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Opioids and non-steroidal anti-inflammatory drugs (NSAIDs) are excellent analgesics, but recent clinical evidence suggests that these drugs might worsen disease severity in Crohn's disease patients, limiting their clinical utility for treating Inflammatory Bowel Disease (IBD). One indicator of change in well-being from conditions such as IBD is behavioral depression and disruption to activities of daily living. Preclinical measures of behavioral depression can provide an indicator of changes in quality of life and subsequent modification by candidate analgesics. In mice, nesting is an adaptive unconditioned behavior that is susceptible to disruption by noxious stimuli, and some types of pain related nesting depression are responsive to opioid and NSAID analgesics. Here we show that a 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) model of experimental colitis depresses nesting behavior in mice, and we evaluated effects of morphine, an opioid, and ketoprofen, a NSAID, on TNBS-induced nesting depression. In Swiss Webster mice, TNBS significantly reduced nesting that peaked on Day 3 and recovered in a time-dependent manner with complete recovery by Day 7. In the absence of colonic inflammation, daily treatment with morphine (1–10 mg/kg) did not decrease nesting except at 10mg/kg/day. However, in TNBS-treated mice 3.2 mg/kg/day morphine significantly exacerbated TNBS-induced nesting depression and delayed recovery. While 3.2 mg/kg/day morphine alone did not alter locomotor activity and TNBS-induced depression of locomotion recovered, the combination of TNBS and 3.2 mg/kg/day morphine significantly attenuated locomotion and prevented recovery. Daily treatment with 3.2 or 10 mg/kg ketoprofen in TNBS-treated mice did not prevent depression of nesting. These data suggest that opioid analgesics but not NSAIDS worsen colonic inflammation-induced behavioral depression. Furthermore, these findings highlight the importance of evaluating analgesic effects in models of colonic inflammation induced depression of behavior.

Herbal teas, which are a rich and diverse source of polyphenols, have been widely consumed due to their association with various health benefits. Preparation techniques can significantly affect the level of polyphenols in a cup of tea. Thus, this study investigated the impact of different preparation techniques, including brewing time in hot water, microwave-assisted extraction with cold and hot water (cold and hot MAE) for both radiation time and power, and laboratory testing condition on extractability of polyphenols in infusion from a teabag. The results showed that brewing time using hot water significantly affected the extractability of polyphenols and antioxidant activity. Cold and hot MAE conditions also significantly affected the extractability of polyphenols and antioxidant activity from a teabag infusion. Hot brewing at 7 min and cold MAE at full power with second boiled (1.93 min on and 1 min off radiation) are recommended for the preparation of herbal tea from a teabag, as these conditions had comparable extractability of polyphenols and antioxidant activity in comparison with other preparation techniques. There are over 20 major chromatogram peaks, of which 7 were identified as gallic acid, catechin, caffeic acid, ferulic acid, epicatechin gallate, quercetin, and kaempferol, revealing potential health benefits of this herbal tea.

The growth and efficiency-related traits of the Boer x Central Highland goats raised semi-intensively were evaluated. Besides, genetic and non-genetic factors affecting growth and efficiency-related traits were identified. Data were collected over a period of nine years in a flock of Boer crossbred goats and analyzed using general linear model procedure of SAS. The overall least-squares mean live weight at birth, 3-, 6-, 9- and 12 months of age were 2.52 ± 0.01, 9.81 ± 0.13, 13.8 ± 0.15, 17.8 ± 0.19, and 20.5 ± 0.29 kg, respectively. The average weight gains from birth to 3 months, 3 to 6, 6 to 9 and 9–12 months of age were 80.3, 37.5, 41.9, and 31.2 g day−1, respectively. Boer blood level, birth type, season and year of kidding had a significant effect on growth traits. The overall least-squares mean of Kleiber ratio from birth to 3 months, 3–6 months, 6–9 months, and 9 month to yearling age were 13.99 ± 0.11, 5.23 ± 0.14, 4.66 ± 0.16, and 2.95 ± 0.18, respectively. Parity, birth type, Boer blood level, season and year of birth had a significant influence on the expression of efficiency-related traits (Kleiber ratios) in different growth phases. In terms of growth performance and efficiency-related traits, F2 and F3 crossbred goats did not show any benefit over F1 crossbred goats. Increasing Boer blood level above 50% would be worthless under the existing minimal input production system. Therefore, besides to improving the management of goats, producing the first filial generation would be suitable for medium to high input production systems.

