Using peritoneal and blood lactate as predictor of condition type, surgical referral and prognosis in equine colic cases / Uso do lactato peritoneal e sanguíneo como preditor do tipo de afecção, encaminhamento cirúrgico e prognóstico em casos de cólica equina

It is important to investigate fast and accurate equine colic syndrome diagnostic forms. Lactate results from anaerobic glycolysis; high levels of it may indicate intestinal disorders with tissue hypoperfusion and hypoxia. The current study aims at investigating whether blood and peritoneal lactate values observed when horses with colic syndrome were hospitalized, were associated with condition type, therapeutic referral, and survival rates. Retrospective analysis was applied to 498 medical records of animals with colic syndrome, at EQUIVET Hospital -SP; 89 cases were herein selected. Based on logistic regression, peritoneal lactate played a more significant part than blood in variables like obstruction type and survival rates. Surgical cases comprised 52.8% of analyzed animals; strangulation changes, 26%; and overall survival, 62.9%. Mean blood and peritoneal lactate level in animals showing strangulation changes reached 5.11 and 7.33mmol/L, whereas non-strangulation cases recorded 3.54 and 3.06mmol/L, respectively. On the other hand, mean blood and peritoneal lactate level recorded for survivors reached 3.43 and 2.42mmol/L, whereas non-survivors recorded 4.84 and 7.13mmol/L, respectively. We concluded that peritoneal lactate measured when horses with colic syndrome were hospitalized was a predictor of condition type, and of animal survival and prognosis. However, blood and peritoneal lactate measurements did not contribute to therapeutic referral.

O estudo de formas diagnósticas rápidas e precisas é importante na síndrome cólica equina. O lactato, produto da glicólise anaeróbica, quando elevado, pode indicar afecções intestinais com hipoperfusão e hipóxia. O objetivo deste estudo foi verificar se os valores de lactato sanguíneo e peritoneal, na admissão de equinos com síndrome cólica, estão relacionados com o tipo de afecção, com o encaminhamento terapêutico e a sobrevida. Uma análise retrospectiva foi realizada em 498 atendimentos em síndrome cólica, no Hospital EQUIVET-SP, quando 89 casos foram selecionados. A regressão logística indicou que o lactato peritoneal obteve maior significância comparado ao sanguíneo, nas variáveis tipo de obstrução e sobrevida. Casos cirúrgicos foram 52,8%; alterações estrangulativas, 26%; e sobrevivência geral, 62,9%. Em alterações estrangulativas, a média do lactato sanguíneo e peritoneal foi de 5,11 e 7,33mmol/L; em não estrangulativos, 3,54 e 3,06mmol/L. Já os sobreviventes obtiveram 3,43 e 2,42mmol/L, e os não sobreviventes 4,84 e 7,13mmol/L, respectivamente. Conclui-se que a mensuração do lactato peritoneal na admissão de cavalos com cólica foi considerada um preditor do tipo de afecção, bem como da sobrevida e do prognóstico. Porém, as mensurações de lactato sanguíneo e peritoneal não auxiliaram no encaminhamento terapêutico.

The sperm mitochondria: clues and challenges

Sperm cells rely on different substrates to fulfil thei energy demand for different functions and diverse moments of their life. Species specific mechanism involve both energy substrate transport and their utilization: hexose transporters, a protein family of facilitative passive transporters of glucose and other hexose, have been identified in spermatozoa of different species and, within the species, their localization has been identified and, in some cases, linked to specific glycilitic enzyme presence. The catabolism of hexose sources for energy purposes has been studied in various species, and recent advances has been made in the knowledge of metabolic strategies of sperm cells. In particular, the importance of aerobic metabolism has been defined and described in horse, boar and even mouse spermatozoa; bull sperm cells demonstrate to have a good adaptability and capacity to switch between glycolysis and oxidative phosphorylation; finally, dog sperm cells have been demonstrated to have a great plasticity in energy metabolism management, being also able to activate the anabolic pathway of glycogen syntesis. In conclusion, the study of energy management and mitochondrial function in spermatozoa of different specie furnishes important base knowledge to define new media for preservation as well as newbases for reproductive biotechnologies.(AU)

Sperm energy metabolism

The development of techniques to increase sperm longevity demands knowledge about cell metabolism. Sperm requires a constant supply of energy to maintain its cell functions. Approximately 500 metabolic reactions take place in somatic cells, and several of them require energy. Most of the produced energy goes for sperm motility, which is an ATP-dependent specialized process. Spermatozoa possess the required mechanisms to produce energy through glycolysis, citric acid cycle (Krebs cycle) and oxidative phosphorylation. Understanding these pathways provides knowledge on the interactions between the semen extender substrates and sperm cells. The aim of this review is to approach the major energy producing pathways of the sperm, as well as the substrates available for this metabolism.

Para o desenvolvimento de técnicas que visam o aumento da longevidade espermática, é necessário o entendimento metabólico desta célula. O espermatozoide exige um fornecimento constante de energia para a manutenção das funções celulares. Aproximadamente 500 reações metabólicas são conhecidas por ocorrer nas células somáticas, sendo que grande parte delas requer energia. Para atingir seu objetivo, grande parte da produção energética do espermatozoide é destinada à motilidade, o qual é um processo especializado e dependente de ATP. O espermatozoide possui os mecanismos necessários para produzir energia através da glicólise, Ciclo do Ácido Cítrico (Ciclo de Krebs) e Fosforilação Oxidativa. O entendimento destas vias torna-se indispensável para a compreensão das interações entre os substratos do meio diluente e o espermatozoide. O objetivo desta revisão é abordar as principais vias de produção energética do espermatozoide, assim como os substratos disponíveis para metabolização.

