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The primate scapula has been studied widely since its shape has been shown to correlate with how the forelimb is used in daily activities. In this study, we expand on the existing literature and use an image-based methodology that was originally developed for orthopaedic practice to quantify and compare the three-dimensional (3D) morphology of the scapula across humans and great apes. We expect that this image-based approach will allow us to identify differences between great apes and humans that can be related to differences in mobility and loading regime of the shoulder. We hypothesize that gorillas and chimpanzees will have a similar scapular morphology, geared towards stability and weight-bearing in knuckle-walking, whilst the scapular morphology of orangutans is expected to be more similar to that of humans given their high glenohumeral mobility associated with their suspensory lifestyle. We made 3D reconstructions of computed tomography scans of 69 scapulae from four hominid genera (Pongo, Gorilla, Pan and Homo). On these 3D bone meshes, the inferior glenoid plane was determined, and subsequently, a set of bony landmarks on the scapular body, coracoid, and acromion were defined. These landmarks allowed us to measure a set of functionally relevant angles which represent acromial overhang, subacromial space and coracoacromial space. The angles that were measured are: the delto-fulcral triangle (DFT), comprising the alpha, beta, and delta angle, the acromion-glenoid angle (AGA), the coracoid-glenoid centre-posterior acromial angle (CGA), the anterior tilt (TA CGA) and the posterior tilt of the CGA (PT CGA). Three observers placed the landmarks on the 3D bone meshes, allowing us to calculate the inter-observer error. The main differences in the DFT were found between humans and the great apes, with small differences between the great apes. The DFT of humans was significantly lower compared to that of the great apes, with the smallest alpha (32.7 degrees), smallest delta (45.7 degrees) and highest beta angle (101.6 degrees) of all genera. The DFT of chimpanzees was significantly higher compared to that of humans (p < 0.01), with a larger alpha (37.6 degrees) and delta angle (54.5 degrees) and smaller beta angle (87.9 degrees). The mean AGA of humans (59.1 degrees) was significantly smaller (p < 0.001) than that of gorillas (68.8 degrees). The mean CGA of humans (110.1 degrees) was significantly higher (p < 0.001) than in orangutans (92.9 degrees). Humans and gorillas showed mainly a posterior tilt of their coracoacromial complex whilst chimpanzees showed mainly an anterior tilt. The coracoacromial complex of the orangutans was not tilted anteriorly or posteriorly. With our image-based method, we were able to identify morphological features of the scapula that differed significantly between hominid genera. However, we did not find an overall dichotomy in scapular morphology geared towards high stability (Pan/Gorilla) or high mobility (Homo/Pongo). Further research is needed to investigate the functional implications of these differences in scapular morphology.

This study investigated the influence of several covariates on the time and sequence of deciduous dentition emergence in puppies. Data were obtained in a longitudinal study, with some cross-sectional observations, of 1001 puppies of 53 dog breeds. A parametric proportional hazards survival model was used to estimate median emergence time and evaluate the effect of the covariates. No significant differences were found between the left and right sides of a puppy's dentition, but differences were statistically significant for the earlier appearance of maxillary incisors and canines and later appearance of maxillary premolars compared with their mandibular counterparts. The tendency for delayed onset and completion of emergence in female compared to male puppies was statistically but not clinically significant. The differences between puppies of breeds of different size or skull type were both statistically and clinically significant, with small and brachycephalic breeds showing later emergence times, longer clinical eruption times and more individual variation. Per quadrant, regardless of dog breed, canines or incisors were usually the first teeth to emerge and fully erupt, followed by premolars in the order Pd3 > 4 > 2. The maxillary canines and incisors usually emerged earlier than mandibular canines. Age estimation standards for breed size groups are presented based on the number of emerged teeth per quadrant. To assess whether a puppy has reached the legally required minimum age of 8 weeks to leave the litter, the best predictive capability using the data from this study is obtained when assessing the emergence status of the deciduous third premolars.

Vascular corrosion casting is a method used to visualize the three dimensional anatomy and branching pattern of blood vessels, guiding insight into health and cardiovascular disease pathogenesis and progression. A polymer resin is injected in the vascular system and, after curing, the surrounding tissue is removed. This corrosion process often deforms or even fractures the fragile cast, resulting in an overall loss of information. Here, we propose a method that does not require corrosion of the tissue, based on in-situ high-resolution computed tomography (micro-CT) scans. Since there is a lack of CT contrast between the polymer cast and the animals’ surrounding soft tissue, we introduce hafnium oxide nanocrystals (HfO2 NCs) as CT contrast agents into the resin. The NCs dramatically improve the overall CT contrast of the cast and allow for straightforward segmentation in the CT scans. We designed the NC surface chemistry to ensure colloidal stability of the NCs in the casting resin, resulting in a homogeneous dispersion that remains stable during casting and curing. Using only 5 m% of HfO2 NCs, high-quality casts of both zebrafish and mouse models could be segmented using CT imaging software, allowing us to differentiate even μm scale details, without having to alter the resin injection method or affecting the resin’s mechanical properties. Our new method of virtual dissection by visualizing casts in-situ using contrast enhanced CT imaging greatly expands the application potential of the technique.

The sesamoid bone in the tendon of the m. abductor digiti primi longus is considered present in most dog breeds and is described to be radiologically detectable at the level of the carpus from the age of 4 months. However, an extensive investigation of this sesamoid bone has not been conducted before. The aim of this study was therefore to determine its prevalence in different dog breeds, to describe its histological development, and to determine the age at which it becomes radiologically visible. The prevalence of the sesamoid bone was assessed on radiographic images of the carpus or by dissection of the carpal region in 743 adult dogs of 115 breeds. Its development was studied by dissection and histological analysis in 45 puppies and its timing of radiological appearance was evaluated in 209 puppies. At least one sesamoid bone was present in all adult dogs, except for 14 dogs of six breeds of predominantly the small breed category. The lowest prevalence rate of 38.46% was exhibited in the French bulldog. The histological development could be divided into five stages. The first radiographic appearance corresponded to the coalescence of smaller ossification centers into one big nucleus (stage 4). The mean time of radiographic appearance was 108.4 days. This study provides extensive data on the prevalence and timing of the radiographic appearance of a sesamoid at the carpus of the dog. The data on radiographic appearance may be helpful in the age estimation of puppies.

