Crystal size, shape, and conformational changes drive both the disappearance and reappearance of ritonavir polymorphs in the mill.

Organic compounds can crystallize in different forms known as polymorphs. Discovery and control of polymorphism is crucial to the pharmaceutical industry since different polymorphs can have significantly different physical properties which impacts their utilization in drug delivery. Certain polymorphs have been reported to 'disappear' from the physical world, irreversibly converting to new ones. These unwanted polymorph conversions, initially prevented by slow nucleation kinetics, are eventually observed driven by significant gains in thermodynamic stabilities. The most infamous of these cases is that of the HIV drug ritonavir (RVR): Once its reluctant form was unwillingly nucleated for the first time, its desired form could no longer be produced with the same manufacturing process. Here we show that RVR's extraordinary disappearing polymorph as well as its reluctant form can be consistently produced by ball-milling under different environmental conditions. We demonstrate that the significant difference in stability between its polymorphs can be changed and reversed in the mill-a process we show is driven by crystal size as well as crystal shape and conformational effects. We also show that those effects can be controlled through careful design of milling conditions since they dictate the kinetics of crystal breakage, dissolution, and growth processes that eventually lead to steady-state crystal sizes and shapes in the mill. This work highlights the huge potential of mechanochemistry in polymorph discovery of forms initially difficult to nucleate, recovery of disappearing polymorphs, and polymorph control of complex flexible drug compounds such as RVR.

Evaluation of Truck Cab Decontamination Procedures following Inoculation with Porcine Epidemic Diarrhea Virus and Porcine Reproductive and Respiratory Syndrome Virus.

This experiment aimed to evaluate commercially available disinfectants and their application methods against porcine epidemic diarrhea virus (PEDV) and porcine reproductive and respiratory syndrome virus (PRRSV) on truck cab surfaces. Plastic, fabric, and rubber surfaces inoculated with PEDV or PRRSV were placed in a full-scale truck cab and then treated with one of eight randomly assigned disinfectant treatments. After application, surfaces were environmentally sampled with cotton gauze and tested for PEDV and PRRSV using qPCR duplex analysis. There was a disinfectant × surface interaction (p < 0.0001), indicating a detectable amount of PEDV or PRRSV RNA was impacted by disinfectant treatment and surface material. For rubber surfaces, 10% bleach application had lower detectable amounts of RNA compared to all other treatments (p < 0.05) except Intervention via misting fumigation, which was intermediate. In both fabric and plastic surfaces, there was no evidence (p > 0.05) of a difference in detectable RNA between disinfectant treatments. For disinfectant treatments, fabric surfaces with no chemical treatment had less detectable viral RNA compared to the corresponding plastic and rubber (p < 0.05). Intervention applied via pump sprayer to fabric surfaces had less detectable viral RNA than plastic (p < 0.05). Furthermore, 10% bleach applied via pump sprayer to fabric and rubber surfaces had less detectable viral RNA than plastic (p < 0.05). Also, a 10 h downtime, with no chemical application or gaseous fumigation for 10 h, applied to fabric surfaces had less detectable viral RNA than other surfaces (p < 0.05). Sixteen treatments were evaluated via swine bioassay, but all samples failed to produce infectivity. In summary, commercially available disinfectants successfully reduced detectable viral RNA on surfaces but did not eliminate viral genetic material, highlighting the importance of bioexclusion of pathogens of interest.

Beak shape and nest material use in birds.

The evolution of behaviour can both influence, and be influenced by, morphology. Recent advances in methods and data availability have facilitated broad-scale investigations of physical form and behavioural function in many contexts, but the relationship between animal morphology and object manipulation-particularly objects used in construction-remains largely unknown. Here, we employ a new global database of nest materials used by 5924 species of birds together with phylogenetically informed random forest models to evaluate the link between beak shape and these nest-building materials. We find that beak morphology, together with species diet and access to materials, can predict nest-material use above chance and with high accuracy (68-97%). Much of this relationship, however, is driven by phylogenetic signal and sampling biases. We therefore conclude that while variation in nest material use is linked with that of beak shape across bird species, these correlations are modulated by the ecological context and evolutionary history of these species. This article is part of the theme issue 'The evolutionary ecology of nests: a cross-taxon approach'.

Impact of storage conditions and premix type on water-soluble vitamin stability.