Sperm energy metabolism

The development of techniques to increase sperm longevity demands knowledge about cell metabolism. Sperm requires a constant supply of energy to maintain its cell functions. Approximately 500 metabolic reactions take place in somatic cells, and several of them require energy. Most of the produced energy goes for sperm motility, which is an ATP-dependent specialized process. Spermatozoa possess the required mechanisms to produce energy through glycolysis, citric acid cycle (Krebs cycle) and oxidative phosphorylation. Understanding these pathways provides knowledge on the interactions between the semen extender substrates and sperm cells. The aim of this review is to approach the major energy producing pathways of the sperm, as well as the substrates available for this metabolism.(AU)

Para o desenvolvimento de técnicas que visam o aumento da longevidade espermática, é necessário o entendimento metabólico desta célula. O espermatozoide exige um fornecimento constante de energia para a manutenção das funções celulares. Aproximadamente 500 reações metabólicas são conhecidas por ocorrer nas células somáticas, sendo que grande parte delas requer energia. Para atingir seu objetivo, grande parte da produção energética do espermatozoide é destinada à motilidade, o qual é um processo especializado e dependente de ATP. O espermatozoide possui os mecanismos necessários para produzir energia através da glicólise, Ciclo do Ácido Cítrico (Ciclo de Krebs) e Fosforilação Oxidativa. O entendimento destas vias torna-se indispensável para a compreensão das interações entre os substratos do meio diluente e o espermatozoide. O objetivo desta revisão é abordar as principais vias de produção energética do espermatozoide, assim como os substratos disponíveis para metabolização.(AU)

Avaliação da concentração do lactato desidrogenase em cadelas portadoras de neoplasias mamárias / Evaluation of the concentration of lactate dehydrogenase in bitches with mammary gland tumor

The increase in lactate dehydrogenase (LDH) activity is associated with hypoxia due to the rapid proliferation of cancer cells and high metabolic demands. Few studies have evaluated the LDH concentration in canine mammary tumor (CMT). Thus, the objective of this work was to evaluate serum LDH concentrations in CMT and its association with prognostic factors. Thirty bitches participated in the work and were divided equally into two groups, with and without CMT. The average concentration of LDH was significantly higher in CMT (424.9±244.4U / L) compared to the control group (299.0±170.3 U / L) and its values were positively correlated with inflammation and / or macroscopic ulceration (r=0.6), tumor size (r=0.5), histopathological grade (r=0.6), metastatic lymph node (r=0.7) and clinical stage (r=0.5).

Avaliação da concentração do lactato desidrogenase em cadelas portadoras de neoplasias mamárias / Evaluation of the concentration of lactate dehydrogenase in bitches with mammary gland tumor

The increase in lactate dehydrogenase (LDH) activity is associated with hypoxia due to the rapid proliferation of cancer cells and high metabolic demands. Few studies have evaluated the LDH concentration in canine mammary tumor (CMT). Thus, the objective of this work was to evaluate serum LDH concentrations in CMT and its association with prognostic factors. Thirty bitches participated in the work and were divided equally into two groups, with and without CMT. The average concentration of LDH was significantly higher in CMT (424.9±244.4U / L) compared to the control group (299.0±170.3 U / L) and its values were positively correlated with inflammation and / or macroscopic ulceration (r=0.6), tumor size (r=0.5), histopathological grade (r=0.6), metastatic lymph node (r=0.7) and clinical stage (r=0.5).(AU)

Behavior and blood profile in Friesian-Holstein dairy cows in the special region of Yogyakarta, Indonesia

The background of this study was blood profile and cows' behavior that could be used as a tool to diagnose reproductive status in the dairy cows. This research was aimed to determine biochemical blood profile, progesterone level, and behaviors of Friesian Holstein crossbreed dairy cow in the tropical climates. Fifteen Friesian-Holstein crossbreeds cows, in early lactation, reared by local farmers were divided into two barn types: individual and groups of cows. Blood samples were collected from the jugular vein using a 22G needle on days 0, 14, and 28. For the determination of blood profile, blood was collected into Vacuntainer® blood tubes containing ethylenediaminetetraacetic acid (EDTA) to arrest glycolysis. The blood was transported to the laboratory with a coolbox. The concentration of progesterone in plasma was assayed byenzyme-linked immunosorbent assay (ELISA) using a commercial kit and the biochemical blood profile serum was measured by using microlab 300. The variables observed were glucose, cholesterol, urea, and non-pregnant progesterone. Data obtained was examined by T-test analysis. The result showed that the Temperature-Humidity Index were 81.3 and 79.6 respectively, there were no significant differences between two observation, in terms of total feeding time (3.06±0.48 and 2.91±0.22 hours/day), total water consumption (86.23±0.58 and 85.37±0.88 l/day), total standing (12.70±0.21 and 12.83±0.34 hours/day), total laying (11.30±0.71 and 11.16±0.62 hours/day). The blood profile and progesterone levels were lower than the normal range in dairy cows. Also, the result indicated that blood parameters were not significantly different (P > 0.05) between individual pen and group. The result indicated that the progesterone level was not significant in two models of management dairy cows. The conclusion of the research is the climate can affect behavior and the blood parameters. Blood profiles [...].(AU)

Behavior and blood profile in Friesian-Holstein dairy cows in the special region of Yogyakarta, Indonesia

The background of this study was blood profile and cows' behavior that could be used as a tool to diagnose reproductive status in the dairy cows. This research was aimed to determine biochemical blood profile, progesterone level, and behaviors of Friesian Holstein crossbreed dairy cow in the tropical climates. Fifteen Friesian-Holstein crossbreeds cows, in early lactation, reared by local farmers were divided into two barn types: individual and groups of cows. Blood samples were collected from the jugular vein using a 22G needle on days 0, 14, and 28. For the determination of blood profile, blood was collected into Vacuntainer® blood tubes containing ethylenediaminetetraacetic acid (EDTA) to arrest glycolysis. The blood was transported to the laboratory with a coolbox. The concentration of progesterone in plasma was assayed byenzyme-linked immunosorbent assay (ELISA) using a commercial kit and the biochemical blood profile serum was measured by using microlab 300. The variables observed were glucose, cholesterol, urea, and non-pregnant progesterone. Data obtained was examined by T-test analysis. The result showed that the Temperature-Humidity Index were 81.3 and 79.6 respectively, there were no significant differences between two observation, in terms of total feeding time (3.06±0.48 and 2.91±0.22 hours/day), total water consumption (86.23±0.58 and 85.37±0.88 l/day), total standing (12.70±0.21 and 12.83±0.34 hours/day), total laying (11.30±0.71 and 11.16±0.62 hours/day). The blood profile and progesterone levels were lower than the normal range in dairy cows. Also, the result indicated that blood parameters were not significantly different (P > 0.05) between individual pen and group. The result indicated that the progesterone level was not significant in two models of management dairy cows. The conclusion of the research is the climate can affect behavior and the blood parameters. Blood profiles [...].