Objective To develop an ultrasound (US)-guided transversus abdominis plane (TAP) block in pigs. Study design Prospective, descriptive, experimental study. Animals A total of 26 hemi-abdomens belonging to 13 Seghers Hybrid cadavers. Methods The study consisted of two parts. In part 1, a preliminary anatomical understanding of the abdominal wall innervation in pigs was established (two hemi-abdomens of one pig). Part 2 was divided into three phases, and a US-guided TAP technique using methylene blue dye (0.3 mL kg–1 for each hemi-abdomen) was developed. In chronological order, a pilot study (phase I) was conducted to establish good injection points (four hemi-abdomens). In phase II a two-point injection technique (10 hemi-abdomens) was performed. A cranial injection was made at two-thirds of the distance between the xyphoid process and the iliac crest, immediately ventral to the rib arch. A caudal injection was performed ventral to the last rib. In phase III a three-point injection technique was performed (10 hemi-abdomens) with an extra injection point halfway between the cranial and caudal injection point. Staining of the different nerves was recorded during dissection. Results In part 1 the nerves innervating the pig’s abdominal wall, thoracic vertebrae 12–16 (T12–16) and lumbar vertebrae 1–3 (L1–3), were identified. In part 2, data from the pilot study, adequate nerve staining with the two- or three-point injection technique, were obtained in T12 (29 versus 45% respectively), T13 (29 versus 100%), T14 (73 versus 45%), T15 (75 versus 78%), T16 (58 versus 78%), L1 (100 versus 75%), L2 (88 versus 84%) and L3 (23 versus 0%). Conclusions and clinical relevance US-guided TAP block can be used in swine but only a moderate success rate for adequate nerve staining was achieved in this study. Further studies are necessary to determine a correct injection volume and assess clinical intra- and postoperative efficacy.

Significance The ever-growing human population faces problems providing sufficient animal proteins. For shellfish, wild catch is supplemented by aquaculture to meet this increasing global demand. However, aquaculture faces serious problems with infectious diseases. One of the main problems for oriented disease control is lack of information on pathogen entry in the host. The present study fully describes the anatomy of the shrimp’s excretory organ, the antennal gland, identifying the organ as a major portal candidate. Additional findings show that pathogens may indeed enter through this organ naturally, infecting shrimp. We also demonstrate involvement in molting. These insights into the molting process and pathogen entry open doors in fundamental biology and the potential development of disease control measures., Viruses, such as white spot syndrome virus, and bacteria, such as Vibrio species, wreak havoc in shrimp aquaculture [C. M. Escobedo-Bonilla et al., J. Fish. Dis. 31, 1–18 (2008)]. As the main portal of entry for shrimp-related pathogens remain unclear, infectious diseases are difficult to prevent and control. Because the cuticle is a strong pathogen barrier, regions lacking cuticular lining, such as the shrimp’s excretory organ, “the antennal gland,” are major candidate entry portals [M. Corteel et al., Vet. Microbiol. 137, 209–216 (2009)]. The antennal gland, up until now morphologically underexplored, is studied using several imaging techniques. Using histology-based three-dimensional technology, we demonstrate that the antennal gland resembles a kidney, connected to a urinary bladder with a nephropore (exit opening) and a complex of diverticula, spread throughout the cephalothorax. Micromagnetic resonance imaging of live shrimp not only confirms the histology-based model, but also indicates that the filling of the diverticula is linked to the molting cycle and possibly involved therein. Based on function and complexity, we propose to rename the antennal gland as the “nephrocomplex.” By an intrabladder inoculation, we showed high susceptibility of this nephrocomplex to both white spot syndrome virus and Vibrio infection compared to peroral inoculation. An induced drop in salinity allowed the virus to enter the nephrocomplex in a natural way and caused a general infection followed by death; fluorescent beads were used to demonstrate that particles may indeed enter through the nephropore. These findings pave the way for oriented disease control in shrimp.

The organogenesis of the digestive system was described in the African lungfish (Protopterus aethiopicus) from 6 days post-hatching (6 DPH) to 17 days post-hatching (17 DPH) reared at 27 degrees C. To elucidate the position of the gastrointestinal tract in relation to the neural tube, notochord and yolk sac at 6 DPH, and to the vertebral column, lungs and kidneys at 17 DPH, larvae were mapped by means of computer-assisted 3D reconstructions starting from histological serial sections. The larvae showed a simple digestive tract, which appeared as a straight undifferentiated and closed tube at 6 DPH. Microscopical observation showed that yolk reserves were not completely depleted by 17 DPH. During the endogenous feeding period at least up to 17 DPH, the larval digestive system experienced a fast transformation with almost complete development of most digestive organs (pharyngeal teeth, intestinal vestibule, intestine and liver). Our findings suggest that, by 17 DPH, African lungfish larvae are ready to start exogenous feeding.

Hondenfokkerij en -handel zijn strikt gereguleerd door EU-wetgeving en nationale en regionale wetgeving. Desondanks wordt aangenomen dat een deel van de verhandelde puppy’s binnen de EU jonger is dan de wettelijke minimumleeftijd. Dit kan zowel de gezondheid en het welzijn van de pup als de volksgezondheid in gevaar brengen. Een juiste leeftijdsbepaling is noodzakelijk om mogelijke inbreuken aan te tonen. Het combineren van meerdere onafhankelijke methoden is essentieel ter compensatie van mogelijke variaties binnen een methode. Het vaststellen van zowel de gebits- als de botleeftijd, meer specifiek aan de hand van de eruptiestatus van het gebit en de radiologische ontwikkeling van ossificatiecentra, komen hiervoor in aanmerking. De huidige beschikbare gegevens zijn echter niet gedetailleerd genoeg. Om de nauwkeurigheid van de leeftijdsschatting te verhogen en fouten te vermijden zijn 1. meer rasspecifieke gegevens noodzakelijk, 2. moeten meer tussenliggende ontwikkelingsstadia beschreven worden en 3. moet rekening gehouden worden met mogelijke ontwikkelingsstoornissen en afwijkingen.

