Sheep naturally infected with Anaplasma ovis - evaluation of cardiac and inflammatory biomarkers

Background: Anaplasmosis, also called gall sickness or tropical bovine ehrlichiosis, is an infectious disease caused by species belonging to the genus Anaplasma in domestic and wild animals in tropical and subtropical regions. Anaplasma ovis and A. phagocytophilum are important pathogens of sheep. A. ovis is considered the most common species affecting sheep. The infection is usually subclinical and progresses with high fever, anaemia, icterus, weight loss and abortions. This study aimed to investigate changes in cardiac damage markers, oxidative stress and antioxidant status, cytokines, and acute phase proteins in sheep naturally infected with A. ovis. Materials, Methods & Results: For this purpose, a total of 40 animals, including 20 healthy sheep and 20 sheep infected with anaplasmosis, were used. A. ovis was diagnosed based on clinical findings and peripheral blood smear. Blood smears were prepared from the ear vein. The smears were stained with Giemsa and examined for the presence of Anaplasma spp. Infection was also confirmed by polymerase chain reaction (PCR) analysis. The genomic DNA was isolated from blood, and the MSP-4 gene region was amplified as A. ovis specific target gene. Twenty clinically healthy sheep of the same age group, reared under the same conditions and testing negative in the molecular assessment were used as controls. Blood samples were collected from the cephalic vein and and centrifuged to obtain serum. The serum stored at -20°C until the analysis stage. Serum samples were used for the analysis of cardiac damage markers [troponin I (cTnI), creatine kinase MB (CK-MB), lactate dehydrogenase (LDH) and aspartate transaminase (AST)], oxidative stress parameters [malondialdehyde (MDA), total antioxidant status (TAS), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)], cytokines [interleukins IL-6, IL-1ß and IL-10, tumour necrosis factor α (TNF-α), and interferon-γ (IFN-γ)] and acute phase proteins [C-reactive protein (CRP), serum amyloid A (SAA) and haptoglobin (Hp)]. cTnI and CK-MB levels were measured using a chemiluminescent immunoassay. MDA, TAS, SOD, CAT, GPx, TNF-α, IL-1ß, IL-6, IL-10, IFN-γ, SAA and Hp levels were measured by an ELISA reader. LDH, AST and CRP levels were measured in an autoanalyzer. cTnI and LDH levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The concentration of AST was decreased in infected animals. MDA, TAS, SOD, CAT and GPx levels were significantly increased in the infected animals compared to the healthy ones (P < 0.05). The levels of the inflammatory parameters such as TNF-α, IL-1ß, IL-10 and IFN-γ were significantly increased in the infected animals compared to the healthy ones (P < 0.05). Hp level were significantly increased in the infected group compared with the control group (P < 0.05). However, there was no significant change in CK-MB, SAA and CRP concentrations in the infected animals (P > 0.05). Discussion: Ovine anaplasmosis is an obligate intracellular arthropod disease that causes widespread changes in haematobiochemical, immune response and oxidative stress parameters. Cardiac damage is often overlooked in field conditions due to the lack of adequate knowledge about the pathophysiology of the disease. Our results showed that A. ovis infection leads to significant changes in cardiac biomarkers and that the parasite can cause cardiac dysfunction. This is the first report on cardiac damage markers in Anaplasma-infected sheep. Additionally, the levels of proinflammatory and oxidative stress markers that may cause functional disorders were also found to be increased. Thus, measuring markers of cardiac function, oxidative stress and inflammation can be a useful tool in the early diagnosis of ovine anaplasmosis.

Citric acid minimizes oxidative stress in Amazonian fish (Colossoma macropomum) when fed plant protein-based diets

This study aimed to evaluate the antioxidant potential of citric acid in plant protein based-diets offered to juvenile tambaqui, Colossoma macropomum. Four isonitrogenous and isoenergetic diets containing different levels of citric acid (0, 10, 20, and 30 g.kg−1) were formulated. Fish (n = 160; 27.56±1.03 g) were randomly distributed in 310-L water tanks (n = 16), with four replicates per treatment. After the experimental period (80 days), animals were anesthetized, and blood was collected for cell count. They were euthanized and dissected to remove livers for subsequent oxidative stress analyses. No significant differences were observed in the levels of thrombocytes, total leukocytes, lymphocytes, positive granular leukocytes, and immature leukocytes among the experimental treatments. However, there were fewer monocytes and neutrophils in fish fed the diet containing 30 g.kg−1 of citric acid. All citric acid diets increased glutathione concentration and reduced lipid peroxidation levels in livers. Therefore, citric acid acts as a potent antioxidant in juvenile tambaqui fed plant protein-based diets and, thus, can improve their maintenance conditions in production systems.

