Factors contributing to high performance of sows in free farrowing systems.

BACKGROUND: Pressure to abolish farrowing crates is increasing, and producers are faced with decisions about which alternative system to adopt. For sow welfare, well designed free farrowing systems without close confinement are considered optimal but producers have concerns about increased piglet mortality, particularly crushing by the sow. Reporting accurate performance figures from commercial farms newly operating such systems could inform the transition process. This study investigated performance on three commercial farms operating four different zero-confinement systems, three of which were newly installed. A total of 3212 litters from 2920 sows were followed from farrowing to weaning over a three-year period with key performance indicators (KPIs) recorded. Mixed Models (LMMs, GLMMs) determined the influence of different factors (e.g. farrowing system, sow parity, management aspects) and litter characteristics on performance, including levels and causes of piglet mortality. RESULTS: Piglet mortality was significantly influenced by farm/system. Live-born mortality ranged from 10.3 to 20.6% with stillbirths ranging from 2.5 to 5.9%. A larger litter size and higher parity resulted in higher levels of mortality regardless of system. In all systems, crushing was the main cause of piglet mortality (59%), but 31% of sows did not crush any piglets, whilst 26% crushed only one piglet and the remaining sows (43%) crushed two or more piglets. System significantly influenced crushing as a percentage of all deaths, with the system with the smallest spatial footprint (m2) compared to the other systems, recording the highest levels of crushing. Time from the start of the study influenced mortality, with significant reductions in crushing mortality (by ~ 4%) over the course of the three-year study. There was a highly significant effect of length of time (days) between moving sows into the farrowing accommodation and sows farrowing on piglet mortality (P < 0.001). The less time between sows moving in and farrowing, the higher the levels of piglet mortality, with ~ 3% increase in total mortality every five days. System effects were highly significant after adjusting for parity, litter size, and days pre-farrowing. CONCLUSION: These results from commercial farms demonstrate that even sows that have not been specifically selected for free farrowing are able, in many cases, to perform well in these zero-confinement systems, but that a period of adaptation is to be expected for overall farm performance. There are performance differences between the farms/systems which can be attributed to individual farm/system characteristics (e.g. pen design and management, staff expertise, pig genotypes, etc.). Higher parity sows and those producing very large litters provide a greater challenge to piglet mortality in these free farrowing systems (just as they do in crate systems). Management significantly influences performance, and ensuring sows have plenty of time to acclimatise between moving in to farrowing accommodation and giving birth is a critical aspect of improving piglet survival.

Piglets' behaviour and performance in relation to sow characteristics.

The importance of maternal care in commercial pig production is largely ignored. The sow has little possibility to interact with her piglets, and piglets are often subjected to early weaning or artificial rearing. This study aimed to investigate aspects of physiological and behavioural maternal provisioning that contribute to offspring outcomes. We hypothesised that better maternal care and nutritional provisioning would relate positively to piglet immunity, growth and behaviour. Nineteen sows and their litters were studied in free-farrowing pens. Oxytocin and tumour necrosis factor-α in colostrum/milk and salivary cortisol were sampled from sows throughout lactation. Sows were assessed for dominance rank, response to handling, maternal defensiveness, suckling initiation and termination, posture and sow-piglet contact. Piglets were weighed, measured for body mass index (BMI) and sampled for blood (Immunoglobulin G; at birth). After weaning, they experienced a human approach test (HAT) and novel object test. Correlations were explored between individual sow characteristics, individual piglet outcomes, and between sow characteristics and piglet outcomes averaged by litter. Significant correlations between sow and piglet factors were analysed at the litter level in mixed models with piglet outcomes as response variables and sow characteristics as predictor variables, while accounting for sow parity, litter size and batch. Litters grew faster when their sow had lower cortisol values (P = 0.03), while sows with lower cortisol levels had more successful suckling bouts and engaged in greater amounts of sow-piglet contact. Litters had a lower BMI at weaning when the sow had a higher milk fat percentage at d3. Litters of the most dominant sows took longer to approach the human in the HAT, while litters of sows with higher cortisol at d0 took longer to approach the novel object when assessed on correlations (r = 0.82, P < 0.001) but not when the model accounted for parity and litter size (P = 0.35). Only some of the measured nutritive and non-nutritive sow factors influenced litter performance and behaviour, with parity and litter size also playing a role. Given the continued increase in litter size, but also the interest in loose-housed lactation pens for sows, further research on sows' maternal investment and how it can be optimised is warranted.

