scholarly journals DNA-based biomonitoring in the tropics: Detection and control of Batrachochytrium dendrobatidis in Ecuadorian ecosystem

2021 ◽  
Vol 4 ◽  
Author(s):  
Lenin Riascos-Flores ◽  
Andrea Carrera ◽  
Leopoldo Naranjo ◽  
Jomira Yanez ◽  
Peter Goethals ◽  
...  
Keyword(s):  
Invasive Species ◽  
Water Samples ◽  
Large Scale ◽  
Batrachochytrium Dendrobatidis ◽  
Action Plan ◽  
Detection Methods ◽  
Population Declines ◽  
Amphibian Species ◽  
Tissue Samples ◽  
And Control

Batrachochytrium dendrobatidis (Bd) is a fungus that parasites vertebrates, and is associated with population declines worldwide in endemic amphibian species. As such, it is one of several invasive species which pose a serious threat to a variety of vertebrate hosts, in casu: amphibians. Detection of such invasive species is generally based on DNA-based methods where, for instance, swabs or tissue samples of candidate hosts are analysed for their presence. Any management strategy of these invasive species would greatly benefit from sensitive and rapid detection methods which can be applied at a large scale. The analysis of eDNA from the habitat of candidate host organisms may hold significant potential for this purpose. In this study, we compare the ability of eDNA from habitat samples with that of swab and/or tissue samples of candidate hosts to detect the presence of Bd in Ecuador. We collected individuals from the amphibians: Pristimantis (Anura: Craugastoridae), Rhinella (Anura: Bufonidae), Gastroteca (Anura: Hemiphractidae), from the endangered toad species of the genus Atelopus (Anura: Bufonidae) as well as water samples from different water bodies in Andean and coastal Ecuadorian areas. Samples were processed using a portable field molecular laboratory. Commercial primers for the internal transcribed spacer (ITS), in combination with a new set of primers designed from Bd sequences from tropical countries, were used. Positive PCR results from both types of samples were obtained within eight hours after sampling. Prevalence of BD was detected in eDNA, swab and tissue samples in four of the six ecosystems monitored -14 out of 26 water samples and 27 out of 43 amphibian of in total 12 species- including three endangered toad species (Atelopus balios, A. nanay, and the rediscovered A. bomolochos). Our results highlight the potential of eDNA-based monitoring to assess the presence and prevalence of Bd in Ecuadorian aquatic ecosystems, in accordance with the National Action Plan for the Conservation of Ecuadorian Amphibians. Furthermore, our field lab approach leads to reliable and fast results for the monitoring of invasive species in a tropical context of a pandemic.

scholarly journals Ancestral chytrid pathogen remains hypervirulent following its long coevolution with amphibian hosts

2019 ◽  
Vol 286 (1904) ◽  
pp. 20190833 ◽  
Author(s):  
Minjie Fu ◽  
Bruce Waldman
Keyword(s):  
Batrachochytrium Dendrobatidis ◽  
Population Declines ◽  
Amphibian Species ◽  
South Korean ◽  
Global Pandemic ◽  
Litoria Caerulea ◽  
Experimental Treatments ◽  
Amphibian Chytrid Fungus ◽  
Genomic Recombination ◽  
Blank Solution

Many amphibian species around the world, except in Asia, suffer morbidity and mortality when infected by the emerging infectious pathogen Batrachochytrium dendrobatidis (Bd). A lineage of the amphibian chytrid fungus isolated from South Korean amphibians (BdAsia-1) is evolutionarily basal to recombinant global pandemic lineages (BdGPL) associated with worldwide amphibian population declines. In Asia, the Bd pathogen and its amphibian hosts have coevolved over 100 years or more. Thus, resilience of Asian amphibian populations to infection might result from attenuated virulence of endemic Bd lineages, evolved immunity to the pathogen or both. We compared susceptibilities of an Australasian amphibian, Litoria caerulea , known to lack resistance to BdGPL, with those of three Korean species, Bufo gargarizans , Bombina orientalis and Hyla japonica , after inoculation with BdAsia-1, BdGPL or a blank solution. Subjects became infected in all experimental treatments but Korean species rapidly cleared themselves of infection, regardless of Bd lineage. They survived with no apparent secondary effects. By contrast, L. caerulea , after infection by either BdAsia-1 or BdGPL, suffered deteriorating body condition and carried progressively higher Bd loads over time. Subsequently, most subjects died. Comparing their effects on L. caerulea , BdAsia-1 induced more rapid disease progression than BdGPL. The results suggest that genomic recombination with other lineages was not necessary for the ancestral Bd lineage to evolve hypervirulence over its long period of coevolution with amphibian hosts. The pathogen's virulence may have driven strong selection for immune responses in endemic Asian amphibian host species.

