Ranaviruses have been associated with mortality of lower vertebrates around the world. Frog virus 3 (FV3)-like ranaviruses have been isolated from different ectothermic vertebrate classes; however, few studies have demonstrated whether this pathogen can be transmitted among classes. Using FV3-like ranaviruses isolated from the American bullfrog Lithobates catesbeianus, eastern box turtle Terrapene carolina carolina, and Pallid Sturgeon Scaphirhynchus albus, we tested for the occurrence of interclass transmission (i.e., infection) and host susceptibility (i.e., percent mortality) for five juvenile fish and three juvenile turtle species exposed to each of these isolates. Exposure was administered via water bath (10(3) PFU/mL) for 3 d and survival was monitored for 28 d. Florida softshell turtles Apalone ferox experienced no mortality, but 10% and 20% of individuals became infected by the turtle and fish isolate, respectively. Similarly, 5% of Mississippi map turtles Graptemys pseudogeographica kohni were subclinically infected with the turtle isolate at the end of the experiment. Channel Catfish Ictalurus punctatus experienced 5% mortality when exposed to the turtle isolate, while Western Mosquitofish Gambusia affinis experienced 10% mortality when exposed to the turtle and amphibian isolates and 5% mortality when exposed to the fish isolate. Our results demonstrated that interclass transmission of FV3-like ranaviruses is possible. Although substantial mortality did not occur in our experiments, the occurrence of low mortality and subclinical infections suggest that fish and aquatic turtles may function as reservoirs for FV3-like ranaviruses. Additionally, our study is the first to report transmission of FV3-like ranaviruses between fish and chelonians.
The bald eagle (Haliaeetus leucocephalus) is a culturally and ecologically vital species in North America that embodies conservation success but continues to face threats that include emerging pathogens. The introduction of A/goose/Guangdong/1/1996 lineage highly pathogenic (HP) clade 2.3.4.4b H5N1 influenza A virus (IAV) in North America in late 2021 resulted in high rates of mortality among bald eagles. Here we show an alarming rate of bald eagle nest failure and mortality attributed to HP IAV. We documented fatal, systemic HP IAV infection in breeding adult and nestling bald eagles along the southeastern U.S. coast. Concurrently, annual bald eagle nest surveys in Georgia and Florida revealed a precipitous drop in success in coastal counties compared with previous years, portending negative impacts on population recruitment. As an apex predator and efficient scavenger, it is likely that bald eagles become infected through consumption of infected waterfowl. These results and similar reports of raptor mortality in Europe, Asia, and Africa, indicate a clear threat to raptor health. The possible long-term persistence of HP H5N1 IAV in North America poses an impending threat to bald eagle populations not only related to direct mortality but also decreased recruitment and warrants continued efforts to understand these potential impacts.
Amphibians are one of the most imperiled vertebrate groups, with pathogens playing a role in the decline of some species. Rare species are particularly vulnerable to extinction because populations are often isolated and exist at low abundance. The potential impact of pathogens on rare amphibian species has seldom been investigated. The dusky gopher frog Lithobates sevosus is one of the most endangered amphibian species in North America, with 100-200 individuals remaining in the wild. Our goal was to determine whether adult L. sevosus were susceptible to ranavirus, a pathogen responsible for amphibian die-offs worldwide. We tested the relative susceptibility of adult L. sevosus to ranavirus (103 plaque-forming units) isolated from a morbid bullfrog via 3 routes of exposure: intra-coelomic (IC) injection, oral (OR) inoculation, and water bath (WB) exposure. We observed 100% mortality of adult L. sevosus in the IC and WB treatments after 10 and 19 d, respectively. Ninety-five percent mortality occurred in the OR treatment over the 28 d evaluation period. No mortality was observed in the control treatment after 28 d. Our results indicate that L. sevosus is susceptible to ranavirus, and if adults in the wild are exposed to this pathogen, significant mortality could occur. Additionally, our study demonstrates that some adult amphibian species can be very susceptible to ranavirus, which has been often overlooked in North American studies. We recommend that conservation planners consider testing the susceptibility of rare amphibian species to ranavirus and that the adult age class is included in future challenge experiments.
