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Staphylococcus aureus is one of the main causative agents of food poisoning. This bacterium is an important component of cheese microbiota and plays an important role in foodborne diseases. Another important component of the microbiota is the lactic acid bacterium, which actively participates in processes that define the physicochemical, sensorial, and microbiological features of cheese. Of the various microbiological interactions in cheese, the interaction between lactic acid bacteria and Staph. aureus is most relevant. To this end, we evaluated the viability of Staph. aureus strains and the expression of their enterotoxins in cheeses produced experimentally, using Weissella paramesenteroides GIR16L4 or Lactobacillus rhamnosus D1 or both as starter cultures. Over 7 d, we observed that the presence of lactic acid bacteria did not impair Staph. aureus growth. However, via qPCR we observed a change in the gene expression of staphylococcal enterotoxins, suggesting that molecular communication exists between Staph. aureus strains and lactic acid bacteria in cheese.

The relationship between inflammatory bowel disease (IBD) and mood disorders is complex and involves overlapping metabolic pathways, which may determine comorbidity. Several studies have been shown that this comorbidity could worsen IBD clinical course. The treatment of ulcerative colitis is complex, and involves traditional therapy to promote the function of epithelial barrier, reducing exacerbated inflammatory responses. Recently, it has been shown that some probiotic strains could modulate gut-brain axis, reducing depressive and anxiety scores in humans, including IBD patients. Accordingly, this study aimed to evaluate the role of Weissella paramesenteroides WpK4 in murine models of ulcerative colitis and chronic stress. It was observed that bacterium ingestion improved health of colitis mice, reducing intestinal permeability, besides improving colon histopathological appearance. In stressed mice, bacterial consumption was associated with a reduced anxiety-like and depressive-like behaviors. In both assays, the beneficial role of W. paramesenteroides WpK4 was related to its immunomodulatory feature. It is possible to state that W. paramesenteroides WpK4 exerted their beneficial roles in gut-brain axis through their immunomodulatory effects with consequences in several metabolic pathways related to intestinal permeability and hippocampal physiology.

Probiotics form a promising strategy to maintain intestinal health. Milks fermented with probiotic strains, such as the Lactobacillus paracasei ST11, are largely commercialized in Brazil and form a low-cost alternative to probiotic pharmaceutical formulations. In this study, we assessed the probiotic effects of milk fermented by L. paracasei ST11 (administered through fermented milk) in a Salmonella typhimurium infection model in BALB/c mice. We observed in this murine model that the applied probiotic conferred protective effects against S. typhimurium infection, since its administration reduced mortality, weight loss, translocation to target organs (liver and spleen) and ileum injury. Moreover, a reduction in the mRNA expression of pro-inflammatory cytokines such as IFN-gamma, IL-6, TNF-alpha and IL-17 in animals that received the probiotic before challenge was observed. Additionally, the ileum microbiota was better preserved in these animals. The present study highlights a multifactorial protective aspect of this commercial probiotic strain against a common gastrointestinal pathogen.

The objective of this work was to evaluate the effect of different disinfection procedures as alternatives to formaldehyde fumigation on eggshell microbial load and quality of eggs from a 42-week-old Cobb commercial breeder flock. A total of 10,080 clean eggs collected from the nests were randomly distributed in a randomized complete block design, among the following treatment groups: 13.33 g m-3 formaldehyde fumigation, 5-10 ppm ozone fumigation, 6.36 mW cm-2 UV-C light irradiation, spraying with 1.56% hydrogen peroxide, spraying with 0.13% peracetic acid, spraying with water (wet control), and no disinfection procedure (dry control). Per treatment, eight samples of four eggs each were collected before and after the disinfection procedure, in order to count the number of Enterobacteriaceae and total aerobic mesophilic bacteria on the eggshell. Only eggs subjected to the formaldehyde and UV treatments showed a significant reduction in total aerobic mesophilic bacterial count on the eggshell, when compared with those of the dry control group. Treatments did not affect eggshell thickness and resistance force. UV light exposure is effective in reducing microbial load on 42-week-old breeder flock eggshells, without affecting their quality, and can be considered an alternative to formaldehyde disinfection.

