A neuroanatomical study of the feline brain using MRI and mulligan staining: functional and pathological considerations.

BACKGROUND: Despite multiple studies describing accurate diagnoses using advanced neuroimaging techniques, low and mid-field magnetic resonance imaging (MRI) are still the most frequent scanners in veterinary clinics. To date, these studies in cats do not show a clear distinction of nerve centres in MRI data. AIMS: The objective of this study is to determine the efficacy of Mulligan histological staining as a tool in facilitating the location and identification of the main structures of the feline brain in MRI. This study aims to facilitate the interpretation of MRI obtained with these types of scanners. METHODS: A total of 10 feline brains were used. One specimen was used for MRI (T2 sequence using a 1.5T scanner). The other 9 brains were sectioned and stained with the three Mulligan staining techniques (Mulligan, Le Masurier and Robert). RESULTS: The uptake of stain by the grey matter in these sections allowed the determination of the location and the limits of these nervous structures within the brain. The histological location of these structures was correlated with the MRI scans, leading to the successful identification of many small, indistinct nuclei. CONCLUSION: Mulligan staining is proposed as a tool that facilitates the location of nerve structures in comparison with data from the most frequently-used MRI scanners in veterinary clinics.

Anatomía veterinaria: Experiencia metodológica en el marco del espacio europeo de educación superior / Veterinary Anatomy: methodology experience within the European Space of Higher Education

Se expone la metodología docente aplicada en Anatomía Veterinaria, de la licenciatura de Veterinaria de la Universidad de Murcia, desde el curso 2005 2006 hasta la actualidad. Durante este periodo la Unidad Docente de Anatomía y Embriología Veterinarias ha desarrollado proyectos de innovación educativa dirigidos a realizar experiencias ECTS (Sistema Europeo de Transferencia y acumulación de Créditos). En estos proyectos hemos incrementado el enfoque práctico de la asignatura, con el fin de que el alumno potencie competencias tales como: autoaprendizaje, trabajo en equipo, destrezas, etc., lo que permite una mayor adaptación al perfil profesional que demanda el mercado de trabajo. Para ello, además de las prácticas convencionales, hemos introducido una nueva modalidad práctica denominada: trabajos prácticos dirigidos. Esta modalidad es también presencial y consiste en la realización de actividades prácticas –diferentes a las prácticas convencionales– que los alumnos han de desarrollar durante cada sesión, en grupos de 4-5 alumnos. Estos trabajos varían en función de la materia. Así, en “Osteología” se realizan diversas actividades en el Museo Anatómico Veterinario, tales como la interpretación de imágenes óseas anatómicas, radiografías, etc., mientras que en “Sistemas Neuromusculares” y “Sistemas Viscerales” se realiza la disección de los diferentes planos musculares y viscerales en el perro, así como la interpretación de secciones anatómicas transversales (anatomía seccional). Con el fin de fomentar el estudio de los alumnos, se introduce un sistema de evaluación continua a lo largo del curso, en las diferentes materias. Los resultados han mostrado una mejora en la motivación y en la participación activa de los alumnos, así como unos mejores resultados académicos en la materia práctica (AU)

This work describes the teaching methodology applied in Veterinary Anatomy since 2005 to 2008. This subject is taught in the Veterinary degree of the University of Murcia. During this period the Department of Veterinary Anatomy has carried out some educative innovation projects, in order to apply ECTS methodology (European credit transfer and accumulation system). These projects increase the practical approach to the subject in order to improve some competences (self-learning, job in small groups, skills, etc.) and to allow a better adaptation to the professional profile of the job market. The methodology includes conventional practices and a new practical model: guided practical jobs. This latter is carried out in groups of 4-5 students whose undertake some practical activities that vary according to the particular subject. On “Osteology”, these activities consisted in the interpretation of some anatomical osseus images, radiographies, etc. in the Anatomical Veterinary Museum. On “neuromuscular and visceral systems” the students dissected different muscle and visceral planes of dog cadavers and interpreted some transversal anatomical sections. In order to improve the study methodology, we have introduced a continuous evaluation throughout the course. Results show a increasing of the motivation and participation, as well as better punctuations in the practical exams (AU)

Revisión: desarrollo y crecimiento de la musculatura axial de la lubina Dicentrarchus labrax L / Review: development and growth of the axial musculature of the sea bass Dicentrarchus labrax, L

Se compendian en este trabajo los estudios realizados sobre el desarrollo y crecimiento de la musculaturaaxial de la lubina Dicentrarchus labrax L., incluyéndose también algunos resultados y conclusiones relevDicentrarchus relevantesen otras especies. En el momento de la eclosión, la larva de lubina presenta dos estratos musculares: músculorojo superfi cial y músculo blanco, ambos con un apreciable grado de inmadurez histológica. Desde la eclosiónhasta el momento de la apertura de la boca las fi bras musculares rojas y blancas crecen únicamente por hipertrofia. Sin embargo, con el inicio de la alimentación externa, la larva de lubina inicia un periodo de intenso crecimientomuscular hipertrófi co e hiperplásico. Hasta el fi nal de la fase larvaria, además de importantes cambiosestructurales y en las isoformas de miosina de las fi bras rojas y blancas, se diferencia progresivamente el músculointermedio o rosa. Durante la fase postlarvaria, los procesos de maduración histológica de la musculaturapersisten hasta el comienzo de la edad juvenil (≈ 6 meses). En alevines y juveniles de lubina, la dinámica delcrecimiento muscular está infl uenciada por la época del año, en función de factores medioambientales como latemperatura y el fotoperiodo. Esta infl uencia afecta de forma muy acentuada a la hiperplasia muscular, la cualdescribe una secuencia alternante con valores máximos durante el verano e inicio del otoño