Age estimation in adult dogs can be performed by the radiographic measurement of the tooth pulp cavity, but the technique has hardly been described. In this study, the application of measuring pulp/tooth width ratios (P/T ratios) of the maxillary canine teeth was investigated. Pulp and tooth widths were measured at two locations on 166 maxillary canine teeth of the heads of 84 dog cadavers, using digital extraoral lateral oblique open mouth radiographs. The dogs belonged to different breeds and sexes and had a known age between 194 and 1907 days (approximately 6 months - 5 years). Both at the cemento-enamel junction (CE) and the half-height of the tooth, a comparable non-linear regression with age was demonstrated. Measuring at the CE location was less hindered by wear or superimposition. No statistically significant difference according to sex and breed size and no clinically significant difference according to skull type was found. The highest predictable capacity was found in the youngest dogs until the age of 448 days, of which 84.4% of the canine teeth had a P/T ratio above 0.39. Our results demonstrate that measuring P/T ratios of canine teeth can be used in practice to assign dogs to age categories, with the highest accuracy in young adult dogs.

Highly sensitive conductive polymer composites for piezoresistive sensing are developed by a design of the formulation variables of extrusion-based manufacturing (filler type/amount, polymer amount) and annealing (a), considering thermoplastic polyurethane (TPU) and/or olefin block copolymer (OBC) as polymer matrix and carbon black (CB) as conductive filler. With ternary composites - based on a CB type with stronger filler-matrix interactions and an appropriate OBC/TPU blend mass ratio (40/60 with CB amount of 5–10 m%; 50/50 with CB amount of 10 m%), the challenging region of both high sensitivity and static strain (maximal gauge factors (GFmax) > 50 and emax > 100%) can be realized: GFmax > 104 and emax = 20–240%. OBC binary composites with a high CB2 amount (e.g. 15 m%) are however needed for ultrahigh static strains (emax > 600%). Well-designed ternary composites (e.g. OBC40-CB/TPU60-7-a and OBC30-CB/TPU70-7-a) possess a large dynamic resistance change, negligible hysteresis and high stability and display strain sensor application potential. Highly CB2 loaded binary (≥12 m%) and ternary composites (10 m%) exhibit a more obvious strain-dependent dynamic hysteretic behavior, as they switch from a dual peak to single peak pattern toward the sensing strain limit, which is interesting for self-diagnose.

Polyacrylics have been considered for a broad range of material applications, including coatings, dental applications, and adhesives. In this experimental study, the casting potential of a group of (co)monomers belonging to the acrylic family has been explored to enable a more sustainable use of these polymer materials in the medical and veterinary science field. The individual contributions of each comonomer have been analyzed, the reaction conversion has been studied via gas chromatography (GC), the rheological behavior has been characterized via stress-controlled measurements, and the final mechanical properties have been obtained from tensile, flexure, and impact tests. The GC results allow assessing the pot life and thus the working window of the casting process. For the rheological measurements, which start from low-viscous mixtures, a novel protocol has been introduced to obtain accurate absolute data. The rheological data reflect the time dependencies of the GC data but facilitate a more direct link with the macroscopic material data. Specifically, the steep increase in the viscosity with increasing reaction time for the methyl methacrylate (MMA)/ethylene glycol dimethyl methacrylate (EGDMA) case (2% crosslinker) allows maximizing several mechanical properties: the tensile/flexure modulus, the tensile/flexure stress at break, and the impact strength. This opens the pathway to more dedicated chemistry design for corrosion casting and polyacrylic material design in general.

The blue mussel Mytilus edulis is an intensely studied bivalve in biomonitoring programs worldwide. The lack of detailed descriptions of hemolymph-withdrawal protocols, particularly with regard to the place from where hemolymph could be perfused from, raises questions regarding the exact composition of aspirated hemolymph and does not exclude the possibility of contamination with other body-fluids. This study demonstrates the use of high resolution X-ray computed tomography and histology combined with 3D-reconstruction using AMIRA-software to visualize some important vascular-related anatomic structures of Mytilus edulis. Based on these images, different hemolymph extraction sites used in bivalve research were visualized and described, leading to new insights into hemolymph collection. Results show that hemolymph withdrawn from the posterior adductor muscle could be extracted from small spaces and fissures between the muscle fibers that are connected to at least one hemolymph supplying artery, more specifically the left posterior gastro-intestinal artery. Furthermore, 3D-reconstructions indicate that puncturing hemolymph from the pericard, anterior aorta, atria and ventricle in a non-invasive way should be possible. Hemolymph withdrawal from the heart is less straightforward and more prone to contamination from the pallial cavity. This study resulted simultaneously in a detailed description and visualization of the vascular-related anatomy of Mytilus edulis.

BACKGROUND Echocardiography is the imaging technique of choice for the equine heart. Nevertheless, knowledge about ultrasonographic identification of dorsally located structures and vessels, related to the atria, in horses is scarce. OBJECTIVES To describe the echocardiographic approach and the identification of structures and vessels in relation to the atria in healthy horses. METHODS CT images from two equine hearts, casted with self-expanding foam, were segmented and used to identify atrial-related structures and vessels. These images were compared with standard and nonstandard ultrasound images from ten healthy horses obtained from a left and right parasternal view optimised to visualise the dorsal cardiac area. RESULTS On new standard ultrasound views, specific atrial anatomical landmarks such as vena cava, pulmonary arteries, intervenous tubercle and oval fossa were identified in all horses. In addition, ultrasound views were defined to visualise the brachiocephalic trunk, pulmonary veins and their ostia. The 3D segmented CT images from casted hearts were used to reconstruct slices that corresponded with the echocardiographic images and allowed correct identification of specific structures. MAIN LIMITATIONS Ultrasound examinations and casts were from different animals. A small number of casts and horses were used; therefore, anatomical variation or individual differences in identifying structures on ultrasound could not be assessed. CONCLUSIONS Important cardiac structures and vessels, even the different pulmonary veins, could be identified on standard and nonstandard ultrasound images in adult horses. This knowledge is important to guide and develop interventional cardiology and might be useful for diagnostic and therapeutic purposes.