Genetic engineering of Theileria parva lactate dehydrogenase gene: a new anti-theilerial target

ABSTRACT: Theileria parva is the causative agent of East Coast Fever (ECF), a tick borne disease, which results in major economic losses in cattle. Major problems in dealing with this illness are the high cost of drugs, development of resistance, and absence of effective vaccines. Thus, exploiting new targets for cost effective and higher therapeutic value drugs are imperative. Glycolysis is the main pathway for generation of ATP in T. parva, given its development inside erythrocytes. Thus, the enzymes of this pathway may prove potential targets for designing new-generation anti-theilerials. Lactate dehydrogenase of T. parva (TpLDH) has the highest activity of all glycolytic enzymes and thus we selected this enzyme as the potential therapeutic target. Our study is the first to report the isolation, removal of introns through directed mutagenesis, and cloning of TpLDH and showing that amino acid insertions or deletions most notably corresponded to a 5-amino acid sequence (Asn-91A, Glu-91B, Glu-91C, Trp-91D, Asn-91E) between Ser-91 ve Arg-92 of the enzyme. This region is also present in other apicomplexan such as Babesia bovis, a pathogen of cattle and Plasmodium falciparum, a human pathogen. Providing as the attachment site for the enzyme inhibitors and not being present in LDH of respective hosts, we propose this site as an attractive drug target. The work here is expected to lead new studies on detailed structural and kinetic aspects of apicomplexan LDHs and development of new inhibitors.

Genetic engineering of Theileria parva lactate dehydrogenase gene: a new anti-theilerial target

Theileria parva is the causative agent of East Coast Fever (ECF), a tick borne disease, which results in major economic losses in cattle. Major problems in dealing with this illness are the high cost of drugs, development of resistance, and absence of effective vaccines. Thus, exploiting new targets for cost effective and higher therapeutic value drugs are imperative. Glycolysis is the main pathway for generation of ATP in T. parva, given its development inside erythrocytes. Thus, the enzymes of this pathway may prove potential targets for designing new-generation anti-theilerials. Lactate dehydrogenase of T. parva (TpLDH) has the highest activity of all glycolytic enzymes and thus we selected this enzyme as the potential therapeutic target. Our study is the first to report the isolation, removal of introns through directed mutagenesis, and cloning of TpLDH and showing that amino acid insertions or deletions most notably corresponded to a 5-amino acid sequence (Asn-91A, Glu-91B, Glu-91C, Trp-91D, Asn-91E) between Ser-91 ve Arg-92 of the enzyme. This region is also present in other apicomplexan such as Babesia bovis, a pathogen of cattle and Plasmodium falciparum, a human pathogen. Providing as the attachment site for the enzyme inhibitors and not being present in LDH of respective hosts, we propose this site as an attractive drug target. The work here is expected to lead new studies on detailed structural and kinetic aspects of apicomplexan LDHs and development of new inhibitors.(AU)

Genetic engineering of Theileria parva lactate dehydrogenase gene: a new anti-theilerial target

Theileria parva is the causative agent of East Coast Fever (ECF), a tick borne disease, which results in major economic losses in cattle. Major problems in dealing with this illness are the high cost of drugs, development of resistance, and absence of effective vaccines. Thus, exploiting new targets for cost effective and higher therapeutic value drugs are imperative. Glycolysis is the main pathway for generation of ATP in T. parva, given its development inside erythrocytes. Thus, the enzymes of this pathway may prove potential targets for designing new-generation anti-theilerials. Lactate dehydrogenase of T. parva (TpLDH) has the highest activity of all glycolytic enzymes and thus we selected this enzyme as the potential therapeutic target. Our study is the first to report the isolation, removal of introns through directed mutagenesis, and cloning of TpLDH and showing that amino acid insertions or deletions most notably corresponded to a 5-amino acid sequence (Asn-91A, Glu-91B, Glu-91C, Trp-91D, Asn-91E) between Ser-91 ve Arg-92 of the enzyme. This region is also present in other apicomplexan such as Babesia bovis, a pathogen of cattle and Plasmodium falciparum, a human pathogen. Providing as the attachment site for the enzyme inhibitors and not being present in LDH of respective hosts, we propose this site as an attractive drug target. The work here is expected to lead new studies on detailed structural and kinetic aspects of apicomplexan LDHs and development of new inhibitors.(AU)

Molecular Cloning and Mrna Expression Analysis of Sichuan White Goose (Anser Cygnoides) Chrebp Gene

The carbohydrate response element-binding protein (ChREBP) is an important nuclear factor that regulates glycolysis and de novo lipogenesis. However, the role of ChREBP in fatty liver development in geese remains unclear. In order to understand the function of ChREBP in lipid metabolism of geese, we first cloned the complete cDNA of the ChREBP of the Sichuan White goose (Anser cygnoides) using RT-PCR, 5 RACE and 3 RACE, and analyzed goose ChREBP expression in nine different tissues using real-time PCR technology. The results showed that the goose ChREBP CDS consists of 945bp nucleotides that encode 314 amino acids, and the sequence has high similarities with the swan goose (Anser cygnoides domesticus) and duck (Anas platyrhynchos) sequences, both at the nucleotide and amino acid levels. The predicted ChREBP protein had a molecular mass of 35.64 kDa with pI value of 5.36. The phylogenetic analysis indicated its evolutionary relationships with corresponding orthologous sequences in swan geese and ducks. The qPCR assays revealed that ChREBP is highly expressed in liver in the Sichuan White goose. Together, these results indicate that goose ChREBP may play an important role in the development of hepatic steatosis.(AU)

Molecular Cloning and Mrna Expression Analysis of Sichuan White Goose (Anser Cygnoides) Chrebp Gene

The carbohydrate response element-binding protein (ChREBP) is an important nuclear factor that regulates glycolysis and de novo lipogenesis. However, the role of ChREBP in fatty liver development in geese remains unclear. In order to understand the function of ChREBP in lipid metabolism of geese, we first cloned the complete cDNA of the ChREBP of the Sichuan White goose (Anser cygnoides) using RT-PCR, 5 RACE and 3 RACE, and analyzed goose ChREBP expression in nine different tissues using real-time PCR technology. The results showed that the goose ChREBP CDS consists of 945bp nucleotides that encode 314 amino acids, and the sequence has high similarities with the swan goose (Anser cygnoides domesticus) and duck (Anas platyrhynchos) sequences, both at the nucleotide and amino acid levels. The predicted ChREBP protein had a molecular mass of 35.64 kDa with pI value of 5.36. The phylogenetic analysis indicated its evolutionary relationships with corresponding orthologous sequences in swan geese and ducks. The qPCR assays revealed that ChREBP is highly expressed in liver in the Sichuan White goose. Together, these results indicate that goose ChREBP may play an important role in the development of hepatic steatosis.

