inhibit the growth of Batrachochytrium dendrobatidis (Bd), a cutaneous fungal pathogen that has caused many amphibian population declines and extinctions. Treatment with beneficial bacteria (probiotics) prevents Bd infection in some amphibian species and creates optimism for conservation of species that are highly susceptible to chytridiomycosis, the disease caused by Bd. In a laboratory experiment, we used Bd-inhibitory bacteria from Bd-tolerant Panamanian amphibians in a probiotic development trial with Panamanian golden frogs, Atelopus zeteki , a species currently surviving only in captive assurance colonies. Approximately 30% of infected golden frogs survived Bd exposure by either clearing infection or maintaining low Bd loads, but this was not associated with probiotic treatment. Survival was instead related to initial composition of the skin bacterial community and metabolites present on the skin. These results suggest a strong link between the structure of these symbiotic microbial communities and amphibian host health in the face of Bd exposure and also suggest a new approach for developing amphibian probiotics.


2021 ◽  
pp. 18-26
Author(s):  
Christopher Michaels

The mountain chicken frog (Leptodactylus fallax) is among the 42 % of amphibians threatened with extinction and is dependent upon ex situ populations to recover in the wild. Amphibian captive husbandry is not fully understood and empirical data are required to optimise protocols for each species in captivity. Calcium metabolism and homeostasis are areas of importance in captive husbandry research and have been identified as a challenge in maintaining ex situ populations of L. fallax. We trialled two frequencies (twice and seven times weekly) of calcium supplementation via dusting of feeder insects in two groups of L. fallax juveniles and measured growth and health effects through morphometrics, radiography, ultrasonography and blood and faecal analysis over 167 days, followed by a further 230 days of monitoring on an intermediate diet informed by the initial dataset. We showed that supplementation treatment did not affect growth or health status as measured through blood analysis, radiography and ultrasonography. More frequent supplementation resulted in significantly more radiopaque endolymphatic sacs and broader skulls. Frogs fed more calcium excreted twice as much calcium in their faeces. The intermediate diet resulted in previously lower supplementation frogs approximating the higher supplementation frogs in morphometrics and calcium stores. Comparison with radiographic data from wild frogs showed that both treatments may still have had narrower skulls than wild animals, but mismatching age class may limit this comparison. Our data may be used to inform dietary supplementation of captive L. fallax as well as other amphibians.


2018 ◽  
Vol 27 (Supplement) ◽  
pp. 81-90
Author(s):  
A.A. Cunningham

The unexplained decline of amphibian populations across the world was first recognised in the late 20th century. When investigated, most of these “enigmatic” declines have been shown to be due to one of two types of infectious disease: ranavirosis caused by infection with FV3-like ranavirus or with common midwife toad virus, or chytridiomycosis caused by infection with Batrachochytrium dendrobatidis or B. salamandrivorans. In all cases examined, infection has been via the human-mediated introduction of the pathogen to a species or population in which it has not naturally co-evolved. While ranaviruses and B. salamandrivorans have caused regionally localised amphibian population declines in Europe, the chytrid fungus, B. dendrobatidis, has caused catastrophic multi-species amphibian population declines and species extinctions globally. These diseases have already caused the loss of amphibian biodiversity, and over 40% of known amphibian species are threatened with extinction. If this biodiversity loss is to be halted, it is imperative that regulations are put in place – and enforced – to prevent the spread of known and yet-to-be discovered amphibian pathogens. Also, it is incumbent on those who keep or study amphibians to take measures to minimise the risk of disease spread, including from captive animals to those in the wild.


2013 ◽  
Vol 82 (4) ◽  
pp. 351-355 ◽  
Author(s):  
Vojtech Baláž ◽  
Martina Kubečková ◽  
Petr Civiš ◽  
Roman Rozínek ◽  
Jiří Vojar

A parasitic fungus,Batrachochytrium dendrobatidisis now recognised as an important factor in the amphibian biodiversity crisis. Toad species of the genusBufoare among those susceptible to infection by the pathogen in Europe. The aim of this study was to observe the presence and impact of infection in adults of two toad species collected for captive breeding. The total number of animals included in the study was 162, but only subsets were used for sampling at different occasions (35 specimens in the initial sampling in summer 2011, 48 post hibernation during winter 2011, and 31 in summer 2012, after all toads in captivity were treated with itraconazole). We performed TaqMan real-time quantification PCR to detect and quantify the pathogen. We found that a large infection load was linked to mortality in a single adult green toad (Bufo viridis). However, low infection loads observed in fiveB. viridisand five natterjack toads (B. calamita)were lost over time, with no apparent adverse effect. Intraconazole treated animals were all clear of infection. As infection in these two toad species either led to mortality or recovery, it seems unlikely they could act as permanent carriers ofB. dendrobatidisand therefore persistence of the pathogen is likely maintained by different host species. This is the first study to date that has detected infection and observed its impact and persistence in wild-infected toads in Europe.


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