Highly sensitive conductive polymer composites (CPCs) are designed employing a facile and low-cost extrusion manufacturing process for both low- and high-strain sensing in the field of, for example, structural health/damage monitoring and human body movement tracking. Focus is on the morphology control for extrusion-processed carbon black (CB)-filled CPCs, utilizing binary and ternary composites based on thermoplastic polyurethane (TPU) and olefin block copolymer (OBC). The relevance of the correct CB amount, kinetic control through a variation of the compounding sequence, and thermodynamic control induced by annealing is highlighted, considering a wide range of experimental (e.g., static and dynamic resistance/scanning electron microscopy/rheological measurements) and theoretical analyses. High CB mass fractions (20 m %) are needed for OBC (or TPU)-CB binary composites but only lead to an intermediate sensitivity as their conductive network is fully packed and therefore difficult to be truly destructed. Annealing is needed to enable a monotonic increase of the relative resistance with respect to strain. With ternary composites, a much higher sensitivity with a clearer monotonic increase results, provided that a low CB mass fraction (10-16 m %) is used and annealing is applied. In particular, with CB first dispersed in OBC and annealing, a less compact, hence, brittle conductive network (10-12 m % CB) is obtained, allowing high-performance sensing.

Background: Recurrence of atrial fibrillation after an ablation procedure remains a major problem which emphasizes the need for improved pulmonary vein isolation techniques. Aims: The aim of this study was to describe an implantation procedure of a pulmonary vein-stent which may possibly serve as an ablation technique in the future and to examine stent safety in a follow-up study in pigs. Methods and results: Eight pigs were catheterized and nine self-expanding nitinol stents were implanted through a transfemoral or transatrial approach into the antra of the pulmonary veins. After 3 months' follow-up, the animals were euthanized for further examination. During the follow-up phase, no complications were observed. Absence of thrombus formation or pulmonary vein wall dissection was noticed during anatomical and histological evaluation of the heart-lung packages. All implants were almost completely covered by neo-intima, of which thickness varied between 0.2 and 3.9mm. Conclusions: Stents can safely be positioned and deployed into the antra of the pulmonary veins without any acute or long-term (3 months) adverse effects. In the future, these implants could function as a permanently implanted ablation device and provide new therapeutic strategies for pulmonary vein isolation in patients with atrial fibrillation.

This work reviews the anatomy of the upper respiratory tract in domestic birds including the chicken and pigeon. Non‐exhaustive additional information on other bird species, illustrating the extraordinary diversity in the biological class Aves, can be found in several footnotes. The described anatomical structures are functionally considered in view of avian sound production. In particular, the Syrinx is invaluable. Its most important structures are the Labia and the lateral and medial tympaniform membranes in non‐songbirds and songbirds, respectively. These structures produce sound by vibrating during expiration and eventually inspiration.

Angiopoietins and their TIE receptors are important regulators of vascular stability and remodeling. These molecules are involved not only in the normal development of kidney glomeruli, but also in disease, thus making them promising targets for therapies. Although TIE receptors are mainly found in endothelial cells, some reports observed TIE2 expression in glomerular podocytes as well. This suggests a role of angiopoietins in the regulation of podocytes. In the present study, we aimed to map the subcellular localization of TIE receptors in metanephric glomeruli of fetal pigs using high-resolution immunogold electron microscopy and the relative labeling index stereological approach. TIE1 and TIE2 antibody labeling was detected on the abluminal side of endothelial cell membranes. In endothelial cells, 4.5% of TIE2 was observed close to cell-cell contacts and 11.9% of TIE2 was found in closely associated pairs, which suggests the presence of homodimers. Interestingly, both receptors were also expressed in podocyte foot processes indicating that TIE1 and TIE2 may play a similar role in podocytes as in endothelial cells.

Objective To investigate the anatomy of the ear innervation in horses and develop a new approach for the block of the internal and external pinna and acoustic meatus. Study design Prospective, descriptive, cadaveric study. Animals A total of 25 heads and necks from horse cadavers. Methods Phase I: anatomical dissection and dye deposition on 19 heads. With the head positioned in lateral recumbency, 10 mL methylene blue (1:1 sterile water) for the internal auricular and 2 mL for the great auricular nerve block were injected in 21 ears (seven left, 10 right and two bilaterally). The anatomy of all the ears (bilaterally) and the accuracy of dye deposition (successful nerve staining defined as > 2 cm) was assessed following dissection. Occurrence of intraparotid, intravascular, or intraneural injection was recorded. Phase II: the same technique and volumes were applied, bilaterally, using a lidocaine 2%:iohexol solution mixture (3:1) in six different cadavers. Computed tomography was performed and the needle position and spread of the mixture evaluated and recorded. Results Phase I: Inter- and intraindividual differences were found in the anatomy of the great auricular nerve, but not in the internal and lateral auricular nerve branches and the caudal auricular nerve. No auriculotemporal nerve was found. Nerve staining was successful in 95% of the cases for the great auricular nerve, 85% of the cases for the internal auricular nerve, and 100% of the cases for the lateral auricular branch and caudal auricular nerve. Failure was observed in horses with abundant periauricular fat. No intravascular, intraparotid or intraneural injections occurred. Phase II: A 100% success rate was recorded from computed tomography images. Conclusions and clinical relevance The described approach seems suitable for blockade of the sensory innervation of the equine ear.

