RESUMO
The aim of this study was to assess in vitro sperm characteristics and pregnancies/AI (P/AI) of conventional and sex-sorted semen at timed-AI of suckled, multiparous Nelore cows. All cows (n=348) were submitted to a traditional estradiol/progesterone (P4)-based protocol. At 48h after P4-device removal, the estrous behavior was recorded, and AI was performed with conventional or sex-sorted semen from two bulls. The following sperm assessments were performed: CASA, Hyposmotic Test, sperm morphometry and chromatin structure by TB staining. P/AI were reduced (P<0.001) for sex-sorted compared to conventional semen in cows expressing estrus (27vs47%) or not (11vs.37%). Membrane integrity (Bull1: 30.3±9.6 vs. 52.3±12.4%, P=0.01; Bull2: 24.5±3.0 vs. 48.7±1.6%, P=0.006) and sperm concentration (Bulll: 23.2±0.6 vs. 43.0±0.8x10 sperm/mL, P<0.001; Bull2: 25.1+2.8 VS. 42.1±0.7x10 sperm/mL; P<0.001) were reduced in sex-sorted compared to conventional semen, for both bulls. Total and progressive motility were reduced in sex-sorted semen for Bull1 (TM: 49.7±15.9 vs. 94.9±1.9%, P=0.007; PM: 16.7±3.4 vs. 44.1±13.2%, P=0.009) and no differences were detected for Bull2 (TM: 45.0±17.5 vs. 68.2±19.1%, P=0.098; PM: 12.8±4.7 vs. 30.0±13.0%, P=0.065). Sperm ellipticity from sex-sorted was lower than conventional semen for Bull2 (0.306±0.01 vs. 0.342±0.02, P=0.02) and no difference was detected for Bull1 (0.332±0.01 vs. 0.330±0.01, P=0.55). Reduced in vivo fertility was observed for sex-sorted semen, regardless of estrous behavior. In vitro sperm quality of sex-sorted semen was compromised for both bulls, but differently affected for each sire.
O estudo teve como objetivo avaliar características espermáticas in vitro e a taxa de concepção (TC) de sêmen convencional e sexado em um programa de IATF tradicional de vacas Nelore pós-parto. Todas as vacas (n=348) foram submetidas ao mesmo protocolo de IATF à base de estradiol e de progesterona. Após 48 horas da retirada do implante, foi determinada a expressão de estro dos animais e a IA foi realizada com sêmen convencional e sexado de dois touros Angus. As seguintes características espermáticas foram avaliadas: análise computadorizada do sêmen, teste hiposmótico, morfometria espermática e estrutura cromatinica por meio da coloração com azul de toluidina. A TC foi menor (P<0,001) para sêmen sexado comparado ao convencional, em vacas que expressaram estro (27 vs. 47%) e que não apresentaram estro (11 vs. 37%). A integridade da membrana plasmática (Touro 1: 30,319,6 vs. 52,3+12,4%, P=0,010; Touro 2: 24,5+3,0 vs. 48,7±1,6%, P=0,006) e a concentração espermática (Touro 1: 23,2±0,6 vs. 43,0±0,8x10 sperm/mL, P<0,001; Touro 2: 25,1+2,8 vs. 42,1 +0,7x10'sperm/mL, P<0,001) foram menores no sêmen sexado comparado ao convencional, para ambos os touros. Motilidades total e progressiva foram menores no sêmen sexado comparado ao convencional para o Touro 1 (MT: 49,7±15,9 vs. 94,9±1,9%, P=0,007; MP: 16,7+3,4 vs. 44,1+13,2%, P=0,009), enquanto diferenças não foram detectadas no Touro 2 (MT: 45,0±17,5 vs. 68,2±19,1%, P=0,098; MP: 12,8±4,7 vs. 30,0±13,0%, P=0,065). Elipticidade espermática do sêmen sexado foi menor do que do sêmen convencional no Touro 2 (0,306±0,01 vs. 0,342±0,02, P=0,020), mas não houve diferença no Touro 1 (0,332±0,01 vs. 0,330±0,01, P=0,552). Reduzida fertilidade in vivo foi observada para o sêmen sexado em relação ao convencional, independentemente da expressão de cio das vacas. A qualidade seminal in vitro do sêmen sexado foi comprometida para ambos os touros, mas diferentemente afetada para cada reprodutor.
