ß-Glucosidase activity and antimicrobial properties of potentially probiotic autochthonous lactic cultures.

Background: The demand for lactic acid bacteria products, especially probiotics, has increased. Bacteria that increase polyphenol bioavailability and act as bio preservatives are sought after. This study aims to identify autochthonous lactic acid cultures from EMBRAPA that demonstrate ß-glucosidase activity and inhibitory effect on microbial sanitary indicators. Methods: Cell-free extracts were obtained by sonicating every 5 s for 40 min. The extracts were mixed with cellobiose and incubated at 50 °C. The reaction was stopped by immersing the tubes in boiling water. The GOD-POD reagent was added for spectrophotometer readings. Antimicrobial activity was tested against reference strains using the agar well diffusion method. Lactic cultures in MRS broth were added to 0.9 cm wells and incubated. The diameter of the inhibition zones was measured to determine the extension of inhibition. Results: Only L. rhamnosus EM1107 displayed extracellular ß-glucosidase activity, while all autochthonous strains except L. plantarum CNPC020 demonstrated intracellular activity for this enzyme. L. plantarum CNPC003 had the highest values. On the other hand, L. plantarum CNPC020, similarly to L. mucosae CNPC007, exhibited notable inhibition against sanitary indicators. These two strains significantly differed from the other five autochthonous cultures regarding S. enterica serovar Typhimurium ATCC 14028 inhibition (P < 0.05). However, they did not differ from at least one positive control in terms of inhibition against S. aureus ATCC 25923 and E. coli ATCC 25922 (P > 0.05). Therefore, it is advisable to consider these cultures separately for different technological purposes, such as phenolics metabolism or bio preservative activity. This will facilitate appropriate selection based on each specific property required for the intended product development.

Safety Evaluation of Goat Milk Added with the Prebiotic Inulin Fermented with the Potentially Probiotic Native Culture Limosilactobacillus mucosae CNPC007 in Co-culture with Streptococcus thermophilus QGE: Analysis of Acute and Repeated Dose Oral Toxicity.

Despite functional goat milk products having emerged due to their importance for human nutrition and health, few studies have assessed the safety of consumption of goat dairy products containing potentially probiotic autochthonous lactic acid bacteria supplemented with prebiotic carbohydrates. Aiming this field, this study evaluated the safety of goat's milk fermented with Streptococcus thermophilus QGE, the autochthonous Limosilactobacillus mucosae CNPC007 culture, and the prebiotic inulin, through single- and repeated-dose oral toxicity tests (SDT and RDT, respectively) in animals. Ten female Swiss Webster mice were used for SDT evaluation - 2 groups, SDTc (20 mL/kg of filtered water) and SDTt (20 mL/kg of fermented milk) - and 40 Wistar rats for RDT - RDT3, RDT6, and RDT12 (treated with fermented milk at doses of 3 mL/kg, 6 mL/kg, and 12 mL/kg, respectively) and also RDTc (12 mL/kg of filtered water). For SDT, no signs of mortality or toxicity were observed, and the animals maintained the expected weight gain and feed intake. The RDT trials did not show mortality or signs of toxicity, as well as no change in body weight and organs, in the hematological and biochemical parameters, and also in relation to morphology and histology. Since the fermented milk did not cause any toxic effect in the conditions evaluated, it can be said that its no-adverse effect level (NOAEL) was considered to be higher than 20 mL/kg/day. Thus, the fermented milk with L. mucosae CNPC007 and inulin was considered to be of low toxicity, safe for use in rodents, and allowed for use in further studies.

Autochthonous adjunct culture of Limosilactobacillus mucosae CNPC007 improved the techno-functional, physicochemical, and sensory properties of goat milk Greek-style yogurt.

