Genomic tailoring of autogenous poultry vaccines to reduce Campylobacter from farm to fork.

Campylobacter is a leading cause of food-borne gastroenteritis worldwide, linked to the consumption of contaminated poultry meat. Targeting this pathogen at source, vaccines for poultry can provide short-term caecal reductions in Campylobacter numbers in the chicken intestine. However, this approach is unlikely to reduce Campylobacter in the food chain or human incidence. This is likely as vaccines typically target only a subset of the high genomic strain diversity circulating among chicken flocks, and rapid evolution diminishes vaccine efficacy over time. To address this, we used a genomic approach to develop a whole-cell autogenous vaccine targeting isolates harbouring genes linked to survival outside of the host. We hyper-immunised a whole major UK breeder farm to passively target offspring colonisation using maternally-derived antibody. Monitoring progeny, broiler flocks revealed a near-complete shift in the post-vaccination Campylobacter population with an ~50% reduction in isolates harbouring extra-intestinal survival genes and a significant reduction of Campylobacter cells surviving on the surface of meat. Based on these findings, we developed a logistic regression model that predicted that vaccine efficacy could be extended to target 65% of a population of clinically relevant strains. Immuno-manipulation of poultry microbiomes towards less harmful commensal isolates by competitive exclusion, has major potential for reducing pathogens in the food production chain.

Seated-Shot-Put Equipment in Para Athletics-A Review and Presentation of Data From the Tokyo 2020 Paralympic Games.

This research provides a review of seated shot put alongside new data from the Tokyo 2020 Paralympic Games with the aim to understand the latest trends in equipment within a recently established rule set and how key equipment variables may impact performance for athletes in different classifications. First, a review of the literature found that the throwing pole is a key equipment aid that is not well understood, in part due to limitations in testing design. New data from the 2020 Paralympic Games showed inconsistent trends for the use of the throwing pole among athletes, particularly in transitionary classes (F33-34 and F54-55). A two-way analysis of variance found a main effect of classification on performance (p < .001), as well as an interaction effect between pole use and classification on performance (p < .05). Notably, pole users are seen to perform better than non-pole users in Class F32 (p < .05).

Prediction of Propulsion Kinematics and Performance in Wheelchair Rugby.

Prediction of propulsion kinematics and performance in wheelchair sports has the potential to improve capabilities of individual wheelchair prescription while minimizing testing requirements. While propulsion predictions have been developed for daily propulsion, these have not been extended for maximal effort in wheelchair sports. A two step-approach to predicting the effects of changing set-up in wheelchair rugby was developed, consisting of: (One) predicting propulsion kinematics during a 5 m sprint by adapting an existing linkage model; and (Two) applying partial least-squares regression to wheelchair set-up, propulsion kinematics, and performance. Eight elite wheelchair rugby players completed 5 m sprints in nine wheelchair set-ups while varying seat height, seat depth, seat angle, and tire pressure. Propulsion kinematics (contact and release angles) and performance (sprint time) were measured during each sprint and used for training and assessment for both models. Results were assessed through comparison of predicted and experimental propulsion kinematics (degree differences) for Step One and performance times (seconds differences) for Step Two. Kinematic measures, in particular contact angles, were identified with mean prediction errors less than 5 degrees for 43 of 48 predictions. Performance predictions were found to reflect on-court trends for some players, while others showed weaker prediction accuracy. More detailed modeling approaches that can account for individual athlete activity limitations would likely result in improved accuracy in propulsion and performance predictions across a range of wheelchair sports. Although this would come at an increased cost, developments would provide opportunities for more suitable set-ups earlier in an athlete's career, increasing performance and reducing injury risk.

Wheelchair Rugby chair configurations: an individual, Robust design approach.

Prescription of wheelchair rugby chairs is difficult due to the range of athlete impairment types and severities in the sport, difficulty in adjusting wheelchair settings, and assessing on-court performance. Currently, elite players rely on experiential knowledge (personal, coaches, and support staff) to select an appropriate set-up. Technological advancements, such as with inertial measurement units and processing algorithms, and representative testing approaches, has improved the potential for assessing set-ups at an individual level. An orthogonal design approach was implemented using an adjustable wheelchair to investigate the effect of seat height, seat depth, seat angle, and tyre pressure on performance, mobility, and propulsion kinematics. Six elite wheelchair rugby players completed testing in nine individually tailored wheelchair set-ups while monitoring both quantitative and qualitative measures of performance. From this testing, a recommended set-up was compared with the current set-up for each individual. A single case-study approach shows how the assessment method identifies parameter settings that can potentially improve performance. Three of six players reported a blind preference for the recommended set-up over the current set-up, whilst remaining players often displayed similar performance between their current and recommended set-ups. This approach can improve upon the current prescription process for rugby wheelchairs.

