Cascade reactions with two non-physiological partners for NAD(P)H regeneration via renewable hydrogen.

An attractive application of hydrogenases, combined with the availability of cheap and renewable hydrogen (i.e., from solar and wind powered electrolysis or from recycled wastes), is the production of high-value electron-rich intermediates such as reduced nicotinamide adenine dinucleotides. Here, the capability of a very robust and oxygen-resilient [FeFe]-hydrogenase (CbA5H) from Clostridium beijerinckii SM10, previously identified in our group, combined with a reductase (BMR) from Bacillus megaterium (now reclassified as Priestia megaterium) was tested. The system shows a good stability and it was demonstrated to reach up to 28 ± 2 nmol NADPH regenerated s-1 mg of hydrogenase-1 (i.e., 1.68 ± 0.12 U mg-1, TOF: 126 ± 9 min-1) and 0.46 ± 0.04 nmol NADH regenerated s-1 mg of hydrogenase-1 (i.e., 0.028 ± 0.002 U mg-1, TOF: 2.1 ± 0.2 min-1), meaning up to 74 mg of NADPH and 1.23 mg of NADH produced per hour by a system involving 1 mg of CbA5H. The TOF is comparable with similar systems based on hydrogen as regenerating molecule for NADPH, but the system is first of its kind as for the [FeFe]-hydrogenase and the non-physiological partners used. As a proof of concept a cascade reaction involving CbA5H, BMR and a mutant BVMO from Acinetobacter radioresistens able to oxidize indole is presented. The data show how the cascade can be exploited for indigo production and multiple reaction cycles can be sustained using the regenerated NADPH.

Positive Reinforcement-Based Training for Self-Loading of Meat Horses Reduces Loading Time and Stress-Related Behavior.

The present work aimed to evaluate the efficacy of a self-loading training using positive reinforcement on stress-related behaviors shown by meat horses during loading procedures into a truck. Thirty-two meat horses (M = 18; F = 14; 6 month-old) were included in the study. All horses had limited interactions with the farmer and were not used to be restrained nor lead by halter. Horses were divided in two groups: Control Group (C; N = 14) and Training Group (T; N = 18). T horses were trained to self-load: in order to teach the horses to enter into the truck, a targeting training technique throughout a shaping process was applied. Training sessions were performed three times a week, from 9:30 a.m. to 1:30 p.m. and from 2:30 p.m. to 4:30 p.m., for 6 weeks; training was then repeated once a week to maintain the memory until the transport toward a slaughterhouse. The loading phase was video-recorded and loading time was directly recorded using a stopwatch. All horses were transported to the same slaughterhouse in 14 different days using the same truck. Behavior was subsequently analyzed with a focal animal continuous recording method. Loading time was shorter in T horses (mean ± SD = 44.44 ± 47.58 s) than in C horses (mean ± SD = 463.09 ± 918.19 s) (T-test; p = 0.019). T horses showed more forward locomotion toward the truck than C horses (T-test; p = 0.029). Our preliminary findings suggest that self-loading training may be useful to mitigate loading-related stress in meat horses, minimally socialized with humans.

Trends and correlates of antimicrobial use in broiler and turkey farms: a poultry company registry-based study in Italy.

BACKGROUND: Antimicrobial usage (AMU) in livestock plays a key role in the emergence and spread of antimicrobial resistance. Analysis of AMU data in livestock is therefore relevant for both animal and public health. OBJECTIVES: To assess AMU in 470 broiler and 252 turkey farms of one of Italy's largest poultry companies, accounting for around 30% of national poultry production, to identify trends and risk factors for AMU. METHODS: Antimicrobial treatments administered to 5827 broiler and 1264 turkey grow-out cycles in 2015-17 were expressed as DDDs for animals per population correction unit (DDDvet/PCU). A retrospective analysis was conducted to examine the effect of geographical area, season and prescribing veterinarian on AMU. Management and structural interventions implemented by the company were also assessed. RESULTS: AMU showed a 71% reduction in broilers (from 14 to 4 DDDvet/PCU) and a 56% reduction in turkeys (from 41 to 18 DDDvet/PCU) during the study period. Quinolones, macrolides and polymyxins decreased from 33% to 6% of total AMU in broilers, and from 56% to 32% in turkeys. Broiler cycles during spring and winter showed significantly higher AMU, as well as those in densely populated poultry areas. Different antimicrobial prescribing behaviour was identified among veterinarians. CONCLUSIONS: This study evidenced a decreasing trend in AMU and identified several correlates of AMU in broilers and turkeys. These factors will inform the design of interventions to further reduce AMU and therefore counteract antimicrobial resistance in these poultry sectors.