This study investigated the relationship between the bacterial communities of bulk tank milk (BTM) and water used in Brazilian dairy farms. We also explored the association between BTM microbiota and its quality parameters (i.e., somatic cell count [SCC] and total bacteria count [TBC]). Water and BTM samples were collected twice for high throughput metataxonomic characterization. Milk samples were collected for SCCs, TBCs, and antimicrobial residue analysis. Water samples were submitted for physico-chemical and microbiological analyses. Overall, the BTM bacterial community was not influenced by the water microbiota. Furthermore, a higher number of Bacillus spp. and a tendency toward a higher number of Lactococcus spp. was associated with a higher TBC, and consequently could be used as an indicator of milk quality. A higher number of Streptococcus spp. and a tendency toward a higher number of Staphylococcus spp. were associated with a higher SCC. Apart from a variation on the content of the Bacillus taxa, no effect of sampling time on BTM bacterial community was observed. Finally, a negative correlation between the number of different species (richness) on BTM and SCC was found.

The microbiota of the Brazilian Minas artisanal cheese, made from raw milk, is not well known and may include probiotic bacteria. This study aimed to assess the in vitro and in vivo probiotic properties of the lactic acid bacteria isolated from these cheeses. Thirty-six samples of the Lactobacillus/Pediococcus group were selected for in vitro investigation. Pediococcus acidilactici (PA2) and Lactobacillus plantarum (LP4) showed the best results and were tested for their ability to protect Salmonella Typhimurium orally infected mice. LP4 showed better probiotic potential than PA2 and allowed higher values of weight gain (P

Minas artisanal cheese, produced in the Campo das Vertentes region of Brazil, is made from raw milk and endogenous starter cultures. Although this cheese is of great historical and socioeconomic importance, little information is available about its microbiological and physical-chemical qualities, or about its beneficial microbiota. This work was aimed at evaluating the qualities of the cheese and the components used for its production, comparing samples collected during the dry and rainy seasons. We also conducted molecular identification and isolated 50 samples of lactic acid bacteria from cheese (n=21), water (n=3), raw milk (n=9), and endogenous starter culture (n=17). The microbiological quality of the cheese, water, raw milk, and endogenous starter culture was lower during the rainy period, given the higher counts of coagulase-positive Staphylococcus and total and thermotolerant coliforms. Enterococcus faecalis was the lactic acid bacteria isolated most frequently (42.86%) in cheese samples, followed by Lactococcus lactis (28.57%) and Lactobacillus plantarum (14.29%). Lactobacillus brevis (5.88%), Enterococcus pseudoavium (5.88%), Enterococcus durans (5.88%), and Aerococcus viridans (5.88%) were isolated from endogenous starter cultures and are described for the first time in the literature. The lactic acid bacteria identified in the analyzed cheeses may inhibit undesirable microbiota and contribute to the safety and flavor of the cheese, but this needs to be evaluated in future research.

The Minas artisanal cheese is a traditional product in its way of producing. Produced in the Minas Gerais state, Brazil, this cheese is made using raw cow's milk with the addition of an endogenous starter culture called "pingo", responsible for inoculating specific microorganisms that could enhance flavor and sensorial aspects. There are seven regions able to produce and commercialize this product - Araxa, Campo das Vertentes, Canastra Cerrado, Serra do Salitre, Serro and Triângulo Mineiro. This study aimed to assess the bacterial community of raw milk, endogenous starter culture and to uncover possible shifts in the bacterial community of the rind and core of cheeses at sixty days of ripening located in the Serra do Salitre region by Illumina MiSeq 16S rRNA gene amplicon sequencing. Raw milk and starter culture are responsible for inoculating specific bacteria into the cheese, with Planococcaceae and Streptococcaceae being prevalent throughout ripening time. The Planococcaceae family seems to develop strong interactions with the Leuconostocaceae family on the surface of these cheeses, and is associated with environmental aspects of the region, probably leading to a microbial signature of these products. Additionally, abiotic factors such as geographical location, moisture and acidity are major drivers in the microbial shift.

