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Many studies about fasciolosis control have been carried out, whether acting on the adult parasite or in Pseudosuccinea columella, compromising the development of the larval stages. The present study aimed to evaluate, under laboratory conditions, the susceptibility of P. columella to Heterorhabditis bacteriophora HP88, during for 24 and 48 hours of exposure. The snails were evaluated for 21 days for accumulated mortality; number of eggs laid; hatchability rate; biochemical changes; and histopathological analysis. We found that exposure induced a reduction in glucose and glycogen levels, characterizing a negative energy balance, due to the depletion of energy reserves as a result of the direct competition established by the nematode/endosymbiont bacteria complex in such substrates. A mortality rate of 48.25% and 65.52% was observed in the group exposed for 24 h and 48 h, respectively, along with significant impairment of reproductive biology in both exposed groups in relation to the respective controls. The results presented here show that P. columella is susceptible to the nematode H. bacteriophora, with the potential to be used as an alternative bioagent in the control of this mollusk, especially in areas considered endemic for fascioliasis, in line with the position expressed by the World Health Organization Health.

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Background: The urinary tract is composed by kidneys, urinary bladder and urethra. The kidneys produce urine that achieve urinary bladder by ureters. These have the origin in the renal pelvis, run through the retroperitoneum, end up at the dorsolateral superficies of the urinary bladder, and empty at the trigone. Ureters abnormalities are the rarest congenital defects in the canine urinary tract and ureteroceles are cystic dilatations of the distal segment of the ureter that could be associated to partial or complete urinary obstructions and could lead to megaureter and hydronephrosis. So, the aim of the present study was to describe a case of megaureter by intramural ureteral ectopia in a bitch. Case: A 1-year-old-and-8-month bitch Akita, weighing 18 kg, was referred to the Uniube Veterinary Hospital with vaginal secretion, prostration, hypodipsia, hyporexia and pyrexia related by the tutor. On physical examination, an increase in vulva volume and a vaginal discharge were observed. Nevertheless, others physical exams, blood count and biochemical tests were considered to be within normal parameters. Urinalysis showed cloudy aspect, proteinuria, occult blood, erythrocytes, pyuria, leucocytes, and discreet presence of bacteria. Abdominal ultrasonography revealed a megaureter with right uterocele and excretory urography showed absence of glomerular filtration by right kidney. The patient was submitted to surgery for right kidney and ureter exeresis. Histopathology evaluation showed intense dilation of the ureter and severe multifocal renal fibrosis. The surgery was well succeeded, and the patient recovered completely. Discussion: Once megaureter are associated with congenital abnormalities like ectopic ureter and ureterocele, it is usually diagnosed in young patients with medium age of 10 months, which is below the age of the patient in this case report. Additionally, in the patient here reported, the unilateral alteration could explain the absence of kidney fail symptoms. In more than 90% of the cases, the ureteral ectopia was associated with multiple anomalies in the urinary tract, as was observed in this patient, that presented besides ectopic ureter, ureterocele, megaureter and renal dystrophy. All these morphological alterations made impossible the complete urine elimination, which predispose to urinary tract infection, that was observed in this report. According to literature, urinary tract anomalies are associated with infection in 64 to 85% and 50% of the cases also present hydronephrosis and hydroureter. It was also described that ureteral ectopia is diagnosed by visualization of hydroureter in abdominal ultrasonography. The findings present in this report differs a little, once the right kidney was atrophic possibly by malformation or even so by a chronic renal lesion due to the difficulty in urine flow. The excretory urography showed no filtration in the right kidney, indicating non-functionality that was confirmed by histopathology, in which was observed small glomerulus and large amount of connective tissue deposition. In cases of unilateral megaureter with ipsilateral kidney commitment, there is indication of nephroureterectomy, that was performed in the patient of the present report. As far as we know, this is the first report of megaureter, ureterocele and ectopia ureteral together in the same patient. In conclusion, the procedure was secure, efficient and promote a better quality of life for the patient and prevent the recurrence of urinary tract infections.

