Flávio Ribeiro Alves

Anatomical and radiographic appearance of the capuchin monkey thoracic cavity (Cebus apella)

The capuchin monkey is widespread both north and south of the Legal Amazon and in the Brazilian cerrado. Ten clinically healthy capuchin monkeys were submitted to an anatomical and radiographic study of their thoracic cavities. The radiographic evaluation allowed the description of biometric values associated with the cardiac silhouette and thoracic structures. Application of the VHS (vertebral heart size) method showed positive correlation (P<0.05) with depth of the thoracic cavity, as well as between the body length of vertebrae T3, T4, T5 and T6 and the cardiac length and width. The lung fields showed a diffuse interstitial pattern, more visible in the caudal lung lobes and a bronchial pattern in the middle and cranial lung lobes. The radiographic examination allowed preliminary inferences to be made concerning the syntopy of the thoracic structures and modification of the pulmonary patterns and cardiac anatomy for the capuchin monkey.

Hematopoietic progenitor constituents and adherent cell progenitor morphology isolated from black-rumped agouti (Dasyprocta prymnolopha, Wagler 1831) bone marrow.

Stem cells are present in the adult tissues of most diverse species. Bone marrow is recognized to be the most exploited site to obtain stem cells and cell progenitors. The objective of the present study was to characterize hematopoietic progenitor (HP) morphology and analyze the performance of adherent cell progenitors (ACPs) cultivated in vitro from black‐rumped agouti bone marrow (Dasyprocta prymnolopha). Bone marrow aspirates were obtained from tibia crest and used to prepare histological slides and identify cell morphology. Cells were also scattered on culture plates for later isolation, expansion, and quantification. Smears obtained from bone marrow demonstrated HPs at different stages of maturity. In culture, these cells showed fibroblastoid morphology and a strong tendency to form colonies, demonstrated by the presence of cell aggregates, cytoplasmic elongations lying side by side. An 80% cell confluence was observed at 18 days in culture and progressive reduction in the percentage of nonadherent mononuclear cells. After eight passes, a mean cell viability of 96.07% was observed, from a pool of 1.6 × 107 cells (ACP). Thirteen 25‐cm2 culture bottles were trypsinized, resuspended in freezing medium, stored in 14 criotubes at a concentration of 1 × 106 cells per milliliter, and placed in liquid nitrogen at −196°C. Agouti bone marrow demonstrated high plasticity, moreover different HP lines, and a population of adherent cells demonstrated morphology similar to mesenchymal stem cells in culture. Microsc. Res. Tech. 2012.

Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium

Alves F.R., Emerson T.F., Ricardo R.G., Juliana C.D., Antonio A.N.M.J., Ambrosio C.E., Irina K. & Miglino M.A. 2010. Establishment of a protocol for obtention of neuronal stem cells lineages from the dog olfactory epithelium. Pesquisa Veteri- naria Brasileira 30(4):363-372. Departamento de Ciencia Animal, Campus Universitario, Bairro Cibrazen, Bom Jesus, PI 64900-000, Brazil. E-mail: flavioribeiro@ufpi.edu.br A morphological and cell culture study from nasal mucosa of dogs was performed in order to establish a protocol to obtain a cell population committed to neuronal lineage, as a proposal for the treatment of traumatic and degenerative lesions in these animals, so that in the future these results could be applied to the human species. Twelve mongrel dogs of 60-day aged pregnancy were collected from urban pound dogs in Sao Paulo. Tissue from cribriform ethmoidal lamina of the fetuses was collected at necropsy under sterile conditions around 1h to 2h postmortem by uterine sections and sections from the fetal regions described above. Isolated cells of this tissue were added in DMEM/F-12 medium under standard conditions of incubation (5% CO 2 , >37°C). Cell culture based on isolated cells from biopsies of the olfactory epithelium showed rapid growth when cultured for 24 hours, showing phase-bright sphere cells found floating around the fragments, attached on culture flasks. After 20 days, a specific type of cells, predominantly ellipsoids or fusiform cells was characterized in vitro. The indirect immunofluorescence examination showed cells expressing markers of neuronal precursors (GFAP, neurofilament, oligodendrocyte, and III â-tubulin). The cell proliferation index showed Ki67 immunostaining with a trend to label cell groups throughout the apical region, while PCNA immunostaining label predominantly cell groups lying above the basal lamina. The transmission electron microscopy from the olfactory epithelium of dogs revealed cells with electron-dense cytoplasm and preserving the same distribution as those of positive cell staining for PCNA. Metabolic activity was confirmed by presence of euchromatin in the greatest part of cells. All these aspects give subsidies to support the hypothesis about resident progenitor cells among the basal cells of the olfactory epithelium, committed to renewal of these cell populations, especially neurons.

