Dietary lipid sources on the fatty acid profile of swine fat: a review / Fontes lipídicas da dieta no perfil de ácidos graxos da gordura suína: revisão

ABSTRACT: Swine production has focused its efforts to reduce the amount and improve the quality of fat deposited on carcass. Due to the association of the lipid profile with cardiovascular diseases, researches have been developed to understand how the deposition occurs and to improve the fatty acid profile of swine fat. Among the factors that affect the fat composition, race, sex, and diet, the third one has the greatest impact. Lipid inclusion seems to be a promising way to modulate the lipid profile of fat in pig carcass, with an effect on saturated, monounsaturated and polyunsaturated fatty acids. The effect is greater in that fatty acid with the highest concentration in the source used. Soybean oil can double the value of linoleic acid, a by-product of olive oil increases oleic acid by 4%, and flaxseed oil has the ability to increase α-linolenic acid from less than 1% to over 18% in total fatty acids. In addition to the source, the duration and concentration influence the modulation of the fatty acid profile.

RESUMO: A cadeia produtiva de suínos tem concentrado esforços para reduzir a quantidade e melhorar a qualidade da gordura depositada na carcaça. Devido a associação do perfil lipídico com doenças cardiovasculares, pesquisas têm sido desenvolvidas para entender como ocorre a deposição e para melhorar o perfil de ácidos graxos na carcaça. Dentre os fatores que afetam a composição dessa gordura estão a raça, o sexo e a dieta dos suínos, sendo que este último é o de maior impacto. A inclusão de ingredientes lipídicos parece uma promissora forma de modular o perfil lipídico da gordura na carcaça dos suínos, sendo observado efeito nos ácidos graxos saturados, monoinsaturados e poli-insaturados. O efeito é maior naquele ácido graxo de maior concentração na fonte utilizada. O óleo de soja pode dobrar o valor do ácido linoleico. Um subproduto do óleo de oliva incrementa em 4% o ácido oleico. E o óleo de linhaça tem a capacidade de aumentar o α-linolênico de menos de 1% para mais de 18% no total de ácidos graxos. Além da fonte, o período e a concentração influenciam na modulação do perfil de ácidos graxos.

Dietary lipid sources on the fatty acid profile of swine fat: a review / Fontes lipídicas da dieta no perfil de ácidos graxos da gordura suína: revisão

Swine production has focused its efforts to reduce the amount and improve the quality of fat deposited on carcass. Due to the association of the lipid profile with cardiovascular diseases, researches have been developed to understand how the deposition occurs and to improve the fatty acid profile of swine fat. Among the factors that affect the fat composition, race, sex, and diet, the third one has the greatest impact. Lipid inclusion seems to be a promising way to modulate the lipid profile of fat in pig carcass, with an effect on saturated, monounsaturated and polyunsaturated fatty acids. The effect is greater in that fatty acid with the highest concentration in the source used. Soybean oil can double the value of linoleic acid, a by-product of olive oil increases oleic acid by 4%, and flaxseed oil has the ability to increase α-linolenic acid from less than 1% to over 18% in total fatty acids. In addition to the source, the duration and concentration influence the modulation of the fatty acid profile.

A cadeia produtiva de suínos tem concentrado esforços para reduzir a quantidade e melhorar a qualidade da gordura depositada na carcaça. Devido a associação do perfil lipídico com doenças cardiovasculares, pesquisas têm sido desenvolvidas para entender como ocorre a deposição e para melhorar o perfil de ácidos graxos na carcaça. Dentre os fatores que afetam a composição dessa gordura estão a raça, o sexo e a dieta dos suínos, sendo que este último é o de maior impacto. A inclusão de ingredientes lipídicos parece uma promissora forma de modular o perfil lipídico da gordura na carcaça dos suínos, sendo observado efeito nos ácidos graxos saturados, monoinsaturados e poli-insaturados. O efeito é maior naquele ácido graxo de maior concentração na fonte utilizada. O óleo de soja pode dobrar o valor do ácido linoleico. Um subproduto do óleo de oliva incrementa em 4% o ácido oleico. E o óleo de linhaça tem a capacidade de aumentar o α-linolênico de menos de 1% para mais de 18% no total de ácidos graxos. Além da fonte, o período e a concentração influenciam na modulação do perfil de ácidos graxos.

