Morphological study and mineral analysis of the lower mandible of adult Atlantic salmon (Salmo salar) from Scotland with mandibular deformation / Estudio morfológico y análisis de minerales de la mandíbula inferior de salmones del Atlántico (Salmo salar) adultos con deformación mandibular, provenientes de Escocia

Mandibular deformity is a condition that affects the jaw bone of adult salmon and has been observed in Norway and Chile, causing weight loss, poor quality of farmed fish and increased mortality. The causes range from high temperatures of the state of eggs, to poor nutrition phosphorus or vitamin C. This work aims to analyze this deformity by histochemical and mineral analysis technique during an episode presented in centers of the Scotia Sea. Jaw and spinal segments of 21 Atlantic salmon in Scotland were used. These samples were classified into three groups: Group 1: Severely deformed. Group 2: Mildly affected. Group 3: Normal controls. Four jaws per group were fixed in 10 % formalin and embedded in Paraplast, sections of 5 microns were performed using a Microm® microtome histochemical technique Von Kossa was used for the detection of calcium deposits, which highlights the calcium osteoid black and red color. For proximate analysis, and in order to obtain and compare levels of calcium, phosphorus, zinc and magnesium in total 9 bone jaws (6 affected with DM and 3 controls) and 9 body sections the Mann-Whitney test was used to compare these values between misshapen salmon and controls. To correlate values, jaw and body segment a Spearman corrrelation was applied. Fish group 1 presented a ventral deviation of the alveolar bone body. In fish group 2 prominence of the visible joint on both sides or unilaterally was observed. Comparing the values of % Ca, % P, % Mg and Zn jaws with DM and healthy ones with Mann Witney method it was found that the values of these minerals vary between salmon and controls affected. There was a significant difference in the percentage of P, which indicates that there is less P in affected fish vertebrae. Spearman correlation noted that the percentages of the minerals studied in dental bone and vertebral segments are uncorrelated. Rather, Von Kossa distribution indicates that Ca/P is not homogeneous in the dental bone, as a result of mineral resorption from the skeleton including the operculum, articular bone and dental towards kype. This paper states that Von Kossa histochemical technique showed significant differences between deformed fish and controls and also showed differences between the various segments of the dental bone. The alveolar bone is a dynamic structure adapted to continuous histological changes may be involved in MD, phosphorus deficient diets, coupled with the initial formation of Kippe.

La deformación mandibular es una patología que afecta al hueso dentario de salmones adultos, se observó en Noruega y en Chile, ocasionando disminución de peso, baja calidad de peces cultivados y aumento de la mortalidad. Las causas varían desde temperaturas elevadas al estado de ovas, hasta alimentación deficitaria en fósforo o vitamina C. Este trabajo tiene como propósito analizar esta deformación mediante una técnica histoquímica y de análisis de minerales durante un episodio presentado en centros de mar de Escocia. Se utilizó la mandíbula y segmento vertebral de 21 salmones del Atlántico de Escocia. Estas muestras se clasificaron en tres grupos Grupo 1: Severamente deformes. Grupo 2: Levemente afectados. Grupo 3: Controles normales. Cuatro mandíbulas por grupo fueron fijadas en formalina al 10 % y se incluyeron en paraplast, se realizaron cortes de 5 µm utilizando un micrótomo Microm®. Se utilizó la técnica histoquímica de Von Kossa para la detección de depósitos de calcio la cual destaca al calcio de color negro y el osteoide de color rojo. Para el análisis químico proximal, y con el propósito de obtener y comparar niveles de calcio, fósforo, zinc y magnesio en los huesos se utilizó un total de 9 mandíbulas (6 afectadas con DM y 3 controles) y sus 9 secciones corporales. Para comparar estos valores entre salmones deformes y controles se utilizó la prueba de Mann-Whitney. Para correlacionar los valores de mandíbula y segmento corporal se hizo una correlación por jerarquías de Spearman. Los peces del grupo 1, presentaron una desviación ventral del cuerpo del hueso dentario. En los peces del grupo 2 se observó la prominencia de la articulación visible en ambos lados o unilateralmente. Al comparar los valores de % Ca, % P, % Mg y Zn de las mandíbulas con DM y sanas con el método de Mann Witney se encontró que los valores de estos minerales no varían entre salmones afectados y controles. Hubo una diferencia significativa en el porcentaje de P, lo cual indica que existe menos P en vértebras de peces afectados. La correlación de Spearman señaló que los porcentajes de los minerales estudiados en huesos dentarios y segmentos vertebrales no están correlacionados. Por el contrario, la técnica Von Kossa mostró que la distribución de Ca/ P no es homogénea en el hueso dental producto de la reabsorción mineral desde el esqueleto incluyendo el opérculo, hueso articular y dental hacia la kype. El presente trabajo establece que la técnica histoquímica de Von Kossa fue la que permitió observar diferencias importantes entre peces deformados y controles, además mostró diferencias entre los distintos segmentos del hueso dentario. El hueso dentario es una estructura dinámica adaptada a continuos cambios histológicos pudiendo estar involucrados en la DM, dietas deficientes de fósforo, sumado a la formación inicial del Kippe.