Quantifying the capacity of protected area networks to shield multiple marine megafauna with diverse life histories is complicated, as many species are wide-ranging, requiring varied monitoring approaches. Yet, such information is needed to identify and assess the potential use of umbrella species and to plan how best to enhance conservation strategies. Here, we evaluated the effectiveness of part of the European Natura 2000 protected area network (western Greece) for marine megafauna and whether loggerhead sea turtles are viable umbrella species in this coastal region. We systematically surveyed inside and outside coastal marine protected areas (MPAs) at a regional scale using aerial drones (18,505 animal records) and combined them with distribution data from published datasets (tracking, sightings, strandings) of sea turtles, elasmobranchs, cetaceans and pinnipeds. MPAs covered 56% of the surveyed coastline (~1500 km). There was just a 22% overlap in the distributions of the four groups from aerial drone and other datasets, demonstrating the value of combining different approaches to improve records of coastal area use for effective management. All four taxonomic groups were more likely to be detected inside coastal MPAs than outside, confirming sufficient habitat diversity despite varied life history traits. Coastal habitats frequented by loggerhead turtles during breeding/non-breeding periods combined overlapped with 76% of areas used by the other three groups, supporting their potential use as an umbrella species. In conclusion, this study showed that aerial drones can be readily combined with other monitoring approaches in coastal areas to enhance the management of marine megafauna in protected area networks and to identify the efficacy of umbrella species.

Capsular polysaccharides enable clinically important clones of Klebsiella pneumoniae to cause severe systemic infections in susceptible hosts. Phage-encoded capsule depolymerases have the potential to provide an alternative treatment paradigm in patients when multiple drug resistance has eroded the efficacy of conventional antibiotic chemotherapy. An investigation of 164 K. pneumoniae from intensive care patients in Thailand revealed a large number of distinct K types in low abundance but four (K2, K51, K1, K10) with a frequency of at least 5%. To identify depolymerases with the capacity to degrade capsules associated with these common K-types, 62 lytic phage were isolated from Thai hospital sewage water using K1, K2 and K51 isolates as hosts; phage plaques, without exception, displayed halos indicative of the presence of capsule-degrading enzymes. Phage genomes ranged in size from 41–348 kb with between 50 and 535 predicted coding sequences (CDSs). Using a custom phage protein database we were successful in applying annotation to 30 - 70% (mean = 58%) of these CDSs. The largest genomes, of so-called jumbo phage, carried multiple tRNAs as well as CRISPR repeat and spacer sequences. One of the smaller phage genomes was found to contain a putative Cas type 1E gene, indicating a history of host DNA acquisition in these obligate lytic phage. Whole-genome sequencing (WGS) indicated that some phage displayed an extended host range due to the presence of multiple depolymerase genes; in total, 42 candidate depolymerase genes were identified with up to eight in a single genome. Seven distinct virions were selected for further investigation on the basis of host range, phage morphology and WGS. Candidate genes for K1, K2 and K51 depolymerases were expressed and purified as his6-tagged soluble protein and enzymatic activity demonstrated against K. pneumoniae capsular polysaccharides by gel electrophoresis and Anton-Paar rolling ball viscometry. Depolymerases completely removed the capsule in K-type-specific fashion from K. pneumoniae cells. We conclude that broad-host range phage carry multiple enzymes, each with the capacity to degrade a single K-type, and any future use of these enzymes as therapeutic agents will require enzyme cocktails for utility against a range of K. pneumoniae infections.

Clinically relevant chronic pain is often associated with functional impairment and behavioral depression as an “affective/motivational” sign of pain; however preclinical animal models of inflammatory and neuropathic pain often produce weak evidence of impaired function. We hypothesized that hindpaw mechanical stimulation produced by a requirement to rear on a textured “NOX” plate would punish operant responding in rats treated with intraplantar complete Freund’s adjuvant (CFA, a model of inflammatory pain) or the chemotherapeutic paclitaxel (PTX, a model of neuropathic pain) and produce sustained pain-related depression of operant behavior. Male Sprague–Dawley rats were trained under a progressive-ratio (PR) schedule of food-maintained operant responding, then treated with CFA (100 µL in left hindpaw), PTX (2.0 mg/kg IP on alternate days for four total injections; 6.6 mg/kg IV on alternate days for three total injections), or saline vehicle. PR break points and mechanical thresholds for paw withdrawal from von Frey filaments were then tracked for 28 days. Subsequently, rats were tested with the opioid receptor antagonist naltrexone to assess latent sensitization and with the kappa opioid receptor (KOR) agonist U69593 to assess KOR function. CFA produced significant mechanical hypersensitivity for 3 weeks but decreased PR breakpoints for only 1 day. Both IP and IV PTX produced mechanical hypersensitivity for at least three weeks; however, only IV PTX decreased PR breakpoints, and this decrease was not alleviated by morphine. After recovery, naltrexone reinstated mechanical hypersensitivity in CFA- but not PTX-treated rats, and it did not reinstate depression of breakpoints in any group. U69593 dose-dependently decreased PR breakpoints in all groups with no difference between control vs. CFA/PTX groups. These results suggest that rearing on a textured NOX plate was not sufficient to punish operant responding in CFA- and PTX-treated rats despite the presence of sustained mechanical hypersensitivity. The rapid recovery of operant responding could not be attributed to latent sensitization, KOR downregulation, or behavioral tolerance. These results extend the range of conditions under which putative chronic pain manipulations produce weak evidence for depression of operant responding as a sign of the “affective/motivational” component of pain in rats.

