The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3
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This document discusses using the wood frog (Rana sylvatica) as a model to study Frog Virus 3 (FV3) in the laboratory. The wood frog is a suitable choice as it is native to North America, can be bred in captivity, and is naturally infected by ranavirus in the wild. The researchers have housed and fed wood frog tadpoles and adults, determined an oral LD50 for FV3, and are collecting blood and tissue samples after infecting frogs to analyze cellular and humoral immune responses, pathogenesis, and virus detection using PCR, virus isolation, and histopathology/immunohistochemistry.
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BT IN Ethiopia
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There are over 100 species of Plasmodium, some of which can infect humans and cause malaria. The four main species that infect humans are P. vivax, P. ovale, P. malariae, and P. falciparum. P. falciparum causes the most severe form of malaria and is responsible for over 1 million deaths per year globally. The life cycle of Plasmodium involves both sexual and asexual reproduction in humans and mosquitoes. Female Anopheles mosquitoes transmit the parasite between humans during blood-feeding. Laboratory diagnosis of malaria is usually done by examining thick and thin blood smears under a microscope to look for the parasite in red blood cells. Treatment depends on the species,
Ewan mollison wp4 april 2018
This document summarizes work to analyze genome evolution in the plant pathogen Phytophthora by comparing genes across available sequenced genomes. The aims are to identify a core set of genes common to all Phytophthora species, species-specific genes, and study gene families involved in virulence. Genomes from 26 Phytophthora species were analyzed, identifying over 2,000 orthologous gene clusters shared among all species. The xylanase gene family, important for degrading plant cell walls, was studied in detail, with four main xylanase genes identified. Variations within this family across species may influence virulence and host range. Further work will expand these comparative analyses to additional gene families and newly sequenced
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Plasmodium is a genus of parasitic protozoa that causes malaria in humans and other animals. There are over 100 Plasmodium species but the four that commonly cause malaria in humans are P. vivax, P. ovale, P. malariae, and P. falciparum. P. falciparum causes the most lethal form of malaria and is responsible for over 1 million deaths per year globally. The life cycle involves sexual reproduction in mosquitoes and asexual reproduction in the liver and red blood cells of mammals. Symptoms in humans include fever, chills, headaches and anemia. Diagnosis is usually by microscopic examination of blood smears to identify the parasite species. Treatment depends on the species
Prevalence and molecular characterisation of Eimeria species in Ethiopian vil...
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Multiple paternity in loggerhead turtle ( caretta caretta
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ABSTRACT- A number of 18 adults male outbred albino rats, weighing between 133-137g were used to investigate the drug susceptibility of Trypanosoma evansi strain isolated from naturally infected dromedary camels in Umbadir area, North Kordofan State, Sudan. The rats were divided into 3 groups (C, D and F) of 6 animals each. Group C and D were infected intraperitoneally with T. evansi (Umbadir stabilate) with 1×104 Trypanosome for the inoculum. Group D rats were given quinapyramine sulphate (20 mg/Kg bwt) after parasitaemia was evident. Group F was left as healthy uninfected control for the stabilate. When parasite counts were one or more parasites per field, counting in haemocytometer were used for exact number of parasite per cubic millimeter using Neubaeur’s counter. Parasites from tail blood were first fixed, stained and diluted in trypanosome diluting reagent. The parasites were diluted to the level that can be easily counted in WBC counting chamber in the haemocytometer. The total number of parasites was expressed as log10 number of parasites per ml of blood. The presence and degree of parasitaemia were determined daily for each rat by examining tail blood. The identity of the local stabilates of Trypanosoma evansi was confirmed through adopting PCR where primers that target the internal transcribed spacer one (ITS1) of the ribosomal DNA were used. There was significant reduction in serum glucose and potassium as well as significant increase in total protein, urea, calcium, albumin and cholesterol in group C. The Umbadir stabilate showed low mortality and high sensitivity to quinapyramine sulphate.
