ABSTRACT
Norovirus (NoV) is a member of the Caliciviridae family and is considered an emerging human enteric pathogen. NoVs are detected in farm animals such as cattle, sheep and pigs. Porcine NoV (PoNoV) is widespread worldwide, but frequency of infection is often low. This study aimed to investigate the natural PoNoV infection from adult animals of an important Brazilian pig-production region. Faecal samples (n = 112) of asymptomatic pigs aged 9 to 24 weeks old were collected from 16 grower-to-finish herds located in Paraná state, Brazilian Southern region, and evaluated for PoNoV presence. A reverse transcription-polymerase chain reaction (RT-PCR) assay was performed using specific primers that target a conserved region of the virus capsid gene (VP1). PoNoV was detected in 58 (51.8%) of the 112 faecal samples and in 14 (87.5%) of the 16 herds evaluated. Six of the obtained amplicons were submitted to phylogenetic genotyping analysis. The higher nucleotide (86.5-97.4%) and amino acid (100%) similarities of the sequences in this study were with the representative strains of the porcine NoV genogroup II genotype 11 (PoNoV GII-11). These results reveal that PoNoV infection is endemic in one of the most important pork production areas of Brazil and that the PoNoV GII-11 is prevalent in this region.
Subject(s)
Caliciviridae Infections/veterinary , Norovirus/classification , Sus scrofa/virology , Swine Diseases/epidemiology , Animals , Brazil/epidemiology , Caliciviridae Infections/epidemiology , Caliciviridae Infections/virology , Capsid Proteins/genetics , DNA Primers , Genotype , Meat , Molecular Epidemiology , Phylogeny , RNA, Viral/genetics , Seasons , Swine , Swine Diseases/virologyABSTRACT
OBJETIVO: Estimar a frequência de história pregressa de chiado, descrevendo as características clínicas e formas de tratamento utilizadas. MÉTODOS: Coorte retrospectiva incluindo lactentes (6-23 meses de vida) com sintomas de infecção de vias aéreas superiores e relato de episódio prévio de chiado. Os dados foram registrados em questionário anteriormente validado. RESULTADOS: Dos 451 avaliados, 164 (36,4%; IC95% = 31,9-41,0) apresentaram história pregressa de chiado, 148 (32,8%; IC95% = 28,5-37,4) no primeiro ano de vida. A média de idade no primeiro episódio de chiado foi de 5,3±3,9 meses. Para quem começou a chiar no primeiro ano de vida, 38,5% referiram entre três e seis episódios e 14,2% > seis episódios. A média da idade no primeiro episódio foi menor para os que apresentaram > três episódios em comparação aos que apresentaram até dois episódios (3,2±2,7 versus 5,7±2,5 meses, p < 0,001). CONCLUSÃO: Um terço dos lactentes apresentou chiado no primeiro ano de vida. Quanto mais cedo ocorre o primeiro episódio, mais frequente é a recorrência do chiado.
OBJECTIVE: To estimate the frequency and describe the clinical characteristics and respective treatments of previous history of wheezing. METHODS: Infants aged 6-23 months with upper respiratory tract complaints and reporting previous wheezing were followed-up retrospectively. Data were registered on a validated standardized form. RESULTS: Out of 451 infants, 164 (36.4%; 95%CI: 31.9-41.0) had a report of prior history of wheezing, 148 (32.8%; 95%CI: 28.5-37.4) during the first year of life. The mean age at the first episode of wheezing was 5.3±3.9 months. Among those who had had their first episode before 12 months of age, 38.5% reported 3 to 6 episodes and 14.2% > 6 episodes. Mean age at first episode was lower for those with > 3 episodes in comparison with those with < 2 episodes (3.2±2.7 vs. 5.7±2.5 months, p < 0.001). CONCLUSION: One third of the infants reported wheezing during the first year of life. The earlier the first episode occurs, the more frequently wheezing recurs.