scholarly journals Body size mediates latitudinal population differences in response to Bd infection in two amphibian species.

2021 ◽  
Author(s):  
Sara Meurling ◽  
Maria Cortazar-Chinarro ◽  
Mattias Siljestam ◽  
David Ahlen ◽  
Erik Agren ◽  
...  
Keyword(s):  
Body Size ◽  
Batrachochytrium Dendrobatidis ◽  
Geographical Origin ◽  
Bufo Bufo ◽  
Northern Region ◽  
Population Declines ◽  
Rana Arvalis ◽  
Amphibian Species ◽  
Survival Difference ◽  
Species Survival

Populations of the same species may differ in their sensitivity to pathogens but the factors behind this variation are poorly understood. Moreover, infections may cause sub-lethal fitness effects even in species resistant or tolerant to disease. The chytrid fungus Batrachochytrium dendrobatidis (Bd), is a generalist pathogen which has caused amphibian population declines worldwide. In many species, Bd infection causes the disease chytridiomycosis, often leading to high mortality. We investigated how geographical origin affects tolerance to Bd by exposing newly metamorphosed individuals of two North European amphibians (moor frog Rana arvalis, common toad Bufo bufo) from two latitudinal regions to two different BdGPL strains. Bd exposure strongly lowered survival in B. bufo, and in both species survival was lower in the northern region, this difference being much stronger in B. bufo. Northern individuals were smaller in both species, and the survival difference between the regions was size-mediated with smaller individuals being more sensitive to Bd. In both species, Bd exposure led to sub-lethal effects in terms of reduced growth suggesting that even individuals surviving the infection may have reduced fitness mediated by smaller body size. Bd strain affected size-dependent mortality differently in the two regions. We discuss the possible mechanisms how body size and geographical origin can contribute to the present results.

scholarly journals Nanomaterials and Nanotechnology-Associated Innovations against Viral Infections with a Focus on Coronaviruses

Nanomaterials ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1072 ◽  
Author(s):  
Mahmoud Nasrollahzadeh ◽  
Mohaddeseh Sajjadi ◽  
Ghazaleh Jamalipour Soufi ◽  
Siavash Iravani ◽  
Rajender S. Varma
Keyword(s):  
Large Scale ◽  
Viral Infections ◽  
Vaccine Development ◽  
Human Life ◽  
Detection Methods ◽  
Functionalized Nanoparticles ◽  
Critical Issues ◽  
Pathogenic Viruses ◽  
And Control ◽  
Diagnostic Detection

Viral infections have recently emerged not only as a health threat to people but rapidly became the cause of universal fatality on a large scale. Nanomaterials comprising functionalized nanoparticles (NPs) and quantum dots and nanotechnology-associated innovative detection methods, vaccine design, and nanodrug production have shown immense promise for interfacing with pathogenic viruses and restricting their entrance into cells. These viruses have been scrutinized using rapid diagnostic detection and therapeutic interventional options against the caused infections including vaccine development for prevention and control. Coronaviruses, namely SARS-CoV, MERS-CoV, and SARS-CoV-2, have endangered human life, and the COVID-19 (caused by SARS-CoV-2) outbreak has become a perilous challenge to public health globally with huge accompanying morbidity rates. Thus, it is imperative to expedite the drug and vaccine development efforts that would help mitigate this pandemic. In this regard, smart and innovative nano-based technologies and approaches encompassing applications of green nanomedicine, bio-inspired methods, multifunctional bioengineered nanomaterials, and biomimetic drug delivery systems/carriers can help resolve the critical issues regarding detection, prevention, and treatment of viral infections. This perspective review expounds recent nanoscience advancements for the detection and treatment of viral infections with focus on coronaviruses and encompasses nano-based formulations and delivery platforms, nanovaccines, and promising methods for clinical diagnosis, especially regarding SARS-CoV-2.