Ganglia and telocytes were identified inside the porcine pulmonary veins wall near myocardial sleeve tissue at the atriopulmonary junction. These structures are reported to play a role in the initiation of pulses from outside the heart, which potentially can cause cardiac conduction disorders such as atrial fibrillation. In-depth knowledge on the fine structure of the pulmonary vein wall is a pre-requisite to better understand the underlying pathophysiology of atrial fibrillation and the origin and conduction of ectopic pulses. The importance of pulmonary vein myocardial sleeves as triggering foci for atrial fibrillation has been shown in human patients. In this context, the fine structure of the pulmonary vein wall was investigated qualitatively by light and transmission electron microscopy in the pig, which is a frequently used animal model for development of new treatment strategies. Additionally, intra and extramural ganglia, containing telocytes that create a network near the neurone cell bodies, were identified in pigs. Detailed illustration of the distribution and organization of tissues and cell types, potentially involved in the origin and propagation of ectopic stimuli originating from the pulmonary veins, might lead to a better insight on the actual composition of the tissues affected by ablation as studied in pigs.

Piglets are considered to be suitable animal models for predicting the pharmacokinetics and pharmacodynamics (PK/PD) of test drugs for potential use in the paediatric population. Such PK/PD studies require multiple blood and urine samplings. The goal of the present study was to determine a suitable blood collection strategy applicable in the youngest age categories of six days, four weeks and eight weeks of age, as well as a urine collection technique for male piglets in the same age categories. Blood was collected either by a surgically-placed jugular vein catheter (six days old [ n = 4] and four weeks old [ n = 2] piglets) or by direct venepuncture of the jugular vein (four weeks old [ n = 2] and eight weeks old [ n = 4] piglets). A non-invasive method for total volume urine collection in male piglets was also developed using a urine pouch. No specific complications were encountered during anaesthesia or surgery for jugular catheter placement. After a 24 h recovery period, urine and blood were easily collected without technical complications. One piglet was humanely killed at week 2 because of septicaemia. Histological analysis of both veins in all four piglets revealed negligible damage to the blood vessel wall. In conclusion, the presented techniques for blood (jugular catheter and direct venepuncture) and urine collection (pouches) are suitable for PK/PD studies in piglets.

Corrosion casting is the technique by which a solid, negative replica is created from a hollow anatomical structure and liberated from its surrounding tissues. For centuries, different types of hardening substances have been developed to create such casts, but nowadays, thermosetting polymers are mostly used as casting medium. Although the principle and initial set-up are relatively easy, producing high-quality casts that serve their intended purpose can be quite challenging. This paper evaluates some of the more popular casting resins that are currently available and provides a step-by-step overview of the corrosion casting procedure, including surface casts of anatomical structures. Hurdles and pitfalls are discussed, along with possible solutions to circumvent them, based on personal experience by the authors.

The development of well-designed polylactic acid (PLA) based composite filaments through fused filament fabrication (FFF) is crucial, as PLA printed parts are characterized by a high brittleness. Here we focus on FFF composite formation using ductile poly (butylene adipate-co-terephthalate) (PBAT), the nanoclay cloisite and the chain extender Joncryl. A high modulus, flexibility, toughness and notched impact strength result upon combining PLA/PBAT (80:20; mass basis) with a sufficiently high cloisite amount (1–5 phr) supported by a suited Joncryl amount (0.3 phr). A fine PBAT dispersion is then ensured, as confirmed by scanning and transmission electron microscopy. For high cloisite amounts (e.g. 10 phr) the crystallinity and vicat softening temperature enhance but cloisite aggregation is unavoidable, leading to a strong reduction in tensile strain and toughness. Annealing increases crystallinity thus material strength, rigidity and impact, at least partially overruling the need of a fine dispersion. Novel radar charts are also reported to select an optimal bio-based blend suitable for different end-user applications by adjusting the composition and possible post-annealing of printed products.

Pediatric drug research is still substandard, not reaching the same quality level as adult drug research. Despite the efforts made by the Food and Drug Administration and European Medicines Agency to reduce off-label use in children, the lack of clinical studies involving the pediatric population still stands. Pharmacokinetic and pharmacodynamics studies (PK/PD) taking growth and maturation into account are necessary to rationalize dosing strategies in children. Currently, traditional animal models such as rats, mice, dogs and primates are used to conduct pharmacokinetic and pharmacodynamic studies, however age-related trials are rather uncommon. Moreover, these species have several shortcomings as animal models, such as a different physiology and anatomy of the gastrointestinal tract in dogs or the ethical aspects for the use of primates. In contrast, piglets might be potential biomedical pediatric animal models because of the good resemblance with humans, anatomically, physiologically and biochemically. This review summarizes the comparative anatomy and physiology and postnatal development of piglets and infants, focusing on six major topics, namely growth, cardiovascular system, gastrointestinal tract, liver, kidney and integument. Furthermore, the application of piglets as animal model in pediatric PK/PD research is discussed.

Objective: To expand current knowledge on the canine omental vasculature and refine the existing lengthening technique of the canine omentum. Study Design: Ex vivo study. Animals: Canine cadavers (n=20). Methods: In 10 canine cadavers the omental arteries were mapped using intravascular latex injection and these results were used to create an omental pedicle flap based on the splenic artery in 10 additional cadavers. The operating range of the flap was recorded with particular attention to the main regions of interest for omental transposition in dogs (axillary and inguinal regions). Results: The superficial and deep omental leaves were each predominantly supplied by a left and a right marginal omental artery that anastomosed near the caudal omental border into a superficial and a deep omental arch, respectively. Anastomoses between arteries of the superficial and the deep omental leaves were weak and inconsistent, except for 1 anastomosis that was found in 8 of 10 dogs. By transposing the intact omentum, the right axilla could be reached in 3 dogs, both axillae in 1 dog, and both groins in all cadavers. In all cases, the omental pedicle reached to and beyond the axillary and inguinal regions. By unfolding the pedicle leaves, the width of the pedicle tip could be doubled. Conclusion: When lengthening the omentum is necessary to reach extra-abdominal structures, the omental pedicle flap based on the splenic artery appears to preserve the omental vascular supply. These observations warrant further clinical trials to evaluate this new omtental flap technique in vivo.

