RESUMO
The microbiological and biochemical properties of a goat cheese produced using Helianthus annuus (sunflower) seed extract as a coagulant and the potentially probiotic autochthonous culture Limosilactobacillus mucosae CNPC007 were examined in comparison to a control cheese devoid of the autochthonous culture. Throughout a 60-day storage period at 6 ± 1 °C, lactobacilli maintained a count of above 8 log CFU/g. Additionally, its viability in cheeses subjected to the in vitro gastrointestinal conditions demonstrated improvement over this period. Specifically, the recovery of lactobacilli above 6 log CFU/g was observed in 16.66% of the samples in the first day, increasing to 66.66% at both 30 and 60 days. While total coliforms were detected in both cheese trials, this sanitary parameter exhibited a decline in L. mucosae cheeses during storage, falling below the method threshold (<3 MPN/g) at 60 days. This observation suggests a potential biopreservative effect exerted by this microorganism, likely attributed to the higher acidity of L. mucosae cheeses at that point (1.80 g/100 g), which was twice that of the control trial (0.97 g/100 g). Furthermore, distinct relative proportions of >30 kDa, 30-20 kDa, and <20 kDa proteins during storage was verified for L. mucosae and control cheeses. Consequently, either the H. annuus seed extract or the L. mucosae CNPC007 autochthonous culture influenced the biochemical properties of the cheese, particularly in terms of proteolysis. Moreover, L. mucosae CNPC007 acidification property resulted in a biopreservative effect throughout the storage period, indicating the potential as a promising source of probiotics for this product.
RESUMO
The objective was to evaluate the effect of detoxified castor bean replacing soybean meal in the concentrate diet or as nitrogen organic fertilizer replacing urea on intake and nutrient digestibility, blood parameters and productive performance of sheep finished on irrigated Tamani grass pasture under continuous stocking and variable stocking rate. The treatments were two concentrate diets: standard (ground corn and soybean meal) and alternative diet (ground corn and detoxified castor bean cake), and two nitrogen fertilizers: chemical (urea) and organic (fresh castor bean cake). The randomized complete block design was used in a 2 × 2 factorial arrangement with four replications (500 m² paddocks). Four sheep (2 castrated males and 2 females) were distributed in each experimental unit, totaling 64 animals with an average initial weight of 19.42 ± 3.6 kg. No effects (P > 0.05) were observed on the variables inherent to the evaluation of the pasture. The average stocking rate (SR) among treatments was 85.50 sheep/ha, equivalent to 9.87 Animal Units (AU)/ha. The alternative diet presented lower dry matter digestibility (62.71%), with no negative effects on nutrient intake and kidney parameters. Animals fed the standard and alternative diet showed average daily gain of 103.75 and 86.76 g/day, respectively. A finishing period of up to 100 days is recommended for sheep selected for production systems in semi-arid regions managed intensively on pasture. Detoxified castor bean cake did not alter nutrient intake, liver and kidney parameters of the sheep and can be used in pasture-based sheep farming.
Assuntos
Fertilizantes , Ricinus communis , Animais , Feminino , Masculino , Suplementos Nutricionais , Glycine max , Nitrogênio , Ovinos , UreiaRESUMO
The purpose of this study was to evaluate castor bean cake as alternative input in the control of gastrointestinal parasites in sheep raised on irrigated pasture under continuous stocking. The treatments consisted of sheep supplemented with standard diet and pasture fertilized with urea; sheep supplemented with alternative diet and pasture fertilized with urea; sheep supplemented with standard diet and pasture fertilized with in natura castor bean cake; and sheep supplemented with alternative diet and pasture fertilized with in natura castor bean cake. A randomized complete block design (CBD) was used, with 16 replications (sheep), with repeated measures over time, the plots being the treatments, and the subplots the collection times. Infective nematode larvae in the pasture (L3.g DM-1), number of eggs per gram of feces (EPG), globular volume (GV), and total plasma protein (TPP) were evaluated. For adult gastrointestinal parasite counts, CBD was also used with six replications. Except for FAMACHA© grade, all variables had effect (P < 0.01) of the time factor. The average number of L3.g DM-1 and EPG were 126 and 841, respectively, with no effect (P > 0.05) of the treatment factor. The values observed for GV and TPP were higher than 25.9% and 6.4 g·dL-1, respectively, which were considered normal. As organic fertilizer, the fractionated application of in natura castor bean cake does not reduce the contamination of pastures by nematode larvae. The evaluated feeds improve the resilience of the sheep to infection by gastrointestinal parasites. The treatments using castor bean cake reduced the adult parasites present in the abomasum of sheep.