Novel psychoactive substances (NPS) threaten public health and safety while also straining the limited resources of forensic laboratories. To efficiently allocate the finite resources available, we propose a new strategy for prioritizing NPS with abuse liability testing using a preclinical behavioral procedure in rats known as intracranial self-stimulation (ICSS). To validate this assay, the recently-scheduled synthetic cathinone α-PHP was compared to cocaine, a mechanistically similar drug of abuse, as a positive control and saline as a negative control. Male Sprague-Dawley rats (n=6) were implanted with electrodes targeting the medial forebrain bundle and trained to respond by lever-press for electrical brain stimulation. The rats were tested with doses of 0.32, 1.0, and 3.2 mg/kg α-PHP as well as 10 mg/kg of cocaine and saline administered by intraperitoneal injection. Neither saline nor 0.32 mg/kg α-PHP altered ICSS response rates compared to baseline levels of responding; however, doses of 1.0 and 3.2 mg/kg α-PHP and 10 mg/kg cocaine facilitated ICSS responding. This ICSS profile suggests that α-PHP has high abuse potential, with a rapid onset of effects and a long duration of action, and supports the decision to schedule this compound. This study demonstrates the ability of ICSS to distinguish between compounds of low and high potential for abuse. A strategy is proposed here to screen NPS using ICSS and classify emerging drugs into four priority categories for further analysis.

Productivity and profitability of sheep farming are highly influenced by lamb survival and ewe reproductive performance. Thus, this study was conducted to evaluate the survival and reproductive performance of crossbred sheep. Data collected from 2009 to 2018 from Sirinka sheep breeding stations were utilized for this study. Survival analysis was conducted by using Survival Kit 6.12 software with the Weibull model and the general linear model of SAS 9.0 was used to analyze reproductive traits. The overall mean survival rate of Dorper x Tumele crossbred lambs at 3, 6 and 12 months of age were 86.0, 76.6, and 67.9%, respectively. About 46.8% of mortality from the total death was observed during the first 120 days of life. Gastrointestinal parasites, pneumonia and septicemia were the major causes of lamb mortality. Birth weight, birth type, sex and year of lambing were the most important risk factors for survival of crossbred lambs. The overall least-squares means for litter size at birth, litter size at weaning, total litter weight at birth and total litter weight at weaning were 1.10 lambs, 0.94 lambs, 3.28 kg and 15.5 kg, respectively. Birth type, sex and year of lambing were the most determinants of ewe productive traits. Tumele and their crossbred sheep had good mothering ability necessary to successfully raise lambs to weaning. The current crossbreeding program which aims to improving growth performance had a positive influence on the survival rate of lambs. Improvement of environmental in the flock, special care for small lambs and indirect selection based on birth weight would lead to further survival improvement.

IntroductionPolypharmacy and potentially inappropriate medication use is common in older adults and is associated with adverse outcomes such as falls and hospitalisations.Methods and analysisThis study is a pharmacist-led medication optimisation initiative using an electronic tool (the Drug Burden Index (DBI) Calculator) in four hospital sites in the Canadian province of Nova Scotia. The study aims to enrol 160 participants between the preintervention and intervention groups. The Standard Protocol Items: Recommendations for Interventional Trials (SPIRIT 2013 checklist) was used to develop the protocol for this prospective interventional implementation study. A preintervention retrospective control cohort and a multiple case study analysis will also be used to assess the effect of intervention implementation. Statistical analysis will involve change in DBI scores and assessment of clinical outcomes, such as rehospitalisation and mortality using appropriate statistical tests including t-test, χ2, analysis of variance and unadjusted and adjusted regression methods.Ethics and disseminationEthics approval has been granted by the Nova Scotia Health Authority Research Ethics Board. The findings of this study will be published in peer-reviewed journals and presented at local, national and international conferences.Trial registration numberNCT03698487.