Background Atrial fibrillation is the most frequent arrhythmia in adults of which the interventional cure is hampered by high recurrence rates. Recurrence after ablation is due to an incomplete isolation of the pulmonary veins. A new ablation technique was performed, in the antra of ovine pulmonary veins, by device implantation, which was heated through a wireless heat-generating system. Methods and Results Implants were placed transatrially in the pulmonary veins of sheep. Using a wireless heating system, the energy was afterward transferred through wires to the implanted device according to a defined protocol. The position of the implant and the applied lesions were macroscopically evaluated. Samples of the ablated tissue of the atrio-pulmonary vein junction were histologically and immunohistochemically examined. Conclusions Six ablation procedures in four sheep were successfully performed without adverse cardiac reactions. Implantation of the device and the wireless heat generation was feasible. Sufficient heat was produced at the level of the antra of the pulmonary veins to create ablation lesions, which were histologically and immunohistochemically confirmed.

During endoscopy (tenoscopy) of the distal aspect of the equine digital flexor tendon sheath (DFTS), the digital manica flexoria can be visualized connecting the distal branches of the superficial digital flexor tendon. However, this structure has been inconsistently described and variably named in the veterinary literature. The objectives of this study were to describe the presence, configuration and variability of the digital manica flexoria in the equine distal limb. Dissection of 144 equine cadaveric limbs revealed the presence of this structure in all the feet, although different types and conformations were identified. In the forelimbs, a membranous digital manica flexoria predominated (94%; P

An Oldenburg colt with wry nose was autopsied after having lived for only 30 min. It presented cyanotic oral mucosae, underdeveloped eyes and a right-sided temporal osseous mass. The applicable nomenclature for the defects is discussed, and the potential etiopathogenesis is explored by describing the normal embryonic development of the affected body parts.

Background: To date, reference values for 25-OH-vitamin D, parathyroid hormone (PTH), and calcium in serum of common marmosets (Callithrix jacchus) based on a large sample size are not available. Methods: Serum reference values for these parameters were determined and correlated with sex, age, season of sampling, and time of long bone epiphyseal closure in captive-housed marmosets. Results and Conclusions: The 90% reference range for serum 25-OH-vitamin D is 47.40-370.4 nmol/L, for PTH 2.10-30.51 pmol/L, and for calcium 2.08-2.63 mmol/L. Lower levels of vitamin D were measured in fall compared with the other seasons. Levels of PTH were higher in males than in females, and calcium levels were lower in younger animals compared with older marmosets. No other effects of age, sex, season, or timing of growth plate closure were found.

Background/Aims: Intussusceptive angiogenesis (IA) is a dynamic process which contributes to vascular expansion and remodeling. Intraluminal pillars have long been the distinctive structural indicator of IA. However, the mechanism of their formation has not been fully elucidated. Methods: Using light and electron microscopy, we studied intussusceptive vascular growth in the developing porcine metanephric kidney. Results: We observed intraluminal pillars formed by endothelial cells in the vasculature of developing glomeruli. Their diameter was < 2.5 µm, consistent with the diameter of nascent pillars. TEM revealed that the majority of these pillars consisted only of endothelium. However, a central core of extracellular matrix (ECM) covered by endothelium, reminiscent of a more mature intussusceptive pillar, was also found in the lumen of a glomerular capillary. Perivascular cells or pericytes were not involved in the pillar structure during these stages of formation. Conclusion: This study shows ECM presence in a mature intussusceptive pillar without any perivascular cell involvement in the structure. This leads to the hypothesis that ECM deposition precedes the participation of these cells in the formation of intraluminal pillars during IA in porcine metanephric glomerular capillaries.

International audience; Cirrhosis represents the end-stage of any persistent chronically active liver disease. It is characterized by the complete replacement of normal liver tissue by fibrosis, regenerative nodules, and complete fibrotic vascularized septa. The resulting angioarchitectural distortion contributes to an increasing intrahepatic vascular resistance, impeding liver perfusion and leading to portal hypertension. To date, knowledge on the dynamically evolving pathological changes of the hepatic vasculature during cirrhogenesis remains limited. More specifically, detailed anatomical data on the vascular adaptations during disease development is lacking. To address this need, we studied the 3D architecture of the hepatic vasculature during induction of cirrhogenesis in a rat model. Cirrhosis was chemically induced with thioacetamide (TAA). At predefined time points, the hepatic vasculature was fixed and visualized using a combination of vascular corrosion casting and deep tissue microscopy. Three-dimensional reconstruction and data-fitting enabled cirrhogenic features to extracted at multiple scales, portraying the impact of cirrhosis on the hepatic vasculature. At the macrolevel, we noticed that regenerative nodules severely compressed pliant venous vessels from 12 weeks of TAA intoxication onwards. Especially hepatic veins were highly affected by this compression, with collapsed vessel segments severely reducing perfusion capabilities. At the microlevel, we discovered zone-specific sinusoidal degeneration, with sinusoids located near the surface being more affected than those in the middle of a liver lobe. Our data shed light on and quantify the evolving angioarchitecture during cirrhogenesis. These findings may prove helpful for future targeted invasive interventions

Necropsy examination of an adult neutered male cat, which was humanely destroyed as it had feline immunodeficiency virus (FIV) infection, demonstrated the presence of a left retrocaval ureter that was entrapped around the left limb of a double caudal vena cava. These associated anomalies originate from the complex embryofetal development of the caudal vena cava. Since no clinical signs had been reported and no gross lesions related to this anomaly were observed, this manifestation should be simply considered as an anatomical variation.

The myocardial sleeve of the porcine and equine pulmonary veins were histologically investigated and reconstructed three dimensionally. Moreover, the localization of neuron cell bodies at the veno-atrial junction and alongside the myocardial sleeve was light microscopically visualized to depict the organization of nerve, myocardial and fat tissue. Finally, the presence of telocytes inside the equine pulmonary veins was demonstrated by use of transmission electron microscopy. These structures are thought to play a role in the induction of atrial fibrillation, which is frequently seen in horses, while pigs are often used as a cardiovascular model in this context. This data fills in remaining gaps in the literature concerning the histological build-up of the pulmonary veins wall in pigs and horses. In-depth knowledge on the myocardial sleeve and its surrounding cell types are important to understand the possible outcome of an ablation therapy as an atrial fibrillation treatment. In pigs and horses, the layout of the pulmonary veins wall concerning these structures is comparable to humans. However, neuron cell bodies were recovered at the veno-atrial junction in both species but not alongside the myocardial sleeve in horses.

