Characterisation of the koala (Phascolarctos cinereus) pouch microbiota in a captive population reveals a dysbiotic compositional profile associated with neonatal mortality.

BACKGROUND: Captive koala breeding programmes are essential for long-term species management. However, breeding efficacy is frequently impacted by high neonatal mortality rates in otherwise healthy females. Loss of pouch young typically occurs during early lactation without prior complications during parturition and is often attributed to bacterial infection. While these infections are thought to originate from the maternal pouch, little is known about the microbial composition of koala pouches. As such, we characterised the koala pouch microbiome across the reproductive cycle and identified bacteria associated with mortality in a cohort of 39 captive animals housed at two facilities. RESULTS: Using 16S rRNA gene amplicon sequencing, we observed significant changes in pouch bacterial composition and diversity between reproductive time points, with the lowest diversity observed following parturition (Shannon entropy - 2.46). Of the 39 koalas initially sampled, 17 were successfully bred, after which seven animals lost pouch young (overall mortality rate - 41.18%). Compared to successful breeder pouches, which were largely dominated by Muribaculaceae (phylum - Bacteroidetes), unsuccessful breeder pouches exhibited persistent Enterobacteriaceae (phylum - Proteobacteria) dominance from early lactation until mortality occurred. We identified two species, Pluralibacter gergoviae and Klebsiella pneumoniae, which were associated with poor reproductive outcomes. In vitro antibiotic susceptibility testing identified resistance in both isolates to several antibiotics commonly used in koalas, with the former being multidrug resistant. CONCLUSIONS: This study represents the first cultivation-independent characterisation of the koala pouch microbiota, and the first such investigation in marsupials associated with reproductive outcomes. Overall, our findings provide evidence that overgrowth of pathogenic organisms in the pouch during early development is associated with neonatal mortality in captive koalas. Our identification of previously unreported, multidrug resistant P. gergoviae strains linked to mortality also underscores the need for improved screening and monitoring procedures aimed at minimising neonatal mortality in future. Video Abstract.

Elevated thalamic glutamate levels and reduced water diffusivity in alcohol use disorder: Association with impulsivity.

Alcohol induces neuroinflammation but its role in cognitive impairment and impulsivity in alcohol use disorder (AUD) has been poorly investigated. We used proton magnetic resonance spectroscopy to measure brain glutamate (Glu) levels and diffusion-weighted imaging to measure functional anisotropy (FA) in the thalamus and ventral anterior cingulate cortex (vACC) in 15 recently detoxified patients with AUD and 14 matched controls. Compared to controls, AUD patients showed higher Glu levels (p = 0.04) and lower FA in the thalamus (p = 0.04) but not in the vACC. In AUD, thalamic Glu levels (r = 0.62, p = 0.019) and FA (r=-0.55, p = 0.034) were associated with severity of drinking (drinks/week). Compared to controls, AUD patients showed higher scores on Conners' Adult ADHD Rating Scale for impulsivity (p = 0.03), which correlated with glutamate levels in the thalamus (r = 0.58, p = 0.03) and vACC (r = 0.55, p = 0.036). In a second cohort of AUD patients (n = 32), Glu in dorsal ACC (dACC) also correlated with Barrett Impulsiveness Scale total score (r = 0.43, p = 0.014). We interpret the elevated thalamic Glu levels and the parallel reduction in FA in AUD-which correlated with drinking severity-as possible evidence of neurotoxicity from neuroinflammation. The association of Glu with impulsivity suggests that neurotoxic effects of chronic alcohol exposure in the thalamus and dACC may contribute to impulsivity.

Intraoperative Laser Speckle Contrast Imaging For Real-Time Visualization of Cerebral Blood Flow in Cerebrovascular Surgery: Results From Pre-Clinical Studies.

Cerebrovascular surgery can benefit from an intraoperative system that conducts continuous monitoring of cerebral blood flow (CBF). Such a system must be handy, non-invasive, and directly integrated into the surgical workflow. None of the currently available techniques, considered alone, meets all these criteria. Here, we introduce the SurgeON™ system: a newly developed non-invasive modular tool which transmits high-resolution Laser Speckle Contrast Imaging (LSCI) directly onto the eyepiece of the surgical microscope. In preclinical rodent and rabbit models, we show that this system enabled the detection of acute perfusion changes as well as the recording of temporal response patterns and degrees of flow changes in various microvascular settings, such as middle cerebral artery occlusion, femoral artery clipping, and complete or incomplete cortical vessel cautery. During these procedures, a real-time visualization of vasculature and CBF was available in high spatial resolution through the eyepiece as a direct overlay on the live morphological view of the surgical field. Upon comparison with indocyanine green angiography videoangiography (ICG-VA) imaging, also operable via SurgeON, we found that direct-LSCI can produce greater information than ICG-VA and that continuous display of data is advantageous for performing immediate LSCI-guided adjustments in real time.