Measurement of serum lactate values in domestic cats (Felis catus) submitted to physical and chemical restraint / Mensuração dos valores de lactato sérico em felinos domésticos (Felis catus) submetidos a contenção física e química

The use of serum lactate has become increasingly common in Veterinary Medicine, since it is an excellent marker of tissue oxygenation, elevated in situations of tissue hypoxia, where anaerobic glycolysis is used as an energy source. Studies point out its use in dogs as a prognostic factor in several affections; however it is not commonly used in domestic felines due to the high stress threshold of the species, which may interfere with their values. The aim of the present study was to evaluate serum lactate values in domestic cats during physical restraint and after chemical restraint using the combination of 8mg/kg ketamine-S, 0.4 mg/kg midazolam, and 2 mg/kg Tramadol. To this end, 20 clinically healthy female cats without defined race, from six months to five years of age, were divided into two groups, GPR (n = 10) represented by felines that had only physical restraint and the GCR (n = 10) by felines that underwent physical restraint (TPR) and after 30 hours by chemical restraint (TCR). It was observed that in both groups (GPR and CPR) during physical restraint presented hyperlactatemia (GPR = 6.23 ± 0.9 mmol/L ­ GCR ­ Tpr = 6.66 ± 1.59 mmol/L) and mean values of 4.42 ± 1.28 mmol/L during chemical restraint (GCR ­ Tcr). It was concluded that physical restraint interferes with serum lactate values, with the reduction of such values with the use of chemical restraint in the feline species.(AU)

A utilização do lactato sérico é cada vez mais comum na Medicina Veterinária, por ser um excelente marcador de oxigenação tecidual, elevando-se em situações de hipóxia tecidual e realizando a glicólise anaeróbica como fonte de energia. Pesquisas científicas em cães o apontam como fator prognóstico em diversas afecções, sendo pouco utilizado em felinos domésticos devido ao elevado limiar de estresse da espécie, podendo interferir em seus valores. Assim, este trabalho avaliou os valores de lactato sérico em felinos domésticos durante contenção física e após contenção química, utilizando a associação de 8 mg/kg de cetamina-S, 0,4 mg/kg de midazolam e 2 mg/kg de cloridrato de tramadol. Foram estudados 20 felinos, fêmeas, sem raça definida, de seis meses a cinco anos de idade, clinicamente saudáveis, divididos em dois grupos, com GPR (n = 10) representado por felinos que passaram apenas por contenção física e com GCR por felinos que passaram por contenção física (TPR) e após 30 horas por contenção química (TCR). Foi observado em ambos os grupos (GPR e GCR) e durante a contenção física a hiperlactatemia sérica (GPR = 6.23 ± 0.9 mmol / L ­ GCR ­ TPR = 6.66 ± 1.59 mmol / L) com valores médios de 4,42 ± 1,28 mmol/L durante a contenção química (GCR ­ TCR). Conclui-se que a contenção física interfere nos valores do lactato sérico, com redução dos valores na utilização da contenção química na espécie felina.(AU)

Measurement of serum lactate values in domestic cats (Felis catus) submitted to physical and chemical restraint / Mensuração dos valores de lactato sérico em felinos domésticos (Felis catus) submetidos a contenção física e química

The use of serum lactate has become increasingly common in Veterinary Medicine, since it is an excellent marker of tissue oxygenation, elevated in situations of tissue hypoxia, where anaerobic glycolysis is used as an energy source. Studies point out its use in dogs as a prognostic factor in several affections; however it is not commonly used in domestic felines due to the high stress threshold of the species, which may interfere with their values. The aim of the present study was to evaluate serum lactate values in domestic cats during physical restraint and after chemical restraint using the combination of 8mg/kg ketamine-S, 0.4 mg/kg midazolam, and 2 mg/kg Tramadol. To this end, 20 clinically healthy female cats without defined race, from six months to five years of age, were divided into two groups, GPR (n = 10) represented by felines that had only physical restraint and the GCR (n = 10) by felines that underwent physical restraint (TPR) and after 30 hours by chemical restraint (TCR). It was observed that in both groups (GPR and CPR) during physical restraint presented hyperlactatemia (GPR = 6.23 ± 0.9 mmol/L ­ GCR ­ Tpr = 6.66 ± 1.59 mmol/L) and mean values of 4.42 ± 1.28 mmol/L during chemical restraint (GCR ­ Tcr). It was concluded that physical restraint interferes with serum lactate values, with the reduction of such values with the use of chemical restraint in the feline species.(AU)

A utilização do lactato sérico é cada vez mais comum na Medicina Veterinária, por ser um excelente marcador de oxigenação tecidual, elevando-se em situações de hipóxia tecidual e realizando a glicólise anaeróbica como fonte de energia. Pesquisas científicas em cães o apontam como fator prognóstico em diversas afecções, sendo pouco utilizado em felinos domésticos devido ao elevado limiar de estresse da espécie, podendo interferir em seus valores. Assim, este trabalho avaliou os valores de lactato sérico em felinos domésticos durante contenção física e após contenção química, utilizando a associação de 8 mg/kg de cetamina-S, 0,4 mg/kg de midazolam e 2 mg/kg de cloridrato de tramadol. Foram estudados 20 felinos, fêmeas, sem raça definida, de seis meses a cinco anos de idade, clinicamente saudáveis, divididos em dois grupos, com GPR (n = 10) representado por felinos que passaram apenas por contenção física e com GCR por felinos que passaram por contenção física (TPR) e após 30 horas por contenção química (TCR). Foi observado em ambos os grupos (GPR e GCR) e durante a contenção física a hiperlactatemia sérica (GPR = 6.23 ± 0.9 mmol / L ­ GCR ­ TPR = 6.66 ± 1.59 mmol / L) com valores médios de 4,42 ± 1,28 mmol/L durante a contenção química (GCR ­ TCR). Conclui-se que a contenção física interfere nos valores do lactato sérico, com redução dos valores na utilização da contenção química na espécie felina.(AU)