Development of key practical skills is fundamental to bioscience courses in higher education. With limitations on access to laboratory time due to the COVID-19 pandemic, a “Bioskills at home” kit was developed to create opportunities for first year undergraduate students to develop these skills using online support resources to guide their activities and build communities of learning. Equipment and activities in this kit enabled students to practice key skills such as pipetting, data handling, experimental design and microscopy, as well as build an online peer learning community through the use of discussion boards and microscopy competitions that encouraged students to explore their local environment. Students who engaged with these activities reported increased confidence in key practical skills. Practical assessment of skills showed that that there was no reduction in the proportion of students who succeeded in achieving the pipetting learning objective compared to previous years, despite a significantly reduced on-campus provision. Although the celebration event to choose the microscopy competition winners was well attended, there was limited use of the discussion boards by students to build a community of learning during the term. Refinement of this initiative will focus on providing greater scaffolding to encourage greater engagement with activities and enhance community building.

Abstract In worldwide conditions of increasingly antibiotic-resistant hospital infections, it is important to research alternative therapies. Bdellovibrio bacteriovorus bacteria naturally prey on Gram-negative pathogens, including antibiotic-resistant strains and so B. bacteriovorus have been proposed as “living antibiotics” to combat antimicrobially-resistant pathogens. Predator-prey interactions are complex and can be altered by environmental components. To be effective B. bacteriovorus predation needs to work in human body fluids such as serum where predation dynamics may differ to that studied in laboratory media. Here we combine mathematical modelling and lab experimentation to investigate the predation of an important carbapenem-resistant human pathogen, Klebsiella pneumoniae, by B. bacteriovorus in human serum versus buffer. We show experimentally that B. bacteriovorus is able to reduce prey numbers in each environment, on different timescales. Our mathematical model captures the underlying dynamics of the experimentation, including an initial predation-delay at the predator-prey-serum interface. Our research shows differences between predation in buffer and serum and highlights both the potential and limitations of B. bacteriovorus acting therapeutically against K. pneumoniae in serum, informing future research into the medicinal behaviours and dosing of this living antibacterial.

This study involved cross-sectional serological and questionnaire-based surveys to investigate the sero-epidemiology of foot and mouth disease (FMD) in domestic ruminants, and farmers' knowledge and practices about the disease in the Amhara region of Ethiopia. A multistage cluster sampling was carried out to select domestic ruminants for serological sampling and for the interview with farmers. A total of 1,672 sera samples were collected and tested using a 3ABC-Enzyme Linked Immunosorbent Assay, and 170 farmers were interviewed. An overall FMD apparent seroprevalence of 11.48% (95% CI: 7.52–17.14%) was recorded in the domestic ruminants. The overall true prevalence was 12.04%. The seroprevalence of FMD was higher in cattle (14.37%) than in goats (7.10%) and sheep (7.07%). The age stratified seroprevalence in the districts showed that 66.67% of the districts studied experienced a FMD outbreak within the preceding year of the study time. A mixed effect logistic regression analysis revealed that agroecology, the production system and the age of the animal was significantly associated with FMD seropositivity in cattle (P < 0.05). A statistically significant (P < 0.05) positive correlation (r = 0.93) was observed between cattle and small ruminant FMD seroprevalences. About 82% of the farmers interviewed knew of FMD and 85% of them had experienced the disease in their own herds before. The farmers mostly employ traditional means to control FMD. In conclusion, the findings of the study indicated that FMD is a prevalent disease in the Amhara region with more importance in the intensive production systems and the lowlands of the region. High correlation in seroprevalence between small and large ruminants indicated a possible cross transmission between these species. Therefore, small ruminants should not be overlooked in FMD control. Farmers in the region have a good level of knowledge about the disease; however, currently they heavily rely on traditional practices primarily focused on treating wounds of infected animals. This calls for extension work on available effective preventive measures of the disease, such as vaccination and movement restriction.

Chronic rejection of solid organ allografts remains the major cause of transplant failure. Donor-derived tissue-resident lymphocytes are transferred to the recipient during transplantation, but their impact on alloimmunity is unknown. Using mouse cardiac transplant models, we show that graft-versus-host recognition by passenger donor CD4 T cells markedly augments recipient cellular and humoral alloimmunity, resulting in more severe allograft vasculopathy and early graft failure. This augmentation is enhanced when donors were pre-sensitized to the recipient, is dependent upon avoidance of host NK cell recognition, and is partly due to provision of cognate help for allo-specific B cells from donor CD4 T cells recognizing B cell MHC class II in a peptide-degenerate manner. Passenger donor lymphocytes may therefore influence recipient alloimmune responses and represent a therapeutic target in solid organ transplantation.