Mitigation options to reduce the risk of foreign animal disease entry into the United States may lead to degradation of some vitamins. The objective of Exp. 1 was to determine the impact of 0, 30, 60, or 90 d storage time on water-soluble vitamin (riboflavin, niacin, pantothenic acid, and cobalamin) stability when vitamin premix (VP) and vitamin trace mineral premix (VTM) were blended with 1% inclusion of medium-chain fatty acid (MCFA) (1:1:1 blend of C6:C8:C10) or mineral oil (MO) with different environmental conditions. Samples stored at room temperature (RT) (approximately 22 °C) or in an environmentally controlled chamber set at 40 °C and 75% humidity, high temperature high humidity (HTHH). The sample bags were pulled out at day 0, 30, 60, and 90 for RT condition and HTHH condition. Therefore, treatments were analyzed as a 2 × 2 × 2 × 3 factorial, with two premix types (vitamin premix vs. VTM), two oil types (MO vs. MCFA), two storage conditions (RT vs. HTHH), and three time points (day 30, 60, and 90). The objective of Exp. 2 was to determine the effect of heat pulse treatment and MCFA addition on water-soluble vitamin (riboflavin, niacin, pantothenic acid, and cobalamin) stability with two premix types. A sample from each treatment was heated at 60 °C and 20% humidity. Therefore, treatments were analyzed as a 2 × 2 factorial, with two premix types (VP vs. VTM) and two oil types (MO vs. MCFA). For Exp. 1, the following effects were significant for riboflavin: main effect of premix type (P < 0.0001), storage condition (P = 0.015), and storage time (P < 0.0001); for pantothenic acid: premix type × storage time × storage condition (P = 0.003) and premix type × oil type (P < 0.0001) interactions; and for cobalamin: premix type × storage condition (P < 0.0001) and storage time × storage condition (P < 0.0001) interactions and main effect of oil type (P = 0.018). The results of Exp. 2 demonstrated that there was an interaction between oil type and premix type for only pantothenic acid (P = 0.021). The oil type did not affect the stability of riboflavin, niacin, or cobalamin and pantothenic acid stability was not different within similar premixes. The only difference in water-soluble vitamin stability between VP and VTM was for pantothenic acid (P < 0.001). The results of this experiment demonstrated that the stability of water soluble vitamins are dependent on the vitamin of interest and the conditions at which it is stored.

Resolving alternative structure determinations of indapamide using 13C solid-state NMR.

The conflict between alternative crystal structures in the Cambridge Structural Database for the diuretic drug indapamide hemihydrate (IND) has been resolved with the aid of 13C solid-state NMR. IND is seen to contain multiple distinct molecules in the asymmetric unit (Z' = 4) rather than exhibiting disorder in the orientation of sulfonamide groups. The NMR crystallographic approach is a more effective tool for distinguishing between alternative structures than naïve judgements of quality based on crystallographic refinement agreement factors.

Sensor-Based Detection of Predator Influence on Livestock: A Case Study Exploring the Impacts of Wild Dogs (Canis familiaris) on Rangeland Sheep.

In Australia, wild dogs are one of the leading causes of sheep losses. A major problem with managing wild dogs in Australia's rangeland environments is that sheep producers are often unaware of their presence until injuries or deaths are observed. One option for earlier detection of wild dogs is on-animal sensors, such as Global Positioning System (GPS) tracking collars, to detect changes in the behaviour of sheep due to the presence of wild dogs. The current study used spatio-temporal data, derived from GPS tracking collars, deployed on sheep from a single rangeland property to determine if there were differences in the behaviour of sheep when in the presence, or absence, of a wild dog. Results indicated that the presence of a wild dog influenced the daily behaviours of sheep by increasing the daily distance travelled. Differences in sheep diurnal activity were also observed during periods where a wild dog was present or absent on the property. These results highlight the potential for on-animal sensors to be used as a monitoring tool for sheep flocks directly impacted by wild dogs, although further work is needed to determine the applicability of these results to other sheep production regions of Australia.

Pathological lesions of lambs infected in utero with bovine viral diarrhoea virus type 1c (BVDV-1c).

BACKGROUND: Acute bovine viral diarrhoea virus (BVDV) infections in sheep are largely sub-clinical although infections of pregnant ewes have shown to result in significant fetal losses and persistently infected lambs. However, the extent and severity of abnormalities in lambs infected with BVDV in utero is still largely unknown. METHODS: Twenty-two ewes were experimentally infected with BVDV-1c between 59 and 69 days of gestation. Fifteen lambs were submitted for pathological examination and the abnormalities observed in lambs and fetuses characterised. RESULTS: Six lambs were identified as BVDV negative, and nine were identified as BVDV positive. Anasarca and cholestatic hepatopathy was observed in four BVDV positive lambs and associated with ewes with early seroconversion. One BVDV positive lamb was born with muscular tremors and a hairy coat associated with primary follicular dysplasia, a developmental abnormality normally associated with border disease infected lambs. CONCLUSION: If similar lambing outcomes are identified in a commercial setting then BVDV should be considered, particularly in areas where sheep regularly come in to contact with cattle. In addition, as far as the authors are aware, this is the first reported case of a 'hairy shaker' lamb born as a result of an infection with BVDV-1c in Australia.