The objective of the present work was to assess the qualitative and quantitative characteristics of semen from the surubim do Iguaçu (Steindachneridion melanodermatum). Induced spermiation was achieved in eleven males with mean weight of 1.76 ± 0.48 kg and average age of two years and semen was collected by stripping. The average volume was 1.34 ± 0.73 mL. The duration of sperm motility was 154.4 ± 72.6 and 149.0 ± 77.5 seconds after activation with hatchery water and distilled water, respectively. The sperm concentration estimated by hemocytometer was 5.423 ± 2.155 x 1010 spermatozoa/mL. The results indicate that S. melanodermatum semen is easily obtained during the spawning season and the seminal characteristics are adequate insemination and subsequent in vitro fertilization. Resumo O objetivo do presente trabalho foi avaliar características qualitativas e quantitativas do sêmen de surubim do Iguaçu (Steindachneridion melanodermatum). Para as análises, utilizaram-se onze machos com peso médio de 1,76±0,48 kg e idade média de dois anos, submetidos ao tratamento de hipofisação. O volume médio de sêmen coletado foi de 1,34 ± 0,73 mL. O tempo médio de motilidade dos espermatozoides após a coleta foi de 154,4 ± 72,6 e 149,0 ± 77,5 segundos para ativação com água do tanque e água destilada, respectivamente. Para a concentração espermática, a média foi de 5,423 ± 2,155 x1010 espermatozoides/mL. Os resultados indicam que o sêmen de S. melanodermatum é facilmente obtido no período reprodutivo analisado, e que as características seminais avaliadas são adequadas para reprodução induzida.

Ingestion of milks fermented by Lactobacillus strains showing probiotic properties is an important tool to maintain gastrointestinal health. In this study, Lactobacillus rhamnosus D1 and Lactobacillus plantarum B7, isolated from Brazilian artisanal cheese, were used as starters for the functional fermented milks to assess their probiotic properties in a gnotobiotic animal model. Male germ-free Swiss mice received a single oral dose of milk fermented by each sample, and were challenged with Salmonella Typhimurium five days afterwards. Milk fermented by both Lactobacillus strains maintained counts above 108 cfu/ml during cold storage. Lactobacillus strains colonised the gut of the germ-free-mice, maintaining their antagonistic effect. This colonisation led to a protective effect against Salmonella challenge, as demonstrated by reduced pathogen translocation and histological lesions, when compared to control group, especially for Lactobacillus rhamnosus D1. Additionally, mRNA expression of inflammatory (interferon gamma, interleukin (IL)-6, tumour necrosis factor alpha) and anti-inflammatory (transforming growth factor β1) cytokines was augmented in animals previously colonised and then challenged, when compared to other experimental groups. Lactobacillus plantarum B7 colonisation also promoted higher expression of IL-17, showing a proper maturation of colonised germ-free-mice immune system. IL-5 was stimulated by both strains’ colonisation and not by S. Typhimurium challenge.

CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Outra Agência Background: both diarrhea in calves and mastitis in cows limit cattle production. The bacteria involved in these diseases have shown multi-resistance to antimicrobials, however plant metabolites therefore can provide an alternative method of control. This study selected and characterized Cerrado plant extracts showing inhibitory effects against Escherichia coli and Staphylococcus spp. from cattle. Thirteen leaf extracts were initially screened and diameters of inhibition zones produced against the pathogens were recorded using an agar disk diffusion method. Total condensed tannin contents were determined and antibacterial activities were analyzed after tannin removal from the five selected extracts. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were evaluated by macro-dilution antimicrobial susceptibility tests, and the extracts were characterized by high performance liquid chromatography. Results: inter- and intra-specific bacterial variations in the susceptibility to the extracts were detected. The aqueous extract (AE) from Caryocar brasiliense Cambess. leaves produced larger inhibition zones against E. coli strains than did other selected extracts. However, the AE from Schinopsis brasiliensis was the most effective against Staphylococcus spp. strains (P