This work summarizes the previous information on the development and growth of the axial musculature ofthe sea bass Dicentrarchus labrax L. Relevant results and conclusions from other teleost species in these subjectsare also included. At hatching, the axial musculature of sea bass larvae has two strata: the superfi cial redand the deep white muscles, both with an evident histological immaturity. Hypertrophy of red and white musclefi bres is the unique mechanism of muscle growth since hatching to mouth opening. After mouth opening, larvae enter a period of intense muscle growth by both hypertrophy and hyperplasia of muscle fi bres. Throughout thelarval phase, red and white muscle fi bres undergo structural changes and a progressive replacement of differentmyosin isoforms. The intermediate pink muscle is evident towards the end of the larval life. Histological maturationof muscle fi bres persist during the postlarval life until the beginning of the juvenile period (≈ 6 month).In fi ngerlings and juveniles, muscle growth dynamics is seasonally infl uenced by environmental factors such astemperature and photoperiod. Muscle fi bre recruitment is particularly affected so that it is maximum in summerand minimum in winter

Effect of two thermal regimes on the muscle growth dynamics of sea bass larvae, Dicentrarchus labrax L.

Muscle growth was studied in larvae of sea bass, Dicentrarchus labrax L., reared at two temperatures: real ambient temperature ( congruent with 15 degrees C during vitelline phase and increased gradually) and 19 degrees C from fertilization until the end of larval development. Muscle cellularity, body length and body weight were measured. Early temperature influenced larval development and so, pre-larval phase finished earlier at 19 degrees C than at ambient temperature (4 and 6 days, respectively). Temperature also affected muscle growth such that at hatching and at mouth opening hypertrophy of muscle fibres was greater at 19 degrees C (P < 0.05), whereas hyperplasia was similar in both groups. After 25 days, the cross-sectional area of the white muscle was greater at 19 degrees C (P < 0.05), which was mainly associated with a higher proliferation of new white muscle fibres. At this stage the body length was also higher at 19 degrees C. Metamorphosis finished earlier in fish reared at 19 degrees C (52 days) than at natural temperature (82 days). At this developmental stage body length and cross-sectional area of the myotome were similar in both groups. However, muscle cellularity differed between groups. Thus, hypertrophy of muscle fibres was higher in fish reared at ambient temperature (P < 0.05), whereas proliferation of new muscle fibres was higher at 19 degrees C (P > 0.05).

The histochemical profiles of fibre types in porcine skeletal muscle.

Using a variety of histochemical methods -mATPase staining after alkaline and acid preincubations, NADH-TR and alpha-MGPDH- we have investigated the fibre types in porcine skeletal muscle. The results reveal that four major fibre types -I, IIA, IIB and II*- can be separated histochemically in Longissimus lumborum muscle of Landrace pigs. The histochemical properties of the muscle fibre type II* are very similar to that of type IIX described in other mammals. The existence of IIX fibres in pig muscle has been recently demonstrated by molecular biology techniques and our results validate the use of histochemistry (mATPase) as an easy methodology to differentiate the three fast myosins (type II fibres) in pig muscle.

Histochemical and morphometric aspects of the lateral musculature of different species of teleost marine fish of the Percomorphi order.

Cross-sections of the lateral musculature of 19 species of teleost fish of the Percomorphi order were studied histochemically and morphometrically. The red, pink and white muscles were observed in all the species except in Seriola dumerilii and Scomber japonicus, in which no pink muscle was found. The red fibres were the smallest due to their minimal diameters. Histochemically and according to their location at least five types were found in the same species: typical red, in rosette, of the border, infiltrated pink and in mosaic. The pink fibres were larger than the red but smaller than the white. The form and disposition of the pink muscle varied according to the species. This muscle is made up by fibres of medium size, moderate m-ATPase and intermediate oxidative activities. Their histochemical properties change near red and white muscles, where they acquire the histochemical characteristics of the typical red fibres or the low m-ATPase activity of white fibres, respectively. The white muscle is the widest muscular layer. All of the species, except Thalassoma pavo are made up by three fibre types (small with high m-ATPase activity, medium with moderate m-ATPase activity, and large with low m-ATPase activity). They are mixed and show a histochemical and morphometric mosaic form. The extent of this mosaic depends on the species. In most of the analysed species it was observed in areas close to the pink muscle, but in three of them (Liza aurata, Seriola dumerilii and Coris julis) it occupied all of the myotome.

Red muscle development of gilthead sea bream Sparus aurata (L.): structural and ultrastructural morphometry.