Characterizing candidate decompression rates for hypobaric hypoxic stunning of pigs. Part 1: Reflexive behavior and physiological responses.

Alternatives to carbon dioxide (CO2) stunning for the commercial slaughter of pigs are urgently needed because there is robust evidence that exposing pigs to hypercapnic environments is associated with pain, fear, and distress. Hypobaric hypoxia (via gradual decompression, also known as Low Atmospheric Pressure Stunning or LAPS) has been validated in poultry as a humane option, but its potential to improve the welfare of pigs at slaughter is unknown. We investigated the potential of hypobaric hypoxia to reliably elicit a non-recovery state in anesthetized weaner-grower pigs within a commercially viable timeframe. We determined the effect of candidate decompression rates (40, 60, 80, 100 ms-1, at two cycle durations 480 s and 720 s) on a range of physiological and reflexive behavioral indicators of hypoxia and death. We found that the decompression rates tested caused a 100% death rate. As expected, the decompression rate had overarching effects on behavioral and physiological markers of hypoxia and death, with faster decompression rates resulting in shorter latencies to cardiac arrest and cessation of breathing. We observed a higher proportion of pigs displaying repeated and prolonged whole-body movements (likely indicative of convulsive activity) at higher frequencies when we applied the slowest decompression rate (40 ms-1) compared to all other rates. Since these responses may impact the carcass and meat quality, the slower rate of decompression (40 ms-1) should be excluded as a candidate decompression rate. Furthermore, given the marginal effects of decompression rate on physiological indicators of death and reflexive behavioral parameters, we also recommend that the fastest rate tested (100 ms-1) is excluded in further study on conscious pigs (to prevent conscious animals from being exposed to unnecessary faster decompression rates which may compromise animal welfare). This work represents a necessary proof of principle step and confirms the potential of gradual decompression for stunning purposes in pigs. Importantly, however, the data presented provide no information on the welfare outcomes associated with decompression in conscious pigs. Subsequent work should focus on the comprehensive welfare assessment of intermediate decompression rates to determine the potential of hypobaric hypoxia to provide a humane stunning method for pigs.

Transitioning from crates to free farrowing: A roadmap to navigate key decisions.

There are animal welfare concerns about the continued use of permanent crating systems for farrowing and lactating sows, which is the most prevalent maternity system in global pig production. Greater societal attention in recent years has culminated in changes (or proposed changes) to regulations as well as market-driven initiatives to move away from crated systems. Transitioning from farrowing crates to systems that allow the sow greater freedom of movement and behavioral expression requires a number of key decisions, with various trade-offs apparent when trying to balance the needs of different stakeholders. This review discusses these decisions based on common questions asked by farmers, policy makers and other stakeholders when deciding on a new system to build/approve. Based on the latest scientific evidence and practical insight, decisions such as: whether to retrofit an existing barn or build a new one, what spatial dimensions are necessary per sow place, whether to adopt free farrowing or temporary crating, how to provide substrate/enrichment and be hygienic and environmentally friendly, and how to optimize the human inputs and transition between systems are considered. The aim of this paper is to provide a roadmap for those interested in uptake of higher welfare systems and practices, as well as to highlight areas requiring further optimization and research.

Characterizing candidate decompression rates for hypobaric hypoxic stunning of pigs. Part 2: Pathological consequences.

