FATAL ACUTE HEMOLYSIS FOLLOWING TRIAZOLE THERAPY IN AFRICAN PENGUINS (SPHENISCUS DEMERSUS).

Aspergillosis is a major cause of morbidity and mortality in penguins, with triazole antifungal drugs being commonly used for prophylaxis and treatment. This report describes 15 cases of fatal hemolysis associated with liquid itraconazole and voriconazole formulations administered to African penguins (Spheniscus demersus) from four institutions. All penguins underwent stressful events (e.g. relocation, induced molt) and were administered commercial liquid itraconazole formulations or compounded voriconazole liquid suspension. Observed clinical signs in affected penguins prior to death included hyporexia, weight loss, lethargy, dyspnea, red-tinged droppings, and obtunded mentation. Intra- and extravascular hemolysis and hemoglobinuric nephrosis were the primary pathologic manifestations on postmortem examination. The concentration-dependent hemolytic potentials of itraconazole, voriconazole, and commercial and compounded vehicle suspensions were evaluated in vitro by exposing chicken whole blood as a surrogate for penguin blood. Hemoglobin content in blood plasma was then measured by spectrophotometry. Neither itraconazole nor voriconazole alone induced hemolysis in vitro. The vehicle ingredients sorbitol and hydromellose induced hemolysis, but not at predicted plasma levels in chicken erythrocytes, suggesting neither the azole antifungals nor their major vehicles alone were likely to contribute to hemolysis in vivo in these penguins. Potential mechanisms of toxicosis include generation of an unmeasured reactive metabolite causing hemolysis, preexisting erythrocyte fragility, or species-specific differences in hemolytic thresholds that were not assessed in the chicken erythrocyte model. More research is needed on the potential for toxicosis of azole antifungal drugs and carrier molecules in this and other avian species.

CRISPR-Cas9-mediated host signal reduction for 18S metabarcoding of host-associated eukaryotes.

Metabarcoding-based methods for identification of host-associated eukaryotes have the potential to revolutionize parasitology and microbial ecology, yet significant technical challenges remain. In particular, highly abundant host reads can mask the presence of less-abundant target organisms, especially for sample types rich in host DNA (e.g., blood and tissues). Here, we present a new CRISPR-Cas9-mediated approach designed to reduce host signal by selective amplicon digestion, thus enriching clinical samples for eukaryotic endosymbiont sequences during metabarcoding. Our method achieves a nearly 76% increased efficiency in host signal reduction compared with no treatment and a nearly 60% increased efficiency in host signal reduction compared with the most commonly used published method. Furthermore, the application of our method to clinical samples allows for the detection of parasite infections that would otherwise have been missed.

Harnessing ecological theory to enhance ecosystem restoration.

Ecosystem restoration can increase the health and resilience of nature and humanity. As a result, the international community is championing habitat restoration as a primary solution to address the dual climate and biodiversity crises. Yet most ecosystem restoration efforts to date have underperformed, failed, or been burdened by high costs that prevent upscaling. To become a primary, scalable conservation strategy, restoration efficiency and success must increase dramatically. Here, we outline how integrating ten foundational ecological theories that have not previously received much attention - from hierarchical facilitation to macroecology - into ecosystem restoration planning and management can markedly enhance restoration success. We propose a simple, systematic approach to determining which theories best align with restoration goals and are most likely to bolster their success. Armed with a century of advances in ecological theory, restoration practitioners will be better positioned to more cost-efficiently and effectively rebuild the world's ecosystems and support the resilience of our natural resources.

Variation in genomic vulnerability to climate change across temperate populations of eelgrass (Zostera marina).

A global decline in seagrass populations has led to renewed calls for their conservation as important providers of biogenic and foraging habitat, shoreline stabilization and carbon storage. Eelgrass (Zostera marina) occupies the largest geographic range among seagrass species spanning a commensurately broad spectrum of environmental conditions. In Canada, eelgrass is managed as a single phylogroup despite occurring across three oceans and a range of ocean temperatures and salinity gradients. Previous research has focused on applying relatively few markers to reveal population structure of eelgrass, whereas a whole-genome approach is warranted to investigate cryptic structure among populations inhabiting different ocean basins and localized environmental conditions. We used a pooled whole-genome re-sequencing approach to characterize population structure, gene flow and environmental associations of 23 eelgrass populations ranging from the Northeast United States to Atlantic, subarctic and Pacific Canada. We identified over 500,000 SNPs, which when mapped to a chromosome-level genome assembly revealed six broad clades of eelgrass across the study area, with pairwise F ST ranging from 0 among neighbouring populations to 0.54 between Pacific and Atlantic coasts. Genetic diversity was highest in the Pacific and lowest in the subarctic, consistent with colonization of the Arctic and Atlantic oceans from the Pacific less than 300 kya. Using redundancy analyses and two climate change projection scenarios, we found that subarctic populations are predicted to be potentially more vulnerable to climate change through genomic offset predictions. Conservation planning in Canada should thus ensure that representative populations from each identified clade are included within a national network so that latent genetic diversity is protected, and gene flow is maintained. Northern populations, in particular, may require additional mitigation measures given their potential susceptibility to a rapidly changing climate.

