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Biology subjects

Meredith, A. L.

Publications and source records attributed to Meredith, A. L..

4 recordsLinked to original sources

Environment shapes the association between the skin microbiome and infection dynamics of the Dermocystid pathogen Amphibiothecum in Palmate newts (Lissotriton helveticus).

Infectious diseases are a leading cause of biodiversity declines, but the factors shaping the prevalence and intensity of wildlife disease outbreaks remain unresolved. Host-associated microbes are likely a key component of host resistance to infection, but a hosts ability to recruit such protective microbes may be restricted by the availability of microbes present in their immediate environment. Here we investigate the associations between the abiotic environment, skin microbiome, and infection with the Dermocystid pathogen Amphibiothecum meredithae in Palmate newts (Lissotriton helveticus) on the Isle of Rum, Scotland. We found more acidic ponds to be associated with higher skin microbiome diversity and lower pathogen prevalence. Predicted functional analysis of microbiota composition identified bacterially-mediated pathways linked to host immunity upregulated in skin microbiotas associated with more acidic ponds and lower infection prevalence. This study highlights the potential importance of the environment in modulating host- microbiome-pathogen relationships and patterns of infection in the wild.

ecology↗

Buruli ulcer surveillance in south-eastern Australian possums: infection status, lesion mapping and internal distribution of Mycobacterium ulcerans

Buruli ulcer (BU) is a neglected tropical disease of skin and subcutaneous tissues caused by Mycobacterium ulcerans. BU-endemic areas are highly focal, and M. ulcerans transmission dynamics vary in different settings. In the south-eastern Australian state of Victoria, BU is an endemic vector-borne zoonosis, with mosquitoes and native possums implicated in transmission, and humans as incidental hosts. Despite the importance of possums as wildlife reservoirs of M. ulcerans, knowledge of BU in these animals is limited. Opportunistic necropsy-based and active trap-and-release surveillance studies were conducted in and around Melbourne and Geelong, Victoria, to investigate BU in possums. Demographic data and biological samples were collected, and when present, cutaneous lesions suggestive of BU were mapped. Samples were tested for the presence of M. ulcerans DNA by IS2404 qPCR. The final dataset included 26 possums: 20 necropsied; 6 trapped and released. Most possums (77%) were common ringtails from inner Melbourne. Nine possums (eight ringtails, one brushtail) had skin lesions, ranging from single and mild, to multiple and severe, exposing bones and tendons in three cases. M. ulcerans was confirmed in 78% (7/9) of clinically affected possums and 65% of possums without lesions (11/17). Possums with moderate and severe disease had widespread systemic internal bacterial dissemination and were shedding M. ulcerans in their faeces. The anatomical distribution of cutaneous lesions and PCR positivity of biological samples suggests possums may be contracting BU from bites of M. ulcerans-harbouring mosquitoes, traumatic skin wounds, ingestion of an unknown environmental source, and/or during early development in the pouch. Ringtail possums appear highly susceptible to infection with M. ulcerans and are important reservoirs of the bacteria in Victoria. A One Health approach is needed to design and implement integrated interventions that reduce M. ulcerans transmission in Victoria, thereby protecting wildlife and humans from this emerging zoonotic disease. Author summaryBuruli ulcer (BU), a neglected tropical skin disease, is emerging as a public health concern in the temperate Australian state of Victoria. Here, BU is spread by mosquitoes, and native possums are wildlife reservoirs of the causative bacterium, Mycobacterium ulcerans. Possums can be infected by BU, but knowledge of infection and disease in these animals is limited. We conducted surveillance studies in the two largest cities of Victoria, examining live and deceased possums. We found skin lesions in a third of examined possums and confirmed presence of M. ulcerans in almost three-quarters of the animals. Mouth swabs were positive for the bacteria in nearly two thirds of possums, as were pouch swabs of almost half the females. We also conducted mapping of the bodily distribution of skin lesions and found that paws and the undersides of abdomens and tails were the most affected areas. Our findings add support to the concept that possums, particularly ringtails, are M. ulcerans reservoirs in Victoria, and suggest several possible routes of infection for free-living possums that warrant further research. Improved understanding of BU in possums may allow development of targeted interventions that reduce disease transmission and protect both animal and human health.