First evidence of fish nocardiosis in Mexico caused by Nocardia seriolae in farmed red drum (Sciaenops ocellatus, Linnaeus).

Between August and December 2013, the offshore cages of a commercial marine farm culturing red drum Sciaenops ocellatus in Campeche Bay Mexico were affected by an outbreak of an ulcerative granulomatous disease with up to 70% cumulative mortality. Thirty-one adults displaying open ulcers on the skin were submitted for diagnosis. At necropsy, multiple white-yellowish nodules (0.1-0.5 cm in diameter) were present in all internal organs, where the kidney and the spleen were the most severely affected. Histopathology evinced typical systemic granulomatous formations. Gram and Ziehl-Neelsen stains on tissue imprints, bacterial swabs and tissue sections revealed Gram-positive, acid-fast, branching beaded long rod filamentous bacteria. Tissue samples resulted positive for nocardiosis with a Nocardia genus-specific nested PCR. Definite identification at the species level and taxonomic positioning of the fastidious pathogen were achieved through a specific Nocardia seriolae PCR and by sequencing the gyrB gene of pure isolates. After administration of antibiotics during fry production, a posterior follow-up monitoring (from 2014 to 2017) detected mild but recurrent outbreaks of the bacteria with no seasonality pattern. To the extent of our knowledge, this is the first report of piscine nocardiosis in Mexico and the first time this disease is detected in red drum.

Placement effects of inertial measurement units on contact identification in wheelchair racing.

Inertial measurement units (IMUs) provide a practical solution for attaining key performance data for wheelchair sports. The effects of IMU placement position on the identification of propulsion characteristics are unknown. The aim of this study was to determine the variability in the reliability of cycle time measurements (time between hand contacts) across IMU locations on the chair frame (axle housings), and wheels (axle, push rim, outer rim), on both the left and right sides (n = 8). Contacts were defined by spikes in the resultant acceleration data, corresponding to impact between the hands and push rim, and verified against motion capture. Five elite wheelchair racing athletes propelled at racing speeds on a treadmill. Excellent inter-rater Intraclass Correlation Coefficient values indicated high reliability and repeatability for both motion capture and IMU signal analysis approaches (R = 0.997, p < 0.001 and R = 0.990, p < 0.001, respectively). The best results were (as determined by the best between method agreement) were observed for IMUs located on the frame. Detection reliability was positively associated with signal-to-noise ratio of the acceleration data. The IMU assessment approach facilitates an automated processing capability, which is an improvement to the currently used video analysis.

First detection of infectious spleen and kidney necrosis virus (ISKNV) associated with massive mortalities in farmed tilapia in Africa.

In late 2018, unusual patterns of very high mortality (>50% production) were reported in intensive tilapia cage culture systems across Lake Volta in Ghana. Samples of fish and fry were collected and analysed from two affected farms between October 2018 and February 2019. Affected fish showed darkening, erratic swimming and abdominal distension with associated ascites. Histopathological observations of tissues taken from moribund fish at different farms revealed lesions indicative of viral infection. These included haematopoietic cell nuclear and cytoplasmic pleomorphism with marginalization of chromatin and fine granulation. Transmission electron microscopy showed cells containing conspicuous virions with typical iridovirus morphology, that is enveloped, with icosahedral and/or polyhedral geometries and with a diameter c.160 nm. PCR confirmation and DNA sequencing identified the virions as infectious spleen and kidney necrosis virus (ISKNV). Samples of fry and older animals were all strongly positive for the presence of the virus by qPCR. All samples tested negative for TiLV and nodavirus by qPCR. All samples collected from farms prior to the mortality event were negative for ISKNV. Follow-up testing of fish and fry sampled from 5 additional sites in July 2019 showed all farms had fish that were PCR-positive for ISKNV, whether there was active disease on the farm or not, demonstrating the disease was endemic to farms all over Lake Volta by that point. The results suggest that ISKNV was the cause of disease on the investigated farms and likely had a primary role in the mortality events. A common observation of coinfections with Streptococcus agalactiae and other tilapia bacterial pathogens further suggests that these may interact to cause severe pathology, particularly in larger fish. Results demonstrate that there are a range of potential threats to the sustainability of tilapia aquaculture that need to be guarded against.

