Does lipid peroxidation and oxidative DNA damage differ in cryopreserved semen samples from young, adult and aged Nellore bulls?

The aims of this study were to evaluate cryopreserved semen of Nellore bulls of different ages and verify whether sperm quality declines with advancing age and whether lipid peroxidation and DNA damage are involved in this process. For this purpose, 40 Nellore bulls were divided into three age groups: Young, aged 1.8-2 years (n = 9); Adult, aged 3.5-7.0 years (n = 19); and Seniors, aged 8.0-14.3 years (n = 12). Three ejaculates were collected from each bull, cryopreserved and evaluated for various parameters including membrane integrity, mitochondrial potential (FITC-PSA and JC1), lipid peroxidation (C-11BODIPY 581 / 591) and oxidative DNA damage (8OHdG) using flow cytometry. The thawed semen of senior bulls was characterized by a low percentage of motile sperm (33.7 ±â€¯6.1%), higher damage to the plasma and acrosomal membrane (37.5 ±â€¯9.8%), and low mitochondrial potential (29.1 ±â€¯13.8%), as well as higher percentages of peroxidated cells (53.6 ±â€¯12.2%) and DNA damage (44.1 ±â€¯11.0%; P < 0.05). Lipid peroxidation was negatively correlated with motility (r = -0.35, P < 0.0002), average mitochondrial potential (r = -0.42; P < 0.0001) and showed a positive correlation with membrane injury and oxidative DNA damage (r = 039; P = 0.0003). Young bulls presented superior thawed sperm quality, possibly due to greater resistance to oxidative stress and, consequently, to cryopreservation. In conclusion, the sperm quality of bull semen declines with advancing age and is strongly associated with increased oxidative damage to both the plasma membrane and DNA.

Evaluation of sperm head dimensions and chromatin integrity of epididymal sperm from domestic cats using the toluidine blue technique.

When using assisted reproductive technologies, there is seldom an evaluation of DNA integrity during sperm analysis, which is an important variable for proper embryo development. The toluidine blue staining technique allows the simultaneous evaluation of sperm chromatin and sperm head dimensions. The objectives of this study were to evaluate the applicability of the toluidine blue staining method for analyzing DNA abnormalities in epididymal sperm (from the caput, corpus, and cauda) of cats and to investigate the correlations among DNA condensation, morphology, and sperm head dimensions. The DNA alteration indexes were obtained using the toluidine blue and acridine orange techniques, and comparisons of these indexes indicated there was a 65.4% (r = 0.654; P < 0.001) correlation. The sperm from the cauda had greater chromatin stability (97.9%) than the sperm from the epididymal head (92.1%; P = 0.0023). There, however, was no difference in chromatin stability between sperm obtained from the corpus and cauda regions, indicating that these sperm were already mature. The sperm head dimension was correlated with chromatin decondensation, and the sperm head size decreased as the sperm were transported through the three epididymal regions (P < 0.0001). In addition, the percentage of sperm that were deficient in chromatin condensation decreased as the sperm were transported through the epididymal caput, corpus and cauda (26.4, 15.7, and 3.4%, respectively; P < 0.0001). Thus, the sperm head size predicts the quality of chromatin condensation in sperm cells.

Filling of extraction sockets of feline maxillary canine teeth with autogenous bone or bioactive glass.

PURPOSE: To evaluate bone healing in the extraction socket of the feline maxillary canine tooth after grafting. METHODS: Eighteen adult cats were submitted to unilateral extraction of maxillary canine tooth and divided into three groups. In group 1 (n=6), control, the extraction socket was left empty. In group 2 (n=6), the extraction socket was filled with autogenous cancellous bone from the iliac crest and in group 3 (n=6), with bioactive glass particulate material. Cats were euthanized at four weeks postoperative. RESULTS: The radiographic examinations performed four weeks after surgery showed that in all groups the healing process converged to a radiopacity similar to that observed in the surrounding bones. Histological examination showed formation of woven bone within the extraction socket. The percentage of newly formed bone within the extraction socket, measured by the histometry, showed no statistically significant difference among the values of the three groups (Kruskal-Wallis'test p>0.05) (group 1: 63.96 ± 5.85, group 2: 66.84 ± 11.67, group 3: 59.28 ± 15.50). CONCLUSION: The bone regeneration observed in the extraction sockets filled with autogenous cancellous bone or bioactive glass was similar to that observed in the control sites, given an observation period of four weeks after extraction of the maxillary canine tooth.

Filling of extraction sockets with autogenous bone in cats.

PURPOSE: To evaluate bone healing in the extraction socket of the feline mandibular canine tooth after grafting. METHODS: Eighteen adult cats were submitted to unilateral extraction of mandibular canine tooth and divided into three groups. In group 1 (n=6), control, the extraction socket was left empty. In group 2 (n=6), the extraction socket was filled with autogenous cancellous bone from the iliac crest and in group 3 (n=6), with cortical bone chips from the iliac crest. Cats were euthanized at 6 weeks postoperative. RESULTS: Immediate postoperative radiographs in dorsoventral view showed a radiolucent area at the extraction wound. A decreased radiolucency was observed on the radiographs taken at 6 weeks postoperative. Histological examination showed formation of woven bone within the extraction socket. The percentage of newly formed bone within the extraction socket, measured by the histometry, showed no statistically significant difference among the values of the three groups (Kruskal-Wallis'test p>0.05) (group 1: 52.54 ± 15.46, group 2: 50.51 ± 5.01, group 3: 51.85 ± 9.52). CONCLUSION: The bone regeneration observed in the extraction sockets filled with autogenous cancellous bone or autogenous cortical bone chips was similar to that observed in the control sites, given an observation period of 6 weeks after extraction of the mandibular canine tooth.

Miniplate fixation for the repair of segmental mandibular defects filled with autogenous bone in cats.

PURPOSE: To evaluate the use of maxillofacial miniplate 1.5 in the repair of segmental mandibular defects filled with autogenous bone in cats. METHODS: Twelve adult cats were divided into two groups. A segmental defect of 4mm was created in one of the hemimandibles and filled with autogenous iliac crest bone graft. The operated hemimandible was fixed with a 1.5mm titanium miniplate. In group 1 (n=6), the defect was performed in the body of the mandible, behind the 1st molar. In group 2 (n=6), the defect was performed between the 4nd premolar and 1st molar, with extraction of the 1st molar. Oral alimentation was reinitiated 24 hours after surgery. Cats were euthanized at 20 weeks postoperative. RESULTS: Incorporation of the graft was suggested by the radiographs taken 20 weeks after surgery. Macroscopic examination confirmed alignment and bone union of operated hemimandibles. Histological examination showed formation of woven bone in rostral and caudal mandible/graft interfaces. The percentage of bone tissue at these areas was measured by the histometry. There was no statistically significant difference between the values of group 1(64.48 ± 4.51) and group 2 (71.69 ± 14.47) (Mann-Whitney's test p= 0.294). CONCLUSION: The use of miniplate 1.5 for the fixation of mandibular defects filled with autogenous bone in cats provided the main goals in the treatment of mandibular fractures: bone union, normal dental occlusion and immediate return to oral alimentation.

Miniplate 1.5 fixation for the repair of mandibular osteotomies in cats.

PURPOSE: To evaluate the application of the maxillofacial miniplate 1.5 in the repair of unilateral mandibular osteotomies in cats. METHODS: Twelve adult cats were divided into two groups. In group 1 (n=6), the osteotomy was performed in the body of the mandible, behind the 1(st) molar. In group 2 (n=6), the osteotomy was performed between the 4(th) premolar and 1(st) molar. The osteotomy was fixed with a titanium miniplate 1.5. Oral alimentation was reinitiated 24 hours after surgery. Cats were euthanized at 12 weeks postoperative. RESULTS: Radiographs taken 1 week after surgery showed a radiolucent line. The osteotomy line was not more visible on the radiographs taken at 12 weeks postoperative. Macroscopic examination confirmed alignment and bone union of operated hemimandibles. Histological examination showed formation of woven bone within the osteotomy line. The percentage of bone tissue at these areas was measured by the histometry. There was no statistically significant difference between the values of group 1(75.07 ± 5.99) and group 2 (74.76 ± 8.54) (Mann-Whitney's test p= 0.469). CONCLUSION: We concluded that the use of miniplate 1.5 for the fixation of mandibular osteotomy in cats provided the main goals in the treatment of mandibular fractures: bone union, normal dental occlusion and immediate return to oral alimentation.

Filling of extraction sockets of feline maxillary canine teeth with autogenous bone or bioactive glass

PURPOSE: To evaluate bone healing in the extraction socket of the feline maxillary canine tooth after grafting. METHODS: Eighteen adult cats were submitted to unilateral extraction of maxillary canine tooth and divided into three groups. In group 1 (n=6), control, the extraction socket was left empty. In group 2 (n=6), the extraction socket was filled with autogenous cancellous bone from the iliac crest and in group 3 (n=6), with bioactive glass particulate material. Cats were euthanized at four weeks postoperative. RESULTS: The radiographic examinations performed four weeks after surgery showed that in all groups the healing process converged to a radiopacity similar to that observed in the surrounding bones. Histological examination showed formation of woven bone within the extraction socket. The percentage of newly formed bone within the extraction socket, measured by the histometry, showed no statistically significant difference among the values of the three groups (Kruskal-Wallis'test p>0.05) (group 1: 63.96 ± 5.85, group 2: 66.84 ± 11.67, group 3: 59.28 ± 15.50). CONCLUSION: The bone regeneration observed in the extraction sockets filled with autogenous cancellous bone or bioactive glass was similar to that observed in the control sites, given an observation period of four weeks after extraction of the maxillary canine tooth.

Filling of extraction sockets with autogenous bone in cats / Preenchimento de alvéolos dentais de gatos com osso autógeno

PURPOSE: To evaluate bone healing in the extraction socket of the feline mandibular canine tooth after grafting. METHODS: Eighteen adult cats were submitted to unilateral extraction of mandibular canine tooth and divided into three groups. In group 1 (n=6), control, the extraction socket was left empty. In group 2 (n=6), the extraction socket was filled with autogenous cancelous bone from the iliac crest and in group 3 (n=6), with cortical bone chips from the iliac crest. Cats were euthanized at 6 weeks postoperative. RESULTS: Immediate postoperative radiographs in dorsoventral view showed a radiolucent area at the extraction wound. A decreased radiolucency was observed on the radiographs taken at 6 weeks postoperative. Histological examination showed formation of woven bone within the extraction socket. The percentage of newly formed bone within the extraction socket, measured by the histometry, showed no statistically significant difference among the values of the three groups (Kruskal-Wallis'test p>0.05) (group 1: 52.54 ± 15.46, group 2: 50.51 ± 5.01, group 3: 51.85 ± 9.52). CONCLUSION: The bone regeneration observed in the extraction sockets filled with autogenous cancellous bone or autogenous cortical bone chips was similar to that observed in the control sites, given an observation period of 6 weeks after extraction of the mandibular canine tooth.

OBJETIVO: Avaliar a regeneração óssea de alvéolos dentais de gatos após enxertia. MÉTODOS: Dezoito gatos adultos, distribuídos em três grupos de 6 animais cada, foram submetidos a extração do canino mandibular direito ou esquerdo. No grupo 1, controle, o alvéolo foi deixado vazio. No grupo 2, o alvéolo foi preenchido com osso esponjoso autógeno do osso ilíaco e no grupo 3, com raspa de osso cortical do osso ilíaco. Os animais foram submetidos à eutanásia 6 semanas após a cirurgia. RESULTADOS: Nas radiografias realizadas no pós-operatório imediato na projeção ventrodorsal observou-se uma área de radiolucência correspondente ao local da alveolectomia e extração dentária. Diminuição da radiolucência foi verificada nas radiografias realizadas seis semanas após a cirurgia. Nos cortes histológicos verificou-se a presença de trabéculas ósseas. A porcentagem de tecido ósseo esponjoso presente nos alvéolos dentais foi quantificada por exames histométricos. Não houve diferença estatisticamente significante entre os grupos (Teste de Kruskal-Wallis p > 0.05) (grupo 1: 52,54 ± 15,4; grupo 2: 50,51 ± 5,01; grupo 3: 51,85 ± 9,52). CONCLUSÃO: Os alvéolos dentais de gatos preenchidos com osso esponjoso autógeno ou raspa de osso cortical autógeno apresentaram regeneração óssea similar àquela observada no grupo controle, após um período de observação de seis semanas.

Uterine evaluation and gestation diagnosis in owl monkey (Aotus azarai infulatus) using the B mode ultrasound.

BACKGROUND: Gynecological and obstetrical ultrasonography has become an indispensable tool in the routine management, health evaluation and research on captive non-human primates. METHODS: Ultrasound was used to evaluate the uterus and estimate the gestation of owl monkeys. Twelve couples were selected, where five were primiparous and seven multiparous females from the National Primate Center reproductive colony, Ananindeua-PA, Brazil. The procedures were carried out using the GE Logiq 100 MP, equipped with a 7.5 MHz linear probe. RESULTS: The females showed a simple uterus, of elongated shape, regular outline and homogeneous echogenic texture. In the uterine measurements craniocaudal diameter, dorsoventral diameter and uterine volume (UV), significant differences were identified (P < 0.05) between ultrasound examinations of primiparous and multiparous females. The UV showed a positive correlation with the number of births. The gestational sac and the embryonic echo were visible between 28 and 38 days after mating. Between 48 and 68 days after mating, embryonic death was identified in all the gestations. CONCLUSIONS: The chemical (use of tranquilizers) and husbandry factors (capture stress) may be related to the prenatal death. The establishing methods of conditioning the female to the ultrasonographic exam may offer a solution to this problem.

Uso de miniplacas na fixação de mandíbulas de gatos com defeitos segmentares preenchidos com osso autógeno / Miniplate fixation for the repair of segmental mandibular defects filled with autogenous bone in cats

PURPOSE: To evaluate the use of maxillofacial miniplate 1.5 in the repair of segmental mandibular defects filled with autogenous bone in cats. METHODS: Twelve adult cats were divided into two groups. A segmental defect of 4mm was created in one of the hemimandibles and filled with autogenous iliac crest bone graft. The operated hemimandible was fixed with a 1.5mm titanium miniplate. In group 1 (n=6), the defect was performed in the body of the mandible, behind the 1st molar. In group 2 (n=6), the defect was performed between the 4nd premolar and 1st molar, with extraction of the 1st molar. Oral alimentation was reinitiated 24 hours after surgery. Cats were euthanized at 20 weeks postoperative. RESULTS: Incorporation of the graft was suggested by the radiographs taken 20 weeks after surgery. Macroscopic examination confirmed alignment and bone union of operated hemimandibles. Histological examination showed formation of woven bone in rostral and caudal mandible/graft interfaces. The percentage of bone tissue at these areas was measured by the histometry. There was no statistically significant difference between the values of group 1(64.48 ± 4.51) and group 2 (71.69 ± 14.47) (Mann-Whitney's test p= 0.294). CONCLUSION: The use of miniplate 1.5 for the fixation of mandibular defects filled with autogenous bone in cats provided the main goals in the treatment of mandibular fractures: bone union, normal dental occlusion and immediate return to oral alimentation.

OBJETIVO: Avaliar o uso de miniplacas na fixação de mandíbulas de gatos com defeitos segmentares preenchidos com osso autógeno. MÉTODOS: Foram utilizados 12 gatos adultos divididos em dois grupos de seis animais cada. Em uma das hemimandíbulas foi criado um defeito segmentar de 4mm, preenchido com enxerto autógeno do osso ilíaco. A hemimandíbula operada foi fixada com uma miniplaca 1.5 de titânio. No grupo 1, o defeito foi realizado caudalmente ao 1º molar e no grupo 2, entre o 4º pré-molar e 1º molar, com extração do 1º molar. A alimentação oral foi reiniciada 24 horas após a cirurgia. Os animais foram submetidos à eutanásia 20 semanas após a cirurgia. RESULTADOS: As radiografias realizadas 20 semanas após a cirurgia sugeriram a incorporação do enxerto. O exame macroscópico confirmou redução cirúrgica satisfatória e união óssea das hemimandibulas operadas. Os exames histológicos demonstraram a presença de tecido ósseo esponjoso nos locais de transição do enxerto com o osso da mandíbula. Os exames histométricos demonstraram que não houve diferença estatisticamente significante nos valores observados de porcentagem de tecido ósseo nas áreas de incorporação do enxerto das hemimandíbulas operadas dos gatos do grupo 1 (64,48 ± 4,51) e grupo 2 (71,69 ± 14,47) (Teste de Mann-Whitney p=0,294). CONCLUSÃO: Os resultados deste experimento permitiram concluir que o uso de miniplacas na fixação de mandíbulas de gatos com defeitos segmentares preenchidos com osso autógeno garante os principais objetivos no tratamento de fraturas da mandíbula: união óssea, oclusão dentária normal e alimentação oral imediata.

Miniplate 1.5 fixation for the repair of mandibular osteotomies in cats / Fixação interna de osteotomias mandibulares com miniplacas 1.5 em gatos

Purpose: To evaluate the application of the maxillofacial miniplate 1.5 in the repair of unilateral mandibular osteotomies in cats. Methods: Twelve adult cats were divided into two groups. In group 1 (n=6), the osteotomy was performed in the body of the mandible, behind the 1st molar. In group 2 (n=6), the osteotomy was performed between the 4th premolar and 1st molar. The osteotomy was fixed with a titanium miniplate 1.5. Oral alimentation was reinitiated 24 hours after surgery. Cats were euthanized at 12 weeks postoperative. Results: Radiographs taken 1 week after surgery showed a radiolucent line. The osteotomy line was not more visible on the radiographs taken at 12 weeks postoperative. Macroscopic examination confirmed alignment and bone union of operated hemimandibles. Histological examination showed formation of woven bone within the osteotomy line. The percentage of bone tissue at these areas was measured by the histometry. There was no statistically significant difference between the values of group 1(75.07 ± 5.99) and group 2 (74.76 ± 8.54) (Mann-Whitney's test p= 0.469). Conclusion: We concluded that the use of miniplate 1.5 for the fixation of mandibular osteotomy in cats provided the main goals in the treatment of mandibular fractures: bone union, normal dental occlusion and immediate return to oral alimentation.

Objetivo: Avaliar a fixação de osteotomias mandibulares unilaterais com miniplacas 1.5 em gatos. Métodos: Foram utilizados 12 gatos adultos divididos em dois grupos de seis animais cada. No grupo 1, a osteotomia foi realizada caudalmente ao 1º molar e no grupo 2, entre o 4º pré-molar e 1º molar. A osteotomia foi fixada com uma miniplaca 1.5 de titânio. A alimentação oral foi reiniciada 24 horas após a cirurgia. Os animais foram submetidos a eutanásia 12 semanas após a cirurgia. Resultados: A osteotomia pode ser notada como uma linha de radiolucência nas radiografias realizadas uma semana após a cirurgia. Essa linha de radiolucência não foi mais vista nas radiografias realizadas 12 semanas após a cirurgia. No exame macroscópico a hemimandíbula operada apresentava conformação semelhante à hemimandíbula contralateral. A união óssea pode ser confirmada por meio dos exames histológicos que demonstraram a presença de tecido ósseo esponjoso no local da osteotomia. Os exames histométricos demonstraram que não houve diferença estatisticamente significante nos valores observados de porcentagem de tecido ósseo na área de osteotomia das hemimandíbulas operadas dos gatos do grupo 1 (75,07 ± 5.99) e grupo 2 (74,76 ±8,56) (Teste de Mann-Whitney p=0,469). Conclusão: Os resultados deste experimento permitiram concluir que a fixação de osteotomias com miniplacas 1.5 em gatos garante os principais objetivos no tratamento de fraturas da mandíbula: união óssea, oclusão dentária normal e alimentação oral imediata.

Histologia e morfometria em cornos uterinos de cadelas nulíparas, multíparas e tratadas com contraceptivos / Histology and morphometry in uterus horns of nulliparous bitches, multiparous ones and bitches treated with contraceptives

Dados histológicos e morfométricos foram obtidos de útero de cadelas nulíparas (n=6), multíparas (n=6) e de cadelas tratadas com contraceptivo (n=6). Para esse fim foram usadas seis amostras de cornos uterinos, em corte médio, para cada grupo. As mensurações das espessuras da parede uterina, endométrio total, miométrio total, miométrio interno, miométrio externo, estrato vascular e diâmetro das glândulas endometriais normais não foram estatisticamente significantes entre as cadelas multíparas e tratadas, com exceção para a altura do epitélio de glândulas normais. As mensurações das espessuras da parede uterina, endométrio total, miométrio total, miométrio interno, diâmetro das glândulas normais e altura do epitélio glandular foram significantes, comparando cadelas nulíparas com as multíparas e/ou tratadas, com exceção para as espessuras do miométrio externo e estrato vascular. Diante dos resultados obtidos, pode-se concluir que os dados numéricos confirmam que (1) o uso de anticoncepcionais e sucessivas gestações afetam a estrutura uterina em seu total; (2) o miométrio externo e estrato vascular foram as regiões que menos sofreram alterações comparando-se os 3 grupos; (3) as variações morfológicas ocorreram com a mesma intensidade no endométrio e miométrio totais para as cadelas tratadas e multíparas, e (4) a presença de glândulas endometriais dilatadas foi o diferencial encontrado nas cadelas tratadas, pois as mesmas estavam ausentes nas multíparas e nulíparas.

Histological and morphometric data were obtained from uterus of nulliparous bitches (n=6), multiparous ones (n=6), and bitches treated with contraceptive (n=6). Six samples of uterine horns, cut in average for each group. Measurements of the thickness of the uterine wall, total endometrium, total myometrium, internal myometrium, external myometrium, vascular layer and diameter of normal endometrial glands were not statistically significant between the treated and multiparous bitches, except for the height of the epithelium of normal glands. Measurements of thickness of the uterine wall, total endometrium, total myometrium, myometrium internal diameter and height of the glands of normal glandular epithelium were significant, comparing nulliparous with multiparous bitches and/or treated, except for thickness of the myometrium and vascular outer layer. It can be concluded that (1) the use of contraceptives and successive pregnancies affected uterine structures in its full, (2) the vascular layer and external myometrium were the regions with less variation, (3) morphological changes occurred with the same intensity in the total endometrium and myometrium of treated and multiparous bitches; and (4) the presence of dilated endometrial glands was the difference found in the treated bitches, because they were absent in nulliparous and multiparous ones.

Testosterone concentration in a bovine Bos indicus with bilateral varicocele: case report

A bovine, Bos indicus, with bilateral varicocele diagnosticed by palpation and ultrasound observation was observed during 24 months measuring scrotal circumference, semen quality, testosterone concentrations comparing with seasonal variation and other animals of the same species. The abnormal morphology of major defects and minor defects didn't change between the bull and the other animals, however, the total defects were higher during the summer in the bull with varicocele (49,9 percent±6,9) when compared with the other bulls (27,9 percent±2,9). The animal showed higher percentage of major defects during the summer, comparing with the other seasons of the year. For the animal with varicocele testosterone levels were significantly higher during the different season's of the year, whereas decreasing levels in the summer were seen in all Bos indicus. The clinical diagnostic was confirmed by necropsy. This pathology, characterized by a bilateral varicocele with thrombosis of the spermatic cord vessels, showed that the thermoregulation suffered establishing severe testicular degeneration. As seric testosterone increased suggesting lack of steroid retention at the testicle by the pampiniform plexus, the sperm production was abnormal