Assuntos
Parasitos , Ricinus communis , Animais , Proteínas Sanguíneas , Fertilizantes , Larva , Óvulo , Ovinos , UreiaRESUMO
Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P<0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.
Assuntos
Criação de Animais Domésticos , Infecções por Nematoides , Doenças dos Ovinos , Solo , Criação de Animais Domésticos/métodos , Animais , Ricinus communis/química , Fezes , Fertilizantes/parasitologia , Hematócrito , Nematoides , Infecções por Nematoides/imunologia , Infecções por Nematoides/prevenção & controle , Infecções por Nematoides/veterinária , Contagem de Ovos de Parasitas/veterinária , Ovinos , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/prevenção & controle , Solo/parasitologia , Aumento de PesoRESUMO
BACKGROUND: In Brazil, over the last few years there has been an increase in the production and consumption of goat cheeses. In addition, there was also a demand to create options to use the whey extracted during the production of cheeses. Whey can be used as an ingredient in the development of many products. Therefore, knowing its composition is a matter of utmost importance, considering that the reference methods of food analysis require time, trained labor and expensive reagents for its execution. METHODS: Goat whey samples produced in winter and summer were submitted to proximate composition analysis (moisture, total solids, ashes, proteins, fat and carbohydrates by difference) using reference methods and near infrared spectroscopy (NIRS). The spectral data was preprocessed by baseline correction and the Savitzky-Golay derivative. The models were built using Partial Least Square Regression (PLSR) with raw and preprocessed data for each dependent variable (proximate composition parameter). RESULTS: The average whey composition values obtained using the referenced methods were in accordance with the consulted literature. The composition did not differ significantly (p > 0.05) between the summer and winter whey samples. The PLSR models were made available using the following figures of merit: coefficients of determination of the calibration and prediction models (R 2cal and R 2pred, respectively) and the Root Mean Squared Error Calibration and Prediction (RMSEC and RMSEP, respectively). The best models used raw data for fat and protein determinations and the values obtained by NIRS for both parameters were consistent with their referenced methods. Consequently, NIRS can be used to determine fat and protein in goat whey.
RESUMO
Cationic peptides found in Moringa oleifera seeds belong to different protein families and are described as the main flocculating agents of the species. In this study we report the identification and isolation of four new flocculant peptides, called Mo-HLPs 1-4, belonging to the family of hevein-like peptides, previously only known for their members' antimicrobial activity. Purification of the peptides followed two sequential membrane ultrafiltration steps and separation by reverse-phase liquid chromatography. Proteomic analyses showed that Mo-HLPs are extremely basic (pI >10) cysteine-rich molecules with molecular masses between 4.5 and 4.8 kDa and with a highly conserved chitin-binding domain. Searches in BLAST revealed high similarity of Mo-HLPs with hevein and other hevein-like peptides and 90% identity with morintides, which are members of the 8C-hevein-like subfamily found in M. oleifera leaves. Mo-HLPs microflocculation assays showed distinct coagulation/flocculation efficiencies, promoting turbidity reduction levels between 67 and 89% in synthetic turbid water. Activity variations were attributed to the substitution of some amino acids among the isoforms, which may have altered the final net charge of the molecules. The identification of Mo-HLPs represents the discovery of a new group of cationic peptides involved in the flocculation properties of M. oleifera seeds. SIGNIFICANCE: The study reveals the presence of hevein-like peptides in Moringa oleifera seeds. It is reported for the first time that members of this family have properties to act as flocculating agents of importance for water treatment processes. The identification of these peptides as well as new functional assignment broadens the horizon for speculation on new species which could act as sources of green coagulants for sustainable water treatment, and contributes to the knowledge about occurrence, distribution, molecular and active diversity of peptides belonging to the hevein-like family.