Noninvasive Assessment of Retinal Blood Flow Using a Novel Handheld Laser Speckle Contrast Imager.

PURPOSE: We assessed the image quality and reproducibility of blood flow measurements from a novel handheld laser speckle imager in handheld and stabilized use cases. METHODS: Eleven dilated human subjects were imaged with the XyCAM Handheld Retinal Imager investigational device (XyCAM HRI) in the handheld and stabilized use case in nine consecutive imaging sessions. Subjects then underwent standard color fundus photography using a Topcon TRC 50DX. The vessel-to-background contrast of the XyCAM HRI red-free photo was compared to the fundus photograph, while the coefficient of variation of blood flow measurements in specific arteries and veins also was determined. RESULTS: Vessel-to-background contrast was statistically greater in the handheld use case when compared to the standard color fundus photographs (P = 0.01). Estimates of mean blood flow velocity (BFV) were highly correlated between the stabilized and handheld use case (r 2 = 0.96). Peak velocity estimates in arteries were significantly higher than those in veins (P < 0.05). CONCLUSIONS: The XyCAM HRI prototype can acquire fundus photographs with the same or better level of clarity as color fundus photographs, and reproducibly acquire functional blood flow information in the handheld use case. TRANSLATIONAL RELEVANCE: To our knowledge, this is the first human study of a handheld laser speckle retinal imaging device. Determination of retinal blood flow has applications to ophthalmic and systemic disease and a portable handheld retinal imager that determines blood flow may be widely adopted at the point of care.

Structural and functional connectivity of the precuneus and thalamus to the default mode network.

Neuroimaging studies have identified functional interactions between the thalamus, precuneus, and default mode network (DMN) in studies of consciousness. However, less is known about the structural connectivity of the precuneus and thalamus to regions within the DMN. We used diffusion tensor imaging (DTI) to parcellate the precuneus and thalamus based on their probabilistic white matter connectivity to each other and DMN regions of interest (ROIs) in 37 healthy subjects from the Human Connectome Database. We further assessed resting-state functional connectivity (RSFC) among the precuneus, thalamus, and DMN ROIs. The precuneus was found to have the greatest structural connectivity with the thalamus, where connection fractional anisotropy (FA) increased with age. The precuneus also showed significant structural connectivity to the hippocampus and middle pre-frontal cortex, but minimal connectivity to the angular gyrus and midcingulate cortex. In contrast, the precuneus exhibited significant RSFC with the thalamus and the strongest RSFC with the AG. Significant symmetrical structural connectivity was found between the thalamus and hippocampus, mPFC, sFG, and precuneus that followed known thalamocortical pathways, while thalamic RSFC was strongest with the precuneus and hippocampus. Overall, these findings reveal high levels of structural and functional connectivity linking the thalamus, precuneus, and DMN. Differences between structural and functional connectivity (such as between the precuneus and AG) may be interpreted to reflect dynamic shifts in RSFC for cortical hub-regions involved with consciousness, but could also reflect the limitations of DTI to detect superficial white matter tracts that connect cortico-cortical regions. Hum Brain Mapp 38:938-956, 2017. © 2016 Wiley Periodicals, Inc.

Socioeconomic status is associated with striatal dopamine D2/D3 receptors in healthy volunteers but not in cocaine abusers.

Positron emission tomography (PET) studies in animals and humans have shown that social status is associated with striatal dopamine D2/D3 receptor (D2/D3R) availability. That is, higher social hierarchy and higher scores on questionnaires assessing social status correlated positively with striatal D2/D3R availability in animals and humans respectively. Furthermore, subordinate monkeys were vulnerable to cocaine self-administration, suggesting that alternations in social hierarchy can change D2/D3R availability and vulnerability to cocaine use. Here, we investigated whether socioeconomic status (SES) measured with the Hollingshead scale is associated with striatal D2D/3R availability using [(11)C]raclopride PET in 38 cocaine abusers and 42 healthy controls matched for age and education. Compared to controls, cocaine abusers showed lower D2/D3R availability in the caudate, putamen and ventral striatum (all p≤0.001). Despite matching groups for education, SES scores were lower in cocaine abusers than controls (p<0.001). In the control group only, SES scores significantly correlated with D2/D3R in caudate (r=0.35, p=0.024) and putamen (r=0.39, p=0.011) but not in ventral striatum (p=0.61); all corrected for age. The study confirms that SES is associated with striatal D2/D3R availability in healthy human volunteers. However, reductions in D2/D3R availability in cocaine abusers may be driven by factors other than SES such as chronic cocaine exposure.