Key-words- Drug susceptibility, T. evansi, Dromedary camels, Sudan
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This document summarizes the findings of a meeting discussing the threat of the pathogenic fungus Batrachochytrium salamandrivorans (Bsal) to salamander species in the Southern Appalachian region. Bsal was first discovered in the Netherlands in 2013 and can cause severe disease and death in salamanders. The meeting evaluated the potential threat of Bsal to different salamander families in the Southern Appalachians. It was found to pose an extremely high threat to salamandrids, a high threat to dusky and climbing salamanders, and a moderate threat to woodland plethodontids and sirenids. More data and conservation strategies are needed due
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The document discusses designing ranavirus surveillance studies in amphibians. It covers important considerations like using random sampling to obtain unbiased data, estimating required sample sizes based on population size and desired confidence levels, and conducting fieldwork in an aseptic manner to avoid pathogen transmission. Specific examples are provided for enclosure sampling, co-housing animals to study transmission, holding containers, area searches, setting up an aseptic processing station, and collecting additional environmental data. Disinfection procedures like using bleach, ethanol or Virkon solutions to clean equipment are also outlined.
Global Ranavirus Consortium
The Global Ranavirus Consortium (GRC) facilitates communication and collaboration among scientists studying ranaviruses, which are pathogens affecting over 120 ectothermic vertebrate species across five continents. The GRC holds international symposia on ranaviruses, maintains a website and reporting system, and produces outreach resources. Its goal is to advance the scientific understanding of ranaviruses through international collaboration. The most recent symposium was in 2015 in Florida, continuing the GRC's work to connect ranavirus researchers globally.
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This study monitored ranavirus-associated mortality in amphibians in a Dutch heathland area in the year following a 2010 outbreak that killed thousands of common midwife toads and dozens of smooth newts. The aims were to document spatial and temporal patterns of ranavirus disease and identify potential enhancing factors. A fixed monitoring protocol was established for ponds in the area. The virus was found to be continuously present in the outbreak pond from 2010 and also present in surrounding ponds, causing ongoing mortality in all life stages of amphibians. However, no clear triggering factors for the outbreaks or continued mortality were identified.
Mazzoni
This document summarizes a study on the surveillance of ranaviruses in farmed American bullfrogs and sympatric amphibian, fish, and reptile species in Brazil. The study found high prevalence of ranaviruses in bullfrog farms with poor management practices and whole lifecycle production. Ranaviruses were detected at low levels in sympatric species near infected farms but not in other areas without bullfrog introduction. While disease was reported in farmed bullfrog tadpoles, no disease or mortality was observed in sympatric species despite some testing positive for ranaviruses. Future work is needed to further characterize the ranaviruses detected and their potential virulence.
Ranavirus in Costa Rica
This document summarizes research on Ranavirus in Costa Rica. Initial surveys found Ranavirus present in four species of frog. Further study found several species co-infected with Ranavirus and the fungus Batrachochytrium dendrobatidis. Sequencing found the Ranavirus strain matches one from North America, raising questions about its origin. Animals confiscated from the illegal pet trade in Costa Rica were found to be infected, indicating this trade could be a pathway. Current work aims to isolate a Costa Rican strain, examine museum samples for historical presence, and analyze samples from relict sites that survived past die-offs.
Brunner
This document summarizes a model exploring the dynamics of ranavirus outbreaks in pond-breeding amphibians. Initial models with waterborne and direct transmission alone could not reproduce key features of ranavirus epidemics. Including stage-specific susceptibility based on experimental data resulted in a model that matched the timing and shape of real epidemics, with a sharp increase in cases occurring later in the season. While waterborne transmission was found to be minor compared to direct contact, environmental heterogeneity and multi-host communities warrant further study.
Warne
Environmental stressors are ubiquitous and can impact disease susceptibility and transmission in complex ways. Stress can alter an individual's phenotype through effects on reproduction, growth, and development, as well as their stress response and immune function. This in turn can influence their susceptibility to disease and ability to transmit disease to others. The timing, duration, and intensity of stress exposure are important factors that may increase or decrease disease risk in complex, nonlinear ways. Research is needed to better understand how stressors like pollution contribute to disease emergence and outbreaks through their effects beyond just susceptibility, and how individual stress profiles impact transmission capacity and recovery from disease.
Mathematical modeling of ranavirus ecology
This document summarizes the development of a mathematical model to study the ecology of ranavirus, which can cause disease and population declines in common frogs. The model examines whether ranavirus could persist if transmitted only between adults, and whether both ulcerative and hemorrhagic forms of ranaviral disease can coexist in a population. The model suggests ranavirus persistence and coexistence are possible under certain transmission rates and conditions. However, many assumptions need validation with additional data on virus transmission, disease progression, and wild prevalence. The model provides a framework to guide future experimental studies of ranaviral ecology.