Pigs are commonly stunned pre-slaughter by exposure to carbon dioxide (CO2), but this approach is associated with significant welfare concerns. Hypobaric hypoxia, achieved with gradual decompression (also known as Low Atmospheric Pressure Stunning or LAPS) may be an alternative, allowing the retention of welfare friendly handling approaches and group stunning. Although validated in poultry, the feasibility and welfare consequences of gradual decompression for pigs are unknown. Here, we characterize pathological changes in 60 pigs resulting from exposure to a range of candidate decompression curves (ranging from 40 to 100 ms-1 ascent equivalent, with two cycle durations 480 and 720 s). To protect welfare, we worked on unconscious, terminally anesthetized pigs which were subject to detailed post-mortem examinations by a specialized porcine veterinary pathologist. All pigs were killed as a result of exposure to decompression, irrespective of cycle rate or length. Pigs showed no external injuries during ante-mortem inspections. Exposing pigs to decompression and the unavoidable subsequent recompression resulted in generalized congestion of the carcass, organs and body cavities including the ears, oral cavity, conjunctivae and sclera, mucosa of other external orifices (anus and vulva), nasal planum, nasal cavities including nasal conchae, frontal sinuses, cranium, meninges, brain, larynx, trachea, lungs, heart, parietal pleura of the thoracic cavity, peritoneum of the abdominal cavity, stomach, small intestine, caecum, colon, liver, spleen and kidneys and representative joint cavities in the limbs (stifles and elbows). Various severities of hemorrhage were observed in the conjunctivae and sclera, mucosa of other external orifices (anus and vulva), nasal cavities including nasal conchae, frontal sinuses, cranium, meninges, brain, larynx, tracheal lumen, lungs, parietal pleura of the thoracic cavity, liver, spleen and kidneys and representative joint cavities in the limbs (stifles and elbows). In general, faster decompression rates produced higher scores, but in the conjunctivae, sclera and kidneys, faster decompression rates were associated with marginally lower congestion scores. There was considerable individual variation in pathological scores across all body regions. The congestion and hemorrhage observed could translate into welfare harms in conscious pigs undergoing this type of stunning, depending when in the cycle the damage is occurring, but no welfare related conclusions can be drawn from the responses of unconscious pigs. Since recompression is always required, its effects cannot be separated from decompression, however cessation of cardiac activity several minutes before recompression should have eliminated any haemodynamic effects relating to cardiac function and blood pressure. This study represents the first systematic attempt to identify candidate rate profiles to underpin future explorations of decompression as a stunning method for pigs. These pathological findings also inform discussions about the likely carcass quality implications of this novel stunning method.

Improving young pig welfare on-farm: The Five Domains Model.

Considering welfare through the "neonatal and nursery pig perspective" is an exciting approach and one that resonates with consumers. Overlaying this with the Five Domains Model, as we suggest in this review, points to practical on-farm improvements that provide each pig the opportunity to experience positive mental states. The Five Domains Model is broken into physical and functional states, which include Domain 1: Nutrition, Domain 2: Physical Environment, Domain 3: Health, and Domain 4: Behavioral Interaction, and Domain 5: Mental State. The Five Domains Model can build on the breadth and depth of swine welfare science to highlight opportunities to improve welfare on-farm. In Domain 1, management of increasingly large litters is considered, with examples of sow vs. artificial rearing, colostrum quality and quantity, and creep feed management strategies. Efforts can result in positive mental states such as feeling full and content and the ability to experience the pleasure of drinking and food tastes and smells. Domain 2 considers space complexity and access to key resources, along with thermal and physical amenities, to promote feelings of physical comfort. Domain 3 considers pig health in three broads, yet inter-linking categories 1) congenital and hereditary health, 2) environmental pathogen load, and 3) colostrum quality and quantity, and its effect on the microbiome. Improvements can result in a pig that displays vitality and feels healthy. Domain 4 provides the pig opportunities to express its rich behavioral repertoire, specifically positive social interactions, play, and exploration. These efforts can result in pigs feeling calm, safe, comfortable, having companionship, engaged, interested, and rewarded. In conclusion, using the Five Domains Model can highlight numerous opportunities to improve current and future housing and management through the "neonatal and nursery pig perspective" with a focus on inducing positive mental states that can result in improved quality of life and welfare state.