METABOLISMO DE CARBOIDRATOS E EFEITO POUPADOR DE PROTEÍNA PARA O TAMBAQUI (Colossoma macropomum)

A utilização de carboidratos na dieta de peixes vem sendo amplamente estudada visando à diminuição dos níveis proteicos utilizados e, consequentemente, a diminuição dos custos e dos impactos ambientais gerados pela piscicultura. O objetivo deste trabalho foi avaliar o efeito poupador de proteína em dietas para o tambaqui (Colossoma macropomum), buscando encontrar uma adequada proporção proteína/carboidratos, que resulte em ótimo desempenho produtivo, mas sem prejuízo aos mecanismos fisiológicos envolvidos. Foram utilizados 216 peixes com peso médio de 30 gramas, distribuídos em 18 caixas, alimentados com dietas contendo: 23/25%, 23/35%, 26/25%, 26/35%, 29/25% e 29/35% PD/CHO. Os seis tratamentos em arranjo fatorial (2x3) foram distribuídos em um delineamento inteiramente casualizado, com três repetições por tratamento. Foram avaliados os parâmetros de desempenho produtivo e determinadas as atividades hepáticas de enzimas do metabolismo intermediário e a concentração dos metabólitos no sangue. Os resultados foram submetidos à ANOVA e teste de comparação de médias pelo Teste de Tukey (5%), utilizando o software SAS v.9.0. Foi possível constatar o efeito poupador de proteínas com a substituição da proteína dietética por carboidratos, já que não houve diferenças nos parâmetros de desempenho produtivo obtidos. Os dados sugerem grande capacidade de plasticidade metabólica para utilização dos carboidratos pelo tambaqui, com melhores resultados em peixes alimentados com dietas contendo 26%PD e 35% CHO.

The use of carbohydrates in the fish diet has been widely studied with a view to decreasing the protein levels used and, consequently, reducing the costs and environmental impacts generated by fish farming. The objective of this work were be to evaluate the protein-sparing effect in diets for tambaqui (Colossoma macropomum), seeking to find an adequate protein / carbohydrate ratio, which results in excellent productive performance, but without prejudice to the physiological mechanisms involved. 216 fish with average weight of 30g were used, distributed in 18 boxes, feed with diets containing PD/CHO: 23/25%, 23/35%, 26/25%, 26/35%, 29/25% and 29/35%. The six treatments in a factorial arrangement (2x3) were distributed in a completely randomized design, with three replicates per treatment. The parameters of productive performance were evaluated and the hepatic activities of enzymes of intermediate metabolism and the concentration of metabolites in the blood were determined. The results were submitted to ANOVA and test of comparison of means by the Tukey Test (5%), using the software SAS v.9.0. It was possible to verify the protein sparing effect with the replacement of dietary protein by carbohydrates, since there were no differences in the parameters of productive performance obtained. The data suggest great metabolic plasticity capacity for the use of carbohydrates by tambaqui, with better results in fish fed diets containing 26% PD and 35% CHO.

PROTEÔMICA DE TECIDO RUMINAL E CECAL DE BOVINOS NELORE CONFINADOS COM DIFERENTES ESTRATÉGIAS NUTRICIONAIS

A literatura apresenta poucos estudos que investigaram o proteoma de ceco e rúmen de bovinos submetidos a estratégias alimentares com uso de aditivos. Essa proposta é pioneira na identificação, expressão e compreensão do perfil proteico ruminal e cecal de bovinos nelore confinados. Artigo 1: Objetivou-se avaliar a expressão proteica, a fim identificar possíveis moléculas candidatas a biomarcadores e suas relações com funções metabólicas e nutricionais aplicados a animais confinados com diferentes níveis de amido (25, 35 e 45%) e aditivos (Monensina × blend de óleos essências + amilase exógena), as dosagens de monensina sódica, blend de óleos essenciais e amilase exógena foram 26, 90 e 560 mg/kg MS, respectivamente. Foram utilizados machos Nelore (n = 210) (Bos taurus indicus), não castrados (Peso Inicial = ± 380 kg). Após o abate foram coletadas amostras de ceco e acondicionadas imediatamente em nitrogênio líquido. Foi feito um pool de amostras utilizando 90 animais (15 animais/tratamento) no processo de precipitação do pellet proteico. A separação e caracterização do proteoma das amostras biológicas foram feitas por eletroforese bidimensional (2D-PAGE) e identificadas por cromatografia líquida acoplada à espectrometria de massas (LC-MS/MS). Os géis de poliacrilamida foram analisados utilizando ImageMaster 2D Platinum 7.0 para verificar as diferenças na expressão de proteínas, para as comparações do % volume normalizado dos spots. Após a identificação das proteínas foram analisadas as vias metabólicas usando a função da Enciclopédia de Genes e Genomas de Kyoto (KEGG), análise de enriquecimento da via Reactome para mapear as expressões das proteínas que codificam enzimas e suas respectivas funções nas vias afetadas. O uso de Blend de Óleo essenciais associado com -Amilase como aditivo alimentar promoveu maior expressão de enzimas na via da glicólise e gliconeogênese e ausência de proteína ligada a inflamação (Leukocyte elastase inhibitor). Por outro lado, o incremento de amido nas dietas promoveu redução de enzimas ligadas a degradação de carboidrato com aumento de respostas atribuídas à injúrias inflamatórias no rúmen de bovinos Nelore confinados. Artigo 2: Objetivou-se mapear o proteoma do epitélio ruminal de bovinos nelore confinados (n=60) com diferentes níveis de amido (Baixo: 25 e Alto: 45%) e aditivos (Monensina × blend de óleos essências + amilase exógena). As separações e fracionamento das proteínas foram através da eletroforese bidimensional em gel de poliacrilamida (2D-PAGE), e posteriormente a identificação por espectrometria de massas acoplada a cromatografia líquida (LC-MS/MS). Após a identificação das proteínas foram utilizados os acessos das proteínas para a classificação em suas funções moleculares, processos biológicos e componentes celulares utilizando Blast2GO, posteriormente a análise de enriquecimento via String para mapear as redes e interação entres proteínas caracterizadas suas respectivas funções no metabolismo de glicose e ácidos graxos. Dietas contendo blend de óleos essenciais associados à amilase exógena promoveram maior expressão de macromoléculas que participam da degradação de carboidratos pela via glicolítica e cetogênica. Foram identificadas 14 proteínas com maior expressão e presentes no tecido epitelial do rúmen envolvidas na oxidação da glicose, a proteína hidroximetilglutaril-CoA liase que catalisa parte do metabolismo metabólico intermediário, uma etapa fundamental na cetogênese. Nossos resultados sugerem que houve aumento da glicólise a partir da oxidação do gliceraldeído-3-fosfato, que participa da primeira etapa da produção de acetato e butirato e da descarboxilação oxidativa no epitélio ruminal de bovinos nelore confinados. A monensina melhora os precursores de propionato, a maior expressão de metilmalonil-CoA mutase sugere síntese de propionato via propionil-CoA que participa do ciclo do ácido cítrico através do succinil-CoA, que pode aumentar a energia metabolizável e reduzir a ingestão de alimentos.

Few studies have investigated the proteome of cecum and rumen of cattle feeding diferente estrategies with the use of additives. This proposal is a pioneer in the identification, expression. and understanding of the ruminal and cecal protein profile of feedlot Nellore cattle. Manuscript 1: The objective was to evaluate protein expression in order to identify possible candidate molecules for biomarkers and their relationships with metabolic and nutritional functions applied to animals fed with different levels of starch (25, 35 and 45%) and additives (Monensin × blend of essential oils + exogenous amylase), the dosages of sodium monensin, blend of essential oils and exogenous amylase were 26, 90 and 560 mg/kg DM, respectively. Nellore bulls (n = 210) (Bos taurus indicus) (initial weight = ) 380 kg) were used. After slaughter, cecum samples were collected and immediately placed in liquid nitrogen. Samples were pooled using 90 animals (15 animals/treatment) in the protein pellet precipitation process. The separation and characterization of the proteome of biological samples were performed by twodimensional electrophoresis (2D-PAGE) and identified by liquid chromatography coupled to mass spectrometry (LC-MS/MS). Polyacrylamide gels were analyzed using ImageMaster 2D Platinum 7.0 to verify differences in protein expression for the % volume normalized spot comparisons. After identifying the proteins, the metabolic pathways were analyzed using the Kyoto Encyclopedia of Genes and Genomes (KEGG) function, enrichment analysis of the Reactome pathway to map the expressions of proteins encoding enzymes and their respective functions in the affected pathways. The use of blend essential oil associated with -Amylase as a feedd additive promoted greater expression of enzymes in the glycolysis and gluconeogenesis pathway and absence of protein linked to inflammation (Leukocyte elastase inhibitor). On the other hand, the increase in starch in the diets reduced enzymes linked to carbohydrate degradation with increased responses attributed to inflammatory injuries in the rumen of feedlot Nellore cattle. Manuscript 2: The objective was to map the proteome of the ruminal epithelium of feedlot Nellore cattle (n=60) with different levels of starch (Low: 25 and High: 45%) and additives (Monensin × blend of essential oils + exogenous amylase) the dosages of sodium monensin, blend of essential oils and exogenous amylase were 26, 90 and 560 mg/kg DM, respectively. Protein separations were performed using two-dimensional polyacrylamide gel electrophores (2D-PAGE), followed by identification by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). After protein identification, protein accessions were used to classify their molecular functions, biological processes and cellular component using Blast2GO, then enrichment analysis pathways String to map the networks and interaction xii between proteins characterized and their respective functions in glucose and fatty acids metabolic. Diets containing blend of essential oils associated with exogenous amylase promoted greater expression of macromolecules that participate in carbohydrate degradation via glycolytic and ketogenic pathways. We identified 14 proteins with greater expression and present in the epithelial tissue of the rumen involved in glucose oxidation, the protein hydroxymethylglutaryl-CoA lyase that catalyzes part of intermediary metabolic metabolism, a fundamental step in ketogenesis. Our results suggest that there was an increase in glycolysis from the oxidation of glyceraldehyde-3-phosphate, which participates in the first stage of acetate and butyrate production and oxidative decarboxylation in the ruminal epithelium of feedlot Nellore cattle. Monensin improves propionate precursors, the increased expression of methylmalonyl-CoA mutase suggests propionate synthesis via propionyl-CoA that participates in the citric acid cycle through succinyl-CoA, which can increase metabolizable energy and reduce feed intake.

Glycolysis and gluconeogenesis in the liver of catfish fed with different concentrations of proteins, lipids and carbohydrates / Glicólise e gliconeogênese no fígado de jundiá alimentado com diferentes concentrações de proteínas, lipídeos e carboidratos

The activities of enzymes from a number of metabolic pathways have been used as a tool to evaluate the best use of nutrients on fish performance. In the present study the catfish Rhamdia quelen was fed with diets containing crude protein-lipid-carbohydrate (%) as follows: treatment (T) T1: 19-19-44; T2: 26-15-39; T3: 33-12-33; and T4: 40-10-24. The fish were held in tanks of re-circulated, filtered water with controlled temperature and aeration in 2000L experimental units. The feeding experiment lasted 30 days. The following enzymes of the carbohydrate metabolism were determined: Glucokinase (GK), Phosphofructokinase 1 (PFK-1), Pyruvate kinase (PK), Fructose-1,6-biphosphatase 1 (FBP-1). The activities of 6 phosphogluconate dehydrogenase (6PGDH) and glucose 6 phosphate dehydrogenase (G6PDH) were also assayed. The influence of nutrient levels on the enzyme activities is reported. The increase of dietary protein plus reduction of carbohydrates and lipids attenuates the glycolytic activity and induces hepatic gluconeogenesis as a strategy to provide metabolic energy from amino acids. The fish performance was affected by the concentrations of protein, lipid and carbohydrates in the diet. The greatest weight gain was obtained in fish fed diet T4 containing 40.14% of crude protein, 9.70% of lipids, and 24.37% of carbohydrate, respectively.(AU)

As atividades de enzimas das vias metabólicas têm sido utilizadas como uma ferramenta para avaliar a melhor utilização dos nutrientes e o desempenho dos peixes. No presente estudo, o jundiá foi alimentado com rações contendo diferentes concentrações de proteína bruta, lipídeos e carboidratos (%), da seguinte forma: tratamento (T) T1: 19-19-44; T2: 26-15-39; T3: 33-12-33; e T4: 40-10-24. Os peixes foram mantidos em tanques de recirculação, com água filtrada, temperatura controlada e aeração em unidades experimentais de 2.000L. O período experimental foi de 30 dias. Foram aferidas as atividades das enzimas glicoquinase (GK), fosfofrutoquinase 1 (PFK-1), piruvato quinase (PK) e frutose-1,6-difosfatase (FBP-1). Também foram aferidas as atividades da 6-fosfogluconato desidrogenase (6PGDH) e glicose-6-fosfato desidrogenase (G6PDH) da via das pentoses. É relatado que níveis de nutrientes influenciam as atividades enzimáticas das vias metabólicas. No presente estudo, o aumento da proteína da dieta e a redução de hidratos de carbono e lipídeos reduziram a atividade glicolítica e induziram a gliconeogênese hepática como uma estratégia para fornecer energia pelos aminoácidos. O desempenho dos peixes foi afetado pelas concentrações de proteínas, lipídeos e carboidratos na dieta. O maior ganho de peso foi obtido em peixes alimentados com dieta T4 contendo 40,14% de proteína bruta, 9,70% de lipídeos, e 24,37% de carboidratos, respectivamente.(AU)

Glycolysis and gluconeogenesis in the liver of catfish fed with different concentrations of proteins, lipids and carbohydrates / Glicólise e gliconeogênese no fígado de jundiá alimentado com diferentes concentrações de proteínas, lipídeos e carboidratos

The activities of enzymes from a number of metabolic pathways have been used as a tool to evaluate the best use of nutrients on fish performance. In the present study the catfish Rhamdia quelen was fed with diets containing crude protein-lipid-carbohydrate (%) as follows: treatment (T) T1: 19-19-44; T2: 26-15-39; T3: 33-12-33; and T4: 40-10-24. The fish were held in tanks of re-circulated, filtered water with controlled temperature and aeration in 2000L experimental units. The feeding experiment lasted 30 days. The following enzymes of the carbohydrate metabolism were determined: Glucokinase (GK), Phosphofructokinase 1 (PFK-1), Pyruvate kinase (PK), Fructose-1,6-biphosphatase 1 (FBP-1). The activities of 6 phosphogluconate dehydrogenase (6PGDH) and glucose 6 phosphate dehydrogenase (G6PDH) were also assayed. The influence of nutrient levels on the enzyme activities is reported. The increase of dietary protein plus reduction of carbohydrates and lipids attenuates the glycolytic activity and induces hepatic gluconeogenesis as a strategy to provide metabolic energy from amino acids. The fish performance was affected by the concentrations of protein, lipid and carbohydrates in the diet. The greatest weight gain was obtained in fish fed diet T4 containing 40.14% of crude protein, 9.70% of lipids, and 24.37% of carbohydrate, respectively.(AU)

As atividades de enzimas das vias metabólicas têm sido utilizadas como uma ferramenta para avaliar a melhor utilização dos nutrientes e o desempenho dos peixes. No presente estudo, o jundiá foi alimentado com rações contendo diferentes concentrações de proteína bruta, lipídeos e carboidratos (%), da seguinte forma: tratamento (T) T1: 19-19-44; T2: 26-15-39; T3: 33-12-33; e T4: 40-10-24. Os peixes foram mantidos em tanques de recirculação, com água filtrada, temperatura controlada e aeração em unidades experimentais de 2.000L. O período experimental foi de 30 dias. Foram aferidas as atividades das enzimas glicoquinase (GK), fosfofrutoquinase 1 (PFK-1), piruvato quinase (PK) e frutose-1,6-difosfatase (FBP-1). Também foram aferidas as atividades da 6-fosfogluconato desidrogenase (6PGDH) e glicose-6-fosfato desidrogenase (G6PDH) da via das pentoses. É relatado que níveis de nutrientes influenciam as atividades enzimáticas das vias metabólicas. No presente estudo, o aumento da proteína da dieta e a redução de hidratos de carbono e lipídeos reduziram a atividade glicolítica e induziram a gliconeogênese hepática como uma estratégia para fornecer energia pelos aminoácidos. O desempenho dos peixes foi afetado pelas concentrações de proteínas, lipídeos e carboidratos na dieta. O maior ganho de peso foi obtido em peixes alimentados com dieta T4 contendo 40,14% de proteína bruta, 9,70% de lipídeos, e 24,37% de carboidratos, respectivamente.(AU)

Efeito do sistema de terminação, da taxa de ganho e do critério de abate sobre as características de qualidade de carne de bovinos Angus x Nelore