Test design and individual analysis in wheelchair rugby.

OBJECTIVES: Use a task vehicle of sprint testing in wheelchair rugby (WCR) to explore the impact of small changes to test design using both group and individual analysis. DESIGN: Exploratory, repeated measures, on-court study METHOD: 25 national or international level wheelchair rugby players completed 5×5m sprints under two conditions: (i) an acceleration from standstill in their own time, and (ii) an 'active' start, simulating a key aspect of performance. Video analysis and accelerometer data were used to measure key kinematic and performance variables with a focus on the first three strokes. Each player was grouped into a high-, mid-, or low-point group based on their sport-specific classification score. Group (paired sample t-tests) and individual (meaningful differences, performance coefficients, and Cohen's d effect sizes) analysis assessed differences between the two conditions. RESULTS: The low-point classification group performed significantly slower in the active start (p<0.05). There were no differences in sprint time for the high- and mid-point groups. Mid-point players achieved greater peak accelerations for strokes two and three in the active start (p<0.05). Individual sprint performances varied substantially, ranging from 8% decrease to 14% increase in sprint time for the active start. Meaningful differences in peak accelerations were demonstrated for 23 out of the 25 players. CONCLUSIONS: Small amendments to test design can lead to significant differences in individual athlete performance. Traditional group analyses masked important individual responses to testing conditions. There is need to further consider representative test design, and individual analysis for monitoring physical and skill performance.

Overground-Propulsion Kinematics and Acceleration in Elite Wheelchair Rugby.

PURPOSE: Maximal acceleration from standstill has been identified as a key performance indicator in wheelchair rugby; however, the impact of classification and kinematic variables on performance has received limited attention. This study aimed to investigate kinematic variables during maximal acceleration, with level of activity limitation accounted for using sport-classification scores. METHODS: Based on their sporting classification scores, which reflect combined trunk, arm, and hand function, 25 elite wheelchair rugby players were analyzed in high-, mid-, and low-point groups before completing five 5-m sprints from a stationary position. Inertial measurement units and video analysis were used to monitor key kinematic variables. RESULTS: Significant differences in kinematic variables were evident across the classification groups, particularly for the first stroke-contact angle (1-way ANOVA F2,122 = 51.5, P < .05) and first stroke time (F2,124 = 18.3, P < .05). High-point players used a first stroke-contact angle that was closer to top dead center of the wheel than either other group, while also using a shorter overall stroke time than low-point players. A linear mixed-effects model was used to investigate how kinematic variables influenced performance, with results suggesting that increased release angles (ie, farther around the wheel) and decreased stroke angles resulted in larger peak accelerations. Further investigation revealed that these results are likely influenced by strong relationships for the high-point group, as there was often no clear trend evident for midpoint and low-point groups. CONCLUSION: Findings show that various propulsion approaches exist across classification groups, with this information potentially informing individual wheelchair setups and training programs.

Discovery and in vivo evaluation of alcohol-containing benzothiazoles as potent dual-targeting bacterial DNA supercoiling inhibitors.

A series of dual-targeting, alcohol-containing benzothiazoles has been identified with superior antibacterial activity and drug-like properties. Early lead benzothiazoles containing carboxylic acid moieties showed efficacy in a well-established in vivo model, but inferior drug-like properties demanded modifications of functionality capable of demonstrating superior efficacy. Eliminating the acid group in favor of hydrophilic alcohol moieties at C(5), as well as incorporating solubilizing groups at the C(7) position of the core ring provided potent, broad-spectrum Gram-positive antibacterial activity, lower protein binding, and markedly improved efficacy in vivo.

Design, synthesis and structure-activity relationships of substituted oxazole-benzamide antibacterial inhibitors of FtsZ.