We evaluated the performance of Limosilactobacillus mucosae CNPC007 as an autochthonous adjunct culture in the production of goat milk Greek-style yogurt. The techno-functional, physicochemical, and sensory characteristics of the control yogurt (containing only starter culture, CY) and the probiotic yogurt (with the probiotic strain added, PY) were assessed during 28 d of refrigerated storage. Furthermore, we determined the survival of the strain throughout the gastrointestinal tract under simulated conditions. The PY yogurt had a lower extent of proteolysis index and a higher depth of proteolysis index. These results indicate that the proteolytic enzymes of L. mucosae may have a possible action in PY. The PY formulation exhibited viscosity almost 1.5 times as high as CY over the refrigeration period, probably due to higher production of exopolysaccharides by the probiotic strain, which directly interferes with the microstructure, texture, and viscosity of the product. The PY formulation received higher scores for color, flavor, and global acceptance at 1 d of storage and higher texture scores at 28 d. The counts of L. mucosae remained high (>7 log cfu/g and >8.5 log cfu/g) throughout mouth-ileum digestion and storage, respectively, in PY. The autochthonous adjunct culture of L. mucosae CNPC007 can be used for production of a novel potentially probiotic goat yogurt without negatively affecting the general characteristics of the product quality, adding value associated with maintaining its functional potential.

Efeito da oligofrutose (FOS) sobre a capacidade de acidificação da cultura nativa Lactobacillus plantarum CNPC 003 isolada ou em co-cultura com Streptococcus thermophilus em leite de cabra / Influence of oligofrutose (FOS) on the acidification capacity of the indigenous culture Lactobacillus plantarum CNPC 003 solely or in coculture with Streptococcus thermophilus in goat milk

Objetivou-se analisar o comportamento fermentativo da cultura Lactobacillus plantarum CNPC 003 em leite de cabra em quatro tratamentos por 6 horas: T1 L. plantarum CNPC 003; T2 L. plantarum + oliogofrutose (FOS); T3 L. plantarum + Streptococcus thermophilus; T4 com L. plantarum + S. thermophilus + FOS. Os tratamentos estudados diferiram significativamente entre si em relação à acidez e ao pH após 6h de fermentação (p<0,05), tendo sido verificada uma influência positiva do uso de S. thermophilus e da adição de FOS sobre esses parâmetros. Do mesmo modo, a presença de FOS melhorou a viabilidade dos microrganismos estudados após 6h de fermentação. A utilização de S. thermophilus e de FOS é uma alternativa viável para o uso em leites fermentados contendo a cepa nativa L. plantarum CNPC 003.(AU)

Efeito da oligofrutose (FOS) sobre a capacidade de acidificação da cultura nativa Lactobacillus plantarum CNPC 003 isolada ou em co-cultura com Streptococcus thermophilus em leite de cabra / Influence of oligofrutose (FOS) on the acidification capacity of the indigenous culture Lactobacillus plantarum CNPC 003 solely or in coculture with Streptococcus thermophilus in goat milk

Objetivou-se analisar o comportamento fermentativo da cultura Lactobacillus plantarum CNPC 003 em leite de cabra em quatro tratamentos por 6 horas: T1 – L. plantarum CNPC 003; T2 – L. plantarum + oliogofrutose (FOS); T3 – L. plantarum + Streptococcus thermophilus; T4 – com L. plantarum + S. thermophilus + FOS. Os tratamentos estudados diferiram significativamente entre si em relação à acidez e ao pH após 6h de fermentação (p<0,05), tendo sido verificada uma influência positiva do uso de S. thermophilus e da adição de FOS sobre esses parâmetros. Do mesmo modo, a presença de FOS melhorou a viabilidade dos microrganismos estudados após 6h de fermentação. A utilização de S. thermophilus e de FOS é uma alternativa viável para o uso em leites fermentados contendo a cepa nativa L. plantarum CNPC 003.

Feeding behavior and physiological parameters of rearing goats fed diets containing detoxified castor cake / Comportamento alimentar e parâmetros fisiológicos de cabritas recriadas consumindo dietas contendo torta de mamona destoxificada

The objective of this study was to evaluate the influence of replacing soybean meal (SM) with castor bean cake (DCC), detoxified using two alkaline products, on the feeding behavior and physiological parameters of Saanen and Anglo-Nubian goat breeds with an initial body weight of 16.22 ± 0.67 kg. The treatments consisted of three diets that were isoproteic and isoenergetic: the first with a standard formulation based on corn and SM, the second with DCC detoxified using calcium hydroxide (Ca(OH)₂ DCC), and the third with DCC detoxified using sodium hydroxide (NaOH DCC). A completely randomized design with a 3 × 2 (diets and breeds) factorial arrangement and four replicates per combination was used. A dietary effect on the intake of dry matter (DM) and NDFap (P = 0.02 and 0.01) was observed, with lower values for goats fed NaOH DCC than the other diets. The diets influenced feeding and rumination times (P = 0.04 and 0.03). An interaction (P = 0.04 and 0.01) between the breeds and the SM-based diet was found for time variables (other activity and idle time). Increased feeding and neutral detergent fiber (NDF) rumination efficiencies were observed for goats fed SM and Ca(OH)₂ DCC (P = 0.03 and 0.01). Goats fed the NaOH DCC diet consumed large amounts of water, while Saanen goats urinated more times than did Anglo-Nubian goats. The diets had an effect on the heart rate and rectal temperature (P = 0.01 and 0.02), while the body surface temperature was influenced by the breed. Thus, Ca(OH)₂ or NaOH DCC has a great potential to replace SM in the feeding of dairy goats. When used as diet for goats in the growth phase, NaOH DCC causes a reduction of DM and NDF intakes. Neither DCC negatively affected the physiological parameters of goats.(AU)

Objetivou-se avaliar a influência da substituição do farelo de soja (FS) pela torta de mamona destoxificada (TMD) por dois produtos alcalinos sobre o comportamento alimentar e parâmetros fisiológicos de cabritas das raças Saanen e Anglo-nubiana, com peso corporal inicial de 16,22 ± 0,67 kg. Os tratamentos consistiram de três dietas, uma com formulação padrão à base de milho e FS e as demais foram formuladas com torta de mamona destoxificada por hidróxido de cálcio (TMD Ca(OH)₂) e outra por hidróxido de sódio (TMD NaOH), as quais foram isoproteicas e isoenergéticas. Utilizou-se um delineamento inteiramente casualizado, com arranjo fatorial 3 x 2 (dieta e raça) com quatro repetições por combinação. Houve efeito das dietas sobre o consumo de MS e FDNcp (P = 0,02 e 0,01), com menores valores para as cabras alimentadas com TMD NaOH. As dietas influenciaram os tempos de alimentação e ruminação (P = 0,04 e 0,03). Houve interação (P = 0,04 e 0,01) entre as raças e dieta à base de FS para as variáveis tempo com outras atividade e ócio. Observou-se maiores eficiências de alimentação e ruminação da FDN para as cabras que consumiram FS e TMD Ca(OH)₂ (P = 0,03 e 0,01). As cabras da dieta TMD NaOH consumiram maiores quantidades de água e as Saanen urinaram mais vezes que as Anglo-nubiana. Houve efeito das dietas sobre a frequência cardíaca e temperatura retal (P = 0,01 e 0,02), já a temperatura corporal foi influenciada pelas raças. A torta de mamona destoxificada pelo hidróxido de cálcio e/ou hidróxido de sódio tem grande potencial para substituir o farelo de soja na alimentação de cabras leiteiras. Quando usado na composição de dietas para caprinos em fase de crescimento, a torta de mamona destoxificada pelo hidróxido de sódio leva à redução do CMS, bem como da FDN. Ambas as tortas não alteram negativamente os parâmetros fisiológicos das cabras.(AU)

Feeding behavior and physiological parameters of rearing goats fed diets containing detoxified castor cake / Comportamento alimentar e parâmetros fisiológicos de cabritas recriadas consumindo dietas contendo torta de mamona destoxificada

The objective of this study was to evaluate the influence of replacing soybean meal (SM) with castor bean cake (DCC), detoxified using two alkaline products, on the feeding behavior and physiological parameters of Saanen and Anglo-Nubian goat breeds with an initial body weight of 16.22 ± 0.67 kg. The treatments consisted of three diets that were isoproteic and isoenergetic: the first with a standard formulation based on corn and SM, the second with DCC detoxified using calcium hydroxide (Ca(OH)₂ DCC), and the third with DCC detoxified using sodium hydroxide (NaOH DCC). A completely randomized design with a 3 × 2 (diets and breeds) factorial arrangement and four replicates per combination was used. A dietary effect on the intake of dry matter (DM) and NDFap (P = 0.02 and 0.01) was observed, with lower values for goats fed NaOH DCC than the other diets. The diets influenced feeding and rumination times (P = 0.04 and 0.03). An interaction (P = 0.04 and 0.01) between the breeds and the SM-based diet was found for time variables (other activity and idle time). Increased feeding and neutral detergent fiber (NDF) rumination efficiencies were observed for goats fed SM and Ca(OH)₂ DCC (P = 0.03 and 0.01). Goats fed the NaOH DCC diet consumed large amounts of water, while Saanen goats urinated more times than did Anglo-Nubian goats. The diets had an effect on the heart rate and rectal temperature (P = 0.01 and 0.02), while the body surface temperature was influenced by the breed. Thus, Ca(OH)₂ or NaOH DCC has a great potential to replace SM in the feeding of dairy goats. When used as diet for goats in the growth phase, NaOH DCC causes a reduction of DM and NDF intakes. Neither DCC negatively affected the physiological parameters of goats.

Objetivou-se avaliar a influência da substituição do farelo de soja (FS) pela torta de mamona destoxificada (TMD) por dois produtos alcalinos sobre o comportamento alimentar e parâmetros fisiológicos de cabritas das raças Saanen e Anglo-nubiana, com peso corporal inicial de 16,22 ± 0,67 kg. Os tratamentos consistiram de três dietas, uma com formulação padrão à base de milho e FS e as demais foram formuladas com torta de mamona destoxificada por hidróxido de cálcio (TMD Ca(OH)₂) e outra por hidróxido de sódio (TMD NaOH), as quais foram isoproteicas e isoenergéticas. Utilizou-se um delineamento inteiramente casualizado, com arranjo fatorial 3 x 2 (dieta e raça) com quatro repetições por combinação. Houve efeito das dietas sobre o consumo de MS e FDNcp (P = 0,02 e 0,01), com menores valores para as cabras alimentadas com TMD NaOH. As dietas influenciaram os tempos de alimentação e ruminação (P = 0,04 e 0,03). Houve interação (P = 0,04 e 0,01) entre as raças e dieta à base de FS para as variáveis tempo com outras atividade e ócio. Observou-se maiores eficiências de alimentação e ruminação da FDN para as cabras que consumiram FS e TMD Ca(OH)₂ (P = 0,03 e 0,01). As cabras da dieta TMD NaOH consumiram maiores quantidades de água e as Saanen urinaram mais vezes que as Anglo-nubiana. Houve efeito das dietas sobre a frequência cardíaca e temperatura retal (P = 0,01 e 0,02), já a temperatura corporal foi influenciada pelas raças. A torta de mamona destoxificada pelo hidróxido de cálcio e/ou hidróxido de sódio tem grande potencial para substituir o farelo de soja na alimentação de cabras leiteiras. Quando usado na composição de dietas para caprinos em fase de crescimento, a torta de mamona destoxificada pelo hidróxido de sódio leva à redução do CMS, bem como da FDN. Ambas as tortas não alteram negativamente os parâmetros fisiológicos das cabras.

Buffalo mozzarella chemical composition and authenticity assessment by electrophoretic profiling / Avaliação da composição química e autenticidade de muçarela de búfala por meio do perfil eletroforético

Buffalo milk mozzarella is often adulterated by the addition of cows milk. The aim of this study was to assess the quality of buffalo milk mozzarella by using electrophoresis (SDS-PAGE) to detect the addition of cows milk. Reference cheeses were produced exclusively from combinations of buffalo and cow milk, and labeled reference treatment buffalo or cow (RTB and RTC, respectively). Standardized cheeses were made by combining buffalo milk and 2.5%, 5.0%, 10%, 20%, 30%, 40% or 50% cows milk. A total of 9 cheese formulations were produced and either frozen immediately (time 0) or after 20 days (time 20). Eighteen commercial samples of buffalo mozzarella were sampled between 020 days of production. The chemical composition (moisture, ash, fat in dry matter, protein, total solids and defatted dry extract) and physicochemical characteristics (acidity) of the cheeses were evaluated. Proteins and water-soluble peptides (WSP) extracted from RTB, RTC and from the commercial samples, and were analyzed by SDS-PAGE electrophoresis under denaturing conditions. It was not possible to detect the inclusion of cows milk by the cheeses chemical and physicochemical properties. However, the separation and detection of peptide and protein fractions of the cheese was possible by electrophoresis. The results of the electrophoretic analysis suggest that 28% of the commercial samples considered here had evidence of the addition of cows milk. The methodology described here is important to identify occurrences of fraud in buffalo mozzarella production.(AU)