Assuntos
Moringa oleifera , Peptídeos Catiônicos Antimicrobianos , Lectinas de Plantas , Proteínas de Plantas , Proteômica , SementesRESUMO
The genus Streptococcus includes various species, remarkably different in their behavior, applications, virulence, and safety. Taxonomically Streptococcus infantarius subsp. infantarius belonging to the Streptococcus bovis group, which includes several pathogen species, however, has been found as predominant species in some African dairy products that are widely consumed and considered to be safe. Streptococcus infantarius subsp. infantarius' safety may be questioned due to the association of this species with clinical cases. In this study, isolates from dairy origin were selected based on their bacteriocinogenic potential and differentiated by their RAPD-PCR profiles. Two strains were identified by 16S rRNA sequencing as St. infantarius subsp. infantarius and investigated regarding their potential beneficial properties and factors related to virulence and safety. A series of in vitro tests included properties related to survival in the gastrointestinal tract and beneficial intestinal activities. Production of bacteriocin/s, detection of related genes, and partial characterization of expressed antimicrobial protein were evaluated. Genes related to folate biosynthesis were detected in both studied strains. Evaluation of physiological tests related to strains virulence, adhesion, and resistance to antibiotics and detections of virulence and biogenic amines production-related genes were also investigated. Taking in consideration all the aspects of the specific nature of St. infantarius subsp. infantarius K1-4 and K5-1 (beneficial properties and virulence characteristics), both strains cannot be considered safe for human or other animals application, even though they have been isolated from dairy products. This study is highlighting the importance of evaluation for presence of potential virulence factors in newly characterized strains in order to be confident in their safety.
Assuntos
Bacteriocinas/farmacologia , Leite/microbiologia , Streptococcus/genética , Streptococcus/patogenicidade , Fatores de Virulência/farmacologia , Animais , Antibacterianos/farmacologia , Aderência Bacteriana/efeitos dos fármacos , Bacteriocinas/biossíntese , Materiais Biomiméticos/farmacologia , Brasil , Farmacorresistência Bacteriana/genética , Escherichia coli/efeitos dos fármacos , Escherichia coli/crescimento & desenvolvimento , Feminino , Ácido Fólico/biossíntese , Suco Gástrico/química , Cabras , Humanos , Listeria/efeitos dos fármacos , Listeria/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Filogenia , RNA Ribossômico 16S/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Salmonella/efeitos dos fármacos , Salmonella/crescimento & desenvolvimento , Staphylococcus/efeitos dos fármacos , Staphylococcus/crescimento & desenvolvimento , Streptococcus/efeitos dos fármacos , Streptococcus/isolamento & purificação , Virulência , Fatores de Virulência/biossínteseRESUMO
Abstract Gastrointestinal parasitism is one of the factors that discourages farmers from raising small ruminants in cultivated pastures. To validate a soil treatment strategy to control the free-living stages of gastrointestinal nematodes (GIN), castor cake (CC) was used as a fertilizer on a pasture where sheep grazed on guinea grass under continuous stocking. On day zero, the pasture was divided into three paddocks, contaminated by GIN and treated, respectively, with CC divided into two applications (2CC1/2), CC in a single application (CC1) and organic compost in a single application (control). On day 21, eight GIN-free sheep were placed in each paddock. On day 58, significant differences (P<0.05) were observed: reduction of up to 66.10% in larvae.g-1 of dry mass in pastures fertilized with CC, decrease of up to 60.72% in infection rates among the animals in the groups treated with CC, higher average daily weight gain (over 185 g.day-1) and packed cell volume (over 26%) in the groups treated with CC, when compared to the control (128 g.day-1; 20.9%). In view of the results, the use of CC, mainly CC1, as a fertilizer for guinea grass pastures, under continuous stocking, proved to be promising, with 63.41% effectiveness in controlling worm infestations.
