Harmful algal blooms (HABs) are a persistent and increasing problem globally, yet we still have limited knowledge about how they affect wildlife. Although semi-aquatic and aquatic amphibians and reptiles have experienced large declines and occupy environments where HABs are increasingly problematic, their vulnerability to HABs remains unclear. To inform monitoring, management, and future research, we conducted a literature review, synthesized the studies, and report on the mortality events describing effects of cyanotoxins from HABs on freshwater herpetofauna. Our review identified 37 unique studies and 71 endpoints (no-observed-effect and lowest-observed-effect concentrations) involving 11 amphibian and 3 reptile species worldwide. Responses varied widely among studies, species, and exposure concentrations used in experiments. Concentrations causing lethal and sublethal effects in laboratory experiments were generally 1 to 100 µg/L, which contains the mean value of reported HAB events but is 70 times less than the maximum cyanotoxin concentrations reported in the environment. However, one species of amphibian was tolerant to concentrations of 10,000 µg/L, demonstrating potentially immense differences in sensitivities. Most studies focused on microcystin-LR (MC-LR), which can increase systemic inflammation and harm the digestive system, reproductive organs, liver, kidneys, and development. The few studies on other cyanotoxins illustrated that effects resembled those of MC-LR at similar concentrations, but more research is needed to describe effects of other cyanotoxins and mixtures of cyanotoxins that commonly occur in the environment. All experimental studies were on larval and adult amphibians; there were no such studies on reptiles. Experimental work with reptiles and adult amphibians is needed to clarify thresholds of tolerance. Only nine mortality events were reported, mostly for reptiles. Given that amphibians likely decay faster than reptiles, which have tissues that resist decomposition, mass amphibian mortality events from HABs have likely been under-reported. We propose that future efforts should be focused on seven major areas, to enhance our understanding of effects and monitoring of HABs on herpetofauna that fill important roles in freshwater and terrestrial environments. Environ Toxicol Chem 2024;00:1-14. Published 2024. This article is a U.S. Government work and is in the public domain in the USA. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Mass mortality events in wildlife can be indications of an emerging infectious disease. During the spring and summer of 2021, hundreds of dead passerines were reported across the eastern US. Birds exhibited a range of clinical signs including swollen conjunctiva, ocular discharge, ataxia, and nystagmus. As part of the diagnostic investigation, high-throughput metagenomic next-generation sequencing was performed across three molecular laboratories on samples from affected birds. Many potentially pathogenic microbes were detected, with bacteria forming the largest proportion; however, no singular agent was consistently identified, with many of the detected microbes also found in unaffected (control) birds and thus considered to be subclinical infections. Congruent results across laboratories have helped drive further investigation into alternative causes, including environmental contaminants and nutritional deficiencies. This work highlights the utility of metagenomic approaches in investigations of emerging diseases and provides a framework for future wildlife mortality events.

In July 2016, a severe coral reef invertebrate mortality event occurred approximately 200 km southeast of Galveston, Texas, at the East Flower Garden Bank, wherein ∼82% of corals in a 0.06-km2 area died. Based on surveys of dead corals and other invertebrates shortly after this mortality event, responders hypothesized that localized hypoxia was the most likely direct cause. However, no dissolved oxygen data were available to test this hypothesis, because oxygen is not continuously monitored within the Flower Garden Banks sanctuary. Here, we quantify microbial plankton community diversity based on four cruises over 2 years at the Flower Garden Banks, including a cruise just 5 to 8 days after the mortality event was first observed. In contrast with observations collected during nonmortality conditions, microbial plankton communities in the thermocline were differentially enriched with taxa known to be active and abundant in oxygen minimum zones or that have known adaptations to oxygen limitation shortly after the mortality event (e.g., SAR324, Thioglobaceae, Nitrosopelagicus, and Thermoplasmata MGII). Unexpectedly, these enrichments were not localized to the East Bank but were instead prevalent across the entire study area, suggesting there was a widespread depletion of dissolved oxygen concentrations in the thermocline around the time of the mortality event. Hydrographic analysis revealed the southern East Bank coral reef (where the localized mortality event occurred) was uniquely within the thermocline at this time. Our results demonstrate how temporal monitoring of microbial communities can be a useful tool to address questions related to past environmental events. IMPORTANCE In the northwestern Gulf of Mexico in July 2016, ∼82% of corals in a small area of the East Flower Garden Bank coral reef suddenly died without warning. Oxygen depletion is believed to have been the cause. However, there was considerable uncertainty, as no oxygen data were available from the time of the event. Microbes are sensitive to changes in oxygen and can be used as bioindicators of oxygen loss. In this study, we analyze microbial communities in water samples collected over several years at the Flower Garden Banks, including shortly after the mortality event. Our findings indicate that compared to normal conditions, oxygen depletion was widespread in the deep-water layer during the mortality event. Hydrographic analysis of water masses further revealed some of this low-oxygen water likely upwelled onto the coral reef.