The effect of Hippophae rhamnoides L. extract on acrylamideinduced brain injury in rats

ABSTRACT Purpose: Reactive oxygen species (ROS), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) have been shown in the pathogenesis of acrylamide neurotoxicity. Hippophae rhamnoides L. extract (HRE) has a cytoprotective effect by stabilizing the production of ROS, IL-1β and TNF-α. The objective of the article was to investigate the effect of HRE on acrylamide-induced brain damage in rats biochemically and histopathologically. Methods: To the HRE+acrylamide only (ACR) group (n=6) of the animals, HRE was administered orally at a dose of 50 mg / kg into the stomach by gavage. The same volume of solvent (olive oil) was administered orally to the ACR (n=6) and healthy (HG) (n=6) groups. One hour after HRE administration, acrylamide was given orally at a dose of 20 mg/kg to HRE+ACR and ACR groups in the same way. This procedure was repeated once a day for 30 days. At the end of this period, brain tissues extracted from animals killed with 50 mg/kg thiopental anesthesia were examined biochemically and histopathologically. Results: It has been shown that HRE prevents the increase of malondialdehyde (MDA), myeloperoxidase (MPO), IL-1β and TNF-α with acrylamide and the decrease of total glutathione (tGSH) and glutathione reductase (GSHRd) levels in brain tissue. Conclusions: HRE may be useful in the treatment of acrylamide-induced neurotoxicity.

Effects of treatment with quercetin on the quality of cryopreserved bovine semen

Background: Semen cryopreservation is one of the most common biotechnologies in the reproduction of animals ofagricultural interest, especially bulls. However, cryopreservation can be harmful to sperm cells, with susceptibility tooxidative stress being one of the causes. The addition of antioxidants such as quercetin may inhibit and/or reduce suchdamage, reducing fertility. Quercetin can increasing sperm motility and interaction capacity between spermatozoa-oocyte,to increase cellular metabolism and reduced DNA fragmentation and oxidation following thawing. Therefore, the objective of this study was to evaluate the protective effect of quercetin on the metabolism of bovine semen following thawing.Materials, Methods & Results: Three Brahman bulls in reproduction age and previously considered fit for reproductionwere used. The semen samples were collected via the electroejaculation method, and the samples were homogenized toform pooled semen from three ejaculates, which was diluted in Tris-yolk egg-glicerol diluent medium. Quercetin was addedto diluent, to final concentrations of 0, 5, 10, 15 and 20 μg.mL-1 in each group. The samples were kept frozen in strawsof 500 μL, with concentration of 40,000,000 spermatozoid / mL for 15 days and were thawed in water at 36°C for 30 s.All the tests was performed in five replicates. The cell metabolism status was evaluated by quantification of superoxideradical production with a nitroblue tetrazolium test (NBT) and scanning spectrophotometry. By spermatic evaluation, thefollowing parameters were evaluated via the computerized system of sperm analysis (CASA): total motility (TM, %),progressive motility (PM, %), velocity curved line (VCL, mm/s), velocity straight line (VSL, mm/s), velocity averagepath (VAP, mm/s), distance curved line (DCL, mm), distance straight line (DSL, mm), distance average path (DAP, mm),amplitude of lateral head displacement (ALH, mm), beat cross frequency (BCF, Hz), wobble...

Effects of treatment with quercetin on the quality of cryopreserved bovine semen

Background: Semen cryopreservation is one of the most common biotechnologies in the reproduction of animals ofagricultural interest, especially bulls. However, cryopreservation can be harmful to sperm cells, with susceptibility tooxidative stress being one of the causes. The addition of antioxidants such as quercetin may inhibit and/or reduce suchdamage, reducing fertility. Quercetin can increasing sperm motility and interaction capacity between spermatozoa-oocyte,to increase cellular metabolism and reduced DNA fragmentation and oxidation following thawing. Therefore, the objective of this study was to evaluate the protective effect of quercetin on the metabolism of bovine semen following thawing.Materials, Methods & Results: Three Brahman bulls in reproduction age and previously considered fit for reproductionwere used. The semen samples were collected via the electroejaculation method, and the samples were homogenized toform pooled semen from three ejaculates, which was diluted in Tris-yolk egg-glicerol diluent medium. Quercetin was addedto diluent, to final concentrations of 0, 5, 10, 15 and 20 μg.mL-1 in each group. The samples were kept frozen in strawsof 500 μL, with concentration of 40,000,000 spermatozoid / mL for 15 days and were thawed in water at 36°C for 30 s.All the tests was performed in five replicates. The cell metabolism status was evaluated by quantification of superoxideradical production with a nitroblue tetrazolium test (NBT) and scanning spectrophotometry. By spermatic evaluation, thefollowing parameters were evaluated via the computerized system of sperm analysis (CASA): total motility (TM, %),progressive motility (PM, %), velocity curved line (VCL, mm/s), velocity straight line (VSL, mm/s), velocity averagepath (VAP, mm/s), distance curved line (DCL, mm), distance straight line (DSL, mm), distance average path (DAP, mm),amplitude of lateral head displacement (ALH, mm), beat cross frequency (BCF, Hz), wobble...(AU)

Splenic ischemic preconditioning attenuates oxidative stress induced by hepatic ischemia-reperfusion in rats