Resumo O parasitismo gastrintestinal é um dos fatores que fragiliza a exploração de pequenos ruminantes em pastagens cultivadas. Objetivando validar a estratégia de tratamento do solo para o controle dos estágios de vida livre de nematoides gastrintestinais (NGI), a torta de mamona (TM) foi utilizada como adubo, com ovinos pastejando em capim-tanzânia sob lotação contínua. No dia zero, o pasto foi dividido em três piquetes, contaminados por NGI e tratados, respectivamente, com TM parcelada em duas aplicações (2TM1/2), TM em uma única aplicação (TM1) e composto orgânico em única aplicação (testemunha). No dia 21, cada piquete recebeu oito ovinos livres de NGI. No dia 58, observaram-se diferenças significativas (P<0,05): redução de até 66,10% de larvas.g-1 de massa seca nas pastagens adubadas com TM; redução de até 60,72% da infecção dos animais nos grupos tratados com TM; ganho de peso médio diário (acima de 185 g.dia-1) e volume globular (acima de 26%) superior nos grupos tratados com TM, quando comparados com a testemunha (128 g.dia-1; 20,9%). Diante dos resultados, o uso da TM, principalmente TM1, como adubo em pasto de capim-tanzânia, sob lotação contínua, mostrou-se promissor, com eficácia de 63,41% para controlar a verminose.
Assuntos
Animais , Doenças dos Ovinos/imunologia , Doenças dos Ovinos/prevenção & controle , Criação de Animais Domésticos/métodos , Infecções por Nematoides/imunologia , Infecções por Nematoides/prevenção & controle , Infecções por Nematoides/veterinária , Contagem de Ovos de Parasitas/veterinária , Solo/parasitologia , Ovinos , Aumento de Peso , Ricinus communis/química , Fezes , Fertilizantes/parasitologia , Hematócrito , NematoidesRESUMO
The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P < 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P < 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.
Assuntos
Produção Agrícola , Sucos de Frutas e Vegetais , Nematoides/efeitos dos fármacos , Óleos de Plantas/farmacologia , Poaceae/parasitologia , Eliminação de Resíduos , Ricinus/química , Animais , RuminantesRESUMO
Abstract The nematicidal effect of different organic materials was evaluated in order to develop a non-chemical alternative soil treatment for control of the free-living stages of small ruminant gastrointestinal nematodes. The selected organic materials were residues from the juice industry of acerola, cashew, grape, guava, papaya and pineapple, as well as castor residue from the biodiesel industry. LC90 results showed that pineapple residue was the most efficient inhibitor of larval development, followed by castor, grape, cashew, acerola, guava and papaya. Castor residue was also a good source of nitrogen and was used in a greenhouse experiment to prevent larval development in contaminated goat faeces that was deposited in pots containing the grasses Brachiaria brizantha (var. Paiaguás) or Megathyrsus maximus x M. infestum (var. Massai). Castor residue caused a significant (P < 0.05) reduction (85.04%) in Paiaguás grass contamination (L3.dry mass-1) and a reduction of 17.35% in Massai grass contamination (P > 0.05), with an increase in the biomass production of Massai (251.43%, P < 0.05) and Paiaguás (109.19%, P > 0.05) grasses. This strategy, called Econemat®, with good results in vitro shows to be promising on pasture increasing phytomass production.
Resumo O efeito nematicida de diferentes materiais orgânicos foi avaliado, com o objetivo de desenvolver um tratamento alternativo não químico do solo para o controle dos estágios de vida livre de nematoides gastrintestinais de pequenos ruminantes. Os materiais orgânicos selecionados foram resíduos da indústria de suco de acerola, caju, uva, goiaba, mamão e abacaxi, além do resíduo de mamona da indústria de biodiesel. Os resultados da LC90 mostraram que o resíduo de abacaxi foi o mais eficiente inibidor do desenvolvimento larval, seguido pela mamona, uva, caju, acerola, goiaba e mamão. O resíduo de mamona também se mostrou uma boa fonte de nitrogênio, sendo usado em experimento em casa de vegetação para prevenir o desenvolvimento larval em fezes contaminadas, depositadas em vasos, contendo as gramíneas Brachiaria brizantha (var. Paiaguás) ou Megathyrsus maximus x M. infestum (var. Massai). O resíduo de mamona reduziu (85,04%) significativamente (P < 0,05) a contaminação no capim-Paiaguás (L3.dry massa-1), 17,35% no capim Massai (P> 0,05), e aumentou a produção de biomassa das gramíneas Massai (251,43%, P <0,05) e Paiaguás (109,19%, P> 0,05). Essa estratégia, chamada Econemat®, com bons resultados in vitro, mostra-se promissora nas pastagens aumentando a produção de fitomassa.
Assuntos
Animais , Ricinus/química , Óleos de Plantas/intoxicação , Eliminação de Resíduos , Sucos de Frutas e Vegetais , Produção Agrícola , Poaceae/parasitologia , Nematoides/efeitos dos fármacos , RuminantesRESUMO
Enabling the use of castor cake in animal feeding is an excellent alternative strategy to reduce feed costs. The cake is a by-product derived from the extraction of the castor oil by the biodiesel industry whose chemical composition is satisfactory despite the presence of antinutritional factors like toxic lectins, which require detoxification before it can be used as a dietary ingredient. The aim of the present study was to evaluate alternative chemical sources in the degradation and inactivation of ricin and Ricinus communis agglutinin (RCA), two lectins from castor cake. Ten chemical compounds were evaluated: sodium hydroxide, monodicalcium phosphate, dicalcium phosphate, calcium oxide, calcium hydroxide, calcitic limestone, magnesian limestone, urea, potassium chloride, and sodium chloride. Gel electrophoresis indicated 100% lectin degradation only in the cakes treated with 90â¯g sodium hydroxide and 2500â¯mL water per kg of cake. The hemagglutination assay was crucial to providing innocuousness to the treated cakes, with total absence of hemagglutinating activity observed in the castor cakes treated with 60 or 90â¯g sodium hydroxide in water volumes equal to or higher than 1500â¯mL/kg of castor cake and in the cakes treated with 90â¯g calcium oxide with 2500 or 3000â¯mL water/kg castor cake. Thus, though depending on the concentration of the chemical compound and on the volume of water per kilogram of treated cake, sodium hydroxide and calcium oxide showed to be promising chemical products for degradation and complete inactivation of the lectins present in castor cake to allow its use as an ingredient in animal diets.
Assuntos
Lectinas de Plantas/química , Ricina/química , Ricinus communis/química , Ração Animal , Animais , Compostos de Cálcio/química , Eritrócitos , Testes de Hemaglutinação , Óxidos/química , Coelhos , Hidróxido de Sódio/químicaRESUMO
The potential for reusability of whey is of concern due to its substantial nutritional value and, second, in view of the need to reduce environmental impact. The use of goat whey powder in the production of a fermented dairy beverage using a Lactobacillus casei culture as adjunct shows a good prospect of a food product with hypotensive activity. This study investigated the microbial viability, proteolysis and angiotensin-converting enzyme (ACE) inhibitory activity of a fermented dairy beverage produced with goat whey powder and a probiotic culture of L. casei BGP93 co-cultured with Streptococcus thermophilus TA-40. The probiotic beverage exhibited no significant difference from the control (absence of L. casei) with regard to titratable acidity, S. thermophilus viability and proteolysis degree during 21 days of storage (P > 0.05). During this period, the beverage maintained L. casei at appropriate levels (> 7 log cfu ml-1), thereby qualifying as a potential probiotic product. Although both control and probiotic beverages exhibited ACE inhibitory activity, as a result of proteolysis of whey proteins during fermentation, significant increased ACE inhibitory activity was found for the beverage with added probiotic L. casei (P < 0.05). The probiotic beverage has potency as a functional food candidate to be included in a dietary strategy aiming at prevention and control of hypertension.
Assuntos
Inibidores da Enzima Conversora de Angiotensina/metabolismo , Produtos Fermentados do Leite/microbiologia , Lacticaseibacillus casei/metabolismo , Streptococcus thermophilus/metabolismo , Proteínas do Soro do Leite/metabolismo , Inibidores da Enzima Conversora de Angiotensina/química , Animais , Bebidas/análise , Bebidas/microbiologia , Técnicas de Cocultura , Produtos Fermentados do Leite/análise , Fermentação , Cabras , Lacticaseibacillus casei/crescimento & desenvolvimento , Leite/química , Leite/microbiologia , Peptidil Dipeptidase A/química , Proteólise , Streptococcus thermophilus/crescimento & desenvolvimento , Proteínas do Soro do Leite/químicaRESUMO
The search for probiotic candidates among lactic acid bacteria (LAB) isolated from food may uncover new strains with promising health and technological properties. Lactobacillus mucosae strains attracted recent research attention due to their ability to adhere to intestinal mucus and to inhibit pathogens in the gastrointestinal tract, both related to a probiotic potential. Properties of interest and safety aspects of three Lb. mucosae strains (CNPC006, CNPC007, and CNPC009) isolated from goat milk were investigated employing in vitro tests. The presence of genetic factors related to bile salt hydrolase production (bsh), intestinal adhesion properties (msa, map, mub, and ef-tu), virulence, and biogenic amine production were also verified. All strains exhibited the target map, mub, and ef-tu sequences; the msa gene was detected in CNPC006 and CNPC007 strains. Some of the searched sequences for virulence factors were detected, especially in the CNPC009 strain; all strains carried the hyl gene, related to the production of hyaluronidase. Lb. mucosae CNPC007 exhibited a high survival rate in simulated gastric and enteric conditions. Besides, all strains exhibited the bsh sequence, and CNPC006 and CNPC007 were able to deconjugate salts of glycodeoxycholic acid (GDC). Regarding technological properties for dairy product applications, a relatively higher milk acidification and clotting capacity, diacetyl production, and proteolytic activity were registered for CNPC007 in comparison to the other strains. Collectively, the results aim at Lb. mucosae CNPC007 as a promising probiotic candidate for application in dairy products, deserving further studies to confirm and explore its potential.
Assuntos
Genes Bacterianos , Cabras , Lactobacillus/fisiologia , Leite/microbiologia , Animais , Antibacterianos/farmacologia , Aderência Bacteriana/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Ácidos e Sais Biliares/metabolismo , Aminas Biogênicas/biossíntese , Brasil , Farmacorresistência Bacteriana Múltipla , Microbiologia de Alimentos , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/microbiologia , Gelatinases/biossíntese , Interações Hidrofóbicas e Hidrofílicas , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Probióticos/isolamento & purificação , Fatores de Virulência/genéticaRESUMO
This study aimed to characterize the safety and technological properties of Enterococcus faecium strains isolated from Brazilian Coalho cheeses. High levels of co-aggregation were observed between Enterococcus faecium strains EM485 and EM925 and both Escherichia coli and Clostridium perfringens . Both strains presented low levels of hydrophobicity. E. faecium EM485 and EM925 were both able to grow in the presence of 0.5% of the sodium salts of taurocholic acid (TC), taurodeoxycholic acid (TDC), glycocholic acid (GC), and glycodeoxycholic acid (GDC), although they showed the ability to deconjugate only GDC and TDC. Both strains showed good survival when exposed to conditions simulating the gastro intestinal tract (GIT). When tested for the presence of virulence genes, only tyrosine decarboxylase and vancomycin B generated positive PCR results.
Assuntos
Queijo/microbiologia , Enterococcus faecium/isolamento & purificação , Enterococcus faecium/fisiologia , Manipulação de Alimentos/métodos , Inocuidade dos Alimentos , Aderência Bacteriana , Brasil , Fenômenos Químicos , Ácidos Cólicos/metabolismo , Ácidos Cólicos/toxicidade , Clostridium perfringens/química , Clostridium perfringens/fisiologia , Enterococcus faecium/química , Escherichia coli/química , Escherichia coli/fisiologia , Trato Gastrointestinal/química , Interações Hidrofóbicas e Hidrofílicas , Inativação Metabólica , Viabilidade Microbiana/efeitos dos fármacos , Reação em Cadeia da Polimerase , Fatores de Virulência/análise , Fatores de Virulência/genéticaRESUMO
This study aimed to characterize the safety and technological properties of Enterococcus faecium strains isolated from Brazilian Coalho cheeses. High levels of co-aggregation were observed between Enterococcus faecium strains EM485 and EM925 and both Escherichia coli and Clostridium perfringens. Both strains presented low levels of hydrophobicity. E. faecium EM485 and EM925 were both able to grow in the presence of 0.5% of the sodium salts of taurocholic acid (TC), taurodeoxycholic acid (TDC), glycocholic acid (GC), and glycodeoxycholic acid (GDC), although they showed the ability to deconjugate only GDC and TDC. Both strains showed good survival when exposed to conditions simulating the gastro intestinal tract (GIT). When tested for the presence of virulence genes, only tyrosine decarboxylase and vancomycin B generated positive PCR results.