Subject(s)
Female , Humans , Infant , Bronchodilator Agents/therapeutic use , Respiratory Sounds/drug effects , Respiratory Sounds/etiology , Asthma/complications , Brazil/epidemiology , Prevalence , Recurrence , Retrospective Studies , Severity of Illness IndexABSTRACT
OBJECTIVE: To estimate the frequency and describe the clinical characteristics and respective treatments of previous history of wheezing. METHODS: Infants aged 6-23 months with upper respiratory tract complaints and reporting previous wheezing were followed-up retrospectively. Data were registered on a validated standardized form. RESULTS: Out of 451 infants, 164 (36.4%; 95%CI:31.9-41.0) had a report of prior history of wheezing, 148 (32.8%; 95%CI:28.5-37.4) during the first year of life. The mean age at the first episode of wheezing was 5.3±3.9 months. Among those who had had their first episode before 12 months of age, 38.5% reported 3 to 6 episodes and 14.2% > 6 episodes. Mean age at first episode was lower for those with ≥ 3 episodes in comparison with those with ≤ 2 episodes (3.2±2.7 vs. 5.7±2.5 months, p < 0.001). CONCLUSION: One third of the infants reported wheezing during the first year of life. The earlier the first episode occurs, the more frequently wheezing recurs.
Subject(s)
Bronchodilator Agents/therapeutic use , Respiratory Sounds/drug effects , Respiratory Sounds/etiology , Asthma/complications , Brazil/epidemiology , Female , Humans , Infant , Male , Prevalence , Recurrence , Retrospective Studies , Severity of Illness IndexABSTRACT
We report on the presence and formation of cholesterol oxidation products (oxysterols) in bovine sperm. Although cholesterol is the most abundant molecule in the membrane of mammalian cells and is easily oxidized, this is the first report on cholesterol oxidation in sperm membranes as investigated by state-of-the-art liquid chromatographic and mass spectrometric methods. First, oxysterols are already present in fresh semen samples, showing that lipid peroxidation is part of normal sperm physiology. After chromatographic separation (by high-performance liquid chromatography), the detected oxysterol species were identified with atmospheric pressure chemical ionization mass spectrometry in multiple-reaction-monitoring mode that enabled detection in a broad and linear concentration range (0.05-100 pmol for each oxysterol species detected). Second, exposure of living sperm cells to oxidative stress does not result in the same level and composition of oxysterol species compared with oxidative stress imposed on reconstituted vesicles from protein-free sperm lipid extracts. This suggests that living sperm cells protect themselves against elevated oxysterol formation. Third, sperm capacitation induces the formation of oxysterols, and these formed oxysterols are almost completely depleted from the sperm surface by albumin. Fourth, and most importantly, capacitation after freezing/thawing of sperm fails to induce both the formation of oxysterols and the subsequent albumin-dependent depletion of oxysterols from the sperm surface. The possible physiological relevance of capacitation-dependent oxysterol formation and depletion at the sperm surface as well as the omission of this after freezing/thawing semen is discussed.
Subject(s)
Cholesterol/chemistry , Spermatozoa/chemistry , Animals , Cattle , Male , Mass Spectrometry , Oxidation-ReductionABSTRACT
The present study describes the effects of exposure of bovine sperm to mild and more intense ROS generating conditions. The membrane integrity of the incubated sperm was assessed and the incubated sperm were used for IVF after which the percentages of cleavage and blastocyst formation were determined for a period up to 9 days. The incubated sperm samples showed increased levels of molecular oxidation in the plasma membrane, the mitochondria, the cytosol and to a lesser extent in the sperm's DNA. The sperm membrane integrity as well as the first cleavage rates obtained with sperm from mild ROS generating conditions (100 microM H2O2) were not different from sperm incubated without pro-oxidants. However, exposure of sperm to more severe oxidative stress (500 mM H2O2 or a combination of 100 microM ascorbic acid, 20 microM FeSO4 and 500 microM H2O2) led to plasma membrane oxidation, reduced percentages of cleaved embryos and a reduction in the percentages of cleaved embryos that developed to the blastocyst stage. From these results, we conclude that the impact of oxidative stress to sperm becomes primarily manifest after the first cleavage of the formed zygote. Importantly, the level of lipid peroxidation in the sperm plasma membrane significantly correlates with blastocyst formation when the corresponding sperm is used for in vitro fertilization of oocytes.