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The objective of this study was to investigate the in vitro action of triiodothyronine (T3) on the chondrogenic differentiation of adipose tissue-derived stem cells (ASCs) of female rats, with different time periods and doses. ASCs were extracted from female Wistar rats and were cultured in chondrogenic medium with and without the presence of T3. Five groups were established: 1) ASCs without T3; and 2,3,4,5) ASCs with 0.01, 1, 100 and 1,000 nM T3, respectively). After 7, 14 and 21 days, cell morphology, chondrogenic matrix formation, and expression of Sox9, aggrecan, collagen II, and collagen X were evaluated. The Student-Newman-Keuls test was used. ASCs showed CD54, CD73, and CD90 before chondrogenic differentiation. The hormone treatment did not alter chondrogenic matrix formation, Sox9 expression at 14 or 21 days, or expression of collagen II or collagen X at any time. However, the 0.01, 1, and 1000 nM T3 doses decreased Sox9 expression at 7 days. In conclusion, chondrogenic differentiation of ASCs of female rats is not influenced by T3.

O objetivo do presente trabalho foi verificar o efeito in vitro da triiodotironina (T3) na diferenciação condrogênica de células tronco mesenquimais do tecido adiposo (CTM-TA) de ratas, durante vários períodos e em várias doses. CTM-TA foram coletadas de ratas Wistar e cultivadas em meio condrogênico com ou sem a presença de T3. Constitui-se cinco grupos: 1) CTM-TA sem T3; e 2,3,4,5) CTM-TA com T3 (0,01; 1; 100 e 1000 nM, respectivamente). Após sete, 14 e 21 dias, foram avaliados morfologia celular, formação de matriz condrogênica e expressão de Sox9, agrecano, colágeno II e colágeno X. Para as análises foi utilizado o teste de Student Newman Keuls. CTM-TA expressaram CD54, CD73 e CD90 antes da diferenciação condrogênica. O tratamento hormonal não alterou a formação de matriz condrogênica e a expressão de Sox9 aos 14 e 21 dias e expressão dos colágenos II e X em nenhum dos períodos avaliados. No entanto, as doses de 0,01; 1 e 1000 nM T3 diminuíram a expressão de Sox9 aos 7 dias. Conclui-se que a diferenciação condrogênica de CTM-TA de ratas não é influenciada pela T3.

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Background: The endothelium is a layer fundamental to maintaining corneal transparency. In ophthalmology, sheep eyes have been used as a model in research related to corneal transplantation. Different techniques have been used to evaluate the corneal endothelium. Concerning vital dyes, corneal endothelial cell analyses have not yet been studied in ovines. The purpose of the present study was to evaluate the morphology of endothelial cells from different regions of the cornea of sheep after staining with alizarin red and trypan blue using an optical microscope. Materials, Methods & Results: Twenty healthy eyes of 10 male sheep obtained from a licensed commercial slaughterhouse were studied. The study was approved by the Research Committee of the Faculty of Veterinary at UFRGS and followed the ethical standards of the Association for Research in Vision and Ophthalmology (ARVO). Immediately after the slaughter, the eyes were enucleated and underwent eye examination. The corneal endothelium was stained with trypan blue and alizarin red and examined and photographed using an optical microscope. The central, superior, inferior, nasal and temporal areas of the cornea were evaluated for cell morphology. Data were compared by t-tests. Differences were considered statistically significant at P < 0.05. Immediately after staining the corneal endothelium, it was possible to examine with an optical microscope, obtain images and analyse the shape of endothelial cells from all regions of the sheep cornea. Polygonal, uniform and continuous cells were observed in all samples studied. Considering all the corneas analysed, cells with 6 sides (75.11%), 5 sides (12.76%) and 4 sides (12.12%) were found. In the central region of the cornea 75.91% of cells with 6 sides, 12.6% of cells with 5 sides and 11.48% with 7 sides were found. In the superior region of the cornea 76.07% of cells with 6 sides, 13.25% with 5 sides and 10.68% with 7 sides were found. In the lower region were found 74.72% of cells with 6 sides, 13% with 5 sides and 12.27% with 7 sides. In the temporal region, 74.14% were 6-sided cells, 11.42% had 5 sides, and 14.43% had 7 sides. Furthermore, in the nasal region, 74.72% of the cells had 6 sides, 13.54% had 5 sides, and 11.73% had 7 sides. No significant differences were found between cell morphology in all corneal regions evaluated. In addition, no significant difference was found when comparing the right eye with the left eye. Discussion: Different methods are used for the analysis of corneal endothelium. For ex vivo research optical microscopy after endothelial staining is an alternative low-cost technique that allows the analysis of all regions of the cornea. Quantitative analyses must characterise the endothelial parameters of the different species. The analysis of the morphology of corneal endothelium with an optic microscope after staining with alizarin red has been described as an effective, rapid and cost-efficient method, since this dye blends with the borated cells, allowing identification. In the present study, using optical microscopy and coloration with alizarin red it was possible to explore and obtain images of the ovine endothelium of all regions of the cornea. In the current study, the endothelium had a predominance of cells will 6 sides in all regions studied. This study allowed us to obtain images of the endothelium as well as quantitative data on the morphology of the different regions of the sheep cornea. This study demonstrated that morphology did not differ between the central and peripheral regions. The findings of this study represent a further source of reproducible data that should be considered when using sheep cornea as ex vivo model for experimental research.

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Dioctophyme renale is the largest parasitic nematode of animals. It is most often found in the right kidney, but can also occur in the urinary system, ears, free abdominal cavity, mammary gland, thoracic cavity, and more rarely in subcutaneous tissue. The genus Dirofilaria has specific parasitic characteristics according to its location, and may parasitize the respiratory tract or even the skin, varying according to species. This report describes a case of Dioctophyme renale and Dirofilaria sp. in the subcutaneous tissue of a cat in the state of Espírito Santo, Brazil. An adult male mongrel cat showed physical enlargement in the left inguinal region, diagnosed as a subcutaneous nodule. Surgical excision and histopathological evaluation of the nodule were performed, and Dioctophyme renale and Dirofilaria sp. were found inside. Dioctophymosis and heartworm disease are present in Brazil and most other countries, but this is the unprecedented case of the association of Dioctophyme renale and Dirofilaria sp. in the subcutaneous tissue of a cat.

RESUMO

The aim of the present study was to perform the macroscopic and microstructural morphological classification of long bone fractures of Cerdocyon thous. Eighteen cadavers of the species were necropsied, and subjected to radiographic and microscopical evaluation when long bone fractures were detected. Among the 18 cadavers, eight (44%) had fractures equally distributed (33.33%) in the femur, humerus, or tibia. More frequently (61.54%), the fractures were simple and affected the diaphysis, and in smaller proportions (23.08%) reached the physeal line. In diaphyseal and metaphyseal fractures, microscopical evaluation revealed cortical bone tissue, with longitudinal osteons that contained longitudinal and intermediate collagen fibres and lamellae with a delamination aspect. On the other hand, in epiphyseal fractures, trabecular bone tissue was more frequently observed, consisting of trabeculae with disorganised collagen fibres and absence of osteons. In both cases low activity, osteocytes, and low coverage of osteoblasts on the bone surface were noted. It was concluded that the frequency of fractures in the long bones of C. thous was 44%, with females being more predisposed. The findings support the hypothesis that fractures in such animals are caused by being run over by automobiles. The present study contributes significantly in alerting clinicians and surgeons to the types of fractures that C. thous is more predisposed to, its places of greatest occurrence, and its microstructure. Thus, there is a need for joint actions aimed at reducing the number of cases of wild animals being run over by automobiles.

Teve-se como objetivo, no presente trabalho, realizar a classificação morfológica macroscópica e microestrutural das fraturas em ossos longos de Cerdocyon thous. Foram utilizados 18 cadáveres da espécie, necropsiados e submetidos à avaliação radiográfica e microscópica quando detectadas fraturas em ossos longos. Dentre os 18 animais, oito (44%) possuíam fraturas igualmente distribuídas (33,33%) em fêmur, úmero ou tíbia. Mais frequentemente (61,54%) as fraturas eram simples e acometiam a diáfise, e em menores proporções (23,08%) atingiam a linha fisária. Nas fraturas em diáfise e metáfise, predominava o tecido ósseo cortical, com ósteons longitudinais que continham fibras colágenas longitudinais e intermediárias, e lamelas com aspecto de delaminação. Por outro lado, nas fraturas fisárias, o tecido ósseo trabecular foi mais frequentemente observado, constituído por trabéculas com fibras colágenas desorganizadas e ausência de ósteons. Em ambos os casos, notou-se baixa atividade de osteócitos e baixa cobertura de osteoblastos na superfície óssea. Conclui-se que, nas condições observadas, a frequência de fraturas em ossos longos de C. thous foi de 44%, sendo as fêmeas mais predispostas, além de os achados embasarem a hipótese da ocorrência de tais lesões estarem relacionadas a atropelamentos. O presente estudo contribui significativamente para alertar clínicos e cirurgiões em relação aos tipos de fraturas as quais C. thous está mais predisposto, seus locais de maior ocorrência e sua microestrutura. Dessa forma, surge a necessidade de implementação de ações conjuntas que visem reduzir o número de casos de atropelamento de animais silvestres.

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Teve-se como objetivo, no presente trabalho, realizar a classificação morfológica macroscópica e microestrutural das fraturas em ossos longos de Cerdocyon thous. Foram utilizados 18 cadáveres da espécie, necropsiados e submetidos à avaliação radiográfica e microscópica quando detectadas fraturas em ossos longos. Dentre os 18 animais, oito (44%) possuíam fraturas igualmente distribuídas (33,33%) em fêmur, úmero ou tíbia. Mais frequentemente (61,54%) as fraturas eram simples e acometiam a diáfise, e em menores proporções (23,08%) atingiam a linha fisária. Nas fraturas em diáfise e metáfise, predominava o tecido ósseo cortical, com ósteons longitudinais que continham fibras colágenas longitudinais e intermediárias, e lamelas com aspecto de delaminação. Por outro lado, nas fraturas fisárias, o tecido ósseo trabecular foi mais frequentemente observado, constituído por trabéculas com fibras colágenas desorganizadas e ausência de ósteons. Em ambos os casos, notou-se baixa atividade de osteócitos e baixa cobertura de osteoblastos na superfície óssea. Conclui-se que, nas condições observadas, a frequência de fraturas em ossos longos de C. thous foi de 44%, sendo as fêmeas mais predispostas, além de os achados embasarem a hipótese da ocorrência de tais lesões estarem relacionadas a atropelamentos. O presente estudo contribui significativamente para alertar clínicos e cirurgiões em relação aos tipos de fraturas as quais C. thous está mais predisposto, seus locais de maior ocorrência e sua microestrutura. Dessa forma, surge a necessidade de implementação de ações conjuntas que visem reduzir o número de casos de atropelamento de animais silvestres.

The aim of the present study was to perform the macroscopic and microstructural morphological classification of long bone fractures of Cerdocyon thous. Eighteen cadavers of the species were necropsied, and subjected to radiographic and microscopical evaluation when long bone fractures were detected. Among the 18 cadavers, eight (44%) had fractures equally distributed (33.33%) in the femur, humerus, or tibia. More frequently (61.54%), the fractures were simple and affected the diaphysis, and in smaller proportions (23.08%) reached the physeal line. In diaphyseal and metaphyseal fractures, microscopical evaluation revealed cortical bone tissue, with longitudinal osteons that contained longitudinal and intermediate collagen fibres and lamellae with a delamination aspect. On the other hand, in epiphyseal fractures, trabecular bone tissue was more frequently observed, consisting of trabeculae with disorganised collagen fibres and absence of osteons. In both cases low activity, osteocytes, and low coverage of osteoblasts on the bone surface were noted. It was concluded that the frequency of fractures in the long bones of C. thous was 44%, with females being more predisposed. The findings support the hypothesis that fractures in such animals are caused by being run over by automobiles. The present study contributes significantly in alerting clinicians and surgeons to the types of fractures that C. thous is more predisposed to, its places of greatest occurrence, and its microstructure. Thus, there is a need for joint actions aimed at reducing the number of cases of wild animals being run over by automobiles.