Connecting families: a qualitative study examining the experiences of parenting young children under financial strain in Ontario, Canada.

BACKGROUND: There is little research investigating the subjective experiences of parenting young children while living in poverty and experiencing financial strain using qualitative methodologies. Therefore, the objective of this study was to employ a qualitative approach to provide a nuanced and balanced view on the topic of parenting young children under financial strain in the Canadian context. METHODS: We conducted a qualitative study using semi-structured interviews between July and August 2021 in Kingston, Ontario, Canada. Sixteen participants aged 20-39 self-identified as living under financial strain while parenting a child aged 2-5 years. A qualitative inductive thematic analysis was undertaken with a focus on describing the contents of the data. RESULTS: Four major themes emerged from the data: experience of being a parent, impact of financial strain on the family unit, impact of financial strain on the children, and impact of financial strain on the parent. Numerous deleterious physical, mental, and material impacts on the family unit and parent were identified, however parent-perceived impacts of financial strain on their children were minimal. Parents described striking levels of resourcefulness and resiliency in providing the necessities for their families, absorbing the most significant impacts of financial strain through the phenomenon of self-sacrifice. CONCLUSION: The impacts of financial strain on families with young children are far reaching. Further research into the impacts of self-sacrifice on parents experiencing financial strain are needed to better understand this issue, and to inform social programming and resources that could help alleviate the deleterious impacts of poverty on parent mental, social, and physical health.

SEROPREVALENCE OF TOXOPLASMA GONDII INFECTION IN PIGS FROM SOUTHWESTERN MISSISSIPPI.

Toxoplasma gondii infection of swine is a potential public health concern because it can be acquired by humans through the handling and consumption of contaminated raw meat. Infections in immunocompromised individuals and fetuses are the most severe and these individuals are most likely to develop clinical toxoplasmosis. Since Mississippians consume a lot of pork, there was a significant need to know the extent to which it poses a health problem in the State. This study focused on the southwestern region of Mississippi. Between July 2003 and March 2004, blood samples were collected from slaughterhouses in southwestern Mississippi and the Alcorn State University swine farm in Churchill, Mississippi. The collected blood samples were centrifuged and the sera were collected, labeled, and stored in a freezer at -20 C. The modified agglutination test was performed at dilutions of 1:25, 1:50, and 1:500. A titer of 25 was considered seropositive. Of a total of 302 samples tested, 48 (16%) were positive at a titer of 25; 29 (10%) were positive at 50; 11 (4%) were positive at 500. The seroprevalence of T. gondii in pigs in southwestern Mississippi is not as high as previous studies done in Mississippi. This could be attributed to the sample size. However, the potential for infection still exists.

Differences in Breastfeeding Duration by Maternal HIV Status: A Pooled Analysis of Nationally Representative Surveys in Sub-Saharan Africa.

BACKGROUND: Breastfeeding improves child survival but is a source of mother-to-child HIV transmission among women with unsuppressed HIV infection. Estimated HIV incidence in children is sensitive to breastfeeding duration among mothers living with HIV (MLHIV). Breastfeeding duration may vary according to maternal HIV status. SETTING: Sub-Saharan Africa. METHODS: We analyzed pooled data from nationally representative household surveys conducted during 2003-2019 that included HIV testing and elicited breastfeeding practices. We fitted survival models of breastfeeding duration by country, year, and maternal HIV status for 4 sub-Saharan African regions (Eastern, Central, Southern, and Western). RESULTS: Data were obtained from 65 surveys in 31 countries. In 2010, breastfeeding in the first month of life ("initial breastfeeding") among MLHIV ranged from 69.1% (95% credible interval: 68-79.9) in Southern Africa to 93.4% (92.7-98.0) in Western Africa. Median breastfeeding duration among MLHIV was the shortest in Southern Africa at 15.6 (14.2-16.3) months and the longest in Eastern Africa at 22.0 (21.7-22.5) months. By comparison, HIV-negative mothers were more likely to breastfeed initially (91.0%-98.7% across regions) and for longer duration (median 18.3-24.6 months across regions). Initial breastfeeding and median breastfeeding duration decreased during 2005-2015 in most regions and did not increase in any region regardless of maternal HIV status. CONCLUSIONS: MLHIV in sub-Saharan Africa are less likely to breastfeed initially and stop breastfeeding sooner than HIV-negative mothers. Since 2020, UNAIDS-supported HIV estimates have accounted for this shorter breastfeeding exposure among HIV-exposed children. MLHIV need support to enable optimal breastfeeding practices and to adhere to antiretroviral therapy for HIV treatment and prevention of postnatal mother-to-child transmission.