Assuntos
Animais , Gravidez , Bovinos , Prenhez/fisiologia , Inseminação Artificial/veterinária , Taxa de Gravidez , Análise do Sêmen/veterinária , Espermatozoides , EstroRESUMO
This study aimed to evaluate the ultrastructural morphometry of bovine embryos produced in vitro grown at different concentrations of antioxidants. After in vitro maturation and fertilization, the presumptive zygotes were assigned into five treatments. T1) without the addition of any antioxidants (negative control); T2) addition of 50µM/mL cysteamine; and T3, T4 and T5) adding 2.5µg/mL, 5.0µg/mL or 10.0µg/mL of the antioxidants derived from the oily extract from Lippia origanoides, respectively. On D7 of culture, the embryos in the blastocyst stage were fixed and prepared for electron transmission microscopy. These were evaluated for the proportion of cytoplasm-to-nucleus, cytoplasm-to-mitochondria, cytoplasm-to-vacuoles, cytoplasm-to-autophagic vacuoles and cytoplasm-to-lipid droplets. Blastocysts cultured in media containing oily extract of Lippia origanoides presented morphological characteristics such as high cell:mitochondria ratio and low cell:vacuoles and cell:autophagic vacuole ratio, possibly been morphological indicators of embryonic quality. Inner cell mass (ICM) from blastocysts cultured in media without any antioxidants had the highest cell:vacuole ratio. Similar results were found in the trophectoderm (TE) cells of blastocysts from treatment 2. Embryo culture media supplemented with antioxidants derived from Lippia origanoides oil produced embryos with a higher cytoplasmic proportion of organelles, such as mitochondria. Also, treatments without any antioxidants or with the addition of cysteamine presented cytoplasmic vacuolization, a characteristic related to production of poor-quality embryos.(AU)
Este estudo teve como objetivo avaliar a morfometria ultraestrutural de embriões bovinos produzidos in vitro e cultivados em diferentes concentrações de antioxidantes. Após a maturação e a fertilização in vitro, os possíveis zigotos foram divididos em cinco tratamentos: T1) sem adição de antioxidantes (controle negativo); T2) adição de 50µM/mL de cisteamina; e T3, T4 e T5) adição de 2,5µg/mL, 5,0µg/mL ou 10,0µg/mL dos antioxidantes derivados do extrato oleoso de Lippia origanoides, respectivamente. No D7 de cultivo, os embriões em estágio de blastocisto foram fixados e preparados para microscopia eletrônica de transmissão. Estes foram avaliados para a proporção entre citoplasma e núcleo, citoplasma e mitocôndria, citoplasma e vacúolos, citoplasma e vacúolos autofágicos e citoplasma e gotículas lipídicas. Blastocistos cultivados em meio contendo extrato oleoso de Lippia origanoides apresentaram características morfológicas como alta relação célula:mitocôndria e baixa relação célula:vacúolos e célula:vacúolo autofágico, possíveis indicadores morfológicos de qualidade embrionária. A massa celular interna (MCI) de blastocistos cultivados em meio sem quaisquer antioxidantes teve a maior razão célula:vacúolo. Resultados semelhantes foram encontrados nas células do trofectoderma (TE) de blastocistos do tratamento 2. Portanto, o meio de cultivo embrionário suplementado com antioxidantes derivados do óleo de Lippia origanoides produziu embriões com maior proporção citoplasmática de organelas, como mitocôndrias. Além disso, tratamentos sem antioxidantes ou com adição de cisteamina apresentaram vacuolização citoplasmática, característica relacionada à produção de embriões de baixa qualidade.(AU)
Assuntos
Blastocisto , Cisteamina , Lippia , Embrião de Mamíferos/ultraestrutura , Técnicas In Vitro/veterinária , AntioxidantesRESUMO
Placentas from pregnant cows with different gestation periods were used. Placental fragments of all groups were processed and evaluated by transmission electron microscopy. After fragment analysis, bovine placenta was observed to be epitheliochorial type in early pregnancy, becoming progressively sinepiteliocorial at the beginning of the second trimester. There are no ultrastructural evidences of inflammation in the region of caruncles throughout gestation, despite the invasion of caruncle proper lamina by trophoblast cells. However, throughout pregnancy and especially at the end, there were evident signs of cell degeneration in both trophoblast and the uterine epithelium. The active trophoblast cells intensely phagocytize cellular debris. There are complex interdigitations between the surface of the trophoblast and the uterine epithelium, which is related to the increase of the exchange surface between mother and fetus. At the end of pregnancy, interdigitations disappear, favoring the detachment and expulsion of the placenta after birth.(AU)
Foram utilizadas placentas de vacas abatidas em frigorífico com diversos tempos gestacionais. Fragmentos de placentomo de todos os grupos foram processados e avaliados em microscopia eletrônica de transmissão. Após análise dos fragmentos, observou-se que a placenta bovina é do tipo epiteliocorial no início da gestação, tornando-se sinepiteliocorial progressivamente a partir do início do segundo mês de gestação. Não existem evidências ultraestruturais de inflamação na região das carúnculas durante toda a gestação, apesar da invasão da lâmina própria caruncular por células trofoblásticas. No entanto, durante toda a gestação e em especial ao seu final, foram observados sinais evidentes de degeneração celular, tanto do trofoblasto como do epitélio uterino. As células trofoblásticas ativas fagocitam intensamente os debris celulares originados dessas degenerações. Existem complexas interdigitações entre a superfície do trofoblasto e do epitélio uterino, o que estaria relacionado com o aumento da superfície de troca entre mãe e feto. Ao final da gestação, praticamente desaparecem essas interdigitações, favorecendo o descolamento e a expulsão da placenta após o parto.(AU)