O objetivo deste estudo foi determinar o potencial probiótico in vitro de Lactobacillus spp. isolados de queijos minas artesanais da Serra da Canastra, considerando-se o antagonismo entre amostras isoladas frente a microrganismos indicadores, a susceptibilidade a antimicrobianos, a sensibilidade ao ácido gástrico e a sensibilidade a sais biliares. Todas as bactérias ácido-lácticas testadas apresentaram resistência ao ácido gástrico (pH 2,0) e aos sais biliares (0,3%), bem como atividade antagonista contra Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Salmonella enterica var. Typhimurium, Enterococcus faecalis e bactérias ácido-lácticas isoladas dos próprios queijos - Lactobacillus plantarum (D27) e Lactobacillus rhamnosus (B25). Todas as amostras foram sensíveis à eritromicina e tetraciclina e resistentes à ciprofloxacina, gentamicina, oxacilina, estreptomicina e vancomicina. L. plantarum(B17) apresentou melhor potencial probiótico, pois obteve resultados satisfatórios em todas as propriedades avaliadas. Mais estudos são necessários para verificar a presença e a capacidade de transmissão de genes de resistência antimicrobiana a outros microrganismos e para avaliar o potencial dos microrganismos in vivo. As bactérias selecionadas poderão ser utilizadas na elaboração de queijos em que sejam mantidos o sabor e a tradição do queijo minas artesanal do estado de Minas Gerais.

Cepas de Staphylococcus spp. molecularmente identificadas foram submetidas à Reação em Cadeia da Polimerase (PCR), utilizando-se iniciadores específicos para a detecção de genes codificadores de enterotoxinas clássicas (SEA, SEB, SEC, SED, SEE) e da Toxina-1 da Síndrome do Choque Tóxico (TSST-1). Foi realizada PCR-Multiplex para detecção dos genes sea, sec, sed e see. Para seb e tst, foram realizadas PCR-Uniplex. Além disso, foi analisado o perfil de susceptibilidade das cepas a antimicrobianos de diferentes classes e foi verificado antagonismo in vitro entre Lactobacillus spp. e as cepas estudadas. Genes codificadores de enteroxinas clássicas, assim como de TSST-1, não foram encontrados. Em relação ao antibiograma, Sulfonamida, Penicilina, Ceftazidima e Oxacilina apresentaram os maiores percentuais de resistência (100, 80, 60 e 40%, respectivamente). Os demais antimicrobianos foram eficientes em percentuais acima de 70%. Lactobacillus spp. foram capazes de inibir o desenvolvimento in vitro de Staphylococcus spp. Conclui-se que as cepas estudadas não possuem genes codificadores da produção de enterotoxinas clássicas e TSST-1, são sensíveis à maioria dos antimicrobianos e são inibidos por bactérias do gênero Lactobacillus.

Brucella ovis infection is a major cause of epididymitis and infertility in rams, resulting in reproductive failure and significant economic losses worldwide. The goal of this study was to develop a PCR test targeting specific B. ovis genomic sequences. Specific primer pairs were designed targeting 12 of those ORFs. Samples of blood, serum, semen, urine, and preputial wash were collected from experimentally infected rams ( n = 9) every other week up to 180 days post infection (dpi), when tissue samples were obtained. Blood, serum, semen, urine, and preputial wash samples were obtained, in weekly intervals for 1 month, from eight rams belonging to a B. ovis -free flock. Semen samples were also obtained from rams belonging to naturally infected flocks ( n = 40). The limit of detection of this PCR protocol was 100, 10, and 1 CFU/mL for semen, urine and prepucial wash samples, respectively. Sensitivity and specificity values obtained with this PCR method were similar to that of bacteriology when evaluating biological samples. Agreement between PCR and bacteriology results was greater than 90%. These results clearly indicate that this species-specific PCR method is highly efficient for the diagnosