Haematologic and immunologic parameters of bullfrogs, Lithobates catesbeianus, fed probiotics

The effects of two probiotics (P(1) - Lactobacillus acidophilus, Bifidobacterium bifidum and Enterococcus faecium and P(2) - Bacillus subtilis) supplemented to commercial feed (40% crude protein) on the haematological and immunological parameters of the bullfrog Lithobates catesbeianus were studied. Two doses of each probiotic (5 and 10 g kg-1 of food) were added to the diets and fed to frogs, totalling five treatments over 112 days. Haematological analyses consisted of total and differential leucocyte counts, erythrocyte and thrombocyte counts, haematocrit, haemoglobin levels and RBC indices (mean corpuscular volume, mean corpuscular haemoglobin - and mean corpuscular haemoglobin concentration) and the immunological parameters included phagocytic capacity and phagocytic index of peritoneal phagocytes. The results showed that the probiotics did not significantly influence any of the haematological parameters measured. However, immunological assays showed that the probiotics had an immunostimulating effect. The greatest effects were seen with probiotic P(1) fed at a dose of 10 g kg-1 of diet and probiotic P(2) fed at 5 g kg-1 of diet.

Efeito do probiótico Bacillus subtilis no crescimento, sobrevivência e fisiologia de rãs-touro (Rana catesbeiana)

The probiotics are live microorganisms, in latency state, that benefices the development of the animals. The objective of this work was to evaluate the effect of the Bacillus subtilis in different doses about the growing, survival and immunological and hematological analyses in the Bullfrog (Rana catesbeiana) froglets. The test was developed in Experimental Frog farm of Agriculture Department, Pindamonhangaba, SP. Three doses of probiotics were tested (T1 - 2.5 g/Kg, T2 - 5 g/Kg and T3 - 10 g/Kg of food). The test was carried out with three simultaneous replicates, plus control group. The products were added to froglet's diet and mixed to the meal. The animals were previously feed with this diet for a period of 14 days and accompanied for about 42 days after metamorphose. Biometrics were performed every 7 days. It was evaluated the final weight, the survival, phagocytic capacity, phagocytic index and hematological analyses as hematocrit, hemoglobin level, erythrocyte number, absolute indices hematimetrycs (MCV and MCHC), differential counting of leucocytes and total counting of leucocytes and trombocytes. The results had shown that the doses of the probiotics had produced none effect on the final weight and survival. The immunological analyses had shown that the probiotics have presented immunostimulator effect, but haven't influenced the hematological parameters of the animals.

Efeito da adição de probiótico em dietas de leitões desmamados sobre as características do sistema digestório e de desempenho

Os objetivos foram avaliar os efeitos da adição de diferentes concentrações de probiótico (0; 200 e 300mg/kg de ração) em rações de leitões recém desmamados e em diferentes idades de abate, sobre as características do sistema digestório e desempenho zootécnico. Foram utilizados 35 leitões desmamados, distribuídos em delineamento em blocos casualizados em esquema fatorial 3x2+1 (três concentrações de probiótico x duas idades de abate + grupo abatido ao desmame). Foram abatidos cinco leitões no dia da desmama e os demais ao 31º e 41º dia de idade. Foram amostrados segmentos do duodeno e jejuno para a contagem de vilosidades, pesados o pâncreas e fígado e calculado o rendimento de carcaça. Para incidência de diarréia e desempenho, foram utilizados 30 animais e as avaliações foram realizadas nos seguintes períodos: P1- 0 a 10, P2- 11 a 20 e Período Total- 0 a 20 dias pós desmame. Não houve efeito das concentrações do probiótico sobre as características do sistema digestório e a incidência de diarréia, no entanto o ganho de peso e a conversão alimentar foram afetados. Concluiu-se que o ganho de peso e a conversão alimentar de leitões suplementados com 200mg de probiótico/kg de ração foi melhor do que o grupo suplementado com 300mg de probiótico/kg de ração. As diferentes concentrações de probiótico adicionadas na ração de leitões não alteraram as características do sistema digestório, mostrando-se alterações apenas em relação à idade do abate.

Avaliação de probióticos na dieta de frangos de corte criados em cama nova ou reutilizada

Três experimentos foram conduzidos com o objetivo de avaliar o efeito da utilização de probióticos na dieta de frangos de corte sobre as características da cama reutilizada e das lesões de peito, joelho e coxim plantar. Foram utilizados em cada experimento 800 pintos machos de um dia, da linhagem comercial Ross, alojados em 20 boxes, em densidade populacional de 10 aves/m². O delineamento foi inteiramente casualizado e os tratamentos distribuídos em esquema fatorial 2 × 2 (cama nova ou reutilizada x rações com ou sem probiótico). O probiótico utilizado nas dietas foi composto por Bacillus subtilis e B. coagulans (2x10(9) e 1x10(7) ufc, respectivamente). Aos 42 dias de idade, quatro aves de cada boxe foram abatidas para análise das lesões de peito, joelho e coxim plantar. A cama de cada boxe foi amostrada para determinação dos teores de MS e nitrogênio, do potencial de volatilização de amônia e do pH. A cama reutilizada apresentou maior teor de MS, maiores valores de pH e potencial de volatilização de amônia e menor teor de nitrogênio. O probiótico usado nas rações resultou em maior potencial de volatilização de amônia. As lesões de coxim plantar e joelho foram maiores quando utilizada cama nova. As camas reutilizadas por dois, três e quatro ciclos causaram menores lesões de joelho e coxim plantar. O probiótico não promoveu efeito benéfico sobre a cama reutilizada.

CARACTERÍSTICAS FÍSICAS E QUÍMICAS DE BEBIDAS LÁCTEAS FERMENTADAS E PREPARADAS COM SORO DE QUEIJO MINAS FRESCAL

Nesta pesquisa procurou-se verificar as características físicas e químicas de bebidas lácteas preparadas com três concentrações de soro de queijo Minas Frescal (30, 40 e 50%), empregando-se dois tipos de culturas lácticas: uma tradicional para iogurte (YC-180) contendo cepas mistas de Streptococcus salivarus subsp. thermophilus, Lactobacillus delbrueckii subsp. lactis e Lactobacillus delbrueckii subsp. bulgaricus e outra (ABY-1) contendo cepas mistas de Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophillus, Bifidobacteria e Streptococcus salivarius subsp. thermophilus. Constatou-se que as bebidas lácteas apresentaram diferença estatística no tempo zero para os teores de gordura e de extrato seco. À medida em que se elevou a proporção de soro em relação ao leite, os teores de gordura e de extrato seco diminuíram. O teor de proteína também diminuiu à medida em que se aumentou o teor de soro nas bebidas lácteas, embora a diferença não tenha sido tão acentuada quanto as observadas para os teores de gordura e de extrato seco. em relação à lactose, não se constatou diferença entre os tratamentos. Os teores de soro não influenciaram o índice de proteólise das bebidas lácteas. Verificou-se todavia que as bebidas elaboradas com a cultura probiótica ABY-1 apresentaram valores superiores para proteólise quando comparadas às bebidas elaboradas com as culturas YC-180. As bebidas lácteas elaboradas com 30% de soro apresentaram maiores valores para viscosidade. As bebidas elaboradas com a cultura YC-180 apresentaram valores superiores para viscosidade durante o período de armazenamento.

Desenvolvimento de sorvete probiótico a base de leite de cabra: estudo da formulação, características físico químicas, sensoriais e viabilidade das bactérias probióticas

This work targetet the caprine ice cream production added with probiotic bacteria Bifidobacterium animalis subsp. lactis. It is divided into two parts. In the first one, four caprine ice cream formulations were evaluated, in which it was used hydrogenated fat (F1 and F3) or fat substitute (F2 and F4) in two different flavors (F1 and F2, passion fruit, F3 and F4, guava). Statistical differences (p<0.05) were detected for their physical-chemical properties, mainly for total solids and fat, but no differences were observed for melting test results. When it went to sensory acceptance, all four ice cream formulations reached high acceptance indexes, mostly formulation F4, which was selected for further studies. In the second part, F4 formulation was prepared with the addition of probiotic bacteria Bifidobacterium animalis subsp. lactis. The growth kinetics was studied and it was observed that the cellular concentration peak was reached after four fermentation hours (10.14 log UFC/g). This time was selected for pre-fermentation procedure and posterior addition at ice cream syrup. In this part of the study, two experimental groups were evaluated: group G1, in which the probiotic addition occurred before the maturation step and group G2, which included a pre-fermentation step and probiotic addition after ice cream maturation. The physical-chemical properties of these two ice cream groups were similar, except for pH, which was higher for group G2 (p<0.05). G1 samples had superior melting rate (3.566 mL/min) and both groups presented microbiological and sanitary results in accordance to current Brazilian legislation. Also, G1 and G2 were considered sensory accepted due to their acceptance indexes higher than 70%. G1 and G2 sensory profiles were similar (p>0.05), and both ice cream samples exhibited high creaminess (6.76 to 6.91) and mouth melting sensation (6.53 to 6.67) scores, while low sandiness scores (0.85 to 0.86) were observed, positive characteristics for this kind of food product. During the first 24 hours after ice cream production, the population of B. animalis subsp. lactis decreased, reaching 7.15 e 6.92 log CFU/g for G1 and G2, respectively. Probiotic bacteria counts fluctuated in ice cream samples during the first 108 days at frozen storage, especially for G2 group. Decreased probiotic viability was observed for G1 samples during the first 35 days of frozen storage, mild variation between 35 and 63 days and stabilized counts were observed after this time. After 21 days at frozen storage, ice cream samples of G1 and G2 groups reached 1.2 x 109 and 1.3 x 109 CFU/portion, respectively. After 108 days under these storage conditions, the survival rate of B. animalis subsp. lactis was 94.26% and 81.10% for G1 and G2 samples, respectively. After simulation of gastroenteric conditions, G2 group reached 9.72 x 105 CFU/portion. Considering the current requirements of Brazilian legislation, which stipulates that functional foods must have minimum probiotic count between 108 and 109 CFU/portion and detectable probiotic bacteria after being submitted to gastroenteric conditions, it is concluded that the ice cream with the addition of Bifidobacterium animalis subsp. lactis made as shown in this work, can be considered as a dairy functional food

Fermented soy product supplemented with isoflavones affects adipose tissue in a regional-specific manner and improves HDL-cholesterol in rats fed on a cholesterol-enriched diet

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Intake of isoflavone-supplemented soy yogurt fermented with Enterococcus faecium lowers serum total cholesterol and non-HDL cholesterol of hypercholesterolemic rats

The aim of this study was to obtain an isoflavone-supplemented soy yogurt, fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus ssp jugurti, with suitable sensory properties and to assess the effects of the final product on blood lipids in hypercholesterolemic rats. Four isoflavone supplementation procedures were tested, in which the isoflavone was added at these stages: (1) before heat-treatment; (2) after heating and before fermentation; (3) after fermentation and (4) in the okara (by-product of soy milk) flour stirred into the fermented product when consumed. The products were subjected to a test of sensory acceptability. To assess their potential hypocholesterolemic properties in vivo, four groups of rats were used: control (C), hypercholesterolemic (H), hypercholesterolemic plus fermented product (HF) and hypercholesterolemic plus isoflavone-supplemented fermented product (HFI). Hypercholesterolemia was induced in rats of groups H, HF and HFI by feeding them on a commercial rat chow to which cholesterol and cholic acid had been added. Total, HDL and non-HDL cholesterol and triglycerides were measured in the blood of the rats. No significant sensorial differences were detected among the samples of soy yogurt supplemented with isoflavones at various processing stages. Rats fed a fermented soy product enriched with isoflavones (HFI group) had significantly (P < 0.05) less serum total cholesterol (15.5%) compared with rats fed a hypercholesterolemic diet (H group). Non-HDL cholesterol was less (P < 0.05) in rats fed a fermented soy product enriched or not with isoflavones (27.4 and 23.2%) compared to H group. The HDL-C and triglyceride concentrations did not differ significantly among the groups. It was possible to obtain an isoflavone-supplemented soy yogurt with satisfactory sensory characteristics. The resulting supplemented soy yogurt was capable of producing a lipid-lowering effect in hypercholesterolemic rats, relative to the animals that did not consume this product.

Effect of the consumption of a new symbiotic shake on glycemia and cholesterol levels in elderly people with type 2 diabetes mellitus

Background: The consumption of foods containing probiotic and prebiotic ingredients is growing consistently every year, and in view of the limited number of studies investigating their effect in the elderly.Objective: The objective of this study was to evaluate the effect of the consumption of a symbiotic shake containing Lactobacillus acidophilus, Bifidobacterium bifidum and fructooligosaccharides on glycemia and cholesterol levels in elderly people.Methods: A randomized, double-blind, placebo-controlled study was conducted on twenty volunteers (ten for placebo group and ten for symbiotic group), aged 50 to 60 years. The criteria for inclusion in the study were: total cholesterol > 200 mg/dL; triglycerides > 200 mg/dL and glycemia > 110 mg/dL. Over a total test period of 30 days, 10 individuals (the symbiotic group) consumed a daily dose of 200 mL of a symbiotic shake containing 10(8) UFC/mL Lactobacillus acidophilus, 10(8) UFC/mL Bifidobacterium bifidum and 2 g oligofructose, while 10 other volunteers (the placebo group) drank daily the same amount of a shake that did not contain any symbiotic bacteria. Blood samples were collected 15 days prior to the start of the experiment and at 10-day intervals after the beginning of the shake intake. The standard lipid profile (total cholesterol, triglycerides and HDL cholesterol) and glycemia, or blood sugar levels, were evaluated by an enzyme colorimetric assay.Results: The results of the symbiotic group showed a non-significant reduction (P > 0.05) in total cholesterol and triglycerides, a significant increase (P < 0.05) in HDL cholesterol and a significant reduction (P < 0.05) in fasting glycemia. No significant changes were observed in the placebo group.Conclusion: The consumption of symbiotic shake resulted in a significant increase in HDL and a significant decrease of glycemia.

Efeito protetor da microbiota cecal congelada e liofilizada sobre a infecção experimental de frangos de corte por Salmonella enterica sorovar Enteritidis

Estudaram-se os efeitos do tratamento de frangos de corte com microbiota cecal anaeróbia liofilizada (MCL) e congelada (MCC) sobre a infecção do trato digestivo das aves por Salmonella enterica sorovar Enteritidis. Foi usada microbiota intestinal sem prévia identificação bacteriana. A infecção foi persistente, em ordem, no ceco, inglúvio e duodeno. A infecção também foi autolimitante nos grupos tratados e no controle. Não ocorreu diferença entre o grupo-controle positivo e os tratados com MCL ou MCC. Houve redução da colonização do ceco no período de 12 dias após o desafio nos grupos tratados com MCL e MCC, o que não ocorreu no grupo-controle positivo. Não houve variação entre os tratamentos com MCL e MCC quanto às características pesquisadas. A S. Enteritidis reduziu o ganho de peso médio nas aves inoculadas. Os tratamentos com MCL e MCC minimizaram a redução de peso nos grupos infectados.

Probiosis: concepts and prospects

The use of probiotics in animal and human feeding has been a subject of increasing interest both for the academia and the industry, mostly due to their potential positive effects on health and profitability. However, the knowledge on the composition of bacterial gastrointestinal communities in humans and animals, as well as its respective nutritional requirements, is far from being elucidated. Due to the ban of the use of antibiotic growth promoters in animal feeds, there has been an increasing interest on the utilization of probiotics to improve animal intestinal health under commercial settings. However, the possibility of horizontal transference of antibiotic-resistant genes between probiotic bacteria and pathogenic species has become a concern of poultry farmers and consumers around the world. Innovative ideas have emerged, such as the addition of essential oils, spices, and other plant extracts to feeds of monogastric animals to promote intestinal health. These natural compounds are considered ecologically adequate and safe for feeding purposes. This new reality will probably change the direction of research and of the use of additives in poultry production.