O objetivo deste estudo foi explorar o impacto da taxa de ganho (TG) e do sistema de terminação sobre as características de crescimento, metabolismo muscular, metabolismo pós morte e qualidade de carne de bovinos abatidos com base no peso vivo (530 kg ± 20 kg) ou no tempo de alimentação (140 dias). Setenta e dois machos castrados cruzados Angus x Nelore foram designados aleatoriamente para um dos quatro tratamentos: 1) confinamento, alta TG (C-A); 2) confinamento, baixa TG (C-B); 3) pasto, alta TG (P-A); e 4) pasto, baixa TG (P-B). Periodicamente, ao longo do estudo, os animais foram pesados e avaliados, por ultrassonografia, quanto à deposição de tecido muscular e de gordura subcutânea. No momento do abate, foram coletados dados sobre as características de carcaça, além de amostragens do músculo Longissimus thoracis (LT) para análises de glicólise in vitro. Após 24 horas de resfriamento da carcaça, foram coletadas amostras do músculo LT para análises de metabólitos e expressão de enzimas relacionadas com o metabolismo energético. Além disso, foram coletadas amostras do músculo LT para avaliar a cor e a força de cisalhamento em 3 tempos de maturação (0, 7 e 14 dias de maturação), na qual as amostras maturadas por 7 dias também foram avaliadas por um painel sensorial de aceitação do consumidor. Todos os dados foram analisados utilizando o procedimento MIXED do software SAS a partir de um delineamento inteiramente casualizado, no qual o tratamento foi considerado como efeito fixo e cada critério de abate foi analisado separadamente. Houve interação entre tratamento e o tempo de alimentação para as características de peso vivo e deposição de músculo e gordura em ambos os critérios de abate (P < 0,05). Houve interação entre tratamento e o tempo de maturação para as características de cor (L*, a* e b*) e força de cisalhamento em ambos os critérios de abate (P < 0,05). Animais C-A e C-B apresentaram carnes não-maturadas mais macia e com maiores valores de L* quando comparada com animais P-A e P-B. No entanto, a carne dos animais C-A apresentou maiores valores de a* e b* quando comparada com a de animais P-A e P-B (P < 0,05), enquanto a carne dos animais CB apresentou valores semelhantes de a* e b* quando comparada com a de animais P-A e P-B. O processo dematuração alterou a cor da carne e os valores de força de cisalhamento, o qual também foi dependente do critério de abate. No critério de abate com base no peso, a carne de animais C-A apresentaram maiores valores de L* quando comparadas com os outros tratamentos (P < 0,05), além de apresentar maiores valores de b* quando comparado com a de animais P-A e P-B (P < 0,05). Entretanto, no critério de abate com base no tempo de alimentação, a carne de animais CA apresentou maiores valores de L* quando comparada com a de animais PA e PB (P < 0,01), além de maiores valores de a* e b* quando comparada com a dos outros tratamentos quando maturadas por 7 e 14 dias (P < 0,05). Além disso, a maturação por 14 dias eliminou a diferença na força de cisalhamento entre os animais terminados a pasto e confinamento quando abatidos com base no peso (P < 0,05), entretanto isto não aconteceu quando abatidos com base no tempo de alimentação (P > 0,05). No painel sensorial, os consumidores preferiram carne de animais terminados em confinamento (CA e CB) quando comparado com animais terminados a pasto (PA e PB) (P < 0,01). Animais terminados a pasto, principalmente animais PB, apresentaram um metabolismo energético muscular mais direcionado para a via oxidativa. Além disso, animais PB quando abatidos com base no tempo de alimentação apresentaram maiores valores de pH quando comparado com animais terminados em confinamento (P < 0,05), o que não foi observado quando os animais foram abatidos com base no peso vivo. Portanto, o sistema de alimentação e a TG têm um efeito pronunciado sobre o metabolismo muscular e pós morte e, consequentemente, na qualidade de carne, no qual o sistema de terminação aparenta ser o principal responsável pelos atributos de luminosidade, maciez e aceitação sensorial da carne, enquanto a TG exerce principalmente um efeito nos atributos cromáticos da cor da carne. Além disso, a carne escura de animais terminados a pasto pode estar relacionada com carcaças leves e magras e/ou com um metabolismo muscular mais oxidativo.

The aim of this study was to explore the impact of the gain rate (GR) and the finishing system on growth characteristics, muscle metabolism, post-mortem metabolism and meat quality of cattle slaughtered based on live weight (530 kg ± 20 kg) or feeding time (140 days). Seventy-two Angus x Nellore crossbred males were randomly assigned to one of four treatments: 1) feedlot, high GR (FH); 2) feedlot, low GR (FL); 3) pasture, high GR (PH); and 4) pasture, low GR (PL). Periodically throughout the study, the animals were weighed and evaluated by ultrasound for the muscle tissue and subcutaneous fat deposition. At slaughter, data on carcass characteristics were collected and Longissimus thoracis (LT) muscle was sampled for further in vitro glycolysis analysis. After 24 h of carcass cooling, LT muscle samples were collected for further analysis of metabolites and enzymes expression related to energy metabolism. In addition, LT muscle samples were collected to evaluate color and shear force at 3 aging times (0-, 7- and 14-days aging), in which samples aged for 7 days were also evaluated by a sensory panel of consumer acceptance. All data were analyzed using the MIXED procedure of the SAS software from a completely randomized design, in which the treatment was considered as a fixed effect and each slaughter criterion was analyzed separately. There was an interaction between treatment and feeding time for live weight and muscle and fat deposition in both slaughter criteria (P < 0.05). The FH and FL animals showed non-aged meat more tenderness with higher L* values when compared to PH and PL animals. However, meat from FH animals showed higher a* and b* values when compared to that from PH and PL animals (P < 0.05), while the meat from FL animals showed similar a* and b* values when compared to that from PH and PL animals. The aging process modified the beef color and shear force values, which also was dependent on the slaughter criterion. In the slaughter criterion based on weight, meat from FH animals showed higher L* values when compared with that from other treatments (P < 0.05), in addition to presenting higher b* values when compared with that from PH animals and PL (P < 0.05). However, in the slaughter criterion based on feeding time, meat from FH animals showed higher L* valueswhen compared with that from PH and PL animals (P < 0.01), in addition to a higher a* and b* value when compared with that from other treatments when aged for 7 and 14 days (P < 0.05). In addition, aging for 14 days mitigated the difference in shear force between animals finished on pasture and feedlot when slaughtered based on weight (P < 0.05); however, this difference was not observed when animals were slaughtered based on feeding time (P > 0.05). In the sensory panel, consumers preferred meat from animals finished in feedlot (FH and FL) when compared to animals finished in pasture (PH and PL) (P < 0.01). Animals finished in pasture, mainly PL animals, showed a muscle energy metabolism more directed towards the oxidative pathway. In addition, PL animals showed higher pH values when compared to animals finished in feedlot when slaughtered based on feeding time (P < 0.05), which was not observed when the animals were slaughtered based on live weight. Therefore, the feeding system and GR have a pronounced effect on muscle metabolism and postmortem and, consequently, on meat quality, in which the finishing system appears to be the main responsible for the attributes of lightness, tenderness and sensory acceptance of the meat, while GR mainly has an effect on the chromatic attributes of meat color. In addition, the dark meat from animals finished on pasture may be related to light and lean carcasses and/or to a more oxidative muscle metabolism.