Considering welfare through the "neonatal and nursery pig perspective" is an exciting approach. Overlaying this with the Five Domains Model, as we suggest in this review, points to practical on-farm improvements that provide each pig the opportunity to experience positive mental states. The first four domains consider physical and functional states; with Domains 1 through 4 being Nutrition, Physical Environment, Health, and Behavioral Interaction, respectively. All interweave with Domain 5: Mental state. A plethora of examples are discussed; Domain 1 critiques optimal colostrum intake, and milk and feed quality that result in feelings of fullness and contentment. Domain 2 considers space complexity, key resource access, and thermal and physical amenities with these efforts resulting in feelings of comfort and agency. Domain 3 discusses congenital and hereditary health, environmental pathogen load, colostrum quality and quantity effects on the microbiome, and how these improve pig vitality and feelings of good health and fitness. Domain 4 discusses opportunities for the pig to express its behavioral repertoire, particularly positive social interactions, play, and exploration with feelings of control and agency. Improvements will result in pigs feeling calm, safe, comfortable, enjoying companionship, engaged, interested, and leading a rewarding life.

Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research.

Mass Spectrometry (MS) allows the analysis of proteins and peptides through a variety of methods, such as Electrospray Ionization-Mass Spectrometry (ESI-MS) or Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS). These methods allow identification of the mass of a protein or a peptide as intact molecules or the identification of a protein through peptide-mass fingerprinting generated upon enzymatic digestion. Tandem mass spectrometry (MS/MS) allows the fragmentation of proteins and peptides to determine the amino acid sequence of proteins (top-down and middle-down proteomics) and peptides (bottom-up proteomics). Furthermore, tandem mass spectrometry also allows the identification of post-translational modifications (PTMs) of proteins and peptides. Here, we discuss the application of MS/MS in biomedical research, indicating specific examples for the identification of proteins or peptides and their PTMs as relevant biomarkers for diagnostic and therapy.

Review of Temporary Crating of Farrowing and Lactating Sows.

Temporary crating (TC) provides lactating sows with the opportunity to move more freely after crate opening a few days after parturition. The aim of this paper was to evaluate whether TC gives overall welfare improvement when compared to permanent crating or free farrowing. This review shows that when pens with TC allow the sows to turn during the majority of time in the farrowing unit, it is the pen design and period of confinement in a crate within it that influence the extent to which different functional and motivated behaviors can be fulfilled. This review also indicates that there are at least short-term benefits to sows when confinement is reduced, as shown by reported increases in motivated behaviors such as exploration and interactions with piglets when not permanently crated. It remains unclear whether there are any longer-term beneficial effects (until or beyond weaning) due to the paucity of studies. Furthermore, it is uncertain whether the observed short-term benefits translate to other welfare indicators. Research findings indicate no reduction in the frequency of stereotypies or body lesions and do not provide a clear answer regarding sow stress response when released from confinement. Compared to free farrowing, TC appears beneficial for reducing piglet mortality. The impact of the time of onset of TC on the farrowing process and piglet mortality have been inconsistent. While confinement before farrowing prevents nest building behavior, consequences of this for sow physiology have been ambiguous. Confining the sow briefly after farrowing may be the best compromise, allowing the sow to perform motivated nest-building behavior, but the risks of crushing during the unconfined farrowing period may increase. Subsequent crate reopening seems to increase piglet mortality but only if done earlier than 3-5 days after farrowing. The review also provides methodological considerations, a proposal for consistent and accurate terminology when describing systems and highlights gaps of knowledge. In conclusion, TC is a step forward to better pig welfare compared to the farrowing crate, as it allows some freedom of movement for sows without impairing piglet welfare. However, more comprehensive research is needed to draw sound conclusions as to whether TC is a viable transition from permanent crating to free farrowing.

Changes in tail posture detected by a 3D machine vision system are associated with injury from damaging behaviours and ill health on commercial pig farms.