Idiopathic subfertility in stallions. Clinical approaches to a complex problem

At least 30-40% of stallions in commercial breeding programs are moderately fertile and 8-12% are subfertile (0.5-3% with severe subfertility). From the total reported cases of the subfertility, in 2-20% of the stallions the cause is unknown or was not established. The objective of this work is to present the concept of subfertile stallion based on the current state of knowledge and advanced molecular diagnostic technologies. Low pregnancy rates have been reported in stallions with normal semen quality after conventional evaluation. Acrosome reaction (AR) is necessary for natural fertilization and impaired acrosome reaction (IAR) leads to subfertility or infertility in horses, however, AR test is not included in routine semen analysis. Genome-wide association study identified FKBP6 as a strong candidate gene responsible for this failure. The gene encodes for FK506 binding protein 6 (FKBP6) which is involved in sperm development and functions. We could conclude that the evaluation of the acrosomal status is essential in cases of stallions with good motility, concentration, morphology and viability but unexplained (idiopathic) subfertility or infertility. It is important to highlight the recent increase in reports of fertility problems in stallions related to disorders of genetic origin.(AU)

Biochemical and immunohistochemical examination of the effects of ephedrine in rat ovary tissue

Purpose: It was aimed to investigate the biochemical and immunohistochemical effects of ephedrine (EPH) in bilateral ovariectomized rats. Methods: Twenty-four Sprague Dawley female rats were divided into three groups: control group: The abdomen was opened and closed without any treatment; ischemia-reperfusion (IR) group: 2 h of ischemia followed by 2 h of reperfusion were allowed to cause IR injury; IR+EPH group: oral EPH solution (5 mg/kg) was administered for 28 days. Results: Biochemical parameters were statistically significant in group comparisons. Increased interleukin-6 (IL-6) expression, degenerative preantral and antral follicle cells and inflammatory cells around blood vessels were seen in IR group. Negative IL-6 expression was observed in seminal epithelial cells, preantral and antral follicle cells in IR+EPH group. While caspase-3 activity increased in granulosa cells and stromal cells in IR group, caspase-3 expression was negative in preantral and antral follicle cells in the germinal epithelium and cortex in IR+EPH group. Conclusion: The effect of apoptosis, which occurs with the signaling that starts in the cell nucleus, caused the cessation of the stimulating effect at the nuclear level after EPH administration, and a decrease in the antioxidative effect in IR damage and inflammation in the apoptotic process.

Effect of Turmeric (Curcuma Longa) on Duodenal Structure in Broiler Chickens

Supplementation of feed with turmeric (Curcuma longa) has been shown to be beneficial in poultry farming. The present study aimed at evaluating the effect of turmeric on the duodenal structure of broiler chickens during the first 42 days of life. A control and three treatment groups (turmeric powder added to feed at the following doses: E1 - 5; E2 - 10; E3 - 20 g/kg feed) were constituted (n = 8). Addition of turmeric powder in the feed resulted in an increase in intestinal villi height and a decrease in crypts depth in case of groups E1 and E2, while the villus height to crypt depth ratio generally did not differ significantly from the control. Turmeric also influenced the histological structure of the duodenum, as well as the presence of IL-6 and TNFα, as evidenced through staining. Addition of 0.5 and 1 % turmeric powder in the feed had evident results on body weight gain.(AU)

O1-conotoxin Tx6. 7 cloned from the genomic DNA of Conus textile that inhibits calcium currents

Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3' UTR elements. The mature peptide of Tx6.7 (DCHERWDW CPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 µM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development.(AU)