Transmission of ranavirus between ectothermic vertebrate hosts
This study tested whether a frog virus 3 (FV3)-like ranavirus could be transmitted between three ectothermic vertebrate hosts: Cope's gray tree frogs, red-eared slider turtles, and mosquito fish. The results showed that (1) turtles could infect frogs and frogs could infect turtles, (2) fish could infect frogs but frogs did not infect fish, and (3) indirect transmission between species occurred but did not always result in mortality. This is the first study to demonstrate interclass transmission of ranavirus between reptiles, amphibians, and fish, providing evidence that these hosts can act as reservoirs and contribute to the persistence and spread of ranavirus infections.
Environmental dependency of ranavirus/amphibian genotypic interactions
1) The document presents research on the environmental dependency of interactions between amphibian hosts and Ranavirus pathogens.
2) Experiments were conducted exposing multiple host species and Ranavirus strains to different temperatures to measure impacts on host life history traits.
3) Results demonstrated significant effects of temperature, host species, and virus strain in shaping traits like growth rate, time to metamorphosis, and mortality rate. Certain virus strains had higher virulence at warmer temperatures.
Water temperatures affects susceptibility to ranavirus
This document summarizes a study that tested how water temperature affects the pathogenicity of ranavirus in four amphibian species. The study found that at higher temperatures (25C vs 10C): 1) Mortality from ranavirus was greater for wood frogs, spotted salamanders, and green frogs; 2) Infection prevalence was higher in all species tested; and 3) Wood frogs experienced 100% infection but no mortality at 10C, whereas 80-90% mortality occurred at 15C. These results support the hypothesis that higher temperatures increase ranavirus replication and pathogenicity, likely by increasing virus replication rates while suppressing immune function in amphibians. Future research should retest other species and temperatures to further elucidate
Amphibian ranavirus disease dynamics in an industrially altered landscape
This document summarizes research on amphibian ranavirus dynamics in wetlands located near industrial operations in northern Alberta, Canada. The researcher established long-term monitoring sites near air quality monitoring stations representing different levels of industrial disturbance. Preliminary results found ranaviruses detected in 44% of wood frog populations tested across the region. Die-offs occurred at both disturbed and remote sites in late June/early July. Virus sequencing identified the pathogen as FV3-like. Data on population characteristics, water quality parameters, and tissue samples were collected to analyze ranavirus prevalence and impacts in relation to habitat disturbance.
Chelonian diagnostics, pathology and treatment
This document summarizes a presentation on ranavirus epidemiology, diagnostics, and treatment in chelonians. Key points include:
- Ranavirus disease events often occur in local epizootics and some locations see annual outbreaks.
- Quantitative PCR was developed to detect ranavirus using a 54 bp segment of the major capsid protein gene.
- A study found a 1.5% prevalence of ranavirus in free-living and rehabilitation chelonian populations.
- An experimental study found higher mortality from ranavirus infection in red-eared sliders housed at 22°C compared to 28°C, indicating environmental temperature impacts disease outcome.
An overview of ranavirus diagnostics, treatment and management
This document discusses diagnostics, treatment, and management of ranavirus. It outlines several diagnostic methods like virus isolation, serology, histopathology, transmission electron microscopy, and PCR. Real-time PCR is commonly used but has limitations. Proper sample selection and testing the right samples is important to accurately detect ranavirus infections, especially subclinical infections. Both mortality events and disease surveillance require considering potential co-infections and the ability to differentiate ranavirus strains. Control of ranavirus involves regulatory practices, vaccination for small populations, and treatment of individuals, which is debated for persistent infections. Reintroduction programs necessitate easy strain differentiation of detected viruses.
Ranavirus outbreak in captive eastern box turtles
This document describes an outbreak of Ranavirus, Herpesvirus, and Mycoplasma infection in a captive population of 22 Eastern Box Turtles. The outbreak was first detected in July 2011, when several turtles were found dead or moribund. Living turtles were quarantined and treated with antibiotics, antivirals, fluids, and nutrition. Molecular monitoring found high prevalence of Ranavirus shedding that decreased with treatment. After 5 months of follow up, all surviving turtles were virus-free. The intensive intervention improved survival compared to previous reports, though co-infection did not affect outcomes. Further study of antivirals and immune response is warranted.
