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Cyathostomins are the most common and highly prevalent parasites of horses worldwide. Historically, the control of cyathostomins has mainly relied on the routine use of anthelmintic products. Increasing reports on anthelmintic resistance (AR) in cyathostomins are concerning. A potential method proposed for detecting emerging AR in cyathostomins has been estimating the egg reappearance period (ERP). This paper reviews the data available for the ERP of cyathostomins against the three major classes of anthelmintics, macrocyclic lactones, tetrahydropyrimidines, and benzimidazoles. Published peer-reviewed original research articles were obtained from three databases (PubMed, CAB Direct and Web of Science) and were evaluated for their inclusion in a systematic review. Subsets of articles were then subjected to a review of ERP data. A total of 54 (of 134) studies published between 1972 and 2022 met the criteria for inclusion in the systematic review. Until the beginning of 2022, there was no agreed definition of the ERP; eight definitions of ERP were identified in the literature, complicating the comparison between studies. Additionally, potential risk factors for the shortening of the ERP, including previous anthelmintic use and climate, were frequently not described. Reports of shortened ERP for moxidectin and ivermectin are frequent: 20 studies that used comparable ERP definitions reported shortened moxidectin and ivermectin ERPs of 35 and 28 days, respectively. It is unclear whether the ERPs of these anthelmintics reduced to such levels are due to the development of AR or some biological factors related to horses, cyathostomin species, and/or the environment. The ERPs for other anthelmintics, such as fenbendazole and pyrantel, were frequently not reported due to established resistance against these drugs. Future research in horses is required to understand the mechanism(s) behind the shortening of ERP for cyathostomins. Based on this systematic review, we propose recommendations for future ERP studies.

We administered the Realness Scale (RS), Celebrity Attitude Scale (CAS), and the modified Authentic Living Subscale (ALS) from the Authenticity Scale (AS) to undergraduate students from four American institutions of higher learning. We sought to further validate the RS by showing that it correlated positively with the ALS and negatively with the CAS. We also hypothesized that African Americans would score lower than Whites on the RS. Our results supported the first hypothesis, but we found only weak or non-existent support for the other two hypotheses. Discussion focused on reasons why our latter two hypotheses yielded mostly negative results and suggested improvements for future research.

Stress responses are believed to involve corticotropin releasing factor (CRF), its two cognate receptors (CRFWe miniaturized a cell-based assay, where CRFBP(10kD) is fused as a chimera with CRFWe found that CRFBP(10kD) potentiates CRF-intracellular CaThese results provide the first evidence of specific roles for CRF

Context Striped skunks (Mephitis mephitis) occur across a wide range of habitats in North America and co-occur with many wildlife that use similar denning structures. Few studies have quantified interspecific activity at striped skunk dens despite the concomitant implications for pathogen transmission due to direct and indirect contact at shared dens. Further, no studies have examined differences in striped skunk denning behaviour across an urban–rural gradient with various forms and degrees of human modification. Aims Our study described striped skunk denning behaviour in the lower Midwestern United States and assessed interspecific activity at dens. Methods We used radiotelemetry and camera traps to observe winter denning behaviour of radiocollared striped skunks and other sympatric species across an urban–rural gradient in southern Illinois, USA, during November–February 2018–2021. We examined correlations between striped skunks and other species captured in photos at dens. We tested for an effect of human modification on the number of dens used by striped skunks, and the effects of weather and landscape variables on striped skunk denning duration. Key results Striped skunks used 3–21 unique dens during a single winter and denned consecutively in one location for 2–59 days. All individuals were observed denning for ≥8 consecutive days. Three striped skunks participated in communal denning, and ≤3 striped skunks were observed at a den concurrently. Eleven mammalian species were observed at striped skunk dens, and other species were present in 35.3% of photos. Striped skunk presence at a den was positively associated with Virginia opossum (Didelphis virginiana) presence. Human modification had no significant effect on the number of dens used by a striped skunk. Human modification, distance to stream/shoreline, and mean daily temperature had significant negative effects on striped skunk denning duration. Conclusions We demonstrated that winter denning behaviour of striped skunks in southern Illinois, USA, fits into a latitudinal gradient of behaviour across North America, and dens are a shared resource where direct and indirect intraspecific and interspecific interactions occur. Implications Weather and landscape features influenced winter denning behaviour of striped skunks and ultimately pathogen transmission potential at dens, trends that could potentially be seen in other urban-adapted species.