Protocol for in vitro rooting of Pyrus comunnis rootstocks / Protocolo de enraizamento in vitro de porta-enxertos de Pyrus comunnis

Effective protocols for in vitro rooting for woody fruit trees are still a challenge for in vitro seedling production, especially when there is a need to insert new cultivars or rootstocks. These protocols are essential to accelerate studies in plant breeding programs and for seedling distribution. This study evaluated the use of 6-Benzylaminopurine (IBA) in in vitro rooting of Pyruscomunnis rootstocks, clones 'OHxF87' and Pyrodwarf. Explant exposure times (0, 24, 48, 72, and 96 hours) to 20 mg L-1 IBA were tested for in vitro rooting. The exposure to IBA resulted in rooting rates above 80%, surpassing some results reported in the literature. The 24-hour treatment provided 81,81% survival, leading to an average growth of five roots with 19 mm length, for 'OHxF87' rootstock. The same exposure time resulted in the highest survival rate (75%) and the highest mean root number, seven roots per plant with 10 mm length, for 'PDW' rootstock. Root formation did not occur in the absence of synthetic auxin. Therefore, it can be concluded that a 24-hour exposure at 20 mg L-1 IBA was sufficient to promote in vitro rooting in 'OHxF87' and Pyrodwarf rootstocks'.

Protocolos eficazes de enraizamento in vitro de frutíferas lenhosas ainda são um desafio para produção de mudas in vitro, especialmente quando há necessidade de inserção de novas cultivares ou porta-enxerto. Esses protocolos são essenciais para acelerar estudos nos programas de melhoramento genético e também para distribuição posterior das mudas. Nesse sentido, o objetivo deste estudo foi avaliar a utilização da 6-Benzilaminopurina no enraizamento in vitro de porta-enxerto Pirus comunnis, clones 'OHxF87' e Pyrodwarf. Para o enraizamento, foi testado o tempo de exposição dos explantes ao AIB. Para tanto, foram utilizados 20 mg L-1 do fitohormônio nas horas 0, 24, 48, 72 e 96 horas. A exposição ao AIB resultou em taxas de enraizamento acima de 80%, superando alguns resultados encontrados na literatura. Para o porta-enxerto 'OHxF87', o tratamento de 24 horas proporcionou 81,81% de sobrevivência, promovendo em média cinco raízes com comprimento de 19 mm. O mesmo tratamento para o porta-enxerto 'PDW' resultou na maior taxa de sobrevivência (75%), bem como no maior número médio de raízes, sete raízes por planta, com comprimento de 10 mm. Na ausência de auxina sintética, a formação de raízes não ocorreu. Assim sendo, podemos concluir que o tempo de exposição de 24 horas a 20 mg L-1 de IBA foi suficiente para promover o enraizamento de porta-enxertos Pyrus communis de 'OHxF87' e 'PDW'.

Different N6: N3 Ratios on Performance, Egg Quality, and Health of 81-Week-Old Laying Hens

This study aimed to evaluate different n6:n3ratios on performance, serum biochemical variables, and egg quality in 81-week-old laying hens. A total of 224 laying hens, 81-week-old Hysex White, were utilized and distributed in a completely randomized design consisting of seven treatments and eight replications of four birds per cage, totaling 56 cages. The experimental treatments consisted of seven different n6:n3fatty acid ratios: 1.0:1.0, 2.0:1.0, 4.0:1.0, 8.0:1.0, 16.0:1.0, 32.0:1.0, and 64.0:1.0. For diet formulation, sunflower oil, rich in omega-6, and linseed oil, rich in omega-3, were used. Productive performance, egg quality and serum biochemical variables of laying hens were evaluated at the end of the cycle at 26, 27, and 28 days. Analysis of variance (ANOVA) was conducted and orthogonal contrasts were used to obtain the sum of squares of the treatment of the analysis of variance in polynomial regression effects. Egg quality variables did not vary significantly between the diets. Therefore, diet supplementation with polyunsaturated fatty acids at different ratios was possible without altering egg quality. The only exception was Haugh unit, which displayed a quadratic effect indicating that the best value was the n6:n3ratio of 32, according to data significance. It was concluded that an increase in n6:n3ratios decreases laying hens' feed intake. The n6:n3 ratio of 34.64 provides greater eggs' Haugh units, decreasing from that value on. The ratios of n6:n3 did not influence the hens' serum biochemical variables.(AU)