Assuntos
Animais , Feminino , Gravidez , Bovinos , Troca Materno-Fetal/fisiologia , Placenta/fisiologia , Placenta/ultraestrutura , Microscopia Eletrônica de Transmissão/veterináriaRESUMO
Placentas from pregnant cows with different gestation periods were used. Placental fragments of all groups were processed and evaluated by transmission electron microscopy. After fragment analysis, bovine placenta was observed to be epitheliochorial type in early pregnancy, becoming progressively sinepiteliocorial at the beginning of the second trimester. There are no ultrastructural evidences of inflammation in the region of caruncles throughout gestation, despite the invasion of caruncle proper lamina by trophoblast cells. However, throughout pregnancy and especially at the end, there were evident signs of cell degeneration in both trophoblast and the uterine epithelium. The active trophoblast cells intensely phagocytize cellular debris. There are complex interdigitations between the surface of the trophoblast and the uterine epithelium, which is related to the increase of the exchange surface between mother and fetus. At the end of pregnancy, interdigitations disappear, favoring the detachment and expulsion of the placenta after birth.(AU)
Foram utilizadas placentas de vacas abatidas em frigorífico com diversos tempos gestacionais. Fragmentos de placentomo de todos os grupos foram processados e avaliados em microscopia eletrônica de transmissão. Após análise dos fragmentos, observou-se que a placenta bovina é do tipo epiteliocorial no início da gestação, tornando-se sinepiteliocorial progressivamente a partir do início do segundo mês de gestação. Não existem evidências ultraestruturais de inflamação na região das carúnculas durante toda a gestação, apesar da invasão da lâmina própria caruncular por células trofoblásticas. No entanto, durante toda a gestação e em especial ao seu final, foram observados sinais evidentes de degeneração celular, tanto do trofoblasto como do epitélio uterino. As células trofoblásticas ativas fagocitam intensamente os debris celulares originados dessas degenerações. Existem complexas interdigitações entre a superfície do trofoblasto e do epitélio uterino, o que estaria relacionado com o aumento da superfície de troca entre mãe e feto. Ao final da gestação, praticamente desaparecem essas interdigitações, favorecendo o descolamento e a expulsão da placenta após o parto.(AU)
Assuntos
Animais , Feminino , Gravidez , Bovinos , Troca Materno-Fetal/fisiologia , Placenta/fisiologia , Placenta/ultraestrutura , Microscopia Eletrônica de Transmissão/veterináriaRESUMO
The aim of this study was to evaluate the chromatin packing and sperm head morphometry of cryopreserved semen of Nelore bulls (Bos taurus indicus) of different ages. Furthermore, the influence of the degree of chromatin compaction on in vitro embryo production (IVP) was investigated. Forty bulls were divided into three groups: young (1.8-2 years), adult (3.5-7 years), and senile (8-14.3 years). The ejaculates were frozen according to standards established by the Artificial Insemination Center located in the Southeast of Brazil. Toluidine blue staining was used for simultaneous evaluation of the sperm chromatin and sperm head morphometry. Chromomycin A3 (CMA3) was applied to analyze sperm protamination and IVP for embryonic development. Spermatozoa of young bulls presented higher values for area (A, pixels), perimeter (P, pixels), and width (W, pixels) compared to adults and senile (young: A = 1848.5 ± 119.79, P = 10.23 ± 0.29, and W = 1.95 ± 0.1; adults: A = 1672.9 ± 104.46, P = 9.86 ± 0.33, and W = 1.81 ± 0.06; senile: A = 1723.1 ± 124.41, P = 9.97 ± 0.33, and W = 1.83 ± 0.09; P < 0.0001) and showed higher protamination deficiency when analyzed by CMA3 (young: 1.57 ± 0.76; adults: 1.09 ± 0.63, and senile: 0.90 ± 0.59; P < 0.05). Likewise, variables of sperm head size (A, P, and W) and protamination assessed by CMA3 showed negative correlation with age and positive correlation with ellipticity, evaluated by toluidine blue method (P < 0.05). Sperm head area was larger in spermatozoa presenting chromatin instabilities than spermatozoa without chromatin alteration (P < 0.0001). There was no difference in IVP when using semen with larger or smaller portions of spermatozoa with chromatin instabilities, indicating that the proportion of sperm with abnormal chromatin compaction (4%-16.15%) did not interfere with early embryonic development. From our results, it can be concluded that sperm of young Nelore bulls have larger heads compared to adults and senile due to reduced protamine content when evaluated by CMA3 and higher proportion of major sperm defects assessed by differential interference contrast microscopy.