To establish whether pig tail posture is affected by injuries and ill health, a machine vision system using 3D cameras to measure tail angle was used. Camera data from 1692 pigs in 41 production batches of 42.4 (±16.6) days in length over 17 months at seven diverse grower/finisher commercial pig farms, was validated by visiting farms every 14(±10) days to score injury and ill health. Linear modelling of tail posture found considerable farm and batch effects. The percentage of tails held low (0°) or mid (1-45°) decreased over time from 54.9% and 23.8% respectively by -0.16 and -0.05%/day, while tails high (45-90°) increased from 21.5% by 0.20%/day. Although 22% of scored pigs had scratched tails, severe tail biting was rare; only 6% had tail wounds and 5% partial tail loss. Adding tail injury to models showed associations with tail posture: overall tail injury, worsening tail injury, and tail loss were associated with more pigs detected with low tail posture and fewer with high tails. Minor tail injuries and tail swelling were also associated with altered tail posture. Unexpectedly, other health and injury scores had a larger effect on tail posture- more low tails were observed when a greater proportion of pigs in a pen were scored with lameness or lesions caused by social aggression. Ear injuries were linked with reduced high tails. These findings are consistent with the idea that low tail posture could be a general indicator of poor welfare. However, effects of flank biting and ocular discharge on tail posture were not consistent with this. Our results show for the first time that perturbations in the normal time trends of tail posture are associated with tail biting and other signs of adverse health/welfare at diverse commercial farms, forming the basis for a decision support system.

A Single Dose of Fat-Based Energy Supplement to Light Birth Weight Pigs Shortly After Birth Does Not Increase Their Survival and Growth.

Low birth weight piglets are at high risk of mortality, because of the rapid depletion of their energy reserves after birth. At 3 h postpartum, 405 piglets weighing <1.1 kg were either dosed orally with 2 mL of (1) coconut oil (CO, 74 kJ/2 mL, n = 107 piglets), (2) commercial product (CP, 71 kJ/2 mL, n = 101 piglets), (3) water (W, 0 kJ/2 mL, n = 100 piglets) or (4) were sham-dosed (S, n = 97 piglets). Treatments were applied within litter (97 sows). Before treatment piglets were weighed, scored for vitality and blood glucose concentration (subset: CO = 45 piglets, CP = 38 piglets, W = 49 piglets and S = 44 piglets) and rectal temperature were measured. Rectal temperature was remeasured 1 h post-treatment (4 h postpartum). At 24 h post-treatment (27 h postpartum), vitality, weight and blood glucose were remeasured. Piglets were weighed on D5, D7, D10, D14, D21 and at weaning (27 ± 0.1 day old). Mortality rate and cause were recorded until 24h period post-treatment and until weaning. Data were analysed using Generalised Linear Mixed Models in SAS. There was no overall effect of treatment on any of the parameters measured. In conclusion, a single oral of fat-based energy supplement dose at birth did not improve growth, survival, rectal temperature or vitality of low birth weight piglets.

Exploratory study of the effects of intra-uterine growth retardation and neonatal energy supplementation of low birth-weight piglets on their post-weaning cognitive abilities.

The present study investigated the effects of intra-uterine growth retardation (IUGR, score 0-3; i.e., "normal" to "severe") level at birth, and the effects of neonatal energy supplementation (dosed with 2 ml of coconut oil, commercial product or water, or sham-dosed), on post-weaning cognitive abilities of low birth-weight piglets (< 1.1 kg). In total, 184 piglets were recruited at weaning (27 ± 0.1 days) for habituation to the test procedures, and were either tested for spatial learning and memory in a T-maze (n = 42; 37 ± 0.5 days) or for short-term memory in a spontaneous object recognition task (SORT; n = 47; 41 ± 0.3 days). Neonatal supplementation did not affect performances of pigs in the T-maze task or SORT. IUGR3 pigs tended to be faster to enter the reward arm and to obtain the reward in the reversal step of the T-Maze task, suggesting a better learning flexibility, compared to IUGR1 (entry t72.8=2.9, P = 0.024; reward t80 = 3.28, P = 0.008) and IUGR2 (entry t70.3=2.5, P = 0.068; reward t73.9 = 2.77, P = 0.034) pigs. However, a higher percentage of IUGR1 pigs tended to approach the novel object first (DSCF-value = 3.07; P = 0.076) and to interact with it more (t40 = 2.19, P = 0.085), relative to IGUR3 pigs. IUGR1 pigs showed a strong preference for the novel object, as they had a greater percentage time difference interacting with the objects when the novel object was presented (t81 = - 3.41, P = 0.013). In conclusion, some low birth-weight piglets are able to perform a spatial task and an object recognition test, but performances in these tests may be modulated by IUGR level.