The equine ascarids, Parascaris spp., are important nematode parasites of juvenile horses and were historically model organisms in the field of cell biology, leading to many important discoveries, and are used for the study of chromatin diminution. In veterinary parasitology, Parascaris spp. are important not only because they can cause clinical disease in young horses but also because they are the only ascarid parasites to have developed widespread anthelmintic resistance. Despite this, much of the general biology and mechanisms of anthelmintic resistance are poorly understood. This review condenses known basic biological information and knowledge on the mechanisms of anthelmintic resistance in Parascaris spp., highlighting the importance of foundational research programs. Although two variants of this parasite were recognized based on the number of chromosomes in the 1870s and suggested to be two species in 1890, one of these, P. univalens, appears to have been largely forgotten in the veterinary scientific literature over the past 100 years. We describe how this omission has had a century-long effect on nomenclature and data analysis in the field, highlighting the importance of proper specimen identification in public repositories. A summary of important basic biology, including life cycle, in vitro maintenance, and immunology, is given, and areas of future research for the improvement of knowledge and development of new systems are given. Finally, the limited knowledge regarding anthelmintic resistance in Parascaris spp. is summarized, along with caution regarding assumptions that resistance mechanisms can be applied across clades.

Large carnivores have been largely extirpated from Southeast Asia due to deforestation, habitat fragmentation, and poaching. Estimating the density of endangered carnivore populations, and identifying relationships between species occupancy and both environmental and anthropogenic factors, is essential for effective conservation planning. Recently, the IUCN conservation status of the Indochinese leopard (Panthera pardus delacouri) was upgraded to "Critically Endangered. " We surveyed Kweekoh Wildlife Sanctuary in Kawthoolei, an area administered by the Karen ethnic group in eastern Myanmar, to quantify (1) leopard population density using spatially explicit mark-resight (SMR) models, (2) leopard occupancy as influenced by important ecological variables, and (3) potential differences in activity between melanistic and spotted leopard morphs. Leopard density was estimated to be 1.39 +/- SE 0.22/100 km(2). Leopard occupancy (psi = 0.43; 95% credible interval: 0.26-0.67) increased further from roads, at relatively higher elevations, and in areas with higher relative abundance of wild boar. Leopard activity was cathemeral, with higher activity during night hours, and significant overlap (delta = 0.84; 95% confidence interval: 0.71-0.96) between melanistic and spotted morphs. However, melanistic leopards were more active during twilight hours than spotted individuals whose activity did not significantly vary throughout the day. Indochinese leopard density estimates in Kweekoh were among the lowest reported from Southeast Asia. Leopard occupancy was highest in the sanctuary's core areas, suggesting the presence of negative anthropogenic impacts along the sanctuary borders. We suggest our low density estimates warrant immediate and decisive conservation action, including better protection for leopards, their habitat, and their prey.

Chinese hamster ovary (CHO) cells are the primary platform for the production of biopharmaceuticals. To increase yields, many CHO cell lines have been genetically engineered to resist cell death. However, the kinetics that governs cell fate in bioreactors are confounded by many variables associated with batch processes. Here, we used CRISPR-Cas9 to create combinatorial knockouts of the three known BCL-2 family effector proteins: Bak1, Bax, and Bok. To assess the response to apoptotic stimuli, cell lines were cultured in the presence of four cytotoxic compounds with different mechanisms of action. A population-based model was developed to describe the behavior of the resulting viable cell dynamics as a function of genotype and treatment. Our results validated the synergistic antiapoptotic nature of Bak1 and Bax, while the deletion of Bok had no significant impact. Importantly, the uniform application of apoptotic stresses permitted direct observation and quantification of a delay in the onset of cell death through Bayesian inference of meaningful model parameters. In addition to the classical death rate, a delay function was found to be essential in the accurate modeling of the cell death response. These findings represent an important bridge between cell line engineering strategies and biological modeling in a bioprocess context.

Knowledge of genetic diversity is important to wildlife conservation because genetically depleted populations experience an increased risk of extinction. Mammalian carnivores are characterized by small and fragmented populations and low dispersal, so that genetic erosion can lead to the fixation of deleterious genes relatively quickly, leading to morphological abnormalities. Kinked tails and cowlicks are indicative of inbreeding depression and have been described in two wild cat species so far, the puma (Puma concolor) and the cheetah (Acinonyx jubatus). Here we report the first records of morphological abnormalities in five populations of the European wildcat (Felis silvestris silvestris) in Italy by using (1) camera-trapping and (2) necropsy of road-killed individuals assessed through genetic analysis. We collected 24,055 trap-nights from 251 cameras and recorded 566 wildcat detections, from which we identified 148 wildcats. Among these, 11 individuals had a kinked tail and four displayed brachyuria, whereas three wildcats from Sicily had cowlicks on the thorax. We recovered 28 road-killed wildcats and two of them (from Sicily and Friuli Venezia Giulia) had a kinked tail. Among these, one female with a kinked tail had a male foetus with a kinked tail, which proved that this characteristic was genetically inherited. We are unsure why brachyuria or cowlicks were not detected across all monitored wildcat populations, given we found kinked tails throughout Italy. The frequencies at which we have detected these abnormalities in wildcats are far lower than reports from Florida panthers (Puma concolor). Future research is needed to verify whether these abnormalities are also associated with low genetic diversity or other morphological defects which might lower fitness. We recommend a nationwide effort, using these techniques within a standardized sampling design, to further understand the status of the wildcat in Italy.