Assuntos
Queijo/microbiologia , Enterococcus faecium/isolamento & purificação , Enterococcus faecium/fisiologia , Inocuidade dos Alimentos , Manipulação de Alimentos/métodos , Aderência Bacteriana , Brasil , Fenômenos Químicos , Ácidos Cólicos/metabolismo , Ácidos Cólicos/toxicidade , Clostridium perfringens/química , Clostridium perfringens/fisiologia , Enterococcus faecium/química , Escherichia coli/química , Escherichia coli/fisiologia , Trato Gastrointestinal/química , Interações Hidrofóbicas e Hidrofílicas , Inativação Metabólica , Viabilidade Microbiana/efeitos dos fármacos , Reação em Cadeia da Polimerase , Fatores de Virulência/análise , Fatores de Virulência/genéticaRESUMO
No presente estudo objetivou-se avaliar a degradação do bagaço de cana-de-açúcar (BCA) integral (BIN) ou hidrolisado (BH) pela microbiota ruminal de caprinos e ovinos de raças naturalizadas do Nordeste brasileiro e o potencial desses animais como fontes de microrganismos e/ou enzimas celulolíticas para degradação da fibra do BCA. Para hidrólise do BCA foi utilizada uma solução de NaOH a 50%, 30% na matéria seca (MS). Foram determinadas as concentrações de MS, proteína bruta (PB), cinzas (CZ), fibra detergente neutro (FDN), fibra detergente ácido (FDA), celulose (Cel), hemicelulose (Hcel) e lignina (Lig). A degradação in situ da FDN do BH foi determinada pela incubação ruminal em sacos de náilon nos tempos: 0, 6, 24 e 96 horas. A técnica de duas etapas preconizada por Tilley e Terry foi utilizada para determinar a digestibilidade in vitro da MS (DIVMS). Foi coletado conteúdo ruminal dos animais quatro horas após a infusão de 100g de BIN via fístula ruminal para a separação de microrganismos associados às fases líquida e sólida, utilizando filtragem e extração com tampão fosfato de sódio 50 mM e pH 6,9, respectivamente. A fração sólida foi submetida ao cultivo in vitro com o substrato BIN, por 96 horas, para determinação da taxa de degradação da MS, FDN e atividade celulolítica. O pH foi determinado nos tempos de cultivo de 24, 48 e 96 horas. O pré-tratamento com NaOH aumentou a DIVMS (P<0,05). Não houve efeito de espécie sobre a DIVMS (P>0,05). A inclusão de NaOH aumentou a degradação in situ da FDN, apresentando os ovinos menor tempo de colonização (TC). Houve solubilização da Hcel, da Cel e da Lig no BCA pré-tratado com NaOH. A atividade celulolítica se concentrou na fração sólida independente da espécie doadora do inóculo, sendo observado crescimento microbiano no cultivo in vitro do BIN a partir dessa fração. O pH aumentou com o tempo de cultivo in vitro. Microrganismos ruminais de caprinos e ovinos naturalizados do Nordeste brasileiro colonizaram e degradaram o BIN e BH. O NaOH pode ser utilizado no pré-tratamento alcalino do BCA.