Feasibility of Structural and Functional MRI Acquisition with Unpowered Implants in Argus II Retinal Prosthesis Patients: A Case Study.

PURPOSE: Magnetic resonance imaging (MRI) can measure the effects of vision loss and recovery on brain function and structure. In this case study, we sought to determine the feasibility of acquiring anatomical and functional MRI data in recipients of the Argus II epiretinal prosthesis system. METHODS: Following successful implantation with the Argus II device, two retinitis pigmentosa (RP) patients completed MRI scans with their implant unpowered to measure primary visual cortex (V1) functional responses to a tactile task, whole-brain morphometry, V1 cortical thickness, and diffusion properties of the optic tract and optic radiation. Measurements in the subjects with the Argus II implant were compared to measurements obtained previously from RP patients and sighted individuals. RESULTS: The presence of the Argus II implant resulted in artifacts that were localized around the patient's implanted eye and did not extend into cortical regions or white matter tracts associated with the visual system. Structural data on V1 cortical thickness and the retinofugal tract obtained from the two Argus II subjects fell within the ranges of sighted and RP groups. When compared to the RP and sighted subjects, Argus II patients' tactile-evoked cross-modal functional MRI (fMRI) blood oxygen level-dependent (BOLD) responses in V1 also fell within the range of either sighted or RP groups, apparently depending on time since implantation. CONCLUSIONS: This study demonstrates that successful acquisition and quantification of structural and functional MR images are feasible in the presence of the inactive implant and provides preliminary information on functional changes in the brain that may follow sight restoration treatments. TRANSITIONAL RELEVANCE: Successful MRI and fMRI acquisition in Argus II recipients demonstrates feasibility of using MRI to study the effect of retinal prosthesis use on brain structure and function.

Correlation of vision loss with tactile-evoked V1 responses in retinitis pigmentosa.

Neuroimaging studies have shown that the visual cortex of visually impaired humans is active during tactile tasks. We sought to determine if this cross-modal activation in the primary visual cortex is correlated with vision loss in individuals with retinitis pigmentosa (RP), an inherited degenerative photoreceptor disease that progressively diminishes vision later in life. RP and sighted subjects completed three tactile tasks: a symmetry discrimination task, a Braille-dot counting task, and a sandpaper roughness discrimination task. We measured tactile-evoked blood oxygenation level dependent (BOLD) responses using functional magnetic resonance imaging (fMRI). For each subject, we quantified the cortical extent of the tactile-evoked response by the proportion of modulated voxels within the primary visual cortex (V1) and its strength by the mean absolute modulation amplitude of the modulated voxels. We characterized vision loss in terms of visual acuity and the areal proportion of V1 that corresponds to the preserved visual field. Visual acuity and proportion of the preserved visual field both had a highly significant effect on the cortical extent of the V1 BOLD response to tactile stimulation, while visual acuity also had a significant effect on the strength of the V1 response. These effects of vision loss on cross-modal responses were reliable despite high inter-subject variability. Controlling for task-evoked responses in the primary somatosensory cortex (S1) across subjects further strengthened the effects of vision loss on cross-model responses in V1. We propose that such cross-modal responses in V1 and other visual areas may be used as a cortically localized biomarker to account for individual differences in visual performance following sight recovery treatments.

Visual cortex activation induced by tactile stimulation in late-blind individuals with retinitis pigmentosa.

The inter-subject variability of visual cortex reorganization was assessed in late-blind subjects suffering from retinitis pigmentosa (RP), a degenerative retinal disease that results in tunnel vision and eventual loss of sight. fMRI BOLD responses were measured as blindfolded RP and blindfolded sighted control groups completed a tactile discrimination task (in which subjects determined the relative roughness of sandpaper discs) during successive scans in a 3T Siemens scanner. Resulting activation patterns were compared between the two groups in a whole-brain analysis. We found that vision deprivation leads to elevated activation of the visual cortex elicited with tactile stimuli, and the degree of activation correlates with the degree of visual field loss: higher visual cortex activation is associated with greater vision loss. The location of vision loss in the visual field also correlates with the location of tactile responses in the visual cortex, with greater peripheral vision loss leading to stronger activation in the peripheral of V1. Visual cortex responses to tactile stimuli may hence be used as a diagnostic marker in determining the extent of an individual's vision loss and tracking sight recovery following treatments.