Arkansas populations of Ozark Hellbenders, Cryptobranchus alleganiensis bishopi have declined precipitously over the past few decades and are now limited to a single river. Biologists have also observed an increase of distal limb lesions with unidentified etiology and unknown role in morbidity and mortality of the species in this location. We documented lesions and collected associated individual size class data and pathogen samples in Ozark Hellbenders of Arkansas (n = 73) from 2011 to 2014 with the following two objectives: (1) document spatiotemporal patterns and severity of lesions present in this last remaining Arkansas Ozark Hellbender population, and (2) determine if host factors and infection status are associated with lesion severity. A scoring system was created from 0 to 7 based on lesion observations. Linear mixed model regressions followed by AICc model evaluation were used to determine associations among infection status for amphibian pathogens Batrachochytrium dendrobatidis (Bd) and Ranavirus as well as individual biometrics on lesion score. We discovered 93.2% of Hellbenders had lesions characterized by digit swelling that often progressed toward toe-tip ulceration. In severe cases we observed digital necrosis progressing to digit loss. Any recaptured individuals had the same or worse lesion score from previous captures. The top predictive model for lesion severity included individual mass and Bd infection status with a significant, positive association of Bd with increased lesion severity (β = 0.87 ± 0.39 S.E., C.I.: 0.11, 1.63). Our findings highlight a widespread and progressive disease that is an important factor to consider for the future of Ozark Hellbenders. This syndrome is presumptively multifactorial, and future studies will benefit from investigating several factors of host, infectious agents, and environment and their roles in disease manifestation for the purpose of developing effective, multi-faceted conservation strategies. A summary of potential etiologies and mechanisms is provided that may explain observed lesion distribution and that will be applicable to future disease and epidemiological investigations.
Wildlife diseases are a major threat for species conservation and there is a growing need to implement disease surveillance programs to protect species of concern. Globally, amphibian populations have suffered considerable losses from disease, particularly from chytrid fungi ( Batrachochytrium dendrobatidis [Bd] and Batrachochytrium salamandrivorans [ Bsal]) and ranavirus. Hellbenders ( Cryptobranchus alleganiensis) are large riverine salamanders historically found throughout several watersheds of the eastern and midwestern United States. Populations of both subspecies (Ozark hellbender, Cryptobranchus alleganiensis bishopi, and eastern hellbender, Cryptobranchus alleganiensis alleganiensis) have experienced precipitous declines over at least the past five decades, and emerging pathogens are hypothesized to play a role. We surveyed Ozark hellbender populations in Arkansas (AR) and eastern hellbender populations in Middle Tennessee (MTN) and East Tennessee (ETN) for both chytrid fungi and ranavirus from swabs and tail tissue, respectively, from 2011 to 2017. Overall, we detected Bd on hellbenders from nine out of 15 rivers, with total prevalence of 26.7% (54/202) that varied regionally (AR: 33%, 28/86; MTN: 11%, 4/36; ETN: 28%, 22/80). Ranavirus prevalence (9.0%, 18/200) was comparatively lower than Bd, with less regional variation in prevalence (AR: 6%, 5/85; MTN: 11%, 4/36; ETN: 10%, 8/79). We did not detect Bsal in any hellbender populations. We detected a significant negative correlation between body condition score and probability of ranavirus infection (β=−0.13, SE=0.06, 95% CL: −0.24, −0.02). Evaluation of infection load of positive individuals revealed different trends than prevalence alone for both ranavirus and Bd, with Middle Tennessee having a significantly greater average ranaviral load than both other regions. We documented a variety of lesions that likely have multiple etiologies on hellbenders located within all geographic regions. Our data represent a multiyear pathogen dataset across several regions of C. alleganiensis, and we emphasize the need for continued pathogen surveillance.
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