Osteocalcin Expression and Mineralization in Developing Tooth of Xenopus laevis

Osteocalcin (OC) is the most abundant noncollagenous protein of extracellular matrix in the bone. In an OC deficient mouse, bone formation rates are increased in cancellous and cortical bones. OC is known as a negative regulator of mineral apposition. OC is also expressed in the tooth of the rat, bovine, and human. However, little is known about OC during tooth development in Xenopus. The purpose of this study is to compare the expression of OC with mineralization in the developing tooth of Xenopus, by using von Kossa staining and in situ hybridization. At stage 56, the developmental stage of tooth germ corresponds to the cap stage, and an acellular zone was apparent between the dental papilla and the enamel organ. From stage 57, calcium deposition was revealed by von Kossa staining prior to OC expression, and the differentiated odontoblasts forming predentin were located at adjoining predentin. At stage 58, OC transcripts were detected in the differentiated odontoblasts. At stage 66, OC mRNA was expressed in the odontoblasts, which was aligned in a single layer at the periphery of the pulp. These findings suggest that OC may play a role in mineralization and odontogenesis of tooth development in Xenopus.

Tooth Calcification in Low-Weight Human Fetuses / Calcificación de Dientes en Fetos Humanos de Bajo Peso

Tooth calcification begins during the intrauterine life and is related to the nutritional status of fetus. The purpose of this study was to evaluate the dental calcification status of 15 low-weight human fetuses aged from the 14th to 28th weeks of intrauterine life through radiographic analyses and von Kossa histochemical technique, which is specific to the detection of calcium and other mineral salts in tissues. After dissection of jaws, the mandibles were submitted to lateral and occlusal radiographies. Three mm thick slide sections of jaws samples were stained by conventional and von Kossa methods. As results, radiographic analysis did not exhibit any suggestive image of tooth mineralization, although rudiments of mandibular ossification could be noted. Hematoxilin and eosin sections revealed tooth germs in advanced phases of odontogenesis, but relative delay in tooth development was observed. The von Kossa staining did not show any positivity to the deposition of mineral salts in tooth germs from maxilla and mandible, supporting our findings related to the delay of dental development that was found in studied sample. Hence, we suggest that there is relation between the fetal low-weight and the calcification and maturation status of teeth.

La calcificación del diente comienza durante la vida intrauterina y se relaciona con el estado nutricional del feto. El propósito de este estudio fue evaluar el estado de calcificación dental de 15 fetos humanos de bajo peso entre las 14 y 28 semanas de vida intrauterina mediante análisis radiográficos y técnica histoquímica von Kossa, específica para la detección de calcio y otras sales minerales en los tejidos. Después de su disección, las mandíbulas fueron sometidas a radiografías laterales y oclusales. Secciones de 3 mm de espesor de las mandibulas fueron teñidas con la técnica Von Kossa. El análisis radiográfico no mostró ninguna imagen sugerente de mineralización dentaria, aunque rudimentos de la osificación mandibular podrían tenerse en cuenta. Las secciones de H-E revelaron la presencia de los gérmenes dentarios en las fases avanzadas de odontogénesis, pero con un retraso relativo en el desarrollo normal. La tinción de von Kossa no mostró positividad a la deposición de sales minerales en gérmenes dentarios, lo que apoya nuestros hallazgos relacionados con el retraso de desarrollo dental que se encontró en la muestra estudiada. Se sugiere que existe relación entre el bajo peso del feto y el estado de calcificación y maduración de los dientes.

Calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus)

This article reports a first case of calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus). Histopathology demonstrated well defined multiple cystic structures containing granular, dark basophilic materials with peripheral granulomatous reaction, characterized by presence of multinucleated giant cells surrounded by a varying amounts of fibrous connective tissues. Special staining with von Kossa revealed black stained deposits confirming the presence of calcium salts.

Calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus)

This article reports a first case of calcinosis circumscripta in a captive African cheetah (Acinonyx jubatus). Histopathology demonstrated well defined multiple cystic structures containing granular, dark basophilic materials with peripheral granulomatous reaction, characterized by presence of multinucleated giant cells surrounded by a varying amounts of fibrous connective tissues. Special staining with von Kossa revealed black stained deposits confirming the presence of calcium salts.

Subepidermal Calcified Nodule

Subepidermal calcified nodule(SCN) is a form of calcinosis cutis which is usually present as a single small, raised, hard nodule with verrucous surface. A 13-year-old girl presented with a 10-month history of a ricegrain-sized, hard, yellowish white papule on both upper eyelids. The lesions were asymptomatic and had increased slowly in size. An excisional biopsy specimen of the left upper eyelid showed acanthosis and narrow pointed rete ridges of the epidermis, and closely aggregated deposition of basophilic material in the uppermost dermis. The material in the dermis did not stain with von Kossa. It was confirmed as calcium deposition by staining with alizarin red S which is far more specific for calcium than the von Kossa stain.