Effects of systemic inflammation due to hepatic ischemia-reperfusion injury upon lean or obese visceral adipose tissue

Purpose: To evaluate how the induction of liver damage by ischemia and reperfusion affects the adipose tissue of lean and obese mice. Methods: Lean and diet-induced obese mice were subjected to liver ischemia (30 min) followed by 6 h of reperfusion. The vascular stromal fraction of visceral adipose tissue was analyzed by cytometry, and gene expression was evaluated by an Array assay and by RT-qPCR. Intestinal permeability was assessed by oral administration of fluorescein isothiocyanate (FITC)-dextran and endotoxemia by serum endotoxin measurements using a limulus amebocyte lysate assay. Results: It was found that, after liver ischemia and reperfusion, there is an infiltration of neutrophils, monocytes, and lymphocytes, as well as an increase in the gene expression that encode cytokines, chemokines and their receptors in the visceral adipose tissue of lean mice. This inflammatory response was associated with the presence of endotoxemia in lean mice. However, these changes were not observed in the visceral adipose tissue of obese mice. Conclusions: Liver ischemia and reperfusion induce an acute inflammatory response in adipose tissue of lean mice characterized by an intense chemokine induction and leukocyte infiltration; however, inflammatory alterations are already present at baseline in the obese adipose tissue and liver ischemia and reperfusion do not injure further.

Possibilities of Using Chia Oil As an Omega-3 Source in Laying Quail Diets Chia Oil Supplementation on Quails

This study aims to investigate the effects of different levels of chia (Salvia Hispanica L.) seed oil supplementation as omega-3 source to the diets of laying quails on yield performance, egg quality, and egg yolk fatty acid profiles. The study was conducted including 4 treatment groups as follows: C0 (0 g/kg diet), C1 (1 g/kg diet), C2 (2 g/kg diet), and C3 (3 g/kg diet). In the study, 96 female quails at the age of 18 weeks were used and the trial was carried out for 4 weeks. As a result, it was determined that chia seed oil supplementation to the diet was not effective on feed consumption, feed conversion ratio, egg yield, or egg weight of quails, but it was effective on the yolk index of egg quality characteristics (p<0.05). The most important effect of chia seed oil supplementation to the diet was determined to be on the omega-3/omega-6 ratio of egg yolk. Dietary chia seed oil increased the omega-3/omega-6 ratio (p<0.01). It can be stated that increasing the level of omega-3 in the diets, positively affects the polyunsaturated fatty acid (PUFA)/saturated fatty acid (SFA) ratio.(AU)

The Unicorn Mouse: Cranial Lipoma in a B6. Cg Mouse

Intracranial lipomas, also called intracranial lipomatous hamartomas, have been reported in some strains of research mice, but are rare in C57BL6 strains. It is presumed based on current publications that this is the first report of an intracranial lipoma in a mouse with this genetic change (B6.Cg-Cnpy2 tm1.2 Zhli Alb-Cre). Grossly, a fur-covered, soft, cylindrical, exophytic mass on the dorsal midline of the cranium was evident. Upon dissection, a soft, white, tubular structure extended through a 1mm defect in the sagittal suture of the skull to the deep surface of the hypodermis. Histologically, the mass consisted of well demarcated proliferation of mature white adipocytes, each containing one large fat droplet. The mass extended from the cerebrum at the level just caudal to the hippocampus in the third ventricle, between the superior colliculus and the caudal portion of the retrosplenial area and through the sagittal suture to the hypodermis and was surrounded by normal brain tissue.(AU)