The intricate (micro)vascular architecture of the liver has not yet been fully unravelled. Although current models are often idealized simplifications of the complex anatomical reality, correct morphological information is instrumental for scientific and clinical purposes. Previously, both vascular corrosion casting (VCC) and immunohistochemistry (IHC) have been separately used to study the hepatic vasculature. Nevertheless, these techniques still face a number of challenges such as dual casting in VCC and limited imaging depths for IHC. We have optimized both techniques and combined their complementary strengths to develop a framework for multilevel reconstruction of the hepatic circulation in the rat. The VCC and micro-CT scanning protocol was improved by enabling dual casting, optimizing the contrast agent concentration, and adjusting the viscosity of the resin (PU4ii). IHC was improved with an optimized clearing technique (CUBIC) that extended the imaging depth for confocal microscopy more than five-fold. Using in-house developed software (DeLiver), the vascular network - in both VCC and IHC datasets - was automatically segmented and/or morphologically analysed. Our methodological framework allows 3D reconstruction and quantification of the hepatic circulation, ranging from the major blood vessels down to the intertwined and interconnected sinusoids. We believe that the presented framework will have value beyond studies of the liver, and will facilitate a better understanding of various parenchymal organs in general, in physiological and pathological circumstances.

The major salivary glands of birds develop by branching or elongation of the epithelial cords. The development of the minor salivary glands in form of the lingual glands has never been described. Among birds, only Anatidae have three types of the lingual glands: rostral, caudo-lateral, and caudo-medial lingual glands. The study aims to characterize the manner and rate of the lingual glands development in the domestic duck and their topographical arrangement relative to the hyoid apparatus. The study reveals that all three types of the lingual glands develop by branching. We describe five stages of the lingual glands development in the domestic ducks: prebud, initial bud, pseudoglandular, canalicular, and terminal bud stage. The pattern of the lingual glands development in birds is similar to that described for mammals, with the exception, that the terminal buds are formed at the same time as the lumen of the glands. Generally, the rostral lingual gland starts to branch earlier than the caudal lingual glands. The 3D-reconstruction shows the location and direction of lingual gland development relative to the entoglossal cartilage and basibranchial bone. Light microscopy and scanning electron microscopy allow to characterize the histogenesis of the embryonic epithelium into glandular epithelium. At a time of hatching only secretory units of caudal lingual glands resemble the secretory units of the adult domestic duck. The rostral and caudo-lateral lingual glands are arranged on the sides of the entoglossal cartilage and basibranchial bone and caudo-madial lingual glands are located over the basibranchial bone. We suggest that such an arrangement of the lingual glands in the domestic duck is important during food intake and responsible for reduction of friction and formation of food bites.

The intricate (micro)vascular architecture of the liver has not yet been fully unravelled. Although current models are often idealized simplifications of the complex anatomical reality, correct morphological information is instrumental for scientific and clinical purposes. Previously, both vascular corrosion casting (VCC) and immunohistochemistry (IHC) have been separately used to study the hepatic vasculature. Nevertheless, these techniques still face a number of challenges such as dual casting in VCC and limited imaging depths for IHC. We have optimized both techniques and combined their complementary strengths to develop a framework for multilevel reconstruction of the hepatic circulation in the rat. The VCC and micro-CT scanning protocol was improved by enabling dual casting, optimizing the contrast agent concentration, and adjusting the viscosity of the resin (PU4ii). IHC was improved with an optimized clearing technique (CUBIC) that extended the imaging depth for confocal microscopy more than five-fold. Using in-house developed software (DeLiver), the vascular network in both VCC and IHC datasets was automatically segmented and/or morphologically analysed. Our methodological framework allows 3D reconstruction and quantification of the hepatic circulation, ranging from the major blood vessels down to the intertwined and interconnected sinusoids. We believe that the presented framework will have value beyond studies of the liver, and will facilitate a better understanding of various parenchymal organs in general, in physiological and pathological circumstances. Key words: 3D modeling; confocal laser scanning; hepatic vasculature; immunohistochemistry; micro-CTscanning; morphological analysis; rat liver; vascular corrosion casting.

Differentiation of endodermal cells into hepatoblasts is well studied, but the remodeling of the vitelline and umbilical veins during liver development is less well understood. We compared human embryos between 3 and 10 weeks of development with pig and mouse embryos at comparable stages, and used A(MIRA) 3D reconstruction and CINEMA 4D remodeling software for visualization. The vitelline and umbilical veins enter the systemic venous sinus on each side via a common entrance, the hepatocardiac channel. During expansion into the transverse septum at Carnegie Stage (CS) 12 the liver bud develops as two dorsolateral lobes or 'wings' and a single ventromedial lobe, with the liver hilum at the intersection of these lobes. The dorsolateral lobes each engulf a vitelline vein during CS13 and the ventromedial lobe both umbilical veins during CS14, but both venous systems remain temporarily identifiable inside the liver. The dominance of the left-sided umbilical vein and the rightward repositioning of the sinuatrial junction cause de novo development of left-to-right shunts between the left umbilical vein in the liver hilum and the right hepatocardiac channel (venous duct) and the right vitelline vein (portal sinus), respectively. Once these shunts have formed, portal branches develop from the intrahepatic portions of the portal vein on the right side and the umbilical vein on the left side. The gall bladder is a reliable marker for this hepatic vascular midline. We found no evidence for large-scale fragmentation of embryonic veins as claimed by the 'vestigial' theory. Instead and in agreement with the 'lineage' theory, the vitelline and umbilical veins remained temporally identifiable inside the liver after being engulfed by hepatoblasts. In agreement with the 'hemodynamic' theory, the left-right shunts develop de novo.