The transverse red and white muscle area, the superficial red muscle fibres area and their percentages of mitochondria (%mit), myofibrils (%myof) and sarcoplasm (%sarc) were determined in the Mediterranean teleost gilthead sea bream, Sparus aurata (L.). Fish aged from hatching to 78 days were studied. The proportional growth of the red and white muscles was higher for the red muscle in the first half of the larval stage (1-35 days). Then the opposite relationship was observed. The hypertrophic growth of the superficial muscle fibres was continuous except in the first week after hatching. The percentage of mitochondria and percentage of myofibrils showed a significant change just after the half of the larval stage. Whereas the %mit of the superficial muscle fibres was higher than %myof from the first week after hatching to 35 days (average 66.64%), then the %mit decreased significantly and at 73-78 days both parameters were close to an average value of 50%. The meaning of these morphological changes is discussed in relation to the functional role of the red muscle of larvae and the onset of the gills respiration.

Muscle development in gilthead sea bream (Sparus aurata, L.) and sea bass (Dicentrarchus labrax, L.): further histochemical and ultrastructural aspects.

The histochemical profiles--mATPase and NADH-TR reactions--of the red and white muscle fibres of gilthead sea bream and sea bass were determined from the first week after hatching. Modifications of the mATPase technique by combinations of pH/time/molarity were carried out in order to compare the sensitivity of the myosin ATPase of each muscle fibre type of the lateral muscle. Results showed that the staining of muscle fibres was independent of small modifications in the technique. The intermediate 'pink' muscle was histochemically defined towards the end of the larval life and is considered to be implicated in the growth of the myotome. A layer of external cells was observed, by electron microscopical examination, between the connective tissue of the skin and the superficial red muscle fibres of larvae and postlarvae. It is suggested that the external cells are unlikely to be a source of red muscle fibres and implicated on the growth of the myotome, but rather a part of the dermatome. The timing, areas and mechanisms of hyperplastic growth of the myotome were defined and discussed.

The post-larval development of lateral musculature in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.).

Fibre-type differentiation of lateral musculature has been studied in gilthead sea bream Sparus aurata (L.) and sea bass Dicentrarchus labrax (L.) during post-larval development using ultrastructural, histochemical and morphometric techniques. The study showed three muscle layers: red, intermediate (or pink) and white. Initially, most of the red muscle showed low myosin ATPase (m-ATPase) activity fibres, whereas near the transverse septum some small high m-ATPase activity fibres appeared and later acquired a rosette aspect. Afterwards, during adult growth the red muscle showed a histochemical mosaic appearance. The pink muscle in sea bass was observed at the beginning of juvenile development by the oxidative technique (NADH-RT reaction) whereas in gilthead sea bream it was also observed at the end of larval development. The pink layer consists of high m-ATPase activity fibres. However, along the muscle development other low and moderate m-ATPase activity fibres were observed close to the red and white muscles, respectively. The white muscle of juvenile fish showed a histochemical mosaic appearance near the pink muscle. In adult specimens the mosaic white muscle spread out occupying the whole of the myotome. Morphometric analysis shows a significant increase in mean fibre diameter during post-larval development, as shown by the Student's t-test (hypertrophic growth). Skewness and kurtosis values of fibre diameters point to the generation of a new fibres from the myosatellite cells (hyperplastic growth).

[Anatomo-radiologic correlation of the head of the goat of the canary breed (Capra hircus L.)]. / Correlación anatomo-radiológica de la Cabeza en la Cabra de Raza Canaria (Capra hircus L.).

A radiologic and tomographic study was made of the cranioencephalic topography of canary goats of medium weight and size. In a first step, the exocranialis references corresponding to an axis designed at the level of the meatus acusticus externus and parallel to the basicranial axis were anatomically and radiologically defined. Conventional stereotaxic techniques were then used to reach the standard position of the head in order to obtain macroscopical cross sections at different levels perpendicular to the base of the cranium. In a second step, a tomographic study was made to confirm the design of the axis.

[Chronology of the postnatal ossification of the thoracic limb in the Siamese cat (Felis catus L.)]. / Cronología de osificación postnatal del autópodo torácico en el gato Siamés (Felis catus L.).

A study was done by radiological techniques to show the chronology of ossification of the thoracic autopodo of the Siam cat from birth up to the 25th week of postnatal development. For this experiment we used 40 little cats (19 males and 21 females) belonging to 11 litters subjected to different controls: pattern race, healthy, feeding, grow up and radioactivity. The time of the appearance and evolution of the centres that form the basipodo, metapodo and acropodo of the thoracic limb is determined. Likewise it is analyzed the phenomenon of fusion during that time to define the main ossification sequence in the carpo. The aspects of the postnatal ossification of the thoracic autopodo in the Siam cat are compared with the literature of different authors about the common cat.

[Histochemical study of the muscle fibers of some marine and fresh water teleost fish]. / Estudio histoquímico de las fibras musculares de algunos teleósteos marinos y de agua dulce.

Histochemical and morphological studies of skeletal muscle of some fresh and salt water fish is presented. Distribution of three muscle types, red, intermediate and white, is discussed.