Here, we describe molecular engineering of monovalent ultra-long acting two-chain insulin-Fc conjugates. Insulin-Fc conjugates were synthesized using trifunctional linkers with one amino reactive group for reaction with a lysine residue of insulin and two thiol reactive groups used for re-bridging of a disulfide bond within the Fc molecule. The ultra-long pharmacokinetic profile of the insulin-Fc conjugates was the result of concertedly slowing insulin receptor-mediated clearance by (1) introduction of amino acid substitutions that lowered the insulin receptor affinity and (2) conjugating insulin to the Fc element. Fc conjugation leads to recycling by the neonatal Fc receptor and increase in the molecular size, both contributing to the ultra-long pharmacokinetic and pharmacodynamic profiles.

Although numerous studies support a dose–effect relationship between Endocrine disruptors (EDs) and the progression and malignancy of tumors, the impact of a chronic exposure to non-lethal concentrations of EDs in cancer remains unknown. More specifically, a number of studies have reported the impact of Aldrin on a variety of cancer types, including prostate cancer. In previous studies, we demonstrated the induction of the malignant phenotype in DU145 prostate cancer (PCa) cells after a chronic exposure to Aldrin (an ED). Proteins are pivotal in the regulation and control of a variety of cellular processes. However, the mechanisms responsible for the impact of ED on PCa and the role of proteins in this process are not yet well understood. Here, two complementary computational approaches have been employed to investigate the molecular processes underlying the acquisition of malignancy in prostate cancer. First, the metabolic reprogramming associated with the chronic exposure to Aldrin in DU145 cells was studied by integrating transcriptomics and metabolomics via constraint-based metabolic modeling. Second, gene set enrichment analysis was applied to determine (i) altered regulatory pathways and (ii) the correlation between changes in the transcriptomic profile of Aldrin-exposed cells and tumor progression in various types of cancer. Experimental validation confirmed predictions revealing a disruption in metabolic and regulatory pathways. This alteration results in the modification of protein levels crucial in regulating triacylglyceride/cholesterol, linked to the malignant phenotype observed in Aldrin-exposed cells.

A multi-omic approach gives a detailed time-resolved insight into the secondary metabolic capacity of Streptomyces hygroscopicus.

We report the complete genome sequence of a potential polyethylene terephthalate (PET)-degrading bacterium, Sphingopyxis sp. strain PET50, isolated from compost.

We analysed food data for rock ptarmigan Lagopus muta collected during a 10 day period at the end of September and beginning of October 2006–2014 in northeast Iceland or for nine years. The total sample was 679 birds, including 455 juveniles and 224 adults; the annual range was 48–88 birds. To describe the importance of items consumed we used percentage by dry biomass (total 1399 g) and percentage by frequency of occurrence. A total of 80+ species were identified, the most important were Betula nana (27% biomass, 62% frequency), Empetrum nigrum (14%, 63%), Dryas octopetala (11%, 64%), and Salix herbacea (10%, 41%). Important plant parts consumed were berries, woody shoots with buds, buds, catkins, seeds and leaves. Difference in diet, both with respect to species and plant parts consumed, depended on year and altitude of collection site but not on ptarmigan sex, age or sex*age interaction. The only significant age difference observed was that juveniles consumed more seeds and less woody parts than did adult birds. Betula spp. dominated the diet in five years and Salix spp. in four years. Betula and Vaccinium spp. were most important at low and medium elevations but Salix spp. showed an opposite pattern. Berries (E. nigrum and Vaccinium spp.) were the dominant plant parts in the diet for six years and woody parts of Salix and Betula spp. in three years. Berries dominated in the diet at low to medium elevations and woody parts showed an opposite pattern.

Background Parasitic nematodes, including large roundworms colloquially known as ascarids, affect the health and well-being of livestock animals worldwide. The equine ascarids, Parascaris spp., are important parasites of juvenile horses and the first ascarids to develop widespread anthelmintic resistance. The microbiota has been shown to be an important factor in the fitness of many organisms, including parasitic nematodes, where endosymbiotic Wolbachia have been exploited for treatment of filariasis in humans. Methods This study used short-read 16S rRNA sequences and Illumina sequencing to characterize and compare microbiota of whole worm small intestinal stages and microbiota of male and female intestines and gonads. Diversity metrics including alpha and beta diversity, and the differential abundance analyses DESeq2, ANCOM-BC, corncob, and metagenomeSeq were used for comparisons. Results Alpha and beta diversity of whole worm microbiota did not differ significantly between groups, but Simpson alpha diversity was significantly different between female intestine (FI) and male gonad (MG) (P= 0.0018), and Shannon alpha diversity was significantly different between female and male gonads (P = 0.0130), FI and horse jejunum (HJ) (P = 0.0383), and FI and MG (P= 0.0001). Beta diversity (Fig. 2B) was significantly different between female and male gonads (P = 0.0006), male intestine (MI) and FG (P = 0.0093), and MG and FI (P = 0.0041). When comparing organs, Veillonella was differentially abundant for DESeq2 and ANCOM-BC (p P = 0.0008), and metagenomeSeq (P = 0.0118), and Sarcina was differentially abundant across four methods (P Parascaris spp. specimens were compared to establish shared microbiota between groups. Conclusions Overall, this study provided important information regarding the Parascaris spp. microbiota and provides a first step towards determining whether the microbiota may be a viable target for future parasite control options. Graphical abstract