Non-Bovine Species and the Risk to Effective Control of Bovine Viral Diarrhoea (BVD) in Cattle.

Bovine viral diarrhoea virus (BVDV) is an economically important and highly prevalent virus of domestic cattle. Infections with BVDV may lead to both, reproductive and immunological effects that can result in widespread calf losses and increased susceptibility to diseases, such as mastitis and respiratory disease. While BVDV is generally considered to be host specific, it and other Pestivirus species, such as Border disease virus (BDV) in sheep, have been shown to be infecting species other than those from which they were originally isolated from. Recently BVDV was placed on the OIE's list of notifiable disease and control and eradication programmes for BVDV have been developed throughout much of Europe, the United States, and the United Kingdom. While some countries, including Sweden and Ireland have successfully implemented eradication programmes, other countries such as New Zealand and Australia are still in the early stages of BVDV control. Despite effective control methods, incursions of BVDV into previously cleared herds still occur. While the cause of these incursions is often due to lapses in control methods, the ability of ruminant pestiviruses to infect species other than cattle poses the question as to whether non-bovine species could be impeding the success of BVDV eradication and control. As such, the aim of this review is to make mention of what is known about the cross-species transmission of BVDV, BDV and other pestiviruses between cattle and non-bovine ungulate species and draw conclusions as to the risk non-bovine species pose to the successful control and eradication of BVDV from cattle.

Effects of conditioning temperature and pellet mill die speed on pellet quality and relative stabilities of phytase and xylanase.

The objective of this experiment was to determine the effect of conditioning temperature and die speed on pellet quality and enzyme stability of phytase and xylanase. Treatments were initially arranged as a 2 × 3 factorial of conditioning temperature (74 and 85 °C) and die speed (127, 190, and 254 rpm); however, when conditioning at 85 °C, it was not possible to pellet at 127 rpm. Thus, data were analyzed in two different segments using the GLIMMIX procedure of SAS. First, linear and quadratic contrasts were utilized to test the response to increasing die speed at 74 °C. Second, the data was analyzed as a 2 × 2 factorial of conditioning temperature (74 and 85 °C) and die speed (190 and 254 rpm). Treatments were arranged in a completely randomized design and replicated three times. Diets were conditioned for approximately 30 s and pelleted with a 4.8-mm-diameter × 44.5-mm-effective length die at a rate of 4.5 MT/h. Pellet durability index (PDI) was determined using the tumble box and Holmen NHP 100 methods. Samples of the unconditioned mash (M), conditioned mash (CM), and pellets (P) were collected and analyzed for phytase and xylanase concentration. Relative enzyme stabilities were expressed as CM:M, P:CM, and P:M. Stabilities expressed as P:M were used an indication of enzyme stability through the entire pelleting process. Diets conditioned at 74 °C showed no evidence of difference in phytase or xylanase P:M stability when decreasing die speed from 254 to 127 rpm. However, when conditioning diets at 74 °C, decreasing die speed increased (linear, P < 0.001) PDI. There was no conditioning temperature × die speed interaction for overall xylanase P:M stability or PDI. However, there was a conditioning temperature × die speed interaction (P < 0.01) for phytase P:M stability. When conditioning diets at 85 °C, increasing die speed decreased phytase P:M stability. However, when conditioning at 74 °C, increasing die speed did not influence phytase P:M stability. For main effects of conditioning temperature, increasing temperature improved (P < 0.001) PDI with no evidence of difference for xylanase P:M stability. For the main effects of die speed (254 vs. 190 rpm), decreasing die speed decreased (P < 0.001) the P:M xylanase stability, but there was no evidence of difference for PDI. The results of this trial indicate that die speed should be taken into consideration when evaluating enzyme stability of both phytase and xylanase as pellet mill models may be operating at different speeds. Additionally, increasing conditioning temperature will improve PDI but may result in decreased phytase stability.

Impact of storage conditions and premix type on fat-soluble vitamin stability.