Subject(s)
Cattle/physiology , Embryo, Mammalian/drug effects , Ferrous Compounds/pharmacology , Hydrogen Peroxide/pharmacology , Oxidants/pharmacology , Spermatozoa/drug effects , Animals , Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Cell Membrane/drug effects , DNA/drug effects , Lipid Peroxidation/physiology , Male , Mitochondria/drug effects , Oxidation-Reduction/drug effects , Statistics as Topic , Zygote/drug effects , Zygote/growth & developmentABSTRACT
Reactive oxygen species have been implicated in sperm aberrations causing multiple pathologies including sub- and infertility. Freeze/thawing of sperm samples is routinely performed in the cattle breeding industries for semen storage prior to artificial insemination but unusual in porcine breeding industries as semen dilution and storage at 17 degrees C is sufficient for artificial insemination within 2-3 days. However, longer semen storage requires cryopreservation of boar semen. Freeze/thawing procedures induce sperm damage and induce reactive oxygen species in mammalian sperm and boar sperm seems to be more vulnerable for this than bull sperm. We developed a new method to detect reactive oxygen species induced damage at the level of the sperm plasma membrane in bull sperm. Lipid peroxidation in freshly stored and frozen/thawed sperm cells was assessed by mass spectrometric analysis of the main endogenous lipid classes, phosphatidylcholine and cholesterol and by fluorescence techniques using the lipid peroxidation reporter probe C11-BODIPY(581/591). Peroxidation as reported by the fluorescent probe, clearly corresponded with the presence of hydroxy- and hydroperoxyphosphatidylcholine in the sperm membranes, which are early stage products of lipid peroxidation. This allowed us, for the first time, to correlate endogenous lipid peroxidation with localization of this process in the living sperm cells. Cytoplasmatic droplets in incompletely matured sperm cells were intensely peroxidized. Furthermore, lipid peroxidation was particularly strong in the mid-piece and tail of frozen/thawed spermatozoa and significantly less intense in the sperm head. Induction of peroxidation in fresh sperm cells with the lipid soluble reactive oxygen species tert-butylhydroperoxide gave an even more pronounced effect, demonstrating antioxidant activity in the head of fresh sperm cells. Furthermore, we were able to show using the flow cytometer that spontaneous peroxidation was not a result of cell death, as only a pronounced subpopulation of living cells showed peroxidation after freeze-thawing. Although the method was established on bovine sperm, we discuss the importance of these assays for detecting lipid peroxidation in boar sperm cells.
Subject(s)
Cryopreservation/veterinary , Lipid Peroxidation , Semen Preservation/veterinary , Spermatozoa/ultrastructure , Swine , Animals , Cell Membrane/chemistry , Cell Membrane/drug effects , Cholesterol/analysis , Cholesterol/chemistry , Cryopreservation/methods , Fluorescent Dyes , Hot Temperature , Lipid Peroxides/analysis , Male , Membrane Lipids/analysis , Membrane Lipids/chemistry , Microscopy, Confocal , Phospholipids/analysis , Phospholipids/chemistry , Reactive Oxygen Species/pharmacology , Semen Preservation/methods , Spermatozoa/chemistryABSTRACT
Objetivo: verificar a prevalência da exposição pré-natal à cocaina em uma amostra de recém-nascidos utilizando dois métodos: a fluorescência polarizada por imunoesaio no mecônio e a entrevista materna. Método: estudo transversal realizado num hospital geral universitário abrangendo todos os recém-nascidos nascidos vivos no centro obstétrico do hospital durante o pereíodo de 23 de março de 199 a 1 de junho de 1999 (847 recém-nascidos). A presença de exposição pré-natal a cocaína foi definida por um teste de fluorescência polarizada por imunoensaio positivo para benzoilecgonina no mecônio do recém-nascido e/ou por uma entrevista materna positiva. Resultados: a taxa da exposição pré-natal à cocaína foi de 16 casos (2,4 por cento), e de 25 casos (3,4 por cento) através da testagem do mecônio. Foram encontrados 34 casos, com uma prevalência de 4,6 por cento, quando os métodos para a detecção foram considerados de forma complementar. Conclusões: através desta estudo, foi observado que a testagem meconial é mais eficaz que a entrevista materna no diagnóstico da exposição pré-natal à cocaína. A entrevista aumentou, em relação à testagem do mecônio, em 26 por cento a possibilidade do diagnóstico da exposição; e a testagem do mecônio, em relação à entrevista, aumentou em 53,4 por cento o diagnóstico da exposição