Objetivou-se com o presente estudo detectar fraudes por adição de leite de vaca em muçarela de búfala. Foram produzidos queijos exclusivamente com leite de búfala (TRB) e vaca (TRV) e com inclusões crescentes de leite bovino ao bubalino (2,5%, 5,0%, 10%, 20%, 30%, 40% e 50%), totalizando 9 formulações que foram congeladas no dia de fabricação (Tempo 0) e com 20 dias após a data de fabricação (Tempo 20). Foram coletadas 18 amostras comerciais de muçarela de búfala com até 20 dias de fabricação sendo congeladas no Tempo 20. Os queijos foram avaliados em relação à composição química (umidade, cinzas, gordura no extrato seco, proteínas, extrato seco total e extrato seco desengordurado) e característica físico-química (acidez titulável). Proteínas dos queijos muçarela produzidos e amostras comerciais e peptídeos solúveis em água (PSA) extraídos de TRB, TRV e amostras comerciais foram analisados por eletroforese sob condições desnaturantes por meio de dodecil sulfato de sódio (SDS-PAGE). Não foi possível detectar adulterações por meio dos resultados de composição química e caracterização físico-química. A técnica SDS-PAGE mostrou-se eficiente na separação e detecção das frações proteicas e peptídeos dos queijos. Em 28% das amostras comerciais de muçarela de búfala houve indícios de adulteração por adição de leite de vaca. Os resultados deste trabalho mostraram-se importantes para alertar sobre a ocorrência de fraudes em muçarela de búfala.(AU)

Functional Properties of Lactobacillus mucosae Strains Isolated from Brazilian Goat Milk.

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.

Buffalo mozzarella chemical composition and authenticity assessment by electrophoretic profiling / Avaliação da composição química e autenticidade de muçarela de búfala por meio do perfil eletroforético

Buffalo milk mozzarella is often adulterated by the addition of cows milk. The aim of this study was to assess the quality of buffalo milk mozzarella by using electrophoresis (SDS-PAGE) to detect the addition of cows milk. Reference cheeses were produced exclusively from combinations of buffalo and cow milk, and labeled reference treatment buffalo or cow (RTB and RTC, respectively). Standardized cheeses were made by combining buffalo milk and 2.5%, 5.0%, 10%, 20%, 30%, 40% or 50% cows milk. A total of 9 cheese formulations were produced and either frozen immediately (time 0) or after 20 days (time 20). Eighteen commercial samples of buffalo mozzarella were sampled between 020 days of production. The chemical composition (moisture, ash, fat in dry matter, protein, total solids and defatted dry extract) and physicochemical characteristics (acidity) of the cheeses were evaluated. Proteins and water-soluble peptides (WSP) extracted from RTB, RTC and from the commercial samples, and were analyzed by SDS-PAGE electrophoresis under denaturing conditions. It was not possible to detect the inclusion of cows milk by the cheeses chemical and physicochemical properties. However, the separation and detection of peptide and protein fractions of the cheese was possible by electrophoresis. The results of the electrophoretic analysis suggest that 28% of the commercial samples considered here had evidence of the addition of cow’s milk. The methodology described here is important to identify occurrences of fraud in buffalo mozzarella production.