Feed ingredients and additives could be a potential medium for foreign animal disease entry into the United States. The feed industry has taken active steps to reduce the risk of pathogen entry through ingredients. Medium chain fatty acid (MCFA) and heat pulse treatment could be an opportunity to prevent pathogen contamination. The objective of experiment 1 was to determine the impact of 0, 30, 60, or 90 d storage time on fat-soluble vitamin stability when vitamin premix (VP) and vitamin trace mineral premix (VTM) were blended with 1% inclusion of MCFA (1:1:1 blend of C6:C8:C10) or mineral oil (MO) with different environmental conditions. Samples stored at room temperature (RT) (~22 °C) or in an environmentally controlled chamber set at 40 °C and 75% humidity, high-temperature high humidity (HTHH). The sample bags were pulled out at days 0, 30, 60 and 90 for RT condition and HTHH condition. The objective of experiment 2 was to determine the effect of heat pulse treatment and MCFA addition on fat-soluble vitamin stability with two premix types. A sample from each treatment was heated at 60 °C and 20% humidity. For experiment 1, the following effects were significant for vitamin A: premix type × storage condition (P = 0.031) and storage time × storage condition (P = 0.002) interactions; for vitamin D3: main effect of storage condition (P < 0.001) and storage time (P = 0.002); and for vitamin E: storage time × storage condition interaction (P < 0.001). For experiment 2, oil type did not affect the stability of fat-soluble vitamins (P > 0.732) except for vitamin A (P = 0.030). There were no differences for fat-soluble vitamin stability between VP and VTM (P > 0.074) except for vitamin E (P = 0.016). Therefore, the fat-soluble vitamins were stable when mixed with both vitamin and VTM and stored at 22 °C with 28.4% relative humidity (RH). When premixes were stored at 39.5 °C with 78.8%RH, the vitamin A and D3 were stable up to 30 d while the vitamin E was stable up to 60 d. In addition, MCFA did not influence fat-soluble vitamin degradation during storage up to 90 d and in the heat pulse process. The vitamin stability was decreased by 5% to 10% after the premixes was heated at 60 °C for approximately nine and a half hours. If both chemical treatment (MCFA) and heat pulse treatment have similar efficiency at neutralizing or reducing the target pathogen, the process of chemical treatment could become a more practical practice.

Impact of storage conditions and premix type on phytase stability.

Potential use of medium-chain fatty acids (MCFA), increased temperatures and exposure time may be implemented to mitigate biological hazards in premixes and feed ingredients. However, there are no data on how these strategies influence phytase stability. For Exp. 1, there were no four- and three-way interactions among premix type (PT), oil type (OT), storage condition (SC), and storage time (ST) for phytase stability (P > 0.111). There were two-way interactions for PT × SC (P < 0.001) and SC × ST (P < 0.001). The OT did not affect phytase stability when premixes-containing phytase were added as either mineral oil (MO) or MCFA (P = 0.382). For Exp. 2, there was no interaction between PT and OT (P = 0.121). There were also no differences for phytase stability between vitamin premix (VP)- and vitamin trace mineral (VTM) premix-containing phytase were heated at 60 °C (P = 0.141) and between premixes-containing phytase were mixed with 1% MO added and 1% MCFA (P = 0.957). Therefore, the phytase was stable when mixed with both VP and VTM premix and stored at 22 °C with 28.4% relative humidity (RH). The phytase stability was dramatically decreased when the phytase was mixed with premixes and stored at 39.5 °C with 78.8% RH. Also, MCFA did not influence phytase degradation during storage up to 90 d and in the heat pulse process. The phytase activity was decreased by 20% after the premixes containing the phytase was heated at 60 °C for approximately 9.5 h. If both MCFA and heat pulse treatment have similar efficiency at neutralizing or reducing the target pathogen, the process of chemical treatment could become a more practical practice.

Global knowledge gaps in the prevention and control of bovine viral diarrhoea (BVD) virus.