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The effects of two concentrations of triiodothyronine (T3; 0.01 and 1,000 nM) on the osteogenic and chondrogenic differentiation abilities of equine adipose-derived mesenchymal stem cells (AD-MSCs) were evaluated. The osteogenic study evaluated the effect of T3 using alkaline phosphatase activity (ALP) assay; cell viability and density; and formation of mineralized nodules at Days 7, 14, and 21 in culture. The chondrogenic study tested the effect of T3 through ALP assay, mitochondrial metabolism, cell density, and periodic acid-Schiff-positive (PAS+) matrix percentage at Days 7 and 14. In both experiments, analysis of variance was used to compare averages through the Student-Newman-Keuls test. In the osteogenic study, no differences in any variable were detected between groups at Day 7. At Day 14, 0.01 nM T3 reduced cell density and the number of mineralized nodules despite the increase in ALP activity and mitochondrial metabolism (P < .05). ALP activity increased at 1,000 nM T3 concentration (P < .05). At Day 21, 0.01 nM T3 treatment increased ALP activity compared with control treatment (P < .05). At 1,000 nM concentration, T3 reduced mitochondrial metabolism and cell density (P < .05). In the chondrogenic study, the two T3 concentrations increased cell density compared with control treatment at Day 7. At Day 14, higher T3 concentration reduced mitochondrial metabolism, ALP activity, cell density, and PAS+ chondrogenic matrix percentage compared with control treatment (P < .05). Thus, T3 addition to equine AD-MSC cultures has no enhancement effect on osteogenic or chondrogenic differentiation and may, in fact, negatively affect cell density and matrix synthesis depending on hormone concentration and culture time.

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Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães. (AU)

The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.(AU)

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Abstract The objective of this study was to establish a protocol for the isolation and culture of mesenchymal stem cells (MSC) from bone marrow and adipose tissue of dogs. Three 6-month-old male dogs were used. Approximately 3.0 cm3 of adipose tissue and 3.0 mL of bone marrow were collected. The samples were processed and the isolated cells were cultured in DMEM. The cells were subjected to phenotypic characterization and to osteogenic, adipogenic, and condrogenic differentiation to confirm the isolation of the MSC. The cells showed elongated and fusiform morphology and they were able to differentiate into osteoblasts, adipocytes, and chondrocytes. Phenotypic characterization revealed high expression of the MSC markers CD90(80.04%) and CD29(96%) in the cells from bone marrow and high expression of CD90(60.94%) and CD29(77.08%) in the cells from adipose tissue. In addition, phenotypic characterization revealed low expression of hematopoietic markers CD45(1.45%) and CD34(1.53%) in the cells from bone marrow and low expression of CD45(1.45%) and CD34(1.53%) in the cells from adipose tissue. Based on these results, the modifications applied to classical protocols simplified the process and proved to be efficient in the isolation, culture, and expansion of MSC isolated from the bone marrow and adipose tissue of dogs.

Resumo Objetivou-se estabelecer um protocolo para extração, cultivo e expansão de células tronco mesenquimais (CTM), utilizando-se 3,0 mL da medula óssea e 3,0 cm3 de tecido adiposo do subcutâneo de três cães machos com seis meses de idade. As amostras foram processadas e as células extraídas e cultivadas em DMEM. Para comprovação do isolamento de CTM, procedeu-se a caracterização fenotípica e a diferenciação osteogênica, adipogênica e condrogênica. As células isoladas apresentaram morfologia alongada e fusiforme e capacidade de se diferenciar em osteoblastos, adipócitos e condrócitos. A caracterização fenotípica revelou alta expressão de marcadores de CTM CD90 (80,04%) e CD29 (96%) nas células de origem medular e CD90 (60,94%) e CD29 (77,08%) nas de origem adiposa. A expressão de marcadores hematopoiéticos foi baixa tanto nas células de origem medular CD45 (1,45%) e CD34 (1,53%), quanto nas de origem adiposa CD45 (1,45%) e CD34 (1,53%). As modificações e adaptações realizadas nos protocolos clássicos simplificaram o processo e foram eficientes, permitindo o isolamento e cultivo de CTM da medula óssea e do tecido adiposo de cães.