The design, synthesis and structure-activity relationships of a series of oxazole-benzamide inhibitors of the essential bacterial cell division protein FtsZ are described. Compounds had potent anti-staphylococcal activity and inhibited the cytokinesis of the clinically-significant bacterial pathogen Staphylococcus aureus. Selected analogues possessing a 5-halo oxazole also inhibited a strain of S. aureus harbouring the glycine-to-alanine amino acid substitution at residue 196 of FtsZ which conferred resistance to previously reported inhibitors in the series. Substitutions to the pseudo-benzylic carbon of the scaffold improved the pharmacokinetic properties by increasing metabolic stability and provided a mechanism for creating pro-drugs. Combining multiple substitutions based on the findings reported in this study has provided small-molecule inhibitors of FtsZ with enhanced in vitro and in vivo antibacterial efficacy.

Design, synthesis and biological evaluation of α-substituted isonipecotic acid benzothiazole analogues as potent bacterial type II topoisomerase inhibitors.

The discovery and optimisation of a new class of benzothiazole small molecules that inhibit bacterial DNA gyrase and topoisomerase IV are described. Antibacterial properties have been demonstrated by activity against DNA gyrase ATPase and potent activity against Staphylococcus aureus, Enterococcus faecalis, Streptococcus pyogenes and Haemophilus influenzae. Further refinements to the scaffold designed to enhance drug-likeness included analogues bearing an α-substituent to the carboxylic acid group, resulting in excellent solubility and favourable pharmacokinetic properties.

Biological evaluation of benzothiazole ethyl urea inhibitors of bacterial type II topoisomerases.

The type II topoisomerases DNA gyrase (GyrA/GyrB) and topoisomerase IV (ParC/ParE) are well-validated targets for antibacterial drug discovery. Because of their structural and functional homology, these enzymes are amenable to dual targeting by a single ligand. In this study, two novel benzothiazole ethyl urea-based small molecules, designated compound A and compound B, were evaluated for their biochemical, antibacterial, and pharmacokinetic properties. The two compounds inhibited the ATPase activity of GyrB and ParE with 50% inhibitory concentrations of <0.1 µg/ml. Prevention of DNA supercoiling by DNA gyrase was also observed. Both compounds potently inhibited the growth of a range of bacterial organisms, including staphylococci, streptococci, enterococci, Clostridium difficile, and selected Gram-negative respiratory pathogens. MIC90s against clinical isolates ranged from 0.015 µg/ml for Streptococcus pneumoniae to 0.25 µg/ml for Staphylococcus aureus. No cross-resistance with common drug resistance phenotypes was observed. In addition, no synergistic or antagonistic interactions between compound A or compound B and other antibiotics, including the topoisomerase inhibitors novobiocin and levofloxacin, were detected in checkerboard experiments. The frequencies of spontaneous resistance for S. aureus were <2.3 × 10(-10) with compound A and <5.8 × 10(-11) with compound B at concentrations equivalent to 8× the MICs. These values indicate a multitargeting mechanism of action. The pharmacokinetic properties of both compounds were profiled in rats. Following intravenous administration, compound B showed approximately 3-fold improvement over compound A in terms of both clearance and the area under the concentration-time curve. The measured oral bioavailability of compound B was 47.7%.

An improved small-molecule inhibitor of FtsZ with superior in vitro potency, drug-like properties, and in vivo efficacy.

The bacterial cell division protein FtsZ is an attractive target for small-molecule antibacterial drug discovery. Derivatives of 3-methoxybenzamide, including compound PC190723, have been reported to be potent and selective antistaphylococcal agents which exert their effects through the disruption of intracellular FtsZ function. Here, we report the further optimization of 3-methoxybenzamide derivatives towards a drug candidate. The in vitro and in vivo characterization of a more advanced lead compound, designated compound 1, is described. Compound 1 was potently antibacterial, with an average MIC of 0.12 µg/ml against all staphylococcal species, including methicillin- and multidrug-resistant Staphylococcus aureus and Staphylococcus epidermidis. Compound 1 inhibited an S. aureus strain carrying the G196A mutation in FtsZ, which confers resistance to PC190723. Like PC190723, compound 1 acted on whole bacterial cells by blocking cytokinesis. No interactions between compound 1 and a diverse panel of antibiotics were measured in checkerboard experiments. Compound 1 displayed suitable in vitro pharmaceutical properties and a favorable in vivo pharmacokinetic profile following intravenous and oral administration, with a calculated bioavailability of 82.0% in mice. Compound 1 demonstrated efficacy in a murine model of systemic S. aureus infection and caused a significant decrease in the bacterial load in the thigh infection model. A greater reduction in the number of S. aureus cells recovered from infected thighs, equivalent to 3.68 log units, than in those recovered from controls was achieved using a succinate prodrug of compound 1, which was designated compound 2. In summary, optimized derivatives of 3-methoxybenzamide may yield a first-in-class FtsZ inhibitor for the treatment of antibiotic-resistant staphylococcal infections.