Ultrasonographic and radiographic determination of egg development of jurarás (Kinosternon scorpioides) in captivity

Kinosternon scorpioides (Linnaeus, 1766), with its common name of jurara, is a quite variable species of turtles, and many different names have been applied to populations throughout its range. Currently, however, four subspecies are considered valid as K. scorpioides arises from southern Panama over most of northern South America and is found in Ecuador, northern Peru, southern Bolivia, northern Argentina, eastern Guyana and Brazil. Thus, an ultrasonographic and radiographic study was performed in order to describe the morphology and development of eggs of 20 female jurara mud turtles K. scorpioides, from September 2005 to August 2006. In the first month, the ovarian cycle was characterized by absence of vitellogenic follicles, atresic follicles or oviduct eggs. From October 2005 to March 2006 on, ultrasonographic scanning allowed to establish the growing vitellogenic follicles. Vitellogenic follicles were observed with spherical to ovoid shapes, with a surrounding echogenic yolk, a nonechogenic albumin layer, and a high echogenic shell. The oviduct eggs were identified by radiography just 180 days after beginning the experiment, when the shell became enough mineralized to impress the radiographic film. This experiment allowed to obtain by means of the 7.5 MHz linear probe images with adequate resolution and penetration for visualization of follicles. Successive ultrasonographic examinations of 20 K. scorpioides females allowed to access initial stages of vitellogenic follicles and oviduct eggs, and radiographic examination revealed to be an easy technique to assess oviduct eggs and to allow evaluation of egg development in juraras, from 6 months on.

Comparative study by computed radiography, histology, and scanning electron microscopy of the articular cartilage of normal goats and in chronic infection with caprine arthritis‐encephalitis virus

In the northeast of Brazil, caprine arthritis‐encephalitis (CAE) is one of the key reasons for herd productivity decreasing that result in considerable economic losses. A comparative study was carried out using computed radiography (CR), histological analysis (HA), and scanning electronic microscopy (SEM) of the joints of CAE infected and normal goats. Humerus head surface of positive animals presented reduced joint space, increased bone density, and signs of degenerative joint disease (DJD). The carpal joint presented no morphological alterations in CR in any of the animals studied. Tarsus joint was the most affected, characterized by severe DJD, absence of joint space, increased periarticular soft tissue density, edema, and bone sclerosis. Histological analysis showed chronic tissue lesions, complete loss of the surface zone, absence of proteoglycans in the transition and radial zones and destruction of the cartilage surface in the CAE positive animals. Analysis by SEM showed ulcerated lesions with irregular and folded patterns on the joint surface that distinguished the limits between areas of normal and affected cartilage. The morphological study of the joints of normal and CAE positive goats deepened understanding of the alteration in the tissue bioarchitecture of the most affected joints. The SEM finding sustained previous histological reports, similar to those found for rheumatoid arthritis, suggesting that the goat infected with CAE can be considered as a potential model for research in this area. Microsc. Res. Tech. 77:11–16, 2014.

CARDIOTHORACIC RATIO AND VERTEBRAL HEART SCALE IN CLINICALLY NORMAL BLACK-RUMPED AGOUTIS (DASYPROCTA PRYMNOLOPHA, WAGLER 1831)