Identifying factors promoting jaguar (Panthera onca) occupancy is crucial for planning effective conservation and management actions that can contribute to long-term population viability. We used camera-trapping and modeled factors affecting detection and occupancy for jaguars in Sonora at 149 sites during August-November 2008 and 2009. We measured 24 covariates presumed to affect detection (n = 6) and occupancy (n = 18) at camera sites, including sampling period and various habitat characteristics. We obtained 29 jaguar detections at 19 sites (naive occupancy = 0.12) in 5455 trap-days of effort. Jaguar detectability (p = 0.16 +/- 0.05) was negatively affected by human presence and varied by study year and sampling period. Jaguar occupancy (psi = 0.30 +/- 0.03) increased as prey richness, the abundance of calves of domestic cattle, and the proportion of subtropical vegetation increased. Jaguar occupancy was lower with increased abundance of adult cattle and peccary, and higher levels of fragmentation. Jaguar occupancy in Sonora was hence driven by a diverse combination of factors which should be considered when planning conservation actions. We suggest that managers mitigate the impact of livestock, especially on subtropical habitats, by employing appropriate fencing. Fencing may increase wildlife prey for jaguars, thereby reducing calf depredation by jaguars, while concurrently protecting habitat from overgrazing. Furthermore, providing financial support for ecotourism and compensation for depredation to offset potential jaguar impacts should be considered for jaguar conservation in Sonora.

This review is aimed to (i) appraise the literature on the use of molecular techniques for the detection, quantification and differentiation of gastrointestinal helminths (GIH) of equids, (ii) identify the knowledge gaps and, (iii) discuss diagnostic prospects in equine parasitology. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines for systematic reviews, we retrieved 54 studies (horses: 50/54; donkeys and zebras: 4/54) from four databases. Polymerase chain reaction (PCR) was employed in all of the studies whereas PCR amplicons were sequenced in only 18 of them. Other techniques used (including modifications of PCR) were reverse line blot, quantitative (q)PCR, restriction fragment length polymorphism, nested-PCR, PCR-directed next-generation sequencing, Southern blotting, single strand conformation polymorphism, PCR-enzyme linked immunosorbent assay, matrix-assisted laser desorption/ionisation-time of flight and random amplification of polymorphic DNA. Most of the studies (53/54) used nuclear ribosomal RNA (including the internal transcribed spacers, intergenic spacer, 5.8 S, 18 S, 28 S and 12 S) as target loci while cytochrome c oxidase subunit 1 and random genomic regions were targeted in only three and one studies, respectively. Overall, to date, the majority of molecular studies have focused on the diagnosis and identification of GIHs of equids (i.e. species of Anoplocephala, Craterostomum, cyathostomins, Oesophagodontus, Parascaris, Strongylus, Strongyloides and Triodontophorus), with a recent shift towards investigations on anthelmintic resistance and the use of high-throughput nemabiome metabarcoding. With the increasing reports of anthelmintic resistance in equid GIHs, it is crucial to develop and apply techniques such as advanced metabarcoding for surveillance of parasite populations in order to gain detailed insights into their diversity and sustainable control. To the best of our knowledge, this is the first systematic review that evaluates molecular investigations published on the diagnosis and quantification of equid GIHs and provides useful insights into important knowledge gaps and future research directions in equid molecular parasitology.

