Histological and morphometric characteristics of the corneal layer of buffalo hoof capsules / Características histológicas y morfométricas de la capa cornea de las cápsulas del casco de búfalo

SUMMARY: This study aimed to evaluate the microstructure of the buffalo's hoof capsule using hystomorphometry and computerized light microscopy. The length, thickness, gap and number of epidermal papillae and the morphology of the corneal tubules of the hoof were described in this paper. This study used 56 hoof capsules from hooves of 14 adults - age ranging from 24 to 60 months old - female buffaloes of the Jafarabadi breed; weighing around 650 kg. Fifty-six samples, 28 from thoracic limb and 28 from pelvic limbs, were analyzed, resulting in a total of 112 digits. Clinical specimens were collected at the coronary corium, laminar corium of the abaxial wall and pre-bulbar soles. The study concluded that the microstructure of the hoof capsule of buffalo hooves are made of epidermal papillae measuring 1,721.59 mm of length, 62.94 mm of thickness and 49.02 mm of gap between the papillae. This study determines that the coronary corium has more epidermal papillae than the laminar corium of the abaxial wall and pre-bulbar soles. The corneal tubules of the hoof capsules exhibit helical fashion and points to the possibility of applying this feature to the hooves of the biungulate species as well. In this research, we have been able to determine morphological parameters not yet described in scientific literature. These findings can be used in future comparative studies of healthy cattle and buffaloes and speculate evidence about the hooves' vulnerability facing different hoof diseases.

RESUMEN: El objetivo del estudio fue evaluar la microestructura de la cápsula del casco del búfalo mediante histomorfometría y microscopía óptica computarizada. Se describe la longitud, el espesor, el espacio y el número de papilas epidérmicas y la morfología de los túbulos corneales del casco. Utilizamos 56 cápsulas de cascos de 14 hembras adultas entre 24 a 60 meses de edad de la raza Jafarabadi, cuyo peso aproximado era de 650 kg. Analizamos 56 cascos, 28 del miembro torácico y 28 del miembro pélvico, con un total de 112 dedos. Se recogieron especímenes clínicos en el corion coronario, corion laminar de la pared abaxial y suelas pre-bulbares. Concluimos que la microestructura de la cápsula de los cascos de los búfalos está formada por papilas epidérmicas de 1.721,59 mm de longitud, 62,94 mm de espesor y 49,02 mm de espacio entre papilas. Se determinó que el corion coronario tiene más papilas epidérmicas que el corion laminar de la pared axial y las suelas pre-bulbares. Los túbulos córneos de las cápsulas de casco presentan una forma helicoidal y existe la posibilidad de que esta característica también se aplique a las pezuñas de otras especies de animales biungulados. También se determinaron algunos parámetros morfológicos no descritos en la literatura científica. Estos hallazgos pueden ser útiles en estudios comparativos de ganado bovino y bubalino saludable y en consideración de la 'vulnerabilidad' frente a diferentes enfermedades del casco.

Microtomographic Parameters and Nanoindentation of the Hoof of Girolando Cattle.

The aim of this study was to describe the microstructure of the pigmented and depigmented hoof capsule of Girolando cattle by bi- and tridimensional microtomography and nanoindentation, analysing the possible relation between these findings and the susceptibility of such animals to podal diseases. To carry out the microtomography and the nanoindentation, duplicated samples were collected from the dorsal wall, abaxial wall and pre-bulbar sole of the hoof capsule. Material collection was performed in 40 medial digits of thoracic limbs and 40 lateral digits of pelvic limbs. The bidimensional microtomography showed that the dorsal wall of the thoracic and pelvic limbs presented higher density, followed by the abaxial wall, and finally by the sole, with the lowest density. Moreover, the hoof capsule of cows of Girolando breed is a compact, non-porous material, and constituted by extratubular and intratubular keratin. By the tridimensional microtomography, it was possible to measure the angles of the corneal tubules in relation to the periople and the claws in the different regions of the hoof capsule, which were 90° for the dorsal wall, 55° for the abaxial wall and 70° for the sole. The tridimensional microtomography also showed corneal tubules of different diameters: 17, 51, 85, 119 and 153 µm. The nanoindentation test, when performed in different regions of the hoof capsule, did not reveal significant difference of Vickers hardness in the evaluated areas. However, we verified a larger elastic module of these regions on the transversal cut of the corneal tubules compared to the longitudinal cut.