The significant economic impacts of bovine viral diarrhoea (BVD) virus have prompted many countries worldwide to embark on regional or national BVD eradication programmes. Unlike other infectious diseases, BVD control is highly feasible in cattle production systems because the pathogenesis is well understood and there are effective tools to break the disease transmission cycle at the farm and industry levels. Coordinated control approaches typically involve directly testing populations for virus or serological screening of cattle herds to identify those with recent exposure to BVD, testing individual animals within affected herds to identify and eliminate persistently infected (PI) cattle, and implementing biosecurity measures such as double-fencing shared farm boundaries, vaccinating susceptible breeding cattle, improving visitor and equipment hygiene practices, and maintaining closed herds to prevent further disease transmission. As highlighted by the recent DISCONTOOLS review conducted by a panel of internationally recognized experts, knowledge gaps in the control measures are primarily centred around the practical application of existing tools rather than the need for creation of new tools. Further research is required to: (a) determine the most cost effective and socially acceptable means of applying BVD control measures in different cattle production systems; (b) identify the most effective ways to build widespread support for implementing BVD control measures from the bottom-up through farmer engagement and from the top-down through national policy; and (c) to develop strategies to prevent the reintroduction of BVD into disease-free regions by managing the risks associated with the movements of animals, personnel and equipment. Stronger collaboration between epidemiologists, economists and social scientists will be essential for progressing efforts to eradicate BVD from more countries worldwide.

Natural transmission of bovine viral diarrhoea virus-1c from a persistently infected neonate lamb to naïve sheep and cattle.

This study investigated the transmission of bovine viral diarrhoea virus (BVDV)-1c from a persistently infected (PI) neonate lamb to naïve sheep and cattle using three treatment groups: four naïve ewes and their five lambs, which were copaddocked with the PI lamb; five steers, which were housed in a paddock adjacent to the PI lamb; and five steers, which had direct but limited exposure to the PI lamb. Serum samples were collected and tested for BVDV-specific antibodies. Serum samples from the PI lamb, from day of birth to eight weeks of age, were tested for BVDV-specific antibodies and antigen and submitted for quantitative PCR to determine the viral load present at each week of age. Only one lamb from the copaddocked group developed BVDV-specific antibodies following comingling while all the steers in both the cattle treatment groups remained BVDV antibody negative. Quantitative PCR results from the PI lamb showed lower viral loads from day of birth to six weeks of age, compared with the results at seven and eight weeks of age. This may reflect maternal colostral BVDV antibody concentrations in the neonate lamb or other viral properties.

Investigation of AGID and two commercial ELISAs for the detection of Bovine viral diarrhea virus-specific antibodies in sheep serum.

Effective control and the eventual eradication of Bovine viral diarrhea virus (BVDV) from cattle populations depend on the accurate identification of infected animals. Although typically a disease agent of cattle, BVDV is known to infect a wide variety of nonbovine species, including sheep. However, validation of serologic tests in these nonbovine species, particularly sheep, is lacking. We analyzed 99 sheep sera (57 samples from Pestivirus-naive sheep, and 42 samples from BVDV-inoculated sheep) in order to investigate 3 serologic tests: the agarose gel immunodiffusion (AGID) and 2 commercial enzyme-linked immunosorbent assays (ELISAs) for detection of BVDV antibodies. At the manufacturer's cutoff thresholds, the AGID performed with 95.2% diagnostic sensitivity; ELISA-A performed with sensitivity of 90.5% and ELISA-B with 69.1%. All 3 tests performed with 100% diagnostic specificity. Two-graph receiver operating characteristic analysis showed that performance characteristics were optimized, such that both diagnostic sensitivity and diagnostic specificity were >95% for both ELISAs, if the thresholds were altered to 34.9% inhibition for ELISA-A and 63.5 signal-to-noise ratio for ELISA-B.

Ignorance is bliss. How parents of preschool children make sense of front-of-package visuals and claims on food.

With growing scrutiny over how the food industry advertises products aimed toward children and fewer consumers using nutrition facts panels and ingredient lists, the fronts of food packages have become an increasingly important marketing tool to understand. Front-of-package (FOP) visual and verbal claims play a critical role in capturing consumers' attention and helping them choose foods that fit their goals. Due to only possessing emergent literacy skills, preschool children are attuned to FOP visuals while parents are able to use the visuals in combination with verbal claims to make food choices for their children. The purpose of this focus group study was to explore how parents of preschool children make sense of FOP visual and verbal claims on packaged food products that are intended for their children. Thematic analysis revealed that parents associated aspects that most appeal to their preschool children - the characters and other playful visuals - with higher sugar content and artificial ingredients. However, parents were also easily led to believe the product was healthier based on visuals of fruit, more realistic pictures, health claims, cross-branding with healthier foods, and visuals suggesting the product is more natural. While parents recognized that the health claims and some visuals may not truly mean the food is healthier, they agreed that they rarely think beyond their initial impression. The food industry needs better regulatory guidance on how to communicate flavors and ingredients on package fronts in a way that helps consumers - particularly parents wanting to encourage healthy eating habits for their young children - better match their nutrition goals.