Bacillus thuringiensis (Bt) is a Gram-positive soil bacterium that is widely and safely applied in the environment as an insecticide for combatting insect pests that damage crops or are disease vectors. Dominant active ingredients made by Bt are insect-killing crystal (Cry) proteins released as crystalline inclusions upon bacterial sporulation. Some Bt Cry proteins, e.g., Cry5B (formally Cry5Ba1), target nematodes (roundworms) and show exceptional promise as anthelmintics (cures for parasitic nematode diseases). We have recently described inactivated bacteria with cytosolic crystal(s) (IBaCC) in which bioactive Bt Cry crystals (containing Cry5B) are fully contained within the cytosol of dead bacterial ghosts. Here, we demonstrate that these IBaCC-trapped Cry5B crystals can be liberated and purified away from cellular constituents, yielding purified cytosolic crystals (PCC). Cry5B PCC contains ~95% Cry5B protein out of the total protein content. Cry5B PCC is highly bioactive against parasitic nematode larvae and adults

Bu makalede postmodern müze kavramı, epistemolojik tanımlar ve gelecek bilim teorileri üzerinden tartışıl-maktadır. Postmodern müzeler de ziyaretçi odaklı etkileşim ve katılım ön plana çıkmaktadır. Bunun yanı sıra, müzelerin artan yeni sorumlulukları karşısında özellikle küratörün rolü de önemli ölçüde değişmektedir. Bu durumun müzelerin işleyişi, yorumlama ve iletişim kurma şekli üzerinde rolü büyütür. Bu makale, postmodern müzenin bazı teorik perspektiflerine ve küratörlerin günlük faaliyetlerinde daha geniş sorumlulukları nasıl ele aldıklarına bakarak, bu değişikliklerin hem müze pratiğini hem de teorisini nasıl etkilediğini belirlemeyi amaçlamaktadır. Dönüşüm ve fütüroloji, müzelerin sürekli olarak bilgiyi dönüştürme ve geleceğin müzeleri olma ihtiyaçları etrafında dönen dönüştürücü güçtür. Bu bağlamda Dönüştürücü Müze kavramı, bize yeni bir müze modeli sunarak aynı zamanda müzelerin karşılaştıkları zorlukları da özetlemektedir.

Fecal egg counts (FECs) are essential for veterinary parasite control programs. Recent advances led to the creation of an automated FEC system that performs with increased precision and reduces the need for training of analysts. However, the variability contributed by analysts has not been quantified for FEC methods, nor has the impact of training on analyst performance been quantified. In this study, three untrained analysts performed FECs on the same slides using the modified McMaster (MM), modified Wisconsin (MW), and the automated system with two different algorithms: particle shape analysis (PSA) and machine learning (ML). Samples were screened and separated into negative (no strongylid eggs seen), 1–200 eggs per gram of feces (EPG), 201–500 EPG, 501–1000 EPG, and 1001+ EPG levels, and ten repeated counts were performed for each level and method. Analysts were then formally trained and repeated the study protocol. Between analyst variability (BV), analyst precision (AP), and the proportion of variance contributed by analysts were calculated. Total BV was significantly lower for MM post-training (p = 0.0105). Additionally, AP variability and analyst variance both tended to decrease for the manual MM and MW methods. Overall, MM had the lowest BV both pre- and post-training, although PSA and ML were minimally affected by analyst training. This research illustrates not only how the automated methods could be useful when formal training is unavailable but also how impactful formal training is for traditional manual FEC methods.

Horses are ubiquitously infected by a diversity of gastro-intestinal parasitic helminths. Of particular importance are nematodes of the family Strongylidae, which can significantly impact horse health and performance. However, knowledge about equine strongyles remains limited due to our inability to identify most species non-invasively using traditional morphological techniques. We developed a new internal transcribed spacer 2 (ITS2) DNA metabarcoding 'nemabiome' assay to characterise mixed strongyle infections in horses and assessed its performance by applying it to pools of infective larvae from fecal samples from an experimental herd in Kentucky, USA and two feral horse populations from Sable Island and Alberta, Canada. In addition to reporting the detection of 33 different species with high confidence, we illustrate the assay's repeatability by comparing results generated from aliquots from the same fecal samples and from individual horses sampled repeatedly over multiple days or months. We also validate the quantitative potential of the assay by demonstrating that the proportion of amplicon reads assigned to different species scales linearly with the number of larvae present. This new tool significantly improves equine strongyle diagnostics, presenting opportunities for research on species-specific anthelmintic resistance and the causes and consequences of variation in mixed infections.

Despite numerous studies support a dose-effect relation between Endocrine disruptors (EDs) and tumor progression and malignancy, the effects of chronic exposure to non-lethal concentrations of EDs in cancer remains unknown. Previous studies demonstrated induction of the malignant phenotype in DU145 prostate cancer (PCa) cells after chronic exposure to Aldrin (an ED). However, the mechanisms underlying this process remain limited. Here, two complementary computational approaches to investigate the molecular processes involved in the observed malignancy acquisition in Pca are applied. Firstly, the metabolic reprogramming associated to the chronic exposure to Aldrin in DU145 cells was studied by integrating transcriptomics and metabolomics via constraint-based metabolic modeling. Secondly, Gene Set Enrichment Analysis was applied to determine: i) altered regulatory pathways and ii) correlation of changes in the transcriptomic profile between the DU145 Aldrin-exposed cells and tumor progression in different tumor types.The experimentally validated predictions unveiled a dis-regulation in metabolic and regulatory pathways, modifying the triacylglyceride/cholesterol ratio in Aldrin-exposed cells, which is associated with the malignant phenotype reported in Aldrin-exposed cells.This approach can be applied to study the molecular mechanisms triggered by the chronic exposure to sub-lethal concentration of other pollutants ina variety of diseases with potential clinical and environmental applications.