Temperature dependence of competitive ability is cold-shifted compared to that of growth rate in marine phytoplankton.

The effect of climate warming on community composition is expected to be contingent on competitive outcomes, yet approaches to projecting ecological outcomes often rely on measures of density-independent performance across temperatures. Recent theory suggests that the temperature response of competitive ability differs in shape from that of population growth rate. Here, we test this hypothesis empirically and find thermal performance curves of competitive ability in aquatic microorganisms to be systematically left-shifted and flatter compared to those of exponential growth rate. The minimum resource requirement for growth, R*-an inverse indicator of competitive ability-changes with temperature following a U-shaped pattern in all four species tested, contrasting from their left-skewed density-independent growth rate thermal performance curves. Our results provide new evidence that exploitative competitive success is highest at temperatures that are sub-optimal for growth, suggesting performance estimates of density-independent variables might underpredict performance in cooler competitive environments.

Social anxiety disorder-associated gut microbiota increases social fear.

Social anxiety disorder (SAD) is a crippling psychiatric disorder characterized by intense fear or anxiety in social situations and their avoidance. However, the underlying biology of SAD is unclear and better treatments are needed. Recently, the gut microbiota has emerged as a key regulator of both brain and behaviour, especially those related to social function. Moreover, increasing data supports a role for immune function and oxytocin signalling in social responses. To investigate whether the gut microbiota plays a causal role in modulating behaviours relevant to SAD, we transplanted the microbiota from SAD patients, which was identified by 16S rRNA sequencing to be of a differential composition compared to healthy controls, to mice. Although the mice that received the SAD microbiota had normal behaviours across a battery of tests designed to assess depression and general anxiety-like behaviours, they had a specific heightened sensitivity to social fear, a model of SAD. This distinct heightened social fear response was coupled with changes in central and peripheral immune function and oxytocin expression in the bed nucleus of the stria terminalis. This work demonstrates an interkingdom basis for social fear responses and posits the microbiome as a potential therapeutic target for SAD.

ADAMTS-13 conformation influences autoimmune recognition in immune thrombotic thrombocytopenic purpura.

BACKGROUND: Patients with immune-mediated thrombotic thrombocytopenic purpura (iTTP) have anti-ADAMTS-13 immunoglobulin G (IgG) autoantibodies that enhance ADAMTS-13 clearance and/or inhibit its function. ADAMTS-13 normally circulates in a closed conformation, which is manifested by the interaction of the CUB domains with the central spacer domain. Disruption of the spacer-CUB interaction opens ADAMTS-13, which augments its proteolytic function but may also expose cryptic autoimmune epitopes that promote further autoantibody recognition. OBJECTIVES: To explore differences in autoantibody binding to ADAMTS-13 in its closed or open conformations in patients with iTTP and to correlate these differences with disease-related parameters. METHODS: We developed a novel assay to measure autoantibodies binding to closed and open ADAMTS-13. Autoantibody titer and IgG subclass binding to open or closed ADAMTS-13 were measured in 70 iTTP first presentation samples and correlated with clinical data, remission, and relapse. RESULTS: In 70 patients with iTTP, the mean autoantibody titer against open ADAMTS-13 was, on average, approximately 2-fold greater than that against closed ADAMTS-13, suggesting that ADAMTS-13 opening increases epitope exposure and immune complex formation. Autoantibody titer against closed/open ADAMTS-13 and IgG subclass did not correlate with ADAMTS-13 antigen at presentation. Two patients with iTTP and persistent autoantibodies lost specificity for closed ADAMTS-13 in remission. Recognition of closed/open ADAMTS-13 and autoantibody IgG subclass between the first and second iTTP episodes were very similar. CONCLUSION: ADAMTS-13 autoantibody binding is highly influenced by ADAMTS-13 conformation. Although this does not appear to modify the pathogenicity of autoantibodies, the autoantibody signature at relapse suggests that relapse represents re-emergence of the original autoimmune response rather than de novo presentation.