Wild rodents, such as the lowland paca (Cuniculus paca), capybara (Hydrochoerus hydrochaeris), rock cavy (Kerodon rupestris), guinea pig (Cavia aperea), and black-rumped agouti (Dasyprocta prymnolopha) are intensely hunted throughout Amazonia and at the semiarid regions of northeastern Brazil. To contribute to the preservation of these species, more information about their anatomy, physiology and pathophysiology is needed. The aim of this study was to standardize the vertebral heart scale (VHS) and cardiothoracic ratio (CTR) in clinically normal black-rumped agouti, as well as to compare the results of these two methods, which are commonly used to evaluate the cardiac silhouette in domestic animals. Twelve healthy black-rumped agoutis, divided into two groups (six males and six females), obtained from the Nucleus for Wild Animal Studies and Conservation at the Federal University of Piauí, were radiographed in right and left lateral and dorsoventral projections. The values of the VHS were 8.00±0.31v (the number of thoracic vertebral length spanned by each dimension, starting at T4) for males and 8.11±0.41v for females, and there was no statistical difference between the decubitus (right and left) or between males and females (P>0.05). The CTR mean values obtained were 0.51±0.03 for males, and 0.52±0.02 for females, and there was no statistical difference between the genders (P>0.05). However, there was positive correlation between VHS and CTR (r=0.77 right decubitus and r=0.82 left decubitus). The thoracic and heart diameter had mean values of 6.72±0.61 and 3.48±0.30 cm (males), and for the females, it was 6.61±0.51 and 3.5±0.30 cm, respectively, and there was statistical difference between the genders. The results demonstrated high correlation between the VHS and CTR producing similar results, indicating similar clinical precision for assessing the size of the cardiac silhouette in the black-rumped agoutis.

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

PURPOSE To characterize bone marrow progenitors cells grown in vitro, using native goats from northeastern Brazil as animal model. METHODS Ten northeastern Brazil native goats of both genders were used from the Piauí Federal University Agricultural Science Centers (UFPI) - Goat Farming Sector. Bone marrow aspirates where taken from the tibial ridge and seeded on culture plates for isolation, expansion and Flow Cytometry (expression markers - Oct-3/4, PCNA, Ck-Pan, Vimentina, Nanog). RESULTS Progenitor cells showed colonies characterized by the presence of cell pellets with fibroblastoid morphology. Cell confluence was taken after 14 days culture and the non-adherent mononuclear cell progressive reduction. After the first passage, 94.36% cell viability was observed, starting from 4.6 x 106 cell/mL initially seeded. Cells that went through flow cytometry showed positive expression for Oct-3/4, PCNA, Ck-Pan, Vimentina, and Nanog. CONCLUSIONS Bone marrow progenitor isolated of native goats from northeastern Brazil showed expression markers also seen in embryonic stem cells (Oct-3/4, Nanog), markers of cell proliferation (PCNA) and markers for mesenchymal cells (Vimentina and Ck-pan), which associated to morphological and culture growth features, suggest the existence of a mesenchymal stem cell (MSC) population in the goat bone marrow stromal cells studied.

Electrocardiographic pattern of agoutis (Dasyprocta prymnolopha) chemically contained by the association of ketamine and xylazine

Background: The agouti (Dasyprocta prymnolopha, Wagler, 1831) is an existing wild rodent in almost all of Brazil, used as a biological model in several scientific studies. Veterinary cardiology has showed great advances in the diagnostic area due to the possibility of cardiac evaluation by non-invasive methods. For the practice of scientific or handling procedures in wild animals, chemical containment is essential. Thus, it is important to know the effect of anesthetic protocols on the cardiovascular system, observed through complementary tests, such as the electrocardiogram. The objective of this study was to describe the computerized electrocardiographic tracing of agoutis chemically contained by the association of Ketamine and Xylazine. Materials, Methods & Results: Eighteen male and female clinically healthy animals, aged among 2 years, submitted to digital electrocardiographic examination, were used. The device used to obtain the tracing was the veterinary electrocardiograph (Electrocardiogram Acquisition Module for Computer) - Brazilian Electronic Technology (TEB). The analysis was always made from derivation II (DII). Chemical containment was performed by the combination of Ketamine and Xylazine, intramuscularly. The results showed that the heart rate for males was on average 113.25 bpm, while for females the value of 124.60 bpm was observed, and there was no significant difference between the genders (P > 0.05). The mean weight of males was 2.31 kg and for females 2.28 kg; there was no statistical difference for this variable (P > 0.05). For the QRS duration of 46.14 ± 5.05 ms (males) and 44.66 ± 5.94 ms (females) and PR interval of 79.94 ± 12.01ms (males) and 84.29 ± 12.37ms (females), there was no statistical difference (P > 0.05). The amplitude of the R wave of 0.42 ± 0.31 mV (males) and 0.36 ± 0.22 mV (females) showed no statistical difference (P > 0.05). The T wave showed itself both negative (10 animals) and positive (8 animals), with amplitude measured at 0.24 ± 0.16 mV for males and 0.25 ± 0.10 mV for females (P > 0.05). The anesthetic protocol was well tolerated by the animals of this experiment, and there were no episodes of arrhythmias during the time of their monitoring. Discussion: Wild animals, in general, need chemical containment, with a view to reducing stress during scientific and handling procedures. The protocols established in this work can be used in future experiments which require a longer handling time. The values found for heart rate (HR) of anesthetized agoutis were lower than those for non-anesthetized agoutis, proving the influence of the stress during retention and the depressant cardiovascular effect of the anesthetic drug used. The electrocardiographic parameters and the tracing morphology of the agoutis were like the results found for anesthetized small rodents. The QT interval was higher in males than in females, justifying the fact that it could be influenced by variations in HR, which also showed superior results for males. Taking into consideration the morphological similarity between different individuals, the QT interval in anesthetized agoutis did not present significant differences between the genders, a characteristic also found in ferrets. The animals of this experiment were submitted to food fasting, a fact that contributed to a better standardization of the experiment, preventing alterations in the morphology of the QRS complex, which could lead to changes in the measurements.