Macrocyclic lactones have been the most widely used drugs for equine parasite control during the past four decades. Unlike ivermectin, moxidectin exhibits efficacy against encysted cyathostomin larvae, and is reported to have persistent efficacy with substantially longer egg reappearance periods. However, shortened egg reappearance periods have been reported recently for both macrocyclic lactones, and these findings have raised several questions: (i) are egg reappearance period patterns different after ivermectin or moxidectin treatment? (ii) Are shortened egg reappearance periods associated with certain cyathostomin species or stages? (iii) How does moxidectin's larvicidal efficacy affect egg reappearance period? To address these questions, 36 horses at pasture, aged 2-5 years old, were randomly allocated to three treatment groups: 1, moxidectin; 2, ivermectin; and 3, untreated control. Strongylid fecal egg counts were measured on a weekly basis, and the egg reappearance period was 5 weeks for both compounds. Strongylid worm counts were determined for all horses: 18 were necropsied at 2 weeks post-treatment (PT), and the remaining 18 at 5 weeks PT. Worms were identified to species morphologically and by internal transcribed spacer-2 (ITS-2) rDNA metabarcoding. Moxidectin and ivermectin were 99.9% and 99.7% efficacious against adults at 2 weeks post treatment, whereas the respective efficacies against luminal L4s were 84.3% and 69.7%. At 5 weeks PT, adulticidal efficacy was 88.3% and 57.6% for moxidectin and ivermectin, respectively, while the efficacy against luminal L4s was 0% for both drugs. Moxidectin reduced early L3 counts by 18.1% and 8.0% at 2 or 5 weeks, while the efficacies against late L3s and mucosal L4s were 60.4% and 21.2% at the same intervals, respectively. The luminal L4s surviving ivermectin treatment were predominantly Cylicocyclus (Cyc.) insigne. The ITS-2 rDNA metabarcoding was in good agreement with morphologic species estimates but suggested differential activity between moxidectin and ivermectin for several species, most notably Cyc. insigne and Cylicocyclus nassatus. This study was a comprehensive investigation of current macrocyclic lactone efficacy patterns and provided important insight into potential mechanisms behind shortened egg reappearance periods.

Species coexistence is governed by availability of resources and intraguild interactions including strategies to reduce ecological overlap. Gray foxes are dietary generalist mesopredators expected to benefit from anthropogenic disturbance, but populations have declined across the midwestern USA, including severe local extirpation rates coinciding with high coyote and domestic dog occurrence and low red fox occurrence. We used data from a large-scale camera trap survey in southern Illinois, USA to quantify intraguild spatial and temporal interactions among the canid guild including domestic dogs. We used a two-species co-occurrence model to make pairwise assessments of conditional occupancy and detection rates. We also estimated temporal activity overlap among species and fit a fixed-effects hierarchical community occupancy model with the four canid species. We partitioned the posterior distributions to compare gray fox occupancy probabilities conditional on estimated state of combinations of other species to assess support for hypothesized interactions. We found no evidence of broadscale avoidance among native canids and conclude that spatial and temporal segregation were limited by ubiquitous human disturbance. Mean guild richness was two canid species at a site and gray fox occupancy was greater when any combination of sympatric canids was also present, setting the stage for competitive exclusion over time. Domestic dogs may amplify competitive interactions by increasing canid guild size to the detriment of gray foxes. Our results suggest that while human activities can benefit some mesopredators, other species such as gray foxes may serve as bellwethers for habitat degradation with trophic downgrading and continued anthropogenic homogenization.

Population declines have been documented in many species within the genus Neotoma. Eastern woodrat, Neotoma floridana, recovery efforts in the Shawnee National Forest (SNF), Illinois USA provided an opportunity to study the long-term population-level genetic changes following an augmentation and reintroduction. We developed 24 microsatellite markers using QDD and genotyped 32 eastern woodrats from a single population. Number of alleles per locus ranged from 3-14 (mean = 7). Observed heterozygosity ranged from 0.375–0.969 per locus and expected heterozygosity from 0.485-0.854. Two loci showed significant deviation from Hardy-Weinberg equilibrium following Bonferroni sequential corrections. These markers will provide valuable information useful for studying population dynamics of eastern woodrats and closely-related species.

The dominant method for generating Chinese hamster ovary (CHO) cell lines that produce high titers of biotherapeutic proteins utilizes selectable markers such as dihydrofolate reductase (Dhfr) or glutamine synthetase (Gs), alongside inhibitory compounds like methotrexate (MTX) or methionine sulfoximine (MSX), respectively. Recent work has shown the importance of asparaginase (Aspg) for growth in media lacking glutamine-the selection medium for Gs-based selection systems. We generated a Gs/Aspg double knockout CHO cell line and evaluated its utility as a novel dual selectable system via co-transfection of Gs-Enbrel and Aspg-Enbrel plasmids. Using the same selection conditions as the standard Gs system, the resulting cells from the Gs/Aspg dual selection showed substantially improved specific productivity and titer compared to the standard Gs selection method, however, with reduced growth rate and viability. Following adaptation in selection medium, the cells improved viability and growth while still achieving ~5-fold higher specific productivity and ~3-fold higher titer than Gs selection alone. We anticipate that with further optimization of culture medium and selection conditions this approach would serve as an effective addition to workflows for the industrial production of recombinant biotherapeutics. This article is protected by copyright. All rights reserved.

The reliable and cost-efficient manufacturing of monoclonal antibodies (mAbs) is essential to fulfil their ever-growing demand. Cell death in bioreactors reduces productivity and product quality, and is largely attributed to apoptosis. In perfusion bioreactors, this leads to the necessity of a bleed stream, which negatively affects the overall process economy. To combat this limitation, death-resistant Chinese hamster ovary cell lines were developed by simultaneously knocking out the apoptosis effector proteins Bak1, Bax, and Bok with CRISPR technology. These cell lines were cultured in fed-batch and perfusion bioreactors and compared to an unmodified control cell line. In fed-batch, the death-resistant cell lines showed higher cell densities and longer culture durations, lasting nearly a month under standard culture conditions. In perfusion, the death-resistant cell lines showed slower drops in viability and displayed an arrest in cell division after which cell size increased instead. Pertinently, the death-resistant cell lines demonstrated the ability to be cultured for several weeks without bleed, and achieved similar volumetric productivities at lower cell densities than that of the control cell line. Perfusion culture reduced fragmentation of the mAb produced, and the death-resistant cell lines showed increased glycosylation in the light chain in both bioreactor modes. These data demonstrate that rationally engineered death-resistant cell lines are ideal for mAb production in perfusion culture, negating the need to bleed the bioreactor whilst maintaining product quantity and quality.

A virgin phase transition is observed in spherical particles of the industrially relevant magnetocaloric material La(Fe,Mn,Si)13H y. Upon initial cooling, the phase transition is observed 2–3 K below the heating transition on all subsequent cooling and heating transitions. This virgin transition has been studied using differential scanning calorimetry and vibrating sample magnetometry. Incremental measurements show not only how the phase transition can be carefully approached but also that the initial full transformation requires cooling of about 6 K below the observed phase transformation. No signs of structural damage due to the thermal cycling were observed, neither macroscopically or by scanning electron microscopy.

This study aimed to collect information on local and systemic inflammatory responses, and goblet cell-associated components, following anthelmintic treatment with moxidectin and ivermectin in horses naturally infected with cyathostomin parasites. Thirty-six horses aged 2-5 years of age were randomly allocated to three groups. Group 1 received ivermectin/praziquantel (0.2 mg/kg), Group 2 received moxidectin/praziquantel (0.4 mg/kg) and Group 3 were untreated controls. Tissue samples from the Cecum, Dorsal and Ventral Colons were used for histopathological evaluation and preserved for RNA isolation and gene expression analysis. Whole blood was collected weekly for gene expression analysis as well. The control group had significantly higher inflammation associated with higher larval scores. The treatment groups displayed no differences in larval counts and inflammatory cell populations (p .05). Mucosal larval counts were positively correlated with goblet cell hyperplasia scores (p = .047). The moxidectin-treated group had a significantly lower expression of IFN-γ (p .05). The data suggest that removal of cyathostomins reduced the pro-inflammatory response associated with cyathostomin infections. Pro-inflammatory reactions associated with anthelmintic treatment were minimal, but lowest for moxidectin-treated horses. Results suggested that cecum, ventral and dorsal colons responded differently to cyathostomin larvae, which may have implications in the disease process.