microbiology↗

Structural mapping of patient-associated KCNMA1 gene variants

KCNMA1-linked channelopathy is a neurological disorder characterized by seizures, motor abnormalities, and neurodevelopmental disabilities. The disease mechanisms are predicted to result from alterations in KCNMA1-encoded BK K+ channel activity; however, only a subset of the patient-associated variants have been functionally studied. The localization of these variants within the tertiary structure or evaluation by pathogenicity algorithms has not been systematically assessed. In this study, 82 nonsynonymous patient-associated KCNMA1 variants were mapped within the BK channel protein. Fifty-three variants localized within cryo-EM resolved structures, including 21 classified as either gain-of-function (GOF) or loss-of-function (LOF) in BK channel activity. Clusters of LOF variants were identified in the pore, the AC region (RCK1), and near the Ca2+ bowl (RCK2), overlapping with sites of pharmacological or endogenous modulation. However, no clustering was found for GOF variants. To further understand variants of uncertain significance (VUS), assessments by multiple standard pathogenicity algorithms were compared, and new thresholds for sensitivity and specificity were established from confirmed GOF and LOF variants. An ensemble algorithm was constructed (KCNMA1 Meta Score), consisting of a weighted summation of this trained dataset combined with a structural component derived from the Ca2+ bound and unbound BK channels. KMS assessment differed from the highest performing individual algorithm (REVEL) at 10 VUS residues, and a subset were studied further by electrophysiology in HEK293 cells. M578T, E656A, and D965V (KMS+;REVEL-) were confirmed to alter BK channel properties in voltage-clamp recordings, and D800Y (KMS-;REVEL+) was assessed as benign under the test conditions. However, KMS failed to accurately assess K457E. These combined results reveal the distribution of potentially disease-causing KCNMA1 variants within BK channel functional domains and pathogenicity evaluation for VUS, suggesting strategies for improving channel-level predictions in future studies by building on ensemble algorithms such as KMS.

biophysics↗

BK channel properties correlate with neurobehavioral severity in three KCNMA1-linked channelopathy mouse models

KCNMA1 forms the pore of BK K+ channels, which regulate neuronal and muscle excitability. Recently, genetic screening identified heterozygous KCNMA1 variants in a subset of patients with debilitating paroxysmal non-kinesigenic dyskinesia, presenting with or without epilepsy (PNKD3). However, the relevance of KCNMA1 mutations and the basis for clinical heterogeneity in PNKD3 has not been established. Here we evaluate the relative severity of three KCNMA1 patient variants in BK channels, neurons, and mice. In heterologous cells, BKN999S and BKD434G channels displayed gain-of-function (GOF) properties, whereas BKH444Q channels showed loss-of-function (LOF) properties. The relative degree of channel activity was BKN999S > BKD434G > WT > BKH444Q. BK currents and action potential firing were increased, and seizure thresholds decreased, in Kcnma1N999S/WT and Kcnma1D434G/WT transgenic mice but not Kcnma1H444Q/WT mice. In a novel behavioral test for paroxysmal dyskinesia, the more severely affected Kcnma1N999S/WT mice became immobile after stress, consistent with stress-induced immobility episodes observed in PNKD3-affected individuals. Homozygous Kcnma1D434G/D434G mice showed similar immobility, but in contrast, homozygous Kcnma1H444Q/H444Q mice displayed hyperkinetic behavior. These data establish the relative pathogenic potential of patient alleles as N999S > D434G > H444Q and validate Kcnma1N999S/WT mice as a model for PNKD3 with increased seizure propensity.

neuroscience↗