Facial expression as a potential measure of both intent and emotion.

Facial expressions convey information on emotion, physical sensations, and intent. The much debated theories that facial expressions can be emotions or signals of intent have largely remained separated in animal studies. Here we integrate these approaches with the aim to 1) investigate whether pigs may use facial expressions as a signal of intent and; 2) quantify differences in facial metrics between different contexts of potentially negative emotional state. Facial metrics of 38 pigs were recorded prior to aggression, during aggression and during retreat from being attacked in a dyadic contest. Ear angle, snout ratio (length/height) and eye ratio from 572 images were measured. Prior to the occurrence of aggression, eventual initiators of the first bite had a smaller snout ratio and eventual winners showed a non-significant tendency to have their ears forward more than eventual losers. During aggression, pigs' ears were more forward orientated and their snout ratio was smaller. During retreat, pigs' ears were backwards and their eyes open less. The results suggest that facial expressions can communicate aggressive intent related to fight success, and that facial metrics can convey information about emotional responses to contexts involving aggression and fear.

Automatic early warning of tail biting in pigs: 3D cameras can detect lowered tail posture before an outbreak.

Tail biting is a major welfare and economic problem for indoor pig producers worldwide. Low tail posture is an early warning sign which could reduce tail biting unpredictability. Taking a precision livestock farming approach, we used Time-of-flight 3D cameras, processing data with machine vision algorithms, to automate the measurement of pig tail posture. Validation of the 3D algorithm found an accuracy of 73.9% at detecting low vs. not low tails (Sensitivity 88.4%, Specificity 66.8%). Twenty-three groups of 29 pigs per group were reared with intact (not docked) tails under typical commercial conditions over 8 batches. 15 groups had tail biting outbreaks, following which enrichment was added to pens and biters and/or victims were removed and treated. 3D data from outbreak groups showed the proportion of low tail detections increased pre-outbreak and declined post-outbreak. Pre-outbreak, the increase in low tails occurred at an increasing rate over time, and the proportion of low tails was higher one week pre-outbreak (-1) than 2 weeks pre-outbreak (-2). Within each batch, an outbreak and a non-outbreak control group were identified. Outbreak groups had more 3D low tail detections in weeks -1, +1 and +2 than their matched controls. Comparing 3D tail posture and tail injury scoring data, a greater proportion of low tails was associated with more injured pigs. Low tails might indicate more than just tail biting as tail posture varied between groups and over time and the proportion of low tails increased when pigs were moved to a new pen. Our findings demonstrate the potential for a 3D machine vision system to automate tail posture detection and provide early warning of tail biting on farm.

A new route to N-aromatic heterocycles from the hydrogenation of diesters in the presence of anilines.

The hydrogenation of dicarboxylic acids and their esters in the presence of anilines provides a new synthesis of heterocycles. [Ru(acac)3] and 1,1,1-tris(diphenylphosphinomethyl)ethane (triphos) gave good to excellent yields of the cyclic amines at 220 °C. When aqueous ammonia was used with dimethyl 1,6-hexadienoic acid, ε-caprolactam was obtained in good yield. A side reaction involving alkylation of the amine by methanol was suppressed by using diesters derived from longer chain and branched alcohols. Hydrogenation of optically pure diesters (dimethyl (R)-2-methylbutanedioate and dimethyl (S)-2-methylbutanedioate) with aniline afforded racemic 3-methyl-1-phenylpyrrolidine in 78% yield.

Connecting defects and amorphization in UiO-66 and MIL-140 metal­organic frameworks: a combined experimental and computational study.