Purpose: To evaluate the effects of splenic ischemic preconditioning (sIPC) on oxidative stress induced by hepatic ischemia-reperfusion in rats. Methods: Fifteen male Wistar rats were equally divided into 3 groups: SHAM, IRI and sIPC. Animals from IRI group were subjected to 45 minutes of partial liver ischemia (70%). In the sIPC group, splenic artery was clamped in 2 cycles of 5 min of ischemia and 5 min of reperfusion (20 min total) prior to hepatic ischemia. SHAM group underwent the same surgical procedures as in the remaining groups, but no liver ischemia or sIPC were induced. After 1h, hepatic and splenic tissue samples were harvested for TBARS, CAT, GPx and GSH-Rd measurement. Results: sIPC treatment significantly decreased both hepatic and splenic levels of TBARS when compared to IRI group (p<0.01). Furthermore, the hepatic and splenic activities of CAT, GPx and GSH-Rd were significantly higher in sIPC group than in IRI group. Conclusion: sIPC was able to attenuate hepatic and splenic IRI-induced oxidative stress.(AU)

Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods

This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.(AU)

Este trabalho teve como objetivos determinar a ocorrência e realizar a caracterização molecular de Cryptosporidium spp. em 498 amostras fecais de canários (Serinus canaria) criados em cativeiro, utilizando três métodos de diagnóstico: análise microscópica pela coloração negativa com verde malaquita, nested PCR seguida de sequenciamento dos fragmentos amplificados e PCR duplex em tempo real específica para detecção de Cryptosporidium galli e Cryptosporidium genótipo III de aves. A positividade total para Cryptosporidium spp. (total de amostras positivas em pelo menos um método de diagnóstico) obtida pela análise microscópica, nested PCR e PCR duplex em tempo real foi de 13,3% (66/498). As taxas de positividade para Cryptosporidium spp. foram 2,0% (10/498) e 4,6% (23/498) por microscopia e nested PCR, respectivamente. O sequenciamento de 20 amostras amplificadas pela nested PCR identificou C. galli (3,0%; 15/498), Cryptosporidium genótipo I de aves (0,8%; 4/498) e Cryptosporidium avium (0,2%; 1/498). A PCR duplex em tempo real revelou positividade de 7,8% (39/498) para C. galli e 2,4% (12/498) para Cryptosporidium genótipo III de aves. A análise microscópica diferiu significativamente da nested PCR para detecção de Cryptosporidium spp. A PCR duplex em tempo real apresentou maior sensibilidade que a nested PCR/sequenciamento para detectar as espécies/genótipos gástricos de Cryptosporidium.(AU)

Transferability of heterologous microsatellite primers in Brycon gouldingi / Transferibilidade de primers microssatélites heterólogos em Brycon gouldingi

Brycon gouldingi is a species of neotropical fish of socioeconomic and environmental importance in the Tocantins-Araguaia Basin. Genetic studies on this species are still limited, making it difficult to evaluate the population structure and genetic diversity in natural and captive stocks. Here, we aimed to evaluate the transferability of heterologous microsatellite primers in B. gouldingi. A total of 30 primers for eight species were evaluated: Brycon hilarii, Brycon opalinus, Brycon cephalus, Brycon orbignyanus, Prochilodus lineatus, Prochilodus argenteus, Piaractus mesopotamicus, and Colossoma macropomum. The primers that showed the best amplification patterns were applied to 20 specimens of B. gouldingi, and their genetic parameters were assessed. Among the 30 primers, seven showed satisfactory transferability, six of which belonged to the genus Brycon: Bh13 (B. hilarii), BoM5, BoM13 (B. opalinus), Borg9, Borg13, and Borg59 (B. orbignyanus), and one belonged to P. argenteus (Par80). The primers for the other species tested showed non-specificity or monomorphism; and were therefore, excluded from the analyses. The number of alleles ranged between two (Borg13 and Borg59) and three (Bh13, BoM5, BoM13, Borg9 and Par80), with sizes varying between 103 bp (BoM5) and 430 bp (Borg9). Four primers showed evidence of null alleles (BoM13, Borg9, Borg13, and Par80), which could probably be attributed to the respective Hardy-Weinberg deviation. Thus, seven primers were validated for cross-amplification in B. gouldingi, which may be used in future studies involving this species.(AU)