Characterization of glutamine synthetase from the ammonium-excreting strain HM053 of Azospirillum brasilense / Caracterização da glutamina sintetase da estipe excretora de amônio HM053 de Azospirillum brasilense

Glutamine synthetase (GS), encoded by glnA, catalyzes the conversion of L-glutamate and ammonium to L-glutamine. This ATP hydrolysis driven process is the main nitrogen assimilation pathway in the nitrogen-fixing bacterium Azospirillum brasilense. The A. brasilense strain HM053 has poor GS activity and leaks ammonium into the medium under nitrogen fixing conditions. In this work, the glnA genes of the wild type and HM053 strains were cloned into pET28a, sequenced and overexpressed in E. coli. The GS enzyme was purified by affinity chromatography and characterized. The GS of HM053 strain carries a P347L substitution, which results in low enzyme activity and rendered the enzyme insensitive to adenylylation by the adenilyltransferase GlnE.

A glutamina sintetase (GS), codificada por glnA, catalisa a conversão de L-glutamato e amônio em L-glutamina. Este processo dependente da hidrólise de ATP é a principal via de assimilação de nitrogênio na bactéria fixadora de nitrogênio Azospirillum brasilense. A estirpe HM053 de A. brasilense possui baixa atividade GS e excreta amônio no meio sob condições de fixação de nitrogênio. Neste trabalho, os genes glnA das estirpes do tipo selvagem e HM053 foram clonados em pET28a, sequenciados e superexpressos em E. coli. A enzima GS foi purificada por cromatografia de afinidade e caracterizada. A GS da estirpe HM053 possui uma substituição P347L que resulta em baixa atividade enzimática e torna a enzima insensível à adenililação pela adenililtransferase GlnE.

Characterization of glutamine synthetase from the ammonium-excreting strain HM053 of Azospirillum brasilense

Abstract Glutamine synthetase (GS), encoded by glnA, catalyzes the conversion of L-glutamate and ammonium to L-glutamine. This ATP hydrolysis driven process is the main nitrogen assimilation pathway in the nitrogen-fixing bacterium Azospirillum brasilense. The A. brasilense strain HM053 has poor GS activity and leaks ammonium into the medium under nitrogen fixing conditions. In this work, the glnA genes of the wild type and HM053 strains were cloned into pET28a, sequenced and overexpressed in E. coli. The GS enzyme was purified by affinity chromatography and characterized. The GS of HM053 strain carries a P347L substitution, which results in low enzyme activity and rendered the enzyme insensitive to adenylylation by the adenilyltransferase GlnE.

Resumo A glutamina sintetase (GS), codificada por glnA, catalisa a conversão de L-glutamato e amônio em L-glutamina. Este processo dependente da hidrólise de ATP é a principal via de assimilação de nitrogênio na bactéria fixadora de nitrogênio Azospirillum brasilense. A estirpe HM053 de A. brasilense possui baixa atividade GS e excreta amônio no meio sob condições de fixação de nitrogênio. Neste trabalho, os genes glnA das estirpes do tipo selvagem e HM053 foram clonados em pET28a, sequenciados e superexpressos em E. coli. A enzima GS foi purificada por cromatografia de afinidade e caracterizada. A GS da estirpe HM053 possui uma substituição P347L que resulta em baixa atividade enzimática e torna a enzima insensível à adenililação pela adenililtransferase GlnE.

Morphophysiological and productive changes of onion under vitamin application / Alterações morfofisiológicas e produtivas da cebola sob aplicação de vitaminas

Studies have been developed to improve cultivation conditions through the introduction of beneficial compounds, among which are vitamins. The present work aimed to investigate the effects of the application of vitamins in the onion crop. For that, five treatments were used, composed of different vitamins (ascorbic acid, thiamine, niacin and pyridoxine), in a concentration of 100 mg L-¹ and a control treatment, with water application, via foliar spray. The characteristics of chlorophyll relative index, number of plant leaves, plant height, fresh leaf mass, leaf dry mass, bulb diameter, bulb mass and bulb yield were evaluated. It was found that the application of ascorbic acid and pyridoxine increased the relative levels of chlorophyll concerning the control treatment and those composed by the application of niacin and thiamine. In addition, for liquid photosynthesis, a superior result was shown when niacin and thiamine were applied, concerning the other treatments. It was concluded that the exogenous application of vitamins alters the physiological and developmental behavior of onion plants, but does not positively affect bulb yield. The increases in photosynthetic activity were not reversed in bulb development, but in the elongation of plant leaves. Finally, the use of the vitamins studied for onion cultivation is not recommended, under conditions similar to those of the present study.

Estudos vêm sendo desenvolvidos a fim melhorar as condições de cultivo através da introdução de compostos benéficos, dentre os quais encontram-se as vitaminas. O presente trabalho objetivou avaliar os efeitos da aplicação de vitaminas na cultura da cebola. Para tanto, utilizaram-se cinco tratamentos, compostos por diferentes vitaminas (ácido ascórbico, tiamina, niacina e piridoxina), em concentração de 100 mg L-¹ e um tratamento controle, com aplicação de água, via pulverização foliar. Foram avaliadas as características de teor relativo de clorofila, número de folhas das plantas, altura de planta, massa fresca de folhas, massa seca de folhas, diâmetro de bulbo, massa de bulbo e a produtividade de bulbos. Verificou-se que a aplicação do ácido ascórbico e da piridoxina elevaram os teores relativos de clorofila em relação ao tratamento controle e aqueles compostos pela aplicação de niacina e tiamina. Em complemento, para a fotossíntese líquida foi evidenciado resultado superior quando aplicadas as vitaminas niacina e tiamina, em relação aos demais tratamentos. Concluiu-se que a aplicação exógena de vitaminas altera o comportamento fisiológico e de desenvolvimento das plantas de cebola, porém não afetando positivamente a produtividade de bulbos. Os incrementos quanto à atividade fotossintética não foram revertidos em desenvolvimento dos bulbos, mas sim em um alongamento das folhas das plantas. Por fim, não se indica a utilização das vitaminas estudadas para a cultura da cebola, em condições semelhantes ao do presente estudo.

Characterization of glutamine synthetase from the ammonium-excreting strain HM053 of Azospirillum brasilense / Caracterização da glutamina sintetase da estipe excretora de amônio HM053 de Azospirillum brasilense

Glutamine synthetase (GS), encoded by glnA, catalyzes the conversion of L-glutamate and ammonium to L-glutamine. This ATP hydrolysis driven process is the main nitrogen assimilation pathway in the nitrogen-fixing bacterium Azospirillum brasilense. The A. brasilense strain HM053 has poor GS activity and leaks ammonium into the medium under nitrogen fixing conditions. In this work, the glnA genes of the wild type and HM053 strains were cloned into pET28a, sequenced and overexpressed in E. coli. The GS enzyme was purified by affinity chromatography and characterized. The GS of HM053 strain carries a P347L substitution, which results in low enzyme activity and rendered the enzyme insensitive to adenylylation by the adenilyltransferase GlnE.(AU)

A glutamina sintetase (GS), codificada por glnA, catalisa a conversão de L-glutamato e amônio em L-glutamina. Este processo dependente da hidrólise de ATP é a principal via de assimilação de nitrogênio na bactéria fixadora de nitrogênio Azospirillum brasilense. A estirpe HM053 de A. brasilense possui baixa atividade GS e excreta amônio no meio sob condições de fixação de nitrogênio. Neste trabalho, os genes glnA das estirpes do tipo selvagem e HM053 foram clonados em pET28a, sequenciados e superexpressos em E. coli. A enzima GS foi purificada por cromatografia de afinidade e caracterizada. A GS da estirpe HM053 possui uma substituição P347L que resulta em baixa atividade enzimática e torna a enzima insensível à adenililação pela adenililtransferase GlnE.(AU)