Assuntos
Envelhecimento/fisiologia , Bovinos/fisiologia , Cromatina/fisiologia , Fertilização in vitro/veterinária , Espermatozoides/citologia , Animais , Masculino , Espermatozoides/fisiologiaRESUMO
ABSTRACT Placentas from pregnant cows with different gestation periods were used. Placental fragments of all groups were processed and evaluated by transmission electron microscopy. After fragment analysis, bovine placenta was observed to be epitheliochorial type in early pregnancy, becoming progressively sinepiteliocorial at the beginning of the second trimester. There are no ultrastructural evidences of inflammation in the region of caruncles throughout gestation, despite the invasion of caruncle proper lamina by trophoblast cells. However, throughout pregnancy and especially at the end, there were evident signs of cell degeneration in both trophoblast and the uterine epithelium. The active trophoblast cells intensely phagocytize cellular debris. There are complex interdigitations between the surface of the trophoblast and the uterine epithelium, which is related to the increase of the exchange surface between mother and fetus. At the end of pregnancy, interdigitations disappear, favoring the detachment and expulsion of the placenta after birth.
RESUMO Foram utilizadas placentas de vacas abatidas em frigorífico com diversos tempos gestacionais. Fragmentos de placentomo de todos os grupos foram processados e avaliados em microscopia eletrônica de transmissão. Após análise dos fragmentos, observou-se que a placenta bovina é do tipo epiteliocorial no início da gestação, tornando-se sinepiteliocorial progressivamente a partir do início do segundo mês de gestação. Não existem evidências ultraestruturais de inflamação na região das carúnculas durante toda a gestação, apesar da invasão da lâmina própria caruncular por células trofoblásticas. No entanto, durante toda a gestação e em especial ao seu final, foram observados sinais evidentes de degeneração celular, tanto do trofoblasto como do epitélio uterino. As células trofoblásticas ativas fagocitam intensamente os debris celulares originados dessas degenerações. Existem complexas interdigitações entre a superfície do trofoblasto e do epitélio uterino, o que estaria relacionado com o aumento da superfície de troca entre mãe e feto. Ao final da gestação, praticamente desaparecem essas interdigitações, favorecendo o descolamento e a expulsão da placenta após o parto.
RESUMO
Campylobacteriosis is a worldwide foodborne zoonosis disease caused by Campylobacter jejuni. This microorganism is considered a commensal bacterium in chicken hosts. C. jejuni produces epithelial cell modifications and induces a cytokine gene transcription innate immunity repertoire. In the present study, we describe the invasiveness, morphological cellular modifications, and transcript level expressions of innate immune cytokines from C. jejuni-inoculated chicken ileum explants. C. jejuni was internalized by epithelial ileum cells at 15 minutes postinoculation (p.i.) and was detected intracellularly for 4hs (p.i.). Inoculated explants displayed significant increases in cell height. C. jejuni induced a significant elevation of Transforming Growth Factor Beta 3 (TGF-b3) and Interleukin-1b (IL-1b) transcripts. In conclusion, C. jejuni is internalized in explanted epithelial ileum cells, produces morphological cell modifications, and induces gene transcription of both anti-inflammatory and pro-inflammatory cytokines.
Assuntos
Animais , Campylobacter jejuni/patogenicidade , Enterócitos/patologia , Galinhas/anormalidades , Infecções por Campylobacter/veterináriaRESUMO
This study aimed at proposing a new technical criteria for condemnation of turkey carcasses due to fowlpox in turkeys as a contribution for the work of the Brazilian Federal Meat Inspection Service. Skin samples from 30 carcasses of a flock of 840 turkeys (Meleagris gallopavo), previously vaccinated for fowlpox and slaughtered in June 2013, were collected. Samples were submitted to histological examination under light microscopy. The virus was identified using standard PCR techniques. The main histological findings were hyperplasia and hydropic degeneration of the epithelium and the presence of intracytoplasmic eosinophilic inclusion bodies. PCR results yielded 83.3% positive and 16.7% negative samples. Fowlpox virus is species specific, and there are no reports of its occurrence in mammals. The macroscopic and microscopic findings of the skin lesions do not justify the total condemnation of carcasses of poultry affected with fowlpox, except in cases of cachexia or repulsive appearance, as established by SIF regulation.