Creating an antibacterial with in vivo efficacy: synthesis and characterization of potent inhibitors of the bacterial cell division protein FtsZ with improved pharmaceutical properties.

3-Methoxybenzamide (1) is a weak inhibitor of the essential bacterial cell division protein FtsZ. Alkyl derivatives of 1 are potent antistaphylococcal compounds with suboptimal drug-like properties. Exploration of the structure-activity relationships of analogues of these inhibitors led to the identification of potent antistaphylococcal compounds with improved pharmaceutical properties.

DNA gyrase (GyrB)/topoisomerase IV (ParE) inhibitors: synthesis and antibacterial activity.

The synthesis and antibacterial activities of three chemotypes of DNA supercoiling inhibitors based on imidazolo[1,2-a]pyridine and [1,2,4]triazolo[1,5-a]pyridine scaffolds that target the ATPase subunits of DNA gyrase and topoisomerase IV (GyrB/ParE) is reported. The most potent scaffold was selected for optimization leading to a series with potent Gram-positive antibacterial activity and a low resistance frequency.

Antibacterial alkoxybenzamide inhibitors of the essential bacterial cell division protein FtsZ.

3-Methoxybenzamide is a weak inhibitor of the essential bacterial cell division protein FtsZ. Exploration of the structure-activity relationships of 3-methoxybenzamide analogues led to the identification of potent anti-staphylococcal compounds.

An inhibitor of FtsZ with potent and selective anti-staphylococcal activity.

FtsZ is an essential bacterial guanosine triphosphatase and homolog of mammalian beta-tubulin that polymerizes and assembles into a ring to initiate cell division. We have created a class of small synthetic antibacterials, exemplified by PC190723, which inhibits FtsZ and prevents cell division. PC190723 has potent and selective in vitro bactericidal activity against staphylococci, including methicillin- and multi-drug-resistant Staphylococcus aureus. The putative inhibitor-binding site of PC190723 was mapped to a region of FtsZ that is analogous to the Taxol-binding site of tubulin. PC190723 was efficacious in an in vivo model of infection, curing mice infected with a lethal dose of S. aureus. The data validate FtsZ as a target for antibacterial intervention and identify PC190723 as suitable for optimization into a new anti-staphylococcal therapy.

Novel inhibitors of bacterial cytokinesis identified by a cell-based antibiotic screening assay.

The continuous emergence of antibiotic resistance demands that novel classes of antibiotics continue to be developed. The division machinery of bacteria is an attractive target because it comprises seven or more essential proteins that are conserved almost throughout the bacteria but are absent from humans. We describe the development of a cell-based assay for inhibitors of cell division and its use to isolate a new inhibitor of FtsZ protein, a key player in the division machinery. Biochemical, cytological, and genetic data are presented that demonstrate that FtsZ is the specific target for the compound. We also describe the effects of more potent analogues of the original hit compound that act on important pathogens, again at the level of cell division. The assay and the compounds have the potential to provide novel antibiotics with no pool of pre-existing resistance. They have provided new insight into cytokinesis in bacteria and offer important reagents for further studies of the cell division machinery.

Rhodanine-3-acetic acid derivatives as inhibitors of fungal protein mannosyl transferase 1 (PMT1).

The first inhibitors of fungal protein: mannosyl transferase 1 (PMT1) are described. They are based upon rhodanine-3-acetic acid and several compounds have been identified, for example, 5-[[3-(1-phenylethoxy)-4-(2-phenylethoxy)phenyl]methylene]-4-oxo-2-thioxo-3-thiazolidineacetic acid (5a), which inhibit Candida albicans PMT1 with IC(50)s in the range 0.2-0.5 microM. Members of the series are effective in inducing changes in morphology of C. albicans in vitro that have previously been associated with loss of the transferase activity. These compounds could serve as useful tools for studying the effects of protein O-mannosylation and its relevance in the search for novel antifungal agents.