Uso de probiótico e antibiótico sobre o desempenho, o rendimento e a qualidade de carcaça de frangos de corte

Dois mil e quatrocentos pintos de corte foram distribuídos em um delineamento inteiramente ao acaso, com esquema fatorial 2 x 2 x 2 (sexo, com e sem probiótico, com e sem antibiótico), totalizando oito tratamentos e seis repetições com 50 aves cada uma. Independente da suplementação recebida, os machos sempre foram mais pesados que as fêmeas. Entretanto, a análise de desempenho apresentou maiores peso corporal e ganho de peso para os machos que receberam suplementação de antibiótico, quando comparados aos não-suplementados. Até 42 dias de idade, o uso do antibiótico determinou aumento do consumo de ração, mas não influiu nas demais características de desempenho. No período de 1 a 21 dias e 1 a 42 dias de idade, a suplementação com probiótico influenciou negativamente o peso corporal, ganho de peso e consumo de ração das aves. Houve interação probiótico x antibiótico para rendimento de carcaça das aves, obtendo-se maior rendimento de carcaça com o uso da associação dos produtos na ração. Nenhum dos produtos afetou o rendimento de partes (cabeça, pés, asas, perna, peito e dorso), o rendimento de vísceras (coração, fígado e vísceras não-comestíveis), gordura abdominal e características sensoriais (intensidade de aroma, aroma estranho, sabor, sabor estranho, maciez, suculência, preferência, cor característica e aparência geral) da carne de peito e pernas. No presente estudo, não foram encontrados efeitos benéficos da suplementação de probiótico para frangos de corte.

Inhibitory effect of essential oils against Lactobacillus rhamnosus and starter culture in fermented milk during its shelf-life period

The use of essential oils in foods has attracted great interest, due to their antagonistic action against pathogenic microorganisms. However, this action is undesirable for probiotic foods, as products containing Lactobacillus rhamnosus. The aim of the present study was to measure the sensitivity profile of L. rhamnosus and a yogurt starter culture in fermented milk, upon addition of increasing concentrations of cinnamon, clove and mint essential oils. Essential oils were prepared by steam distillation, and chemically characterised by gas chromatography-mass spectrometry (GC-MS) and determination of density. Survival curves were obtained from counts of L. rhamnosus and the starter culture (alone and in combination), upon addition of 0.04% essential oils. In parallel, titratable acidity was monitored over 28 experimental days. Minimum inhibitory concentration values, obtained using the microdilution method in Brain Heart Infusion medium, were 0.025, 0.2 and 0.4% for cinnamon, clove and mint essential oils, respectively. Cinnamon essential oil had the highest antimicrobial activity, especially against the starter culture, interfering with lactic acid production. Although viable cell counts of L. rhamnosus were lower following treatment with all 3 essential oils, relative to controls, these results were not statistically significant; in addition, cell counts remained greater than the minimum count of 10(8)CFU/mL required for a product to be considered a probiotic. Thus, although use of cinnamon essential oil in yogurt makes starter culture fermentation unfeasible, it does not prevent the application of L. rhamnosus to probiotic fermented milk. Furthermore, clove and mint essential oil caused sublethal stress to L. rhamnosus.

Cell enumeration and visualisation by transmission electron microscopy of Lactobacillus rhamnosus treated with cinnamon (Cinnamomum zeylanicum B.) essential oil

The use of essential oils (EOs) in functional foods containing probiotic microorganisms must consider the antimicrobial activity of these oils against beneficial bacteria such as Lactobacillus rhamnosus. This study aimed to evaluate the sensitivity of L. rhamnosus cultures treated with cinnamon EO through viable cell counts and visualisation by transmission electron microscopy. Cinnamon EO at a concentration of 0.04% had a bacteriostatic activity after 2 h of incubation. Although slight alterations were detected in the cell structure, this concentration was considered to be bactericidal, since it led to a significant reduction in cell numbers after 24 h. on the other hand, cinnamon EO at a 1.00% concentration decreased cell counts by 3 log units after 2 h incubation and no viable cell count was detected after 24 h. Transmission electron microscopy indicated that cells treated with 1.00% cinnamon EO were severely damaged and presented cell membrane disruption and cytoplasmic leakage.