Microstructure and Hardness of Buffalo's Hoofs.

The aim of this study was to describe the microstructure of hoof capsules of the buffalo. In addition, the study emphasized the morphometric aspects of the horn tubules, the Vickers nanohardness of the dorsal and abaxial walls and sole of the digits of the thoracic and pelvic limbs of the buffalo. The abaxial wall in the thoracic and pelvic digits showed larger diameter of the horn tubules when compared to all dorsal wall and sole. In addition, the abaxial wall of the thoracic digits showed larger diameter of the horn tubules when compared with the pelvic digits. According to the three-dimensional microtomography, the dorsal wall was higher in density compared with the abaxial wall. The latter exhibited an intermediate density, while the sole showed the lowest density. The Vickers nanohardness test showed that there was no difference in hardness and resistance between the experienced regions. However, the elastic modulus was greater on the transversal section of the hoof capsule. In conclusion, the results of the current study show that modern technologies such as microtomography and subsequent imaging can be used to investigate details of the basic morphology in different regions of the buffalo's hoof.

Biochemical composition of the hoof capsule of buffaloes and its influence on hoof quality / Composição bioquímica do estojo córneo de bubalinos e sua influência na qualidade do casco

The purpose of this study was to establish the biochemical parameters of the abaxial wall, dorsal wall and sole of the hoof of the medial thoracic, lateral, and medial pelvic digits of buffalos. The hoof samples were subjected to destructive biochemical analyses to identify the dry material (DM), mineral matter (MM), organic matter (OM), crude protein (CP) and ether extract (EE) contents. Sulfur (S), calcium (Ca), potassium (K), phosphorus (P), zinc (Zn) and copper (Cu) levels were determined based on nondestructive biochemical analyses. The parameters of dry material, mineral matter, organic matter, crude protein and ether extract of hoof capsule of the digits of buffalos can be determined by means of both destructive and nondestructive biochemical analysis. In addition, this study revealed that the highest concentrations of DM, CP and minerals such as, K, Zn and Cu are concentrated in the digits that bear the greatest body mass weight, suggesting that there is a positive correlation between the aforementioned parameters and the strength and growth of the hoof capsule in the digits. As for the element S, this study demonstrated that its highest concentration is located in the lateral digits of the pelvic members.(AU)

O presente estudo objetivou estabelecer os parâmetros bioquímicos da muralha abaxial, muralha dorsal e sola do casco dos dígitos torácico medial e pélvico lateral dos bubalinos. Foram realizadas analises bioquímicas destrutivas das amostras dos cascos para se obterem os teores de matéria seca (MS), matéria mineral (MM), matéria orgânica (MO), proteína bruta (PB) e extrato etéreo (EE). As análises bioquímicas não destrutivas foram empregadas para se avaliarem os níveis de enxofre (S), cálcio (Ca), potássio (K), fósforo (P), zinco (Zn) e cobre (Cu). Concluiu-se que os parâmetros matéria seca, matéria mineral, matéria orgânica, proteína bruta e extrato etéreo do estojo córneo dos dígitos de bubalinos podem ser definidos por análises bioquímicas destrutivas e não destrutivas. Os maiores valores dos elementos minerais estão concentrados nos dígitos que suportam o maior peso do animal, sugerindo que existe uma relação positiva entre esses parâmetros. Além disso, este estudo revelou que as maiores concentrações de MS, PB e minerais como, K, Zn e Cu estão nos dígitos que carregam o maior peso de massa corporal, o que indica que há uma correlação positiva entre os parâmetros acima referidos, resistência e crescimento do estojo córneo dos dígitos. Em relação ao elemento de S, este estudo demonstrou que a sua maior concentração situa-se nos dígitos laterais dos membros pélvicos.(AU)

Insertion of the LINE-1 element in the C-MYC gene and immunoreactivity of C-MYC, p53, p21 and p27 proteins in different morphological patterns of the canine TVT / Inserção do elemento LINE - 1 no gene C-MYC e imunorreatividade das proteinas C-MYC, p53, p21 e p27 nos diferentes padrões morfológicos do tumor TVT