Assuntos
Animais , Perus/anormalidades , Perus/classificação , Varíola Aviária , Reação em Cadeia da Polimerase/veterináriaRESUMO
Campylobacteriosis is a worldwide foodborne zoonosis disease caused by Campylobacter jejuni. This microorganism is considered a commensal bacterium in chicken hosts. C. jejuni produces epithelial cell modifications and induces a cytokine gene transcription innate immunity repertoire. In the present study, we describe the invasiveness, morphological cellular modifications, and transcript level expressions of innate immune cytokines from C. jejuni-inoculated chicken ileum explants. C. jejuni was internalized by epithelial ileum cells at 15 minutes postinoculation (p.i.) and was detected intracellularly for 4hs (p.i.). Inoculated explants displayed significant increases in cell height. C. jejuni induced a significant elevation of Transforming Growth Factor Beta 3 (TGF-b3) and Interleukin-1b (IL-1b) transcripts. In conclusion, C. jejuni is internalized in explanted epithelial ileum cells, produces morphological cell modifications, and induces gene transcription of both anti-inflammatory and pro-inflammatory cytokines.(AU)
Assuntos
Animais , Infecções por Campylobacter/veterinária , Campylobacter jejuni/patogenicidade , Enterócitos/patologia , Galinhas/anormalidadesRESUMO
This study aimed at proposing a new technical criteria for condemnation of turkey carcasses due to fowlpox in turkeys as a contribution for the work of the Brazilian Federal Meat Inspection Service. Skin samples from 30 carcasses of a flock of 840 turkeys (Meleagris gallopavo), previously vaccinated for fowlpox and slaughtered in June 2013, were collected. Samples were submitted to histological examination under light microscopy. The virus was identified using standard PCR techniques. The main histological findings were hyperplasia and hydropic degeneration of the epithelium and the presence of intracytoplasmic eosinophilic inclusion bodies. PCR results yielded 83.3% positive and 16.7% negative samples. Fowlpox virus is species specific, and there are no reports of its occurrence in mammals. The macroscopic and microscopic findings of the skin lesions do not justify the total condemnation of carcasses of poultry affected with fowlpox, except in cases of cachexia or repulsive appearance, as established by SIF regulation. (AU)
Assuntos
Animais , Varíola Aviária , Perus/anormalidades , Perus/classificação , Reação em Cadeia da Polimerase/veterináriaRESUMO
Regardless of species, advances in preantral follicle culture and cryopreservation and transplant of ovarian tissue techniques are dependent on the number and density of preantral follicles in the ovary. This study tested the effect of different histological section thicknesses on number, classification, and density of equine preantral follicles. An ovarian fragment was obtained from 5- to 10-year-old mares (n = 14) after slaughter, and each fragment was submitted to three histological section thickness treatments: 3, 5, and 7 µm. The area (cm(2)) of each ovarian fragment was measured, and the sections were evaluated by light microscopy. The percentage of morphologically normal follicles (89%) was similar (P > 0.05) among primordial, transitional, and primary follicles and also among histological section thicknesses. A greater (P < 0.05) number of preantral follicles per histological section were seen in the 7-µm (8.0 ± 2.2) than that in the 3-µm (3.4 ± 0.7) treatment. Furthermore, a linear regression analysis reported that the number of preantral follicles increased (P < 0.05) when a thicker section treatment was used. However, no association (P > 0.05) between follicular density and treatment was observed. The mean number of preantral follicles per fragment (45.3 ± 18.8) and the follicular density (3.0 ± 0.5 follicles per cm(2)) were different (P < 0.05) among mares. In conclusion, this study on equine preantral follicles reported that (1) a 7-µm histological section thickness might be recommended because it allowed identification of a greater number of preantral follicles per sample, (2) a large individual variation in follicle population and density was detected regardless of histological section thickness, and (3) mares have a low number and density of preantral follicles when compared with those reported for other species.
Assuntos
Técnicas Histológicas/veterinária , Cavalos/anatomia & histologia , Folículo Ovariano/anatomia & histologia , Animais , Feminino , Cavalos/fisiologiaRESUMO
The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.
Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/crescimento & desenvolvimento , Farmacorresistência Bacteriana , Antibacterianos/farmacologia , Brasil , Campylobacter jejuni/isolamento & purificação , Genes Bacterianos , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , TemperaturaRESUMO
Ehrlichia canis is an obligate intracellular microorganism and the etiologic agent of canine monocytic ehrlichiosis. The invasion process has already been described for some bacteria in this genus, such as E. muris and E. chaffeensis, and consists of four stages: adhesion, internalisation, intracellular proliferation and intercellular spreading. However, little is known about the spreading process of E. canis. The aim of this study was to analyse the role of the actin cytoskeleton, calcium, iron and lysosomes from the host cell in the spreading of E. canis in dog macrophages in vitro. Different inhibitory drugs were used: cytochalasin D (actin polymerisation inhibitor), verapamil (calcium channel blocker) and deferoxamine (iron chelator). Our results showed a decrease in the number of bacteria in infected cells treated with all drugs when compared to controls. Lysosomes in infected cells were cytochemically labelled with acid phosphatase to allow the visualisation of phagosome-lysosome fusion and were further analysed by transmission electron microscopy. Phagosome-lysosome fusion was rarely observed in vacuoles containing viable E. canis. These data suggest that the spreading process of E. canis in vitro is dependent on cellular components analysed and lysosomal evasion.
Assuntos
Citoesqueleto de Actina/metabolismo , Cálcio/metabolismo , Ehrlichia canis/crescimento & desenvolvimento , Ferro/metabolismo , Macrófagos/microbiologia , Citoesqueleto de Actina/imunologia , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Células Cultivadas , Citocalasinas/farmacologia , Desferroxamina/farmacologia , Doenças do Cão/tratamento farmacológico , Doenças do Cão/imunologia , Doenças do Cão/microbiologia , Cães , Ehrlichia canis/efeitos dos fármacos , Ehrlichia canis/imunologia , Ehrlichiose/tratamento farmacológico , Ehrlichiose/imunologia , Ehrlichiose/veterinária , Lisossomos/metabolismo , Lisossomos/microbiologia , Lisossomos/ultraestrutura , Macrófagos/metabolismo , Macrófagos/ultraestrutura , Microscopia Eletrônica de Transmissão , Monócitos/metabolismo , Monócitos/microbiologia , Monócitos/ultraestrutura , Sideróforos/farmacologia , Verapamil/farmacologiaRESUMO
The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.
Assuntos
Humanos , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/crescimento & desenvolvimento , Farmacorresistência Bacteriana , Antibacterianos/farmacologia , Brasil , Campylobacter jejuni/isolamento & purificação , Genes Bacterianos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , TemperaturaRESUMO
The objective of this study was to characterize the C. jejuni IAL2383 strain isolated from humans in Brazil. Transcripts for the racR, dnaJ and ciaB genes were found and flaA, plda and cadF genes were present in the genome and bacteria was sensitive to most of the important antimicrobials used to treat humans. C. jejuni IAL2383 is a good experimental model to analyze the interactions with cells.(AU)
Assuntos
Humanos , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/efeitos dos fármacos , Campylobacter jejuni/crescimento & desenvolvimento , Farmacorresistência Bacteriana , Antibacterianos/farmacologia , Brasil , Campylobacter jejuni/isolamento & purificação , Genes Bacterianos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , TemperaturaRESUMO
Há algumas décadas, pensava-se que espermatozoides maduros possuíssem uma cromatina não funcional, inerte, sem a capacidade de transcrição. Acreditava-se que a matriz nuclear não existisse e que a presença de histonas no núcleo seria um erro no processo de compactação cromatínica, o que poderia interferir na fertilidade do macho. Hoje se sabe que, entre as estruturas toroidais, unidades básicas da cromatina espermática altamente compactada, há algumas poucas regiões que contêm sequências de nucleossomos, e estas geralmente estão anexadas a uma matriz nuclear proteica. Os espermatozoides possuem cromatina altamente organizada e são condutores metabolicamente funcionais do genoma masculino, carreando RNAs de diferentes tipos, os quais, tanto quanto os nucleossomos, são importantes sinalizadores epigenéticos paternos e, logo,influenciam o desenvolvimento embrionário inicial. Portanto, alterações cromatínicas podem não somente interferir no processo de fecundação, como principalmente no desenvolvimento embrionário, o que reforça a necessidade da análise da cromatina na avaliação de reprodutores machos.(AU)
Decades ago, it was thought thatspermatozoa had a chromatin nonfunctional, inert, without the abilityto transcription. It was believed in the absence of nuclear matrix and the presence of the core histones would be an error in the chromatin packaging, which could interfere with male fertility. Today we know that between thetoroidal structures, basic units of highly compacted sperm chromatin, there are a few regions containing nucleosome sequences, these usually being attached to a nuclear protein matrix. The sperm chromatin is highly organized and is metabolically functional carrier of male genome, carrying different RNA types, which, along with the nucleosomes are important paternal epigenetic signaling, and influencing early embryonic development. Therefore, chromatin alterations not only interfere with the fertilization process, but also influencethe embryonic development, which reinforces the need for chromatin analysis in the evaluation of breeding males.(AU)
Assuntos
Cromatina Sexual/genética , Espermatozoides , Matriz Nuclear/genéticaRESUMO
Há algumas décadas, pensava-se que espermatozoides maduros possuíssem uma cromatina não funcional, inerte, sem a capacidade de transcrição. Acreditava-se que a matriz nuclear não existisse e que a presença de histonas no núcleo seria um erro no processo de compactação cromatínica, o que poderia interferir na fertilidade do macho. Hoje se sabe que, entre as estruturas toroidais, unidades básicas da cromatina espermática altamente compactada, há algumas poucas regiões que contêm sequências de nucleossomos, e estas geralmente estão anexadas a uma matriz nuclear proteica. Os espermatozoides possuem cromatina altamente organizada e são condutores metabolicamente funcionais do genoma masculino, carreando RNAs de diferentes tipos, os quais, tanto quanto os nucleossomos, são importantes sinalizadores epigenéticos paternos e, logo,influenciam o desenvolvimento embrionário inicial. Portanto, alterações cromatínicas podem não somente interferir no processo de fecundação, como principalmente no desenvolvimento embrionário, o que reforça a necessidade da análise da cromatina na avaliação de reprodutores machos.
Decades ago, it was thought thatspermatozoa had a chromatin nonfunctional, inert, without the abilityto transcription. It was believed in the absence of nuclear matrix and the presence of the core histones would be an error in the chromatin packaging, which could interfere with male fertility. Today we know that between thetoroidal structures, basic units of highly compacted sperm chromatin, there are a few regions containing nucleosome sequences, these usually being attached to a nuclear protein matrix. The sperm chromatin is highly organized and is metabolically functional carrier of male genome, carrying different RNA types, which, along with the nucleosomes are important paternal epigenetic signaling, and influencing early embryonic development. Therefore, chromatin alterations not only interfere with the fertilization process, but also influencethe embryonic development, which reinforces the need for chromatin analysis in the evaluation of breeding males.
Assuntos
Cromatina Sexual/genética , Espermatozoides , Matriz Nuclear/genéticaRESUMO
To evaluate the therapeutic activity of low-power laser (InGaAlP: 670 nm/30 mW), at doses of 90 J/cm(2), on the process of acute and chronic-phase repair of bone lesions of Wistar rats. Sixty-three adult males were divided into nine groups subjected to bone injury, in order to form the following treatments: T1 (control); T2 (acute-phase); T3 (chronic-phase) which were subdivided into three subgroups (n=7), analyzed on the 9th, 17th and 28th days post-surgery, after a period of daily treatment with laser. The animals with acute-phase treatment presented a more extensive endochondral ossification process. Laser-treated animals showed significant increases in serum alkaline phosphatase levels and had an effect on biomechanical property, resulting in a gradual increase in bone stiffness. Laser therapy aided the bone consolidation process and favored the physiopathologic mechanisms involved in bone tissue repair, and its effects were more prominent when treatment started during the acute phase of the injury.
Assuntos
Osso e Ossos/cirurgia , Fraturas Ósseas/veterinária , Terapia a Laser/veterinária , Fosfatase Alcalina/sangue , Animais , Fraturas Ósseas/cirurgia , Terapia a Laser/métodos , Masculino , Osteogênese , Osteotomia/métodos , Osteotomia/veterinária , Ratos , Ratos WistarRESUMO
Foram utilizados 14 coelhos da raça Nova Zelândia, adultos, hígidos, machos e fêmeas, para avaliar os aspectos histológicos e clínicos após o implante de tubo de teflon associado a mitomicina C. Dez olhos direitos formaram o grupo I e dez olhos esquerdos formaram o grupo II avaliados por 30 e 60 dias de pós-operatório (PO), respectivamente. Os outros oito olhos formaram o grupo III observados por 48 horas. Foi constituído uma bolsa na conjuntiva bulbar e dentro dela foi aplicado mitomicina C. Um tubo de teflon foi introduzido na câmara anterior e fixado na esclera, a bolsa conjuntival foi aproximada no limbo sobre o implante. Houve diferença significativa nas médias das pressões intra-oculares (PIO) entre o pré-operatório e o pós-operatório de 30 e 60 dias. Os animais dos grupos I e II apresentaram hipotonia e câmara anterior rasa nos primeiros 15 dias de PO. Na avaliação histológica da bolsa conjuntival foi verificado descamação das células superficiais do epitélio e degeneração do tecido conjuntivo, o que indicou alterações na cicatrização mediante ação da mitomicina C. O implante de teflon associado com mitomicina C demonstrou ser um método eficaz por diminuir e manter a PIO dos 30 aos 60 dias de PO.(AU)
Fourteen healthy, adult New Zealand rabbits, males and females, were used to evaluate the histological and clinical aspects after the implant of Teflon tube associated to mytomicin C. The ten right eyes of group I and ten left eyes of group II were evaluated for 30 and 60 days of postoperative (PO), respectively. The other eight eyes of group III were observed for 48 hours. A bag was constituted in the bulbar conjunctive and inside of it mytomicin C was applied. A Teflon tube was introduced in the anterior chamber and fastened in the sclera, the conjunctive bag was approximated in the limbus over the implant. There was significant difference in the averages of the intraocular pressures (IOP) between the preoperative and the postoperative of 30 and 60 days. The animals of groups I and II presented hypotonia and shallow anterior chamber in the first 15 days of PO. In the histological evaluation of the conjunctive bag, descaling of the epithelium superficial cells and degeneration of the conjunctive tissue was verified, which indicated healing changes due to the action of mytomicin C. The implant of Teflon associated with mytomicin C demonstrated to be an effective method to decrease and to maintain the IOP from 30 to 60 days PO.(AU)
Assuntos
Animais , Masculino , Feminino , Mitomicina/análise , Mitomicina , Mitomicina/uso terapêuticoRESUMO
Ehrlichia canis é o agente etiológico de Erliquiose Monocítica Canina (EMC), uma das mais importantes doenças infecciosas de cães no Brasil. Bactérias intracelulares obrigatórias de monócitos e macrófagos, como E. canis, desenvolvem diversos mecanismos celulares para assegurar a sua evasão das defesas do hospedeiro, que envolvem distintas adaptações para sobrevivência nos compartimentos intracelulares. No entanto, estudos sobre essas estratégias ainda são escassos. Dessa forma, o presente estudo teve como objetivo investigar se a E. canis possui a capacidade de evitar a fusão de lisossomos às inclusões citoplasmáticas, o que favoreceria sua multiplicação. A fusão de lisossomos com endossomos contendo E. canis foi investigada utilizando cultivo celular de histiócitos caninos (DH82) infectados com a cepa São Paulo de E. canis, em uma momento no qual 50% das células estavam parasitadas. Para esse fim as células infectadas foram marcadas citoquimicamente para fosfatase ácida, uma enzima presente nos compartimentos fusionados a lisossomos. Após o processo de inclusão em resina, os cortes ultrafinos foram analisados e documentados fotograficamente em um microscópio eletrônico de transmissão. Foram reazaliadas comparações entre três pares de controle e grupos experimentais em relação a número de bactérias por célula, número de lisossomos marcados por célula e número de vacúolos marcados contendo E. canis por célula. A análise estatística foi realizada pelo teste de Mann-Whitney, sendo considerados significativos valores p<0,05. A marcação citoquímica da enzima lisossomal não foi observada em vacúolos contendo E. canis íntegras indicando que essas bactérias são capazes de inibir a fusão lisossomal aos endossomos, como um mecanismo de evasão imune. A evasão da fusão lisossomal por inclusões erliquiais é fundamental para a sobrevivência e reprodução do patógeno, apesar de ser baseado em diferentes estratégias de acordo para as espécies de Ehrlichia spp. A demonstração da evasão lisossomal durante a propagação de E. canis serve como base para o desenvolvimento e elaboração de novas estratégias terapêuticas.(AU)
Ehrlichia canis is the etiologic agent of canine monocytic ehrlichiosis (CME), one of the most important infectious diseases of dogs in Brazil. Obligate intracellular bacteria of monocytes and macrophages like E. canis develop various cellular mechanisms to avoid host defenses, which involve different adaptations for survival in intracellular compartments. However, studies about these strategies are scarce. Thus, the present study aimed to investigate whether E. canis has the ability to prevent the fusion of lysosomes to cytoplasmic inclusions, which would favor its multiplication. The fusion of lysosomes with endosomes containing E. canis was investigated using cell culture of canine histiocytes (DH82) infected with the São Paulo strain of E. canis when 50% of the cells were parasitized. To this end, infected cells were cytochemistrically labeled for acid phosphatase, an enzyme present in the fused lysosome compartments. After resin embedding process, the ultrathin sections were analyzed and photographically documented in a transmission electron microscope (TEM). We compared three control pairs and experimental groups in relation to the number of bacteria per cell, number lysosomes marked per cell and the number of labeled vacuoles containing E. canis per cell. Statistical analysis was performed using the Mann-Whitney test, at p<0.05 significance level. The cytochemical staining of the lysosomal enzyme was not observed in vacuoles containing intact E. canis indicating that these bacteria are able to inhibit lysosomal fusion to endosomes, as a mechanism of immune evasion. The evasion of lysosomal fusion by erlichia inclusions is critical to the survival and reproduction of the pathogen, despite being based on different strategies according to the species of Ehrlichia spp. Demonstration of lysosomal escape during the propagation of E. canis serves as a basis for the development and design of new therapeutic strategies.(AU)