The purpose of this study was to evaluate the biodegradation of sugar cane bagasse (SCB), whole (WB) or hydrolyzed (HB) by ruminal microorganisms from Brazilian naturalized goat and sheep breeds, and the animals' potential as a source of microorganisms and/or cellulolytic enzymes for the biodegradation of SCB fiber. SCB hydrolysis was performed using 50% sodium hydroxide solution on 30% of dry matter (DM). The concentrations of DM, crude protein (CP), ether extract (EE), ash, neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose (Cel), hemicellulose (Hcel) and lignin (Lig) were determined in the WB and HB. For the in situ biodegradation of NDF, the HB was placed in nylon bags and incubated in rumen for 0, 6, 24 and 96 hours. The two-step technique recommended by Tilley and Terry was used to determine the in vitro digestibility of DM (IVDDM). Four hours after infusion of WB (100 g) through the ruminal fistula, the rumen content was collected from the animals and the microorganisms associated with the liquid and solid phases were separated, the former by filtration and the latter by extraction with 50 mM sodium phosphate buffer, pH 6.9. The solid fraction was subjected to in vitro culture with WB substrate for 96 hours to determine the biodegradation rate of DM, NDF and cellulolytic activity. The pH was measured after culture times of 24, 48 and 96 hours. Pretreatment with NaOH increased the IVDDM (P <0.05). No species effect on the IVDDM was detected (P> 0.05). The addition of NaOH increased the in situ biodegradation rate of NDF, with sheep showing a lower lag time (LT). There was a solubilization of Hcel, Cel and Lig in SCB pretreated with NaOH. Cellulolytic activity was higher in the solid phase, irrespective of the inoculum source, and microbial growth was observed in in vitro cultures using microorganisms from the solid phase and WB as substrate. The pH of the in vitro culture increased over time. Ruminal microorganisms from Brazilian naturalized goat and sheep breeds successfully colonized and biodegraded both WB and HB. Sodium hydroxide proved useful as an alkaline pretreatment for SCB.
Assuntos
Hidróxido de Sódio , Ruminantes , Ovinos , Saccharum , EnzimasRESUMO
Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (<12 kDa) for Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P<0.05, Bonferroni test). The two fractions of Crotalaria spectabilis showed the same ovicidal activity (P>0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.
Assuntos
Haemonchus/efeitos dos fármacos , Proteínas Hemolisinas/farmacologia , Lectinas/farmacologia , Óvulo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Inibidores de Proteases/farmacologia , Sementes , AnimaisRESUMO
Bioactive molecules of plant species are promising alternatives for the chemical control of gastrointestinal nematodes in ruminants. Extracts of native and exotic seed species from Brazil's semi-arid region were tested in vitro in an egg hatch assay and the bioactivity of their proteins was investigated. Each seed species was subjected to three extractions with three types of solvents. All the seeds showed ovicidal activity, which varied according to the solvents. Higher ovicidal activity was found in the molecule fractions of low molecular weight (<12 kDa) for Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera and Ricinus communis (P<0.05, Bonferroni test). The two fractions of Crotalaria spectabilis showed the same ovicidal activity (P>0.05, Bonferroni test). Hemagglutinating activity was detected in the fractions of C. spectabilis and M. oleifera fractions, hemolysin activity in the A. lebbeck and M. oleifera fractions, serine protease inhibitory activity in the A. lebbeck, I. asarifolia, J. curcas, M. oleifera and R. communis fractions, cysteine protease inhibitor activity in the M. oleifera fraction, and no protein activity in the L. ferrea fraction. The results of this work reveal new plant species with a potential for use in controlling nematode parasites in goats, thus opening a new field of research involving plant protein molecules with ovicidal properties.
Moléculas bioativas de espécies vegetais são alternativas promissoras ao controle químico dos nematoides gastrintestinais em ruminantes. Extratos de sementes de espécies nativas e exóticas do Semiárido Brasileiro foram testados in vitro em ensaio de eclosão de ovos e investigada a natureza proteica da bioatividade. Três extrações com três solventes foram feitas para cada semente estudada. Todas as sementes apresentaram atividade ovicida, variando com o solvente utilizado. Maior taxa de inibição da eclosão concentrou-se nas frações de moléculas de baixa massa molecular (<12 kDa) para Albizia lebbeck, Ipomoea asarifolia, Jatropha curcas, Libidibia ferrea, Moringa oleifera e Ricinus communis (P<0,05, teste de Bonferroni). Crotalaria spectabilis mostrou atividade nas duas frações, sem diferença entre elas (P>0,05, teste de Bonferroni). Observou-se atividade hemaglutinante nas frações de C. spectabilis e M. oleifera, de hemolisina em A. lebbeck e M. oleifera, de atividade inibidora de protease da serina em A. lebbeck, I. asarifolia, J. curcas, M. oleifera e R. communis, de atividade inibidora de protease da cisteína em M. oleifera e nenhuma atividade proteica na fração de L. ferrea. Os resultados revelaram novas espécies botânicas com potencial de controle de nematoides em caprinos e um novo campo de pesquisa, o estudo de moléculas de origem proteica com atividade ovicida.