Brycon gouldingi é uma espécie de peixe neotropical com importância socioeconômica e ambiental na Bacia do Tocantins-Araguaia. Estudos genéticos nessa espécie ainda são escassos, dificultando o conhecimento sobre a estrutura populacional e a diversidade genética nos estoques naturais e em cativeiro. O objetivo do presente estudo foi avaliar a transferibilidade de primers microssatélites heterólogos em B. gouldingi. Foram avaliados um total de 30 primers de oito espécies: Brycon hilarii, Brycon opalinus, Brycon cephalus, Brycon orbignyanus, Prochilodus lineatus, Prochilodus argenteus, Piaractus mesopotamicus e Colossoma macropomum. Os primers que demonstraram melhores padrões de amplificação foram aplicados em 20 espécimes de B. gouldingi para os cálculos dos parâmetros genéticos. Sete dos 30 primers apresentaram resultados satisfatórios de transferibilidade, sendo seis oriundos do gênero Brycon: Bh13 (B. hilarii), BoM5, BoM13 (B. opalinus), Borg9, Borg13 e Borg59 (B. orbignyanus), e um oriundo de P. argenteus (Par80). Os primer das outras espécies testados mostraram inespecificidade ou monomorfismo, sendo excluídos das análises. O número de alelos variou de dois (Borg13 e Borg59) a três (Bh13, BoM5, BoM13, Borg9 e Par80), com tamanhos entre 103 pb (BoM5) e 430 pb (Borg9). Quatro primers apresentaram evidências de alelos nulos (BoM13, Borg9, Borg13 e Par80), o que provavelmente inferiu sobre o desvio de Hardy-Weinberg nos mesmos. Concluindo, sete primers foram validados para a amplificação cruzada em B. gouldingi e poderão ser utilizados em futuros estudos com essa espécie.(AU)

Assessment of oxidative status in foot diseases of sheep

Background: Oxidative stress plays an important role in the etiology and/or progression of various diseases in the livestock. The aim of this study was to investigate the relationship between foot diseases and oxidant and antioxidant levels, determining catalase (CAT) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA), nitric oxide (NO) and glutathione (GSH) concentrations in sheep with foot disease such as interdigital dermatitis, footrot and interdigital pouch infection (biflex canal inflammation).Materials, Methods & Results: In the study, a total of 70 sheep aged between 1 and 3, 20 sheep with interdigital dermatitis, 20 with foot-rot, 10 with interdigital pouch inflammation and 20 clinically healthy Akkaraman sheep, were used. The blood samples collected from the V. Jugularis of the sheep were centrifuged and their serums were obtained. MDA and NO levels of the sheep with interdigital dermatitis, footrot and interdigital pouch infection were found to be significantly higher than those of animals in the control group (P < 0.001). CAT activity was determined to be significantly lower in all diseased animals than in the animals in the control group (P < 0.001). GSH-Px and GSH levels were significantly lower in the animals with interdigital dermatitis than in the control group (respectively P < 0.001, P < 0.05).Discussion: Oxidative stress regulates the metabolic activity of some organs and plays an important role in productivity in livestock. The oxidative stress is reported that caused by the increase in the production of excessive free radicals resulting from the exposure of animals to various stress factors and by the weakening of the antioxidant defense has an important role in the pathogenesis of numerous diseases such as foot diseases. In the presence of various stress factors, the level of malondialdehyde (MDA) and nitric oxide (NO) increases. MDA is determine the degree of cellular damage in tissues.[...]

Assessment of oxidative status in foot diseases of sheep

Background: Oxidative stress plays an important role in the etiology and/or progression of various diseases in the livestock. The aim of this study was to investigate the relationship between foot diseases and oxidant and antioxidant levels, determining catalase (CAT) and glutathione peroxidase (GSH-Px) activities and malondialdehyde (MDA), nitric oxide (NO) and glutathione (GSH) concentrations in sheep with foot disease such as interdigital dermatitis, footrot and interdigital pouch infection (biflex canal inflammation).Materials, Methods & Results: In the study, a total of 70 sheep aged between 1 and 3, 20 sheep with interdigital dermatitis, 20 with foot-rot, 10 with interdigital pouch inflammation and 20 clinically healthy Akkaraman sheep, were used. The blood samples collected from the V. Jugularis of the sheep were centrifuged and their serums were obtained. MDA and NO levels of the sheep with interdigital dermatitis, footrot and interdigital pouch infection were found to be significantly higher than those of animals in the control group (P < 0.001). CAT activity was determined to be significantly lower in all diseased animals than in the animals in the control group (P < 0.001). GSH-Px and GSH levels were significantly lower in the animals with interdigital dermatitis than in the control group (respectively P < 0.001, P < 0.05).Discussion: Oxidative stress regulates the metabolic activity of some organs and plays an important role in productivity in livestock. The oxidative stress is reported that caused by the increase in the production of excessive free radicals resulting from the exposure of animals to various stress factors and by the weakening of the antioxidant defense has an important role in the pathogenesis of numerous diseases such as foot diseases. In the presence of various stress factors, the level of malondialdehyde (MDA) and nitric oxide (NO) increases. MDA is determine the degree of cellular damage in tissues.[...](AU)

Estresse oxidativo em cães com doença periodontal: comparação dos biomarcadores plasmáticos e salivares / Oxidative stress in dogs with periodontal disease: comparison of plasmatic and salivary biomarkers