The mechanism and products of the structural collapse of the metal­organic frameworks (MOFs) UiO-66, MIL-140B and MIL-140C upon ball-milling are investigated through solid state 13C NMR and pair distribution function (PDF) studies, finding amorphization to proceed by the breaking of a fraction of metal­ligand bonding in each case. The amorphous products contain inorganic­organic bonding motifs reminiscent of the crystalline phases. Whilst the inorganic Zr6O4(OH)4 clusters of UiO-66 remain intact upon structural collapse, the ZrO backbone of the MIL-140 frameworks undergoes substantial distortion. Density functional theory calculations have been performed to investigate defective models of MIL-140B and show, through comparison of calculated and experimental 13C NMR spectra, that amorphization and defects in the materials are linked.

Positive and negative gestational handling influences placental traits and mother-offspring behavior in dairy goats.

Dairy animals are subjected to a number of potential stressors throughout their lives, including daily interactions with humans. The quality of these interactions may have direct consequences for the animal undergoing the experience, but if such events occur during gestation it may also affect the developing fetus. This study examined the effects of differential handling during mid-gestation in 40 twin-bearing Saanen×Toggenburg primiparous goats. Between days 80 and 115 of gestation (gestation=150 days), goats were subjected to aversive (AVS, n=13), gentle (GEN, n=13) or minimal (M, n=14) handling protocols for 10 minute periods twice daily. The control (M) group did not receive handling treatments and all goats received normal husbandry procedures outside treatment periods. Salivary cortisol measured during the treatment period was higher in AVS goats (mean cortisol (sem) in pg/µl: AVS: 176.7 (18.2), GEN: 119.6 (11.1), M: 126.5 (13.7); P=0.007). Data collection was focussed on mother-offspring behaviors 2h post-partum, placental morphology and colostrum quality. AVS goats were the only treatment group to suffer fetal loss (16% loss vs 0% in GEN and M, P=0.05). Treatment also influenced placental morphology with a tendency for fewer cotyledons evident in placentae from the aversive treatment (AVS: 87.9 (7.8), GEN: 107.1 (7.9), M: 112.1 (9.3), P=0.093), and significantly fewer medium sized cotyledons (AVS: 67.6 (7.8), GEN: 89.3 (6.4), M: 84.3 (5.4), P=0.042). GEN goats displayed more grooming and nosing behaviors towards their young during the first 2h post-partum (grooming: GEN: 89.3% (7.1), AVS: 72.6% (7.7), M: 63.4% (9.0), P=0.045; nosing frequency: GEN: 58.8 (12.5), AVS: 28.6 (11.1), M: 34.7 (6.5), P=0.021). There was an overall trend for kids from mothers experiencing the AVS treatment to take longer to stand, reach the udder and suck compared to kids from GEN and M treatment groups. Treatment significantly affected latency to perform play behavior, with kids from AVS goats taking on average 25 min longer to play for the first time than kids from GEN and M treatment groups (P<0.001). The results show that handling during gestation affects placental morphology, fetal survival and post-partum maternal behaviors, and influences kid behavioral development. Such results have important animal welfare implications, demonstrating that negative handling of pregnant females results in poorer placental quality with potential for fetal loss. It also demonstrates the beneficial effects of positive handling on enhancement of maternal behaviors.

A comparison of the amorphization of zeolitic imidazolate frameworks (ZIFs) and aluminosilicate zeolites by ball-milling.

X-ray diffraction has been used to investigate the kinetics of amorphization through ball-milling at 20 Hz, for five zeolitic imidazolate frameworks (ZIFs) - ZIF-8, ZIF-4, ZIF-zni, BIF-1-Li and CdIF-1. We find that the rates of amorphization for the zinc-containing ZIFs increase with increasing solvent accessible volume (SAV) in the sequence ZIF-8 > ZIF-4 > ZIF-zni. The Li-B analogue of the dense ZIF-zni amorphizes more slowly than the corresponding zinc phase, with the behaviour showing a correlation with their relative bulk moduli and SAVs. The cadmium analogue of ZIF-8 (CdIF-1) amorphizes more rapidly than the zinc counterpart, which we ascribe primarily to its relatively weak M-N bonds as well as the higher SAV. The results for the ZIFs are compared to three classical zeolites - Na-X, Na-Y and ZSM-5 - with these taking up to four times longer to amorphize. The presence of adsorbed solvent in the pores is found to render both ZIF and zeolite frameworks more resistant to amorphization. X-ray total scattering measurements show that amorphous ZIF-zni is structurally indistinguishable from amorphous ZIF-4 with both structures retaining the same short-range order that is present in their crystalline precursors. By contrast, both X-ray total scattering measurements and (113)Cd NMR measurements point to changes in the local environment of amorphous CdIF-1 compared with its crystalline CdIF-1 precursor.