Patient specific implants versus conventional implants in primary total knee arthroplasty: No significant difference in patient reported outcomes at 5 years.

Background: Patient specific implants (PSI) represent a novel innovation aimed to improve patient satisfaction and function after total knee arthroplasty (TKA); however, longitudinal patient reported outcome measures (PROMs) for PSI are not well described. We sought to primarily evaluate PROMs of patients undergoing TKA with either PSI or off-the-shelf (OTS) implants at mid-term follow-up. Methods: A retrospective review was performed on a prospectively collected cohort of 43 primary, cruciate-retaining TKAs performed with PSI (n = 23) and OTS implants (n = 20) by a single surgeon. Patient demographics, operative characteristics, range of motion (ROM) return, reoperations, and outcomes [Patient-Reported Outcomes Measurement Information System (PROMIS) T-score, Knee Injury and Osteoarthritis outcome score (KOOS), and Knee Society Score-Function (KSS-F)] were compared. Mean follow-up was 5 years. Results: TKA performed with either PSI and OTS implants demonstrated no difference in obtaining ROM by 3 months (extension 3° short of full extension vs. 0°, p = 0.16) or flexion (114° vs. 115°, p = 0.99) and final ROM was identical [0° extension to 120° flexion (p = 1)]. Although not significant (p = 0.42), 5 (22%) PSI TKA and 2 (10%) OTS implant patients required manipulation under anesthesia. KSS-F and PROMIS T-scores were higher in the PSI versus OTS TKA patients, respectively (90 vs. 73, p = 0.002; 51.6 vs. 44.5, p = 0.01). However, after multivariable analysis, none of these continuous outcome measures were significantly different (p = 0.28 for KSS and p = 0.45 for PROMIS T-score) between the groups. Conclusion: In a series of TKAs performed with PSI, no difference existed in postoperative ROM, reoperations, or patient-reported outcomes compared to OTS implants at 5 years. Surgeons may utilize the equivocal midterm results during TKA preoperative patient discussion of implant technologies.

Telemedical Interdisciplinary Care Team Evaluation and Treatment of People With Low Back Pain: A Retrospective Observational Study.

Objective: To evaluate the effects of an interdisciplinary care team (ICT) model delivered by telemedicine on patients with low back pain (LBP). Design: Retrospective analysis of deidentified pre-existing data. Setting: Retrospective observational study of patients presenting with LBP to a nationwide telemedicine practice using an ICT model. Participants: Over a 9-month period all patients with a diagnosis related to LBP and who had an ICT evaluation (medical doctor, advanced practice provider, health coach, and physical therapist) were included in the study (n=36). A minimum of 2 follow-up physical therapy visits were required for inclusion. Interventions: Patients were evaluated for LBP, received a diagnosis, and were offered a multidisciplinary treatment plan. Additional real-time audio visual medical, health coaching, registered dietician, and physical therapy services were received as deemed clinically appropriate. Main Outcome Measures: Baseline, 30 day, and final pain (mean 81 day) measurements via numerical pain rating scale (NPRS). Baseline and final Patient-Reported Outcomes Measurement Information System (PROMIS)-10 Global Mental/Physical domains. Secondary Outcome Measures: Use of prescription medication, referral for imaging, need for injections, or surgery. Results: 36 patients met criteria. Pain levels included mild (n=6, 16.7%), moderate (n=19, 52.8%), and severe (n=11, 30.6%). Clinically significant pain improvements were noted in 83.3% (n=25) of those with moderate or severe pain. PROMIS Mental and Physical Health categorization from Fair/Poor to Good/Excellent significantly improved over time. The initial 20% (n=7) in Fair/Poor Mental Health improved to 6.3% (n=2) at finish, while the 80% (n=28) in Good/Excellent Mental Health at start improved to 93.8% (n=30) at finish. Regarding Physical Health, 51.4% (n=18) rated Fair/Poor at start and 31.3% (n=10) at finish, while the 48.6% (n=17) rated Good/Excellent at start improved to 68.8% (n=22) at finish. The need for prescription medication was low (n=6, 16.7%) and spinal imaging orders were negligible (n=1, 2.8%). Injections were warranted in 11.4% (n=4) of patients and surgical referral with operative treatment in 2.8% (n=1). Conclusion: Interdisciplinary care delivered through telemedicine can significantly improve pain and support improved health-related quality of life in patients with LBP, with low rates of imaging, prescription, and interventional use.