Objetivou-se com o presente estudo detectar fraudes por adição de leite de vaca em muçarela de búfala. Foram produzidos queijos exclusivamente com leite de búfala (TRB) e vaca (TRV) e com inclusões crescentes de leite bovino ao bubalino (2,5%, 5,0%, 10%, 20%, 30%, 40% e 50%), totalizando 9 formulações que foram congeladas no dia de fabricação (Tempo 0) e com 20 dias após a data de fabricação (Tempo 20). Foram coletadas 18 amostras comerciais de muçarela de búfala com até 20 dias de fabricação sendo congeladas no Tempo 20. Os queijos foram avaliados em relação à composição química (umidade, cinzas, gordura no extrato seco, proteínas, extrato seco total e extrato seco desengordurado) e característica físico-química (acidez titulável). Proteínas dos queijos muçarela produzidos e amostras comerciais e peptídeos solúveis em água (PSA) extraídos de TRB, TRV e amostras comerciais foram analisados por eletroforese sob condições desnaturantes por meio de dodecil sulfato de sódio (SDS-PAGE). Não foi possível detectar adulterações por meio dos resultados de composição química e caracterização físico-química. A técnica SDS-PAGE mostrou-se eficiente na separação e detecção das frações proteicas e peptídeos dos queijos. Em 28% das amostras comerciais de muçarela de búfala houve indícios de adulteração por adição de leite de vaca. Os resultados deste trabalho mostraram-se importantes para alertar sobre a ocorrência de fraudes em muçarela de búfala.

Buffalo mozzarella chemical composition and authenticity assessment by electrophoretic profiling / Avaliação da composição química e autenticidade de muçarela de búfala por meio do perfil eletroforético

Buffalo milk mozzarella is often adulterated by the addition of cows milk. The aim of this study was to assess the quality of buffalo milk mozzarella by using electrophoresis (SDS-PAGE) to detect the addition of cows milk. Reference cheeses were produced exclusively from combinations of buffalo and cow milk, and labeled reference treatment buffalo or cow (RTB and RTC, respectively). Standardized cheeses were made by combining buffalo milk and 2.5%, 5.0%, 10%, 20%, 30%, 40% or 50% cows milk. A total of 9 cheese formulations were produced and either frozen immediately (time 0) or after 20 days (time 20). Eighteen commercial samples of buffalo mozzarella were sampled between 020 days of production. The chemical composition (moisture, ash, fat in dry matter, protein, total solids and defatted dry extract) and physicochemical characteristics (acidity) of the cheeses were evaluated. Proteins and water-soluble peptides (WSP) extracted from RTB, RTC and from the commercial samples, and were analyzed by SDS-PAGE electrophoresis under denaturing conditions. It was not possible to detect the inclusion of cows milk by the cheeses chemical and physicochemical properties. However, the separation and detection of peptide and protein fractions of the cheese was possible by electrophoresis. The results of the electrophoretic analysis suggest that 28% of the commercial samples considered h

Objetivou-se com o presente estudo detectar fraudes por adição de leite de vaca em muçarela de búfala. Foram produzidos queijos exclusivamente com leite de búfala (TRB) e vaca (TRV) e com inclusões crescentes de leite bovino ao bubalino (2,5%, 5,0%, 10%, 20%, 30%, 40% e 50%), totalizando 9 formulações que foram congeladas no dia de fabricação (Tempo 0) e com 20 dias após a data de fabricação (Tempo 20). Foram coletadas 18 amostras comerciais de muçarela de búfala com até 20 dias de fabricação sendo congeladas no Tempo 20. Os queijos foram avaliados em relação à composição química (umidade, cinzas, gordura no extrato seco, proteínas, extrato seco total e extrato seco desengordurado) e característica físico-química (acidez titulável). Proteínas dos queijos muçarela produzidos e amostras comerciais e peptídeos solúveis em água (PSA) extraídos de TRB, TRV e amostras comerciais foram analisados por eletroforese sob condições desnaturantes por meio de dodecil sulfato de sódio (SDS-PAGE). Não foi possível detectar adulterações por meio dos resultados de composição química e caracterização físico-química. A técnica SDS-PAGE mostrou-se eficiente na separação e detecção das frações proteicas e peptídeos dos queijos. Em 28% das amostras comerciais de muçarela de búfala houve indícios de adulteração por adição de leite de vaca. Os resultados deste trabalho mostraram-se importantes para ale

Biodegradação do bagaço de cana-de-açúcar por microrganismos ruminais de caprinos e ovinos / Biodegradation of sugarcane bagasse by ruminal microorganisms from sheep or goats

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.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus.

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.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus / Lectina, hemolisina e inibidores de protease em frações de sementes com atividade ovicida contra Haemonchus contortus

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.

Lectin, hemolysin and protease inhibitors in seed fractions with ovicidal activity against Haemonchus contortus / Lectina, hemolisina e inibidores de protease em frações de sementes com atividade ovicida contra Haemonchus contortus

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.