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The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3
- 1. Rana sylvatica (Lithobates sylvaticus) as a model to study FV3 María J. Forzán1,2, Kathleen M. Jones2 1Canadian Cooperative Wildlife Health Centre 2Atlantic Veterinary College 2nd Ranavirus Symposium Tennessee, July 2013
- 2. Laboratory vs. field research • Laboratory vs. field research Source: Yale University Historical Library
- 3. Species selection • Native species • Widespread • Sympatric • Suitable for captive breeding
- 4. • Native species • Widespread • Sympatric • Suitable for captive breeding Species selection the wood frog
- 5. Wood frog • Fam. Ranidae (Rana or Lithobates) • 9 species Canada • Terrestrial • Spread across NA • Broadly sympatric • Naturally host of ranavirus • Life span >3-5 y
- 6. Wood frogs in the lab • Housing/feeding of pre- & post-metamorphic frogs • breeding in captivity? • Dosing of infective virus (FV3) • Collection and evaluation of blood samples • cellular and humoral responses • Collection of tissues • pathogenesis • PCR • virus isolation • histopathology • immunohistochemistry
- 7. Eggs for tadpoles and metamorphs
- 8. Eggs for tadpoles and metamorphs
- 9. Eggs for tadpoles and metamorphs
- 14. Adults
- 15. Adults @AVC
- 18. Housing • Temperature 17-23°C • Adults fed • CK, MW, EW • Froglets fed • Dm, Dh, CK, MW • Tadpoles fed • FF, Blet Extremely hard! • 75% adults (44/59, 6mo) • 72% froglets (81/112, 8mo) Date (mm-dd) Cage Frogs Food left Fed Comments Alive Dead C01 4x 1CK Ø 8MW Not hungry C02 C04 C05 C06 C07 C08 C09 C10 C03
- 19. Oral dosing 107.7 pfu / ml
- 20. Oral dosing 0.25ml / adult 0.05ml / 1-y-old
- 21. LD50 1-year-old frogs (3.5 cm SV) • raised from tadpoles • oral infection – 0.05ml • euthanized when terminal (*) Dose (pfu/frog) 105.4 104.4 103.4 102.4 101.4 100.4 Control
- 22. LD50 1-year-old frogs (3.5 cm SV) • raised from tadpoles • oral infection – 0.05ml • euthanized when terminal (*) Dose (pfu/frog) % (*) 105.4 100 104.4 100 103.4 80 102.4 20 101.4 0 100.4 0 Control 0 103.5 pfu/frog or 104.8 pfu/ml
- 23. Pathogenesis • Adults (wild-caught) • 5.3 cm SV • Killed at 0.25, 0.5, 1, 2, 4, 9 and 14 days post- infection • Blood, tissues and abdominal fluid collected
- 24. Blood samples
- 25. Blood cells: N, Eo, B, Lym, Th, Ery
- 26. End day
- 27. Shedding – fecal swab (PCR)
- 28. Immunity – Abs, IF-α/β
- 29. Pathogenesis – PCR, H&E , IHC
- 30. Pathogenesis – PCR, H&E , IHC
- 31. Pathogenesis – PCR, H&E , IHC
- 32. Spleens: Ctrl(-), 9 dpi, 14 dpi
- 33. Done / In progress Housing and feeding of pre- and post-metamorphic frogs Dosing of infective virus (FV3) Collection and evaluation of blood samples • cellular and humoral responses • Pathogenesis • PCR, virus isolation, histopathology, immunohistochemistry
- 34. Acknowledgments Unaccredited images from: Snailtail.com, Silkwormshop.com, Flukefarms.com, Animal Planet , CALA, Acadia University and Env. Canada websites
- 36. Rana sylvatica (Lithobates sylvaticus) as a model to study FV3 • Native to North America • Fam. Ranidae (Rana or Lithobates) • Widespread (Canada, Eastern USA) • 9 species Canada • broadly sympatric • Suitable for captive breeding (possibly) • Naturally host of ranavirus • Life span >3-5 y
- 37. Wood frogs in the laboratory • Housing/feeding of pre- and post-metamorphic frogs • recommendations • breeding in captivity (?) • Dose of infective virus (FV3) • LD50 • oral dosing • Blood samples • cellular & humoral responses • Tissues • pathogenesis • PCR, virus isolation • histopathology, immunohistochemistry
Editor's Notes
- Images:Snailtail.com – D hydeiSilkwormshop.com – D melanogsterAnimal Planet website (how stuff works) – Earth wormFlukefarms.com – meal wormClipart - cricket
- 0.25ml(104.1 pfu)/adult
- C03-10 dead ranavirus Apr 22, 2013
- Spleens T7CTL – T6 – T7