A mortality event involving at least 14 Western Gulls (Larus occidentalis) was observed on 10 October 2019 on Huntington State Beach, Orange County, California, US. Clinical signs of affected gulls included generalized weakness and difficulty standing and flying. Six additional Western Gulls with similar clinical signs were admitted for rehabilitation between 24 October and 7 November, including birds from Newport Beach and Laguna Beach, south of Huntington Beach. Eleven carcasses were submitted for postmortem examination, including nine gulls collected on 10 October from Huntington Beach, one collected on 24 October from Laguna Beach, and one collected on 6 November from Newport Beach. Six of seven gulls tested were positive for Clostridium botulinum toxin type A by mouse bioassay, including five collected on 10 October from Huntington Beach and one from Laguna Beach, approximately 23 km south, on 24 October, suggesting the toxin was available to scavenging birds for nearly 2 wk following the original exposure. Botulism type C, and less commonly type E, are most frequently documented in wild birds, including waterfowl and fish-eating birds, respectively. In contrast, botulism type A is the most common cause of foodborne botulism in humans, acquired from food contaminated with C. botulinum spores, but it has not previously been associated with mortality in free-ranging wild birds.

Eurasian collared doves (Streptopelia decaocto) were introduced into Florida in the 1980s and have since established populations throughout the continental United States. Pigeon paramyxovirus-1 (PPMV-1), a species-adapted genotype VI Avian orthoavulavirus 1, has caused periodic outbreaks among collared doves in the U.S. since 2001 with outbreaks occasionally involving native doves. In California, PPMV-1 mortality events were first documented in Riverside County in 2014 with subsequent outbreaks in 23 additional counties from southern to northern California between 2015 and 2019. Affected collared doves exhibited torticollis and partial paralysis. Pale kidneys were frequently visible on gross necropsy (65.4%; 51/78) while lymphoplasmacytic interstitial nephritis often with acute tubular necrosis (96.0%; 24/25) and pancreatic necrosis (80.0%; 20/25) were common findings on histopathology. In total, PPMV-1 was confirmed by rRT-PCR and sequence analysis from oropharyngeal and/or cloacal swabs in 93.0% (40/43) of the collared doves tested from 16 California counties. In 2017, Avian orthoavulavirus 1 was confirmed in a native mourning dove (Zenaida macroura) found dead during a PPMV-1 outbreak in collared doves by rRT-PCR from formalin-fixed paraffin-embedded (FFPE) tissues, after the initial rRT-PCR from swabs failed to detect the virus. Molecular sequencing of the fusion protein of isolates collected from collared doves during outbreaks in 2014, 2016, and 2017 identified two distinct subgenotypes, VIa and VIn. Subgenotype VIn has been primarily isolated from collared doves in the southern U.S., while VIa has been isolated from mixed avian species in the northeastern U.S., indicating two independent introductions into California. While populations of collared doves are not expected to be substantially impacted by this disease, PPMV-1 may pose a threat to already declining populations of native columbids. This threat could be assessed by monitoring native and non-native columbids for PPMV-1. Based on our study, swab samples may not be sufficient to detect infection in native columbids and may require the use of non-traditional diagnostic approaches, such as FFPE tissues, to ensure virus detection.