Comparative B-mode and Doppler renal ultrasonography with histopathological findings in dogs positive for canine visceral leishmaniasis.

Morphological parameters and renal flow were assessed and measured in six clinically healthy dogs with negative serological and parasitological examinations for canine visceral leishmaniasis (CVL) and six dogs with CVL‐positive serological and parasitological tests and asymptomatic condition of the disease. Kidney length and diameter were measured and compared with the aorta/kidney ratio. Alterations in flow were measured using the flow velocity (cm/s) and resistance index (RI). Renal function was assessed by serum biochemical examination (urea, creatine, total proteins, albumin, and globulin) and urinalysis. CVL‐positive animals presented heterogeneous kidney echotexture, reduced vascular flow, hypoperfusion, lesser visualization of the arcuate arteries, high RI values (1.06 ± 0.28 for the right kidney and 1.10 ± 0.30 for the left kidney), reduced diastolic flow, and high systolic peak. They also presented positive creatine/RI correlation and between the renal artery vascular flow and kidney diameter. Histopathological analysis of CVL‐positive animals was compatible and confirmed hemodynamic changes observed by Doppler ultrasound. The findings in the present study showed that Doppler ultrasonography is a valid method to assess and detect alterations in flow in renal pathological processes with kidney vascular damage, as in the case of leishmaniasis, and can help in the diagnosis of animals with this infection. Microsc. Res. Tech. 79:637–645, 2016.

Análise quantitativa da ecogenicidade testicular pela técnica do histograma de ovinos da baixada ocidental maranhense

Ultrasound is a diagnostic imaging method that allows a no invasive evaluation of different organs and body structures. However, the personal evaluation of images is one of the major hampers in using this technique. In order to reduce this drawback, it is necessary to find out tools that can help diminish the divergence in the interpretation of sonographic findings. Therefore, this study aimed to identify features of the parenchymas and mediastinums testis of sheep echogenicity using the technique of gray scale histogram. 30 animals were divided into three groups according to age (FE): three to six months (FE1), seven to 12 months (FE2), 13 to 18 months (FE3). The testicular scans were performed in the frontal, sagital and transverse views, and the gray scale histograms were analyzed. It was observed that such parenchymal as the mediastinum testis echogenicity gradually increased with the aging of the animals, with mean and standard deviation of 95.00±19.05 and 94.35±18.82 for the echogenicity parenchyma of the right and left antimere, respectively, and 127.95±12.97 to 126.59±11.78, for the right and the left mediastinum. The technique of gray scale histogram was an efficient method for determination of testicular echogenicity, enabling the establishment of normal standards that may assist future research in monitoring testicular development as well as for detecting pathologies. For exclusive regimes of extensive farming, as the western lowland region of Maranhao, is a valuable tool for use in social projects of the State to attend the family farm.