Although the omentum remains an enigmatic organ, research during the last decades has revealed its fascinating functions including fat storage, fluid drainage, immune activity, angiogenesis and adhesion. While clinicians both in human and veterinary medicine are continuously exploring new potential omental applications, detailed anatomical data on the canine omentum are currently lacking, and information is often retrieved from human medicine. In this study, the topographic anatomy of the canine greater and lesser omentum is explored in depth. Current nomenclature is challenged, and a more detailed terminology is proposed. Consistent arteries that are contained within folds of the superficial omental wall are documented, described and named, as they can provide the anatomical landmarks that are necessary for unambiguous scientific communication on the canine omentum. In an included dissection video, the conclusions and in situ findings described in this study are demonstrated.

Summary Reasons for performing study There are no previous studies correlating the anatomy of the equine epiploic foramen and its defining structures with laparoscopic images. Objectives The purpose of this study was to describe the precise anatomy of the epiploic foramen and omental vestibule, to quantify the dimensions of the epiploic foramen and to relate these findings to laparoscopic images. Study design Descriptive study of cadaver material and laparoscopic images. Methods Thirty-two horses subjected to euthanasia for reasons unrelated to colic were studied. Two cadavers were used to make vascular casts of the coeliac artery and portal vein. In 30 cadavers the epiploic foramen and omental vestibule were cast with a polyurethane prepolymer immediately after euthanasia. The cast served as a landmark during dissection and the circumference of the epiploic foramen was measured from these casts. Histology was performed on structures defining the epiploic foramen in 2 horses. Laparoscopic images from 6 standing right flank procedures were reviewed. Results The defining structures of the epiploic foramen and omental vestibule are the hepatoduodenal ligament, hepatogastric ligament and the gastropancreatic and hepatopancreatic folds. The hepatoduodenal ligament has a secondary fold which forms the ventral border of the epiploic foramen, consisting of a central connective tissue core of mainly elastin fibres. The hepatic artery for part of its course is incorporated in the hepatoduodenal ligament. All these structures are clearly visible laparoscopically. The mean circumference of the epiploic foramen is 11.6 ± 2.6 cm and its circumference is positively correlated with body weight but is unrelated to age or gender. Conclusions Several clinically relevant structures delineate the epiploic foramen. Its defining structure consists, in part, of elastin fibres. Anatomical and laparoscopic knowledge may assist surgeons in developing interventions to treat diseases involving the epiploic foramen.

Department of Morphology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, BelgiumCorresponding AuthorIlona Schwarzkopf, DVM, DAC Malpertuus,Leenstraat 2a, 9070 Heusden, O.VL,Belgium.E‐mail: ilona.schwarzkopf@gmail.comSubmitted September 2012Accepted March 2013DOI:10.1111/j.1532-950X.2014.12226.x

Persisterende ductus arteriosus (PDA) is één van de meest vastgestelde congenitale hartaandoeningen bij de hond. Indien de aandoening niet behandeld wordt, sterft meer dan de helft van de honden in hun eerste levensjaar. Het sluiten van de PDA via occlusie of ligatie is dan ook aanbevolen en heeft een goede langetermijnprognose. De belangrijkste complicatie bij de nietinvasieve technieken is het optreden van embolie terwijl bloeding na ruptuur van de ductus of een naburig bloedvat het meest voorkomende probleem is bij ligatietechnieken. Twee gevallen van PDA worden hier besproken. Bij de eerste hond werd de PDA succesvol chirurgisch gecorrigeerd. Bij de tweede hond daarentegen ontstonden er tijdens de chirurgische ingreep meermaals ernstige bloedingen te wijten aan de uitzonderlijke aanwezigheid van een persisterende vijfde aortaboog (PFAA). Er werd intraoperatief besloten om tot euthanasie over te gaan.

Although a full understanding of the hepatic circulation is one of the keys to successfully perform liver surgery and to elucidate liver pathology, relatively little is known about the functional organization of the liver vasculature. Therefore, we materialized and visualized the human hepatic vasculature at different scales, and performed a morphological analysis by combining vascular corrosion casting with novel micro-computer tomography (CT) and image analysis techniques. A human liver vascular corrosion cast was obtained by simultaneous resin injection in the hepatic artery (HA) and portal vein (PV). A high resolution (110 μm) micro-CT scan of the total cast allowed gathering detailed macrovascular data. Subsequently, a mesocirculation sample (starting at generation 5; 88 × 68 × 80 mm³) and a microcirculation sample (terminal vessels including sinusoids; 2.0 × 1.5 × 1.7 mm³) were dissected and imaged at a 71-μm and 2.6-μm resolution, respectively. Segmentations and 3D reconstructions allowed quantifying the macro- and mesoscale branching topology, and geometrical features of HA, PV and hepatic venous trees up to 13 generations (radii ranging from 13.2 mm to 80 μm; lengths from 74.4 mm to 0.74 mm), as well as microvascular characteristics (mean sinusoidal radius of 6.63 μm). Combining corrosion casting and micro-CT imaging allows quantifying the branching topology and geometrical features of hepatic trees using a multiscale approach from the macro- down to the microcirculation. This may lead to novel insights into liver circulation, such as internal blood flow distributions and anatomical consequences of pathologies (e.g. cirrhosis). ispartof: Journal of Anatomy vol:224 issue:4 pages:509-17 ispartof: location:England status: published

The layout of the porcine atriopulmonary junction and immediately adjacent structures was investigated by gross anatomical and vascular corrosion casting studies to meet the need for more in-depth anatomical insights when using the pig as an animal model in the development of innovative approaches for surgical cardiac ablation in man. The veins from the right cranial and middle lung lobes drain through a common ostium in the left atrium, whereas a second ostium receives the blood returning from all other lung lobes, although limited variation to this pattern was observed. Surrounding anatomical structures that are most vulnerable to ablation damage as reported in man are located at a safer distance from the pulmonary veins in pigs, yet a certain locations, comparable risks are to be considered. Additionally, it was histologically confirmed that myocardial sleeves extend to over a centimetre in the wall of the pulmonary veins.