We report a mortality event of Red Phalaropes (Phalaropus fulicarius) that occurred from October to November 2016 on the north coast of British Columbia, Canada. All individuals were severely underweight and showing signs of physiological stress. The guts of all carcasses contained ingested plastics (100%, n = 9). Distribution modelling from pelagic bird surveys (1990-2010) indicated that Red Phalaropes are not typically found in the study area during fall months. Ocean conditions during fall 2016 were unusually warm, coinciding with reduced upwelling in the study area. eBird records since 1980 indicated Red Phalaropes are observed closer to shore during periods associated with reduced upwelling. These results suggest that distribution shifts of Red Phalaropes closer to shore, where plastic debris occurs in higher concentrations, may lead phalaropes to feed on plastic debris while in a weakened state, resulting in a combination of two adverse circumstances.

In August 2015, a gopher tortoise ( Gopherus polyphemus) mortality event was documented on a 40-ha area of Lake Louisa State Park, Lake County, Florida, US. To quantify the extent of the die-off and the seroprevalence of Mycoplasma agassizii and Mycoplasma testudineum, two causative agents of mycoplasmal upper respiratory tract disease (URTD), we conducted a tortoise shell survey on 25 and 26 August 2015 and collected blood samples from live tortoises on 2-4 September 2015 and 1-13 August 2016 within the area of documented mortality. We discovered 94 shells and measured the degree of disarticulation to estimate time since mortality. Results indicated that most mortalities likely occurred more than 3 yr before discovery. Of the 42 blood samples collected in 2016, 31% ( n=13) tested positive, 17% ( n=7) were suspect, and 52% ( n=22) were negative for M. agassizii antibodies; all blood samples were negative for M. testudineum. Sixty-nine percent (9/13) of seropositive tortoises exhibited clinical signs of URTD, and seropositive individuals were more likely to express clinical signs than seronegative tortoises. However, 32% (7/22) of seronegative individuals also exhibited some clinical signs, including naris and eye abnormalities. We suggest additional research to determine causality of this mortality event, as well as examine risks associated with its spread.

In January 2017, an estimated 3,700 (93%) of 4,000 Khaki Campbell ducks (Anas platyrhynchos domesticus) died in Kampong Thom Province, Cambodia. We detected low pathogenicity avian influenza A(H7N3) virus and anatid herpesvirus 1 (duck plague) in the affected flock; however, the exact cause of the mortality event remains unclear.

BACKGROUND: An unusual mortality event in Johnstone River snapping turtles (Elseya irwini) in Far North Queensland, Australia, occurred during the summer months of December 2014 and January 2015. We report the data collected during the mortality event, including counts of sick and dead animals, clinical appearance and one necropsy.
UNASSIGNED: Moribund animals appeared lethargic with variable degrees of necrotising dermatitis. Postmortem investigation of one freshly dead animal revealed bacterial and fungal involvement in the skin lesions as well as multifocal fibrinous hepatitis and splenitis and necrotising enteritis with vascular thrombosis. Aeromonas hydrophila was isolated from liver, spleen and skin lesions. All samples tested negative for ranavirus, and water and soil testing for environmental contaminants were negative. All affected E. irwini either died or were euthanased and no other species of animals in the river were affected.
CONCLUSIONS: Aeromonas hydrophila is ubiquitous in the freshwater environment and although it caused septicaemia in the one individual that was submitted for laboratory diagnosis, the primary aetiology of the outbreak may not have been identified.

Summer mortality episodes in adult Pacific oysters have been described since the 1950s in various farming areas. Starting in 2012, a recrudescence of mortalities in commercial-sized oysters was first observed in France and then in Italy, with seasonality extension and translation later in the year. Moribund individuals collected during an event in Italy in December 2014 showed yellowish lesions of the mantle and adductor muscle. Histological examination revealed filamentous bacteria associated with necrotic areas. Quantitative PCRs targeting OsHV-1 and Vibrio aestuarianus detected only high loads of the pathogenic bacteria in tissues of symptomatic individuals. A lower diversity of the hemolymph microbiota was also evidenced in moribund individuals, with a predominance of Vibrio and Arcobacter species. A strain of Flavobacteriaceae was isolated from all the symptomatic individuals. Sequence analysis of the 16S rRNA gene identified the strain as Tenacibaculum soleae. When strain pathogenicity was tested by injection in adult individuals, it induced mortality rates of up to 45%, even in the absence of V. aestuarianus. As mortality occurred only 11 days post-infection, further investigation is needed to determine its effective virulence in natural conditions. This is the first description of a Tenacibaculum strain associated with bivalve mortalities.