Drug delivery and controlled release from biocompatible metal-organic frameworks using mechanical amorphization.

We have used a family of Zr-based metal-organic frameworks (MOFs) with different functionalized (bromo, nitro and amino) and extended linkers for drug delivery. We loaded the materials with the fluorescent model molecule calcein and the anticancer drug α-cyano-4-hydroxycinnamic acid (α-CHC), and consequently performed a mechanical amorphization process to attempt to control the delivery of guest molecules. Our analysis revealed that the loading values of both molecules were higher for the MOFs containing unfunctionalized linkers. Confocal microscopy showed that all the materials were able to penetrate into cells, and the therapeutic effect of α-CHC on HeLa cells was enhanced when loaded (20 wt%) into the MOF with the longest linker. On one hand, calcein release required up to 3 days from the crystalline form for all the materials. On the other hand, the amorphous counterparts containing the bromo and nitro functional groups released only a fraction of the total loaded amount, and in the case of the amino-MOF a slow and progressive release was successfully achieved for 15 days. In the case of the materials loaded with α-CHC, no difference was observed between the crystalline and amorphous form of the materials. These results highlight the necessity of a balance between the pore size of the materials and the size of the guest molecules to accomplish a successful and efficient sustained release using this mechanical ball-milling process. Additionally, the endocytic pathway used by cells to internalize these MOFs may lead to diverse final cellular locations and consequently, different therapeutic effects. Understanding these cellular mechanisms will drive the design of more effective MOFs for drug delivery applications.

Combined experimental and computational NMR study of crystalline and amorphous zeolitic imidazolate frameworks.

Zeolitic imidazolate frameworks (ZIFs) have attracted great interest in recent years due to their high chemical and thermal stability with promising applications in gas storage and separations. We investigate the structures of three different crystalline ZIFs - ZIF-4, ZIF-8, ZIF-zni - and their amorphous counterparts using high field (13)C and (15)N CP MAS NMR. The high field (20 T) allows for the observation of all crystallographically independent carbon and nitrogen atoms in the crystalline ZIFs. Combining our experimental results with density functional theory calculations enabled the assignment of all chemical shifts. The crystalline spectra reveal the potential of high field NMR to distinguish between two ZIF polymorphs, ZIF-4 and ZIF-zni, with identical [Zn(C3H3N2)2] chemical compositions. (13)C and (15)N CP MAS NMR data obtained for the amorphous ZIFs clearly showed signal broadening upon amorphization, confirming the retention of chemical composition and the structural similarity of amorphous ZIF-4 and ZIF-zni. In the case of amorphous ZIF-8, we present evidence for the partial de-coordination of the 2-methyl imidazole linker.

Hybrid glasses from strong and fragile metal-organic framework liquids.

Hybrid glasses connect the emerging field of metal-organic frameworks (MOFs) with the glass formation, amorphization and melting processes of these chemically versatile systems. Though inorganic zeolites collapse around the glass transition and melt at higher temperatures, the relationship between amorphization and melting has so far not been investigated. Here we show how heating MOFs of zeolitic topology first results in a low density 'perfect' glass, similar to those formed in ice, silicon and disaccharides. This order-order transition leads to a super-strong liquid of low fragility that dynamically controls collapse, before a subsequent order-disorder transition, which creates a more fragile high-density liquid. After crystallization to a dense phase, which can be remelted, subsequent quenching results in a bulk glass, virtually identical to the high-density phase. We provide evidence that the wide-ranging melting temperatures of zeolitic MOFs are related to their network topologies and opens up the possibility of 'melt-casting' MOF glasses.