Globally, 13 million people (4 million in USA) inject insulin with single use disposable syringes daily. Compared to syringe and vials, pens are known to increase adherence and diabetes control and to reduce healthcare utilization costs. Yet, pens are largely unavailable to low-income groups, due to prohibitive cost. GO-Pen has designed a new affordable pen for use with vial insulin and conducted usability testing in collaboration with Médecins Sans Frontières (MSF) , an international medical humanitarian organization working to improve diabetes care for people living in difficult circumstances. Here we report the results of two usability studies conducted among insulin pen users (n=19, f/m: 11/8, age: 23-64) and syringe users (n=20, f/m: 10/10, age: 24-70) in refugee populations in Malaysia and Jordan. A convenience sample of the first 20 to accept enrollment was set up after informed consent. After a short introduction, participants were asked to demonstrate the use of the insulin pen in front of a camera, without revealing their faces or any identifying information, while an interviewer noted their correct use and confidence level. Injection was simulated in a sponge. 100% (n=19) of pen users and 75% (n=15) of syringe users performed >90% of the tasks correctly. Among pen users, the most common failed tasks were priming the needle (n=4) . Among syringe users, the most common failed tasks were failing to check the vial label (n=9) and priming the needle (n=6) . 5% (n=1) of pen users preferred the new insulin pen over the pen they were already familiar with. 75% (n=15) of syringe users preferred the new insulin pen over syringes. The most common complaint among pen users was that there were too many steps in the process. In conclusion, we find current pen users require little training and prefer the pen they are already familiar with. Syringe users who are unfamiliar with pen usage require more training than pen users, however, they have largely favorable attitudes towards this new insulin pen. Disclosure O.K.Nielsen: Stock/Shareholder; GO-Pen ApS. G.Huisman: None. J.A.Qasem: None. C.Eng: None. A.F.Jørgensen: None. M.Torrone: None. B.S.Larsen: None. A.Reddy: None.

Essentially all grazing horses are infected with cyathostomin parasites. Adult cyathostomins reside in the large intestine of the horse and larval stages encyst within intestinal mucosa. Manual worm collection from aliquots of intestinal content is the current gold standard for retrieval and enumeration of luminal parasites, however, no research has been conducted to standardize specific parameters for processing and storage of samples. The aims of this study were (1) to evaluate the precision of current standard operating procedures for enumeration of luminal adult cyathostomin populations, (2) investigate the influence of chosen fixative, either 70 % ethanol or 10 % buffered formalin, as well as storage duration, immediately post necropsy vs. stored for eight weeks, on the magnitude and precision of worm counts, and (3) compare the luminal count magnitude between the three intestinal segments (cecum, ventral colon, dorsal colon). Ten miniature horses were enrolled in this study for euthanasia and necropsy over a four-week period. Luminal worm counts were conducted for 2 % aliquots of the cecum, ventral colon, and dorsal colon and samples were allocated to the two fixatives and the two storage durations. Precision was evaluated by coefficient of variation (CV) and was 13.04 % for total cyathostomin counts. Mean CV for large intestinal segments ranged from 15.31 % to 52.50 % irrespective of fixative used or storage duration. cecum worm counts were significantly lower compared to the ventral colon (p = 0.008) and dorsal colon (p = 0.01). Fixative and storage duration were not statistically associated with count precision or magnitude. This study demonstrated moderate to high precision estimates for luminal cyathostomin worm counts but did not identify any effects of fixative and storage duration within the framework of the study. This is the first study to determine cyathostomin worm count precision, and results will be useful for power analyses in the future.

Bacillus thuringiensis or Bt is a Gram-positive soil bacterium, widely and safely applied in the environment as an insecticide for combatting insect pests that damage crops and vector diseases. Dominant active ingredients made by Bt are insect-killing crystal (Cry) proteins released as crystalline inclusions upon bacterial sporulation. Some Bt Cry proteins, e.g., Cry5B, target nematodes (roundworms) and show exceptional promise as anthelmintics (cures for parasitic nematode diseases). We have recently described IBaCC (for Inactivated Bacteria with Cytosolic Crystal(s)) in which bioactive Bt Cry crystals (containing Cry5B) are fully contained within the cytosol of dead bacterial ghosts. Here we demonstrate that these IBaCC-trapped Cry5B crystals can be liberated and purified away from cellular constituents yielding Purified Cytosolic Crystals (PCC). Cry5B PCC contains ∼95% Cry5B protein out of the total protein content. Cry5B PCC is highly bioactive against parasitic nematode larvae and adults in vitro. Cry5B PCC is also highly active in vivo against experimental human hookworm and Ascaris infections in rodents. The process was scaled up to the 100 liter scale to produce PCC for a pilot study to treat two foals infected with the Ascarid, Parascaris spp. Single dose Cry5B PCC brought the fecal egg counts of both foals to zero. These studies describe the process for the scalable production of purified Bt crystals and define a new active pharmaceutical ingredient form of Bt Cry proteins.NON-TECHNICAL IMPORTANCE PARAGRAPHBacillus thuringiensis crystal proteins are widely and safely used as insecticides. Recent studies show they also can cure gastrointestinal parasitic worm (nematode) infections when ingested. However, reproducible, scalable, and practical techniques for purifying these proteins have been lacking. Here, we address this severe limitation and present scalable and practical methods for large-scale purification of potently bioactive B. thuringiensis crystals and crystal proteins. The resultant product, called Purified Cytosolic Crystals (PCC), is highly compatible with ingestible drug delivery and formulation. Furthermore, there are growing applications in agriculture and insect control where access to large quantities of purified crystal proteins are desirable and where these methods will find great utility.