Characterization of glutamine synthetase from the ammonium-excreting strain HM053 of Azospirillum brasilense / Caracterização da glutamina sintetase da estipe excretora de amônio HM053 de Azospirillum brasilense

Glutamine synthetase (GS), encoded by glnA, catalyzes the conversion of L-glutamate and ammonium to L-glutamine. This ATP hydrolysis driven process is the main nitrogen assimilation pathway in the nitrogen-fixing bacterium Azospirillum brasilense. The A. brasilense strain HM053 has poor GS activity and leaks ammonium into the medium under nitrogen fixing conditions. In this work, the glnA genes of the wild type and HM053 strains were cloned into pET28a, sequenced and overexpressed in E. coli. The GS enzyme was purified by affinity chromatography and characterized. The GS of HM053 strain carries a P347L substitution, which results in low enzyme activity and rendered the enzyme insensitive to adenylylation by the adenilyltransferase GlnE.

A glutamina sintetase (GS), codificada por glnA, catalisa a conversão de L-glutamato e amônio em L-glutamina. Este processo dependente da hidrólise de ATP é a principal via de assimilação de nitrogênio na bactéria fixadora de nitrogênio Azospirillum brasilense. A estirpe HM053 de A. brasilense possui baixa atividade GS e excreta amônio no meio sob condições de fixação de nitrogênio. Neste trabalho, os genes glnA das estirpes do tipo selvagem e HM053 foram clonados em pET28a, sequenciados e superexpressos em E. coli. A enzima GS foi purificada por cromatografia de afinidade e caracterizada. A GS da estirpe HM053 possui uma substituição P347L que resulta em baixa atividade enzimática e torna a enzima insensível à adenililação pela adenililtransferase GlnE.

Use of pyridoxine hydrochloride in the interruption of lactation in female dogs with pseudopregnancy

Abstract The aim of this research was to evaluate the use of pyridoxine hydrochloride and its associated side effects in the treatment of pseudopregnancy in female dogs. A total of 40 female dogs, with no defined breed, in non-gestational diestrus, with clinical complaint of milk production were selected. The female dogs were divided into four experimental groups of 10 animals each, treated orally for 20 days with 10mg/kg/day (G1) and 50mg/kg/day (G2) of pyridoxine hydrochloride (vitamin B6), 5μg/kg/day of cabergoline (G3), and with a placebo, in the case of the control group (G4). The effects of the treatments on milk production were investigated, as well as possible systemic side effects, macroscopic uterine and ovarian alterations, and uterine histology. During the investigated period, G2 and G3 were equally efficient (P>0.05) in lactation suppression, differing (P>0.05) from the other groups. There were no systemic side effects or uterine changes associated with administration of the studied drug. Vitamin B6 (50mg/kg) has shown to be a safe and economically viable alternative for lactation interruption in female dogs with pseudopregnancy.

Use of pyridoxine hydrochloride in the interruption of lactation in female dogs with pseudopregnancy

The aim of this research was to evaluate the use of pyridoxine hydrochloride and its associated side effects in the treatment of pseudopregnancy in female dogs. A total of 40 female dogs, with no defined breed, in non-gestational diestrus, with clinical complaint of milk production were selected. The female dogs were divided into four experimental groups of 10 animals each, treated orally for 20 days with 10mg/kg/day (G1) and 50mg/kg/day (G2) of pyridoxine hydrochloride (vitamin B6), 5μg/kg/day of cabergoline (G3), and with a placebo, in the case of the control group (G4). The effects of the treatments on milk production were investigated, as well as possible systemic side effects, macroscopic uterine and ovarian alterations, and uterine histology. During the investigated period, G2 and G3 were equally efficient (P>0.05) in lactation suppression, differing (P>0.05) from the other groups. There were no systemic side effects or uterine changes associated with administration of the studied drug. Vitamin B6 (50mg/kg) has shown to be a safe and economically viable alternative for lactation interruption in female dogs with pseudopregnancy.(AU)

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus)

ABSTRACT: This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.

RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)