The canine transmissible venereal tumor (TVT) affects the external genitalia of dogs by the natural transplant of viable tumor cells. Thus, this research aimed to diagnose and characterize TVT morphological patterns, identify the insertion of the LINE-1 element in C-MYC gene, by means of the polymerase chain reaction (PCR), and evaluate the immunohistochemical expression of C-MYC, p53, p21 and p27 proteins. The relationship between C-MYC and p53 proteins and their interference on the expression of p21 and p27 were also studied. For that, 20 samples of naturally occurring TVT were used, subjected to cytopathological, histopathological and immunohistochemical analysis, and to molecular diagnosis of neoplasia. The increased tissue expression and the correlation among C-MYC, p53, p21 and p27 proteins indicate reduction and/or loss of their functionality in the TVT microenvironment, with consequent apoptotic suppression, maintenance of cell growth and progression of neoplasia.(AU)

O tumor venéreo transmissível canino (TVT) afeta a genitália externa de cães pelo transplante natural de células tumorais viáveis. Assim, esta pesquisa teve como objetivo diagnosticar e caracterizar TVT em padrões morfológicos, identificar a inserção do elemento LINE-1 em gene C-MYC, por meio da reação em cadeia da polimerase (PCR), e avaliar a expressão imuno-histoquímica do C-MYC, p53, p21 e p27. A relação entre C-MYC e as proteínas p53 e a sua interferência na expressão de p21 e p27 foram também estudadas. Para isso, foram utilizadas 20 amostras de ocorrência natural de TVT, submetido a exame citopatológico, histopatológica e imuno-histoquímica e ao diagnóstico molecular de neoplasia. A expressão aumentada do tecido e a correlação entre a C-MYC e as proteínas p53, p21 e p27 indicam redução e/ou perda de funcionalidade na TVT em seu microambiente, com consequente supressão apoptótica, manutenção do crescimento celular e progressão da neoplasia.(AU)

Histomorphological evaluation of the digital coronary region at different fetal development stages of Holstein cattle / Avaliação histomorfológica da região coronária digital em diferentes fases de desenvolvimento fetal de bovinos da raça Holandesa

The scientific literature lacks detailed morphological descriptions of the histological development and cell differentiation of fetal bovine hoof. In this study, 40 extremity members of Holstein bovine fetuses were collected and divided into four groups (G1 to G4) based on the estimated age. Fragments were removed from wall and sole, processed and stained with hematoxylin - eosin (HE) for light microscopy observation. In G1, it was found that the epidermis was very thin, including keratinocyte layers and clusters of mesenchymal cells. In group G2 it was observed that the thickness of the epidermis covering the limbs remained variable and laminar corium developed in the germinal layer. In group G3 it was noted that in the germinal epithelium there were papillae in little advanced development and cells of the stratum corneum in the initial process of keratinization. In G4, the epidermis was well developed with layers distributed homogeneously, containing symmetrical and long papillae and intense production of keratin. In this work, the most important cellular events for the formation of the fetal hoof in Holstein cattle were first described in different stages of their formation.

A literatura científica carece de descrições morfológicas detalhadas sobre o desenvolvimento histológico e diferenciação celular do casco fetal bovino. Neste estudo, 40 extremidades de membros de fetos bovinos da raça Holandesa foram coletados e divididos em quatro grupos (G1 a G4) com base na estimativa da idade. Desses membros, foram retirados fragmentos da parede e sola, processados e corados por hematoxilina-eosina (HE) para observação em microscopia óptica. No grupo G1, observou-se que a epiderme era bastante delgada, incluindo camadas de queratinócitos e conjuntos de células mesenquimais. No grupo G2, foi observado que a espessura da epiderme, cobrindo as extremidades dos membros, permaneceu variável, e o cório laminar desenvolveu-se na camada germinativa. No grupo G3, notou-se, no epitélio germinal, papilas em desenvolvimento pouco avançado e células da camada córnea no processo inicial de queratinização. No grupo G4, a epiderme apresentava-se bem desenvolvida, com camadas distribuídas de forma homogênea, contendo papilas simétricas e longas e com intensa produção de queratina. Neste trabalho, os eventos celulares mais importantes para a formação do casco fetal em bovinos da raça Holandesa foram descritos pela primeira vez, em diferentes etapas da sua formação.