scholarly journals Composition of symbiotic bacteria predicts survival in Panamanian golden frogs infected with a lethal fungus

2015 ◽  
Vol 282 (1805) ◽  
pp. 20142881 ◽  
Author(s):  
Matthew H. Becker ◽  
Jenifer B. Walke ◽  
Shawna Cikanek ◽  
Anna E. Savage ◽  
Nichole Mattheus ◽  
...  
Keyword(s):  
Batrachochytrium Dendrobatidis ◽  
Population Declines ◽  
Amphibian Species ◽  
Amphibian Population ◽  
New Approach ◽  
Skin Bacteria ◽  
Probiotic Treatment ◽  
Host Health ◽  
The Face ◽  
Inhibitory Bacteria

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.

scholarly journals Long distance (>20 km) downstream detection of endangered stream frogs suggests an important role for eDNA in surveying for remnant amphibian populations

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12013
Author(s):  
Cecilia Villacorta-Rath ◽  
Conrad J. Hoskin ◽  
Jan M. Strugnell ◽  
Damien Burrows
Keyword(s):  
Water Samples ◽  
Survey Methods ◽  
Environmental Dna ◽  
Small Population ◽  
Population Declines ◽  
Long Distance ◽  
Amphibian Species ◽  
Filtration Method ◽  
North East ◽  
Water Volumes

Background Globally, amphibian species have suffered drastic population declines over the past 40 years. Hundreds of species are now listed as Critically Endangered, with many of these considered “possibly extinct”. Most of these species are stream-dwelling frogs inhabiting remote, montane areas, where remnant populations are hard to find using traditional surveys. Environmental DNA (eDNA) could revolutionize surveys for ‘missing’ and endangered amphibian populations by screening water samples from downstream sections to assess presence in the upstream catchments. However, the utility of this survey technique is dependent on quantifying downstream detection probability and distances. Methods Here we tested downstream detection distances in two endangered stream frogs (Litoria lorica and L. nannotis) that co-occur in a remote stream catchment in north-east Australia, and for which we know precise downstream distributional limits from traditional surveys. Importantly, the two last populations of L. lorica persist in this catchment: one small (~1,000 frogs) and one very small (~100 frogs). We conducted eDNA screening at a series of sites kilometers downstream from the populations using precipitation from two fixed water volumes (15 and 100 mL) and via water filtering (mean 1,480 L). Results We detected L. nannotis and the small L. lorica population (~1,000 frogs) at most sampling sites, including 22.8 km downstream. The filtration method was highly effective for far-downstream detection, as was precipitation from 100 mL water samples, which also resulted in consistent detections at the far-downstream sites (including to 22.8 km). In contrast, we had limited downstream detection success for the very small L. lorica population (~100 frogs). Discussion The ecological aspects of our study system, coupled with thorough traditional surveys, enabled us to measure downstream eDNA detection distances with accuracy. We demonstrate that eDNA from a small population of approximately 1,000 frogs can be detected as far as 22.8 km downstream from the population. Water filtration is considered best for eDNA detection of rare aquatic species—indeed it was effective in this study—but we also achieved far-downstream detections when precipitating eDNA from 100 mL water samples. Collecting small water volumes for subsequent precipitation in the lab is more practical than filtration when surveying remote areas. Our downstream detection distances (>20 km) suggest eDNA is a valuable tool for detecting rare stream amphibians. We provide recommendations on optimal survey methods.

scholarly journals Prevalence of Batrachochytrium dendrobatidis in Amphibians From 2000 to 2021: A Global Systematic Review and Meta-Analysis

2021 ◽  
Vol 8 ◽  
Author(s):  
Zhongle Li ◽  
Qi Wang ◽  
Keping Sun ◽  
Jiang Feng
Keyword(s):  
Large Scale ◽  
Batrachochytrium Dendrobatidis ◽  
Detection Method ◽  
Meta Analysis ◽  
Current Status ◽  
Population Declines ◽  
Habitat Types ◽  
China National Knowledge Infrastructure ◽  
Chemical Methods ◽  
Knowledge Infrastructure

Chytridiomycosis is an amphibian fungal disease caused by Batrachochytrium dendrobatidis (Bd), which has caused large-scale death and population declines on several continents around the world. To determine the current status of Bd infection in amphibians, we conducted a global meta-analysis. Using PubMed, ScienceDirect, SpringerLink, China National Knowledge Infrastructure (CNKI) and Wanfang database searches, we retrieved a total of 111 articles from 2000 to 2021. Based on these, we estimated the Bd prevalence to be 18.54% (95% CI: 13.76–20.52) in current extent amphibians. Among these populations, the prevalence of Bd in Asia was the lowest at 7.88% (95% CI: 1.92–8.71). Further, no Bd infection was found in Vietnam. However, the prevalence of Bd in Oceania was the highest at 36.34% (95% CI: 11.31–46.52). The Bd prevalence in Venezuela was as high as 49.77% (95% CI: 45.92–53.62). After 2009, the global Bd prevalence decreased to 18.91% (95% CI: 13.23–21.56). The prevalence of Bd in epizootic populations was significantly higher than enzootic populations. The highest prevalence of Bd was detected with real-time PCR at 20.11% (95% CI: 13.12–21.38). The prevalence of Bd in frogs was the highest at 20.04% (95% CI: 13.52–21.71), and this different host was statistically significant (P < 0.05). At the same time, we analyzed the geographic factors (longitude, latitude, elevation, rainfall and temperature) that impacted the fungal prevalence in amphibians. Our meta-analysis revealed that factors including region, disease dynamic, detection method, host and climate may be sources of the observed heterogeneity. These results indicate that chytridiomycosis was a consistent threat to amphibians from 2000 to 2021. Based on different habitat types and geographical conditions, we recommend formulating corresponding control plans and adopting reasonable and efficient biological or chemical methods to reduce the severity of such diseases.

scholarly journals Mosquitoes as a potential vector for the transmission of the amphibian chytrid fungus

2019 ◽  
Vol 29 (1) ◽  
pp. 39-44 ◽  
Author(s):  
John Gould ◽  
Jose W. Valdez ◽  
Michelle P. Stockwell ◽  
Simon Clulow ◽  
Michael J. Mahony
Keyword(s):  
Batrachochytrium Dendrobatidis ◽  
Agar Plate ◽  
Insect Host ◽  
Chytrid Fungus ◽  
Amphibian Species ◽  
Potential Vector ◽  
Amphibian Chytrid Fungus ◽  
And Control ◽  
Zoospore Suspension

The amphibian chytrid fungus, Batrachochytrium dendrobatidis (Bd), is an infectious disease responsible for the worldwide decline of amphibian species. To mitigate these declines, it is necessary to identify the various vectors by which the fungus can be transmitted between individuals and populations. The objective of this study was to determine whether adult female mosquitoes can carry and transfer Bd fungal cells. Mosquitoes were exposed to netting soaked in a live Bd zoospore suspension to determine whether they are able to externally acquire the fungus. Another group was placed into containers with a sterile and Bd-inoculated agar plate to determine whether mosquitoes could transfer Bd between these surfaces. Bd DNA was found to be present on mosquito legs exposed to inoculated netting and agar plates suggesting that Bd can be transmitted by the mosquito over short distances. This is the first study to demonstrate that an insect host may be a mechanical vector of Bd and suggests that we should begin to consider the role of mosquitoes in the dissemination and control of the fungus.

scholarly journals Infectious disease threats to amphibian conservation

2018 ◽  
Vol 27 (Supplement) ◽  
pp. 81-90
Author(s):  
A.A. Cunningham
Keyword(s):  
Infectious Disease ◽  
Batrachochytrium Dendrobatidis ◽  
Biodiversity Loss ◽  
Amphibian Conservation ◽  
Disease Spread ◽  
Population Declines ◽  
Amphibian Species ◽  
Late 20Th Century ◽  
Amphibian Population ◽  
In The Wild

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.

scholarly journals Non-native Amphibian Pet Trade via Internet in Poland

2018 ◽  
Vol 4 (1) ◽  
pp. 30-40
Author(s):  
Mikołaj Kaczmarski ◽  
Krzysztof Kolenda
Keyword(s):  
Invasive Species ◽  
Native Species ◽  
Batrachochytrium Dendrobatidis ◽  
Amphibian Species ◽  
Monthly Basis ◽  
Pet Trade ◽  
Exotic Animals ◽  
American Bullfrog ◽  
Rare And Endangered ◽  
Online Stores

Abstract Overharvesting and trade in amphibian populations is one of the causes of their global decline. Online trade not only encourages the exploitation of an increasing number of rare and endangered amphibian species from all over the world but also influences the spread of invasive species. The aim of our research was to investigate the amphibian pet trade conducted in online stores and portals in Poland and determine its potential impact on native species. Between November 2013 and October 2014, we regularly (on a monthly basis) checked sale offers on the websites of the 18 biggest pet shops in the country specialised in exotic animals, on a nationwide auction portal and on three exotic pet fan portals. During the study, we reported 486 offers of 112 amphibian species in online stores and on portals. Most of the offers involved one of the four families of amphibians: poison dart frogs (Dendrobatidae), tree frogs (Hylidae), true toads (Bufonidae) and true salamanders (Salamandridae). Our data show increased interest in amphibians as pets in Poland. At least half of the offered species are possible hosts for the chytrid fungus Batrachochytrium dendrobatidis. However, only one species, the American bullfrog Lithobates catesbeianus (Shaw, 1802), appears to be a potential invasive species. To summarise, the species offered in Poland that are characterised as threatened are predominantly those that are relatively easy to breed and that are popular as pets. Further studies are required to investigate the real threat to wild amphibian populations caused by the pet trade

scholarly journals eDNA Increases the Detectability of Ranavirus Infection in an Alpine Amphibian Population

Viruses ◽  
2019 ◽  
Vol 11 (6) ◽  
pp. 526 ◽  
Author(s):  
Claude Miaud ◽  
Véronique Arnal ◽  
Marie Poulain ◽  
Alice Valentini ◽  
Tony Dejean
Keyword(s):  
Water Samples ◽  
Rana Temporaria ◽  
Clinical Signs ◽  
Environmental Dna ◽  
Diagnostic Techniques ◽  
Mitigation Measures ◽  
Activity Period ◽  
Amphibian Species ◽  
Tissue Samples ◽  
Species Activity

The early detection and identification of pathogenic microorganisms is essential in order to deploy appropriate mitigation measures. Viruses in the Iridoviridae family, such as those in the Ranavirus genus, can infect amphibian species without resulting in mortality or clinical signs, and they can also infect other hosts than amphibian species. Diagnostic techniques allowing the detection of the pathogen outside the period of host die-off would thus be of particular use. In this study, we tested a method using environmental DNA (eDNA) on a population of common frogs (Rana temporaria) known to be affected by a Ranavirus in the southern Alps in France. In six sampling sessions between June and September (the species’ activity period), we collected tissue samples from dead and live frogs (adults and tadpoles), as well as insects (aquatic and terrestrial), sediment, and water. At the beginning of the breeding season in June, one adult was found dead; at the end of July, a mass mortality of tadpoles was observed. The viral DNA was detected in both adults and tadpoles (dead or alive) and in water samples, but it was not detected in insects or sediment. In live frog specimens, the virus was detected from June to September and in water samples from August to September. Dead tadpoles that tested positive for Ranavirus were observed only on one date (at the end of July). Our results indicate that eDNA can be an effective alternative to tissue/specimen sampling and can detect Ranavirus presence outside die-offs. Another advantage is that the collection of water samples can be performed by most field technicians. This study confirms that the use of eDNA can increase the performance and accuracy of wildlife health status monitoring and thus contribute to more effective surveillance programs.

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