Summary: MHC alloantigen is recognized by two pathways: “directly,” intact on donor cells, or “indirectly,” as self-restricted allopeptide. The duration of each pathway, and its relative contribution to allograft vasculopathy, remain unclear. Using a murine model of chronic allograft rejection, we report that direct-pathway CD4 T cell alloresponses, as well as indirect-pathway responses against MHC class II alloantigen, are curtailed by rapid elimination of donor hematopoietic antigen-presenting cells. In contrast, persistent presentation of epitope resulted in continual division and less-profound contraction of the class I allopeptide-specific CD4 T cell population, with approximately 10,000-fold more cells persisting than following acute allograft rejection. This expanded population nevertheless displayed sub-optimal anamnestic responses and was unable to provide co-stimulation-independent help for generating alloantibody. Indirect-pathway CD4 T cell responses are heterogeneous. Appreciation that responses against particular alloantigens dominate at late time points will likely inform development of strategies aimed at improving transplant outcomes. : The T cell allorecognition pathways governing rejection of organ allografts are poorly understood. Using a model of chronic heart graft rejection, Ali et al. show that continual presentation of MHC class I allopeptide drives late division in the responding alloreactive CD4 T cell population, resulting in a markedly augmented maintenance phase.

Hot springs are characterised by water temperatures above 36.7 °C. Temperature decreases with distance in flow away from spring vents; this natural gradient provides a unique opportunity to investigate the influence of water temperature on aquatic biota. This study investigated the relationship between water temperature and the aquatic invertebrates and benthic diatoms in outflows from a hot spring complex in tropical north Queensland, Australia. Water temperature ranged from 62.7 °C at the vents to 26.0 °C at the location furthest downstream. Richness of benthic diatoms and aquatic invertebrates increased linearly in response to decreasing temperature, with no species present in the hot vents. Multivariate analysis showed that both community assemblages had a response to the temperature gradient. A drop in aquatic invertebrate richness and a change in assemblage composition occurred between 40 °C and 42 °C, indicating a threshold at this temperature. The nearby Einasleigh River has experienced several contemporary peaks in water temperature over 40 °C, which corresponds to this threshold level. The relationships indicate that consistent increases in water temperature expected under climate change could decrease biological richness and precipitate changes in the aquatic invertebrate and benthic diatom taxa of tropical aquatic ecosystems.

Improved fertility following artificial insemination with frozen-thawed spermatozoa would offer rabbit producers faster genetic improvement. Previous work investigating cryoprotectants for rabbit spermatozoa have reported inconsistent results. Semen was collected from three rabbit bucks by artificial vagina and frozen using a standard procedure with varied cryodiluent components. Post-thaw analysis encompassed motility, sperm kinematic parameters and acrosome and membrane integrity. Spermatozoa were evaluated at 0, 2 and 4 h after thawing. Experiment 1 compared diluents with 3.5% dimethyl sulfoxide (DMSO), 1.5% acetamide, 1.75% DMSO + 0.75% acetamide or 3.5% DMSO + 1.5% acetamide. The treatment that resulted in the highest post-thaw motility (P

Although Escherichia coli K1 strains are benign commensals in adults, their acquisition at birth by the newborn may result in life-threatening systemic infections, most commonly sepsis and meningitis. Key features of these infections, including stable gastrointestinal (GI) colonization and age-dependent invasion of the bloodstream, can be replicated in the neonatal rat. We previously increased the capacity of a septicemia isolate of E. coli K1 to elicit systemic infection following colonization of the small intestine by serial passage through two-day-old (P2) rat pups. The passaged strain, A192PP (belonging to sequence type 95), induces lethal infection in all pups fed 2-6 x 106 CFU. Here we use whole-genome sequencing to identify mutations responsible for the threefold increase in lethality between the initial clinical isolate and the passaged derivative. Only four single nucleotide polymorphisms (SNPs), in genes (gloB, yjgV, tdcE) or promoters (thrA) involved in metabolic functions, were found: no changes were detected in genes encoding virulence determinants associated with the invasive potential of E. coli K1. The passaged strain differed in carbon source utilization in comparison to the clinical isolate, most notably its inability to metabolize glucose for growth. Deletion of each of the four genes from the E. coli A192PP chromosome altered the proteome, reduced the number of colonizing bacteria in the small intestine and increased the number of P2 survivors. This work indicates that changes in metabolic potential lead to increased colonization of the neonatal GI tract, increasing the potential for translocation across the GI epithelium into the systemic circulation.