A doença periodontal (DP) é a enfermidade inflamatória mais comum da cavidade oral dos cães. A quantificação de biomarcadores do plasma e da saliva tem sido utilizada para avaliar o estresse oxidativo sistêmico (EOS) e local (EOL) da DP humana. Na DP canina, os mecanismos do estresse oxidativo não estão bem caracterizados e estabelecidos. O objetivo do presente estudo foi investigar a hipótese de que o EOS ocorre na DP canina e de que a saliva pode ser utilizada para avaliar o EOL. Analisou-se, também, a hipótese de que a ativação do metabolismo oxidativo dos neutrófilos contribui para EOS na DP dos cães. Para tal, foram selecionados 20 cães adultos portadores de DP, agrupados de acordo com o grau de lesão: gengivite (n=6), periodontites leve (n=8) e avançada (n=6). O grupo controle foi composto pelos mesmos 20 cães, 30 dias após o tratamento periodontal. Para avaliar o metabolismo oxidativo dos neutrófilos circulantes foi quantificada a produção de superóxido pelo teste de redução do tetrazólio nitroazul (NBT). As concentrações de oxidante total (TOC) e de espécies reativas ao ácido tiobartbitúrico (TBARS) no plasma foram quantificadas para avaliar o EOS. Para a avaliação do estresse oxidativo local, foi quantificado o TOC salivar e a concentração dos principais antioxidantes da saliva (albumina, ácido úrico e bilirrubina total). O EOS na DP foi confirmado pelo aumento da produção de superóxido dos neutrófilos circulantes, TOC e TBARS plasmático. Foi possível quantificar todos os biomarcadores na saliva de cães, porém nenhum foi capaz de expressar o EOL da DP canina. Esta é uma das primeiras evidências de que o EOS ocorre em cães com DP e que a ativação do metabolismo oxidativo dos neutrófilos pode contribuir para desequilíbrio entre antioxidantes e oxidantes. Este estudo ressalta a importância da higiene bucal dos cães para a prevenção da DP e de lesões degenerativas crônicas de diversos tecidos causadas pelo EOS.(AU)

Periodontal disease (PD) is the most common inflammatory disease of the oral cavity of dogs. Quantitation of plasma and salivary biomarkers have been used to assess the systemic oxidative stress (SOS) and local (LOS) of human PD. In canine PD, oxidative stress mechanisms are not well characterized and established. Our objective was to investigate the hypothesis that SOS occurs in dog PD and saliva can be used to evaluate the LOS. We also investigated the hypothesis that the activation of neutrophil oxidative metabolism contributes to SOS in dog SD. For this purpose, 20 adult dogs were selected PD patients, grouped according to the degree of injury: gingivitis (n=6), light periodontitis (n=8) and advanced periodontitis (n=6). The control group was composed of the same 20 dogs, 30 days after periodontal treatment. To assess oxidative metabolism of circulating neutrophils superoxide production was measured by test nitroblue tetrazolium reduction (NBT). The total oxidant concentrations (TOC) and reactive species to tiobartbitúrico acid (TBARS) in plasma were quantified to evaluate SOS. For the evaluation of local oxidative stress were quantified salivary TOC and concentration of the main antioxidant in saliva (albumin, uric acid, and total bilirubin). EOS in dogs with PD was confirmed by increased superoxide production of circulating neutrophils, TOC, and plasma TBARS. It was possible to quantify all the biomarkers in the saliva of dogs, but none was able to express the LOS canine PD. This is the first evidence that SOS occurs in dogs with PD and that activation of the oxidative metabolism of neutrophils may contribute to an imbalance between oxidants and antioxidants. This study highlights the importance of oral hygiene of dogs to prevent PD and chronic degenerative lesions of various tissues caused by SOS.(AU)

Estresse oxidativo em cães com doença periodontal: comparação dos biomarcadores plasmáticos e salivares / Oxidative stress in dogs with periodontal disease: comparison of plasmatic and salivary biomarkers

A doença periodontal (DP) é a enfermidade inflamatória mais comum da cavidade oral dos cães. A quantificação de biomarcadores do plasma e da saliva tem sido utilizada para avaliar o estresse oxidativo sistêmico (EOS) e local (EOL) da DP humana. Na DP canina, os mecanismos do estresse oxidativo não estão bem caracterizados e estabelecidos. O objetivo do presente estudo foi investigar a hipótese de que o EOS ocorre na DP canina e de que a saliva pode ser utilizada para avaliar o EOL. Analisou-se, também, a hipótese de que a ativação do metabolismo oxidativo dos neutrófilos contribui para EOS na DP dos cães. Para tal, foram selecionados 20 cães adultos portadores de DP, agrupados de acordo com o grau de lesão: gengivite (n=6), periodontites leve (n=8) e avançada (n=6). O grupo controle foi composto pelos mesmos 20 cães, 30 dias após o tratamento periodontal. Para avaliar o metabolismo oxidativo dos neutrófilos circulantes foi quantificada a produção de superóxido pelo teste de redução do tetrazólio nitroazul (NBT). As concentrações de oxidante total (TOC) e de espécies reativas ao ácido tiobartbitúrico (TBARS) no plasma foram quantificadas para avaliar o EOS. Para a avaliação do estresse oxidativo local, foi quantificado o TOC salivar e a concentração dos principais antioxidantes da saliva (albumina, ácido úrico e bilirrubina total). O EOS na DP foi confirmado pelo aumento da produção de superóxido dos neutrófilos circulantes, TOC e TBARS plasmático. Foi possível quantificar todos os biomarcadores na saliva de cães, porém nenhum foi capaz de expressar o EOL da DP canina. Esta é uma das primeiras evidências de que o EOS ocorre em cães com DP e que a ativação do metabolismo oxidativo dos neutrófilos pode contribuir para desequilíbrio entre antioxidantes e oxidantes. Este estudo ressalta a importância da higiene bucal dos cães para a prevenção da DP e de lesões degenerativas crônicas de diversos tecidos causadas pelo EOS.(AU)

Periodontal disease (PD) is the most common inflammatory disease of the oral cavity of dogs. Quantitation of plasma and salivary biomarkers have been used to assess the systemic oxidative stress (SOS) and local (LOS) of human PD. In canine PD, oxidative stress mechanisms are not well characterized and established. Our objective was to investigate the hypothesis that SOS occurs in dog PD and saliva can be used to evaluate the LOS. We also investigated the hypothesis that the activation of neutrophil oxidative metabolism contributes to SOS in dog SD. For this purpose, 20 adult dogs were selected PD patients, grouped according to the degree of injury: gingivitis (n=6), light periodontitis (n=8) and advanced periodontitis (n=6). The control group was composed of the same 20 dogs, 30 days after periodontal treatment. To assess oxidative metabolism of circulating neutrophils superoxide production was measured by test nitroblue tetrazolium reduction (NBT). The total oxidant concentrations (TOC) and reactive species to tiobartbitúrico acid (TBARS) in plasma were quantified to evaluate SOS. For the evaluation of local oxidative stress were quantified salivary TOC and concentration of the main antioxidant in saliva (albumin, uric acid, and total bilirubin). EOS in dogs with PD was confirmed by increased superoxide production of circulating neutrophils, TOC, and plasma TBARS. It was possible to quantify all the biomarkers in the saliva of dogs, but none was able to express the LOS canine PD. This is the first evidence that SOS occurs in dogs with PD and that activation of the oxidative metabolism of neutrophils may contribute to an imbalance between oxidants and antioxidants. This study highlights the importance of oral hygiene of dogs to prevent PD and chronic degenerative lesions of various tissues caused by SOS.(AU)

The genetic diversity among strawberry breeding resources based on SSRs

Cultivated strawberry (Fragaria × ananassa Duch.) is a high value horticultural crop. In this study, the genetic diversity of 160 strawberry accessions was determined using five highly polymorphic simple sequence repeat (SSR) markers. Sixty different alleles were identified, with allele frequencies in the range of 0.006 to1. Similarity scores were in the range of 0.034 to 0.963 (average: 0.507). The accessions were categorized into five groups. Group 1 contained two diploid Fragaria vesca species and one unknown accession. Group 2 contained one accession (F x ananassa). Group 3 contained 20 F × ananassa accessions and six unknown accessions. Group 4 contained 48 F. × ananassa accessions, one octaploid Fragaria chiloensis species, and six unknown accessions while Group 5 contained 69 F. × ananassa accessions and six unknown accessions. Accessions within a pedigree were frequently grouped together. A total of 30 novel accessions were categorized alongside existing accessions. These results will allow breeders to develop strategies which incorporate more genetic diversity into new cultivars.

The genetic diversity among strawberry breeding resources based on SSRs

Cultivated strawberry (Fragaria × ananassa Duch.) is a high value horticultural crop. In this study, the genetic diversity of 160 strawberry accessions was determined using five highly polymorphic simple sequence repeat (SSR) markers. Sixty different alleles were identified, with allele frequencies in the range of 0.006 to1. Similarity scores were in the range of 0.034 to 0.963 (average: 0.507). The accessions were categorized into five groups. Group 1 contained two diploid Fragaria vesca species and one unknown accession. Group 2 contained one accession (F x ananassa). Group 3 contained 20 F × ananassa accessions and six unknown accessions. Group 4 contained 48 F. × ananassa accessions, one octaploid Fragaria chiloensis species, and six unknown accessions while Group 5 contained 69 F. × ananassa accessions and six unknown accessions. Accessions within a pedigree were frequently grouped together. A total of 30 novel accessions were categorized alongside existing accessions. These results will allow breeders to develop strategies which incorporate more genetic diversity into new cultivars.(AU)

Cross-amplification of heterologous microsatellite markers in Piracanjuba / Amplificação cruzada de marcadores microssatélites heterólogos em piracanjuba

Brycon orbignyanus, popularly known in Brazil as piracanjuba, is a fish with great economic value but whose natural population drastically decreased in number during the last years. In this context, genetic variability studies of natural stocks and in restocking programs are fundamental for the adoption of conservation measures. Current analysis verifies the cross-amplification of heterologous primers in B. orbignyanus. Fifty-two primers of the species Brycon opalinus, Brycon hilarii, Brycon insignis, Prochilodus sp., Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus were tested. Primers with the best reproducibility were applied to a sample of 20 individuals and the genetic parameters were calculated. Nine primers provided good results for cross-amplification with B. orbignyanus, involving (BoM5 and BoM13) of Brycon opalinus, (Bh5, Bh6, Bh8, Bh13 and Bh16) of Brycon hilarii, (Bc48-10) of Brycon insignis and (Par80) of Prochilodus argenteus. Primers of Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus failed to provide amplification or provided non-specificity. Results demonstrated the possibility of using primers of different species and genera of B. orbignyanus, facilitating genetic studies on the species.

A piracanjuba (Brycon orbignyanus) é um peixe de grande valor econômico que nos últimos anos tem apresentado uma redução drástica em suas populações naturais. Nesse contexto, estudos de variabilidade genética dos estoques naturais e nos programas de repovoamento são fundamentais para adoção de medidas conservacionistas. O objetivo do presente trabalho foi verificar a amplificação cruzada de primers heterologos em B. orbignyanus. Foram avaliados um total de 52 primers das espécies Brycon opalinus, Brycon hilarii, Brycon insignis, Prochilodus sp., Piaractus mesopotamicus, Colossoma macropomum e Oreochromis niloticus. Os primers com melhor reprodutibilidade foram aplicados a uma amostra de 20 indivíduos e os parâmetros genéticos foram calculados. Nove primers apresentaram resultados satisfatórios de amplificação cruzada com B. orbignyanus, sendo das espécies Brycon opalinus (BoM5 e BoM13), Brycon hilarii (Bh5, Bh6, Bh8, Bh13 e Bh16), Brycon insignis (Bc48-10) e Prochilodus argenteus (Par80). Os primers de Piaractus mesopotamicus, Colossoma macropomum e Oreochromis niloticus não apresentaram amplificação ou apresentaram inespecificidade. Os resultados revelaram a possibilidade da utilização de primers de diferentes espécies e gênero em B. orbignyanus, o que facilita a realização de estudos genéticos nessa espécie.

Cross-amplification of heterologous microsatellite markers in Piracanjuba / Amplificação cruzada de marcadores microssatélites heterólogos em piracanjuba

Brycon orbignyanus, popularly known in Brazil as piracanjuba, is a fish with great economic value but whose natural population drastically decreased in number during the last years. In this context, genetic variability studies of natural stocks and in restocking programs are fundamental for the adoption of conservation measures. Current analysis verifies the cross-amplification of heterologous primers in B. orbignyanus. Fifty-two primers of the species Brycon opalinus, Brycon hilarii, Brycon insignis, Prochilodus sp., Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus were tested. Primers with the best reproducibility were applied to a sample of 20 individuals and the genetic parameters were calculated. Nine primers provided good results for cross-amplification with B. orbignyanus, involving (BoM5 and BoM13) of Brycon opalinus, (Bh5, Bh6, Bh8, Bh13 and Bh16) of Brycon hilarii, (Bc48-10) of Brycon insignis and (Par80) of Prochilodus argenteus. Primers of Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus failed to provide amplification or provided non-specificity. Results demonstrated the possibility of using primers of different species and genera of B. orbignyanus, facilitating genetic studies on the species.(AU)

A piracanjuba (Brycon orbignyanus) é um peixe de grande valor econômico que nos últimos anos tem apresentado uma redução drástica em suas populações naturais. Nesse contexto, estudos de variabilidade genética dos estoques naturais e nos programas de repovoamento são fundamentais para adoção de medidas conservacionistas. O objetivo do presente trabalho foi verificar a amplificação cruzada de primers heterologos em B. orbignyanus. Foram avaliados um total de 52 primers das espécies Brycon opalinus, Brycon hilarii, Brycon insignis, Prochilodus sp., Piaractus mesopotamicus, Colossoma macropomum e Oreochromis niloticus. Os primers com melhor reprodutibilidade foram aplicados a uma amostra de 20 indivíduos e os parâmetros genéticos foram calculados. Nove primers apresentaram resultados satisfatórios de amplificação cruzada com B. orbignyanus, sendo das espécies Brycon opalinus (BoM5 e BoM13), Brycon hilarii (Bh5, Bh6, Bh8, Bh13 e Bh16), Brycon insignis (Bc48-10) e Prochilodus argenteus (Par80). Os primers de Piaractus mesopotamicus, Colossoma macropomum e Oreochromis niloticus não apresentaram amplificação ou apresentaram inespecificidade. Os resultados revelaram a possibilidade da utilização de primers de diferentes espécies e gênero em B. orbignyanus, o que facilita a realização de estudos genéticos nessa espécie.(AU)

Universal tail sequence-SSR applied to molecular characterization of tropical maize hybrids

The development of efficient and low-cost genotyping methods is essential to precise genetic characterization of cultivars. Here, we present a system based on fluorescently labeled universal tail sequence primers (UTSP) to resolve microsatellite (SSR) markers as an alternative for molecular fingerprinting of maize. A set of 20 SSRs using the UTSP presented an average polymorphic information content of 0.84, which provided a probability of random identity ranging from 10−7 to 10−14, and a minimum exclusion power of 99.99998 % in a group of 48 tropical maize single-cross hybrids traded in Brazil. The genetic diversity analysis based on multidimensional scaling explained approximately 28 % of the total variance for the first two coordinates, tending to group the hybrids according to their respective origin. Additionally, this genotyping system presented a high distinctiveness capacity, which is widely recommended for genetic purity and fingerprinting analyses. Thus, this marker system has a strong potential as a tool for complementary analysis of distinguishability, uniformity and stability required for cultivar registration.

Universal tail sequence-SSR applied to molecular characterization of tropical maize hybrids

The development of efficient and low-cost genotyping methods is essential to precise genetic characterization of cultivars. Here, we present a system based on fluorescently labeled universal tail sequence primers (UTSP) to resolve microsatellite (SSR) markers as an alternative for molecular fingerprinting of maize. A set of 20 SSRs using the UTSP presented an average polymorphic information content of 0.84, which provided a probability of random identity ranging from 10−7 to 10−14, and a minimum exclusion power of 99.99998 % in a group of 48 tropical maize single-cross hybrids traded in Brazil. The genetic diversity analysis based on multidimensional scaling explained approximately 28 % of the total variance for the first two coordinates, tending to group the hybrids according to their respective origin. Additionally, this genotyping system presented a high distinctiveness capacity, which is widely recommended for genetic purity and fingerprinting analyses. Thus, this marker system has a strong potential as a tool for complementary analysis of distinguishability, uniformity and stability required for cultivar registration.(AU)

Benefits of a prepartum anionic diet on the health of dairy cows in the transition period: prevented subclinical hypocalcemia and minimizing oxidative stress

Background: There are many metabolic diseases that affect dairy cows during the transition period, among them, the postparturient hypocalcemia, also known as milk fever, puerperal paresis syndrome, or fallen cow. This disease may affect animals 72 h after calving due to a nutritional imbalance in the calcium metabolism, which occurs after a sudden demand of this mineral at the beginning of lactation. This problem is more common in animals with high milk production, and usually after the second lactation. Therefore, this study evaluated whether the anionic diet on the prepartum phase would be beneficial for dairy cows in the transition period. Materials, Methods & Results: Eight Holstein cows on their second and third pregnancy with body condition score between 3.5 and 4 were used. During the experiment, the animals remained in paddock with grazing Tifton 85 (Cynodon spp.) and water ad libitum. Two groups of four animals each were used: the group A (control) received a dietary cation-anion difference (DCAD) diet of 2.57 mEq kg-1, and the second group, called the group B (treatment), received a DCAD diet of -112.2 mEq kg-1. To evaluate animal health, we performed gynecological examinations, in addition to blood samplings on days 20, 10 and 5 prepartum and 3, 7, 12 and 20 postpartum to assess seric levels of calcium (Ca), total protein, albumin, globulin, cholesterol, triglyceride, urea, and hepatic function (AST, ALT and GGT). It was also measured the production of free radicals (ROS) and lipid peroxidation (TBARS) in the transition period of the cows. No statistical differences were observed between groups regarding body condition score, uterine tone, dominant follicle diameter, size and presence of the corpus luteum and the presence of genital discharge up to 30 days postpartum. However, the anionic diet prevented a dramatic reduction in calcium levels, unlike what occurred in the control cows (the group A) where a subclinical hypocalcemia was observed.[...]

The protective effects of an adsorbent against oxidative stress in quails fed aflatoxin-contaminated diet

Background: Contamination of crops with aflatoxin is considered a serious global threat to food safety, since potent carcinogenic, teratogenic, mutagenic and immunosuppressive effects of aflatoxins are well recognized. Recently, the use of adsorbents has been linked with protective effects against oxidative stress in several diseases. Thus, the aim of this study was to assess the occurrence of oxidative stress in quails (Coturnix coturnix) fed with aflatoxin-contaminated diet, as well as the protective effect of an adsorbent.Materials, Methods & Results: Twenty-eight quails were divided into four groups (n = 7): diet without additives (control; the group A), diet and adsorbent containing aluminosilicates (the group B), aflatoxin-contaminated diet (200 ppb) (the group C), and aflatoxin-contaminated diet (200 ppb) and adsorbent containing aluminosilicates (the group D). The composition of the adsorbent containing aluminosilicates was 0.3% based on yeast cell wall, silymarin, and bentonite. The animals received feed and water ad libitum during 20 days. At the end of the experimental period, total blood was collected by cardiac puncture in tubes without anticoagulant to obtain serum (centrifuged at 3500 g during 10 min) for later determination of biochemical parameters. The liver was placed in a solution of Tris–HCl 10 mM, pH 7.4 for TBARS (Thiobarbituric acid reactive substances), ROS (Reactive oxygen species), SOD (Superoxide dismutase) and CAT (Catalase) analysis. The hepatic tissue was gently homogenized in a glass potter in specific buffer, homogenated, and centrifuged at 10.000 g at 4ºC for 10 min to yield a supernatant (S1) used for analyses. Homogenate aliquots were stored at -80°C until utilization. Fragments of liver and intestine (5 cm) were collected for histopathological analyses. Between days 15 to 20 of the experiment, group C quails showed clinical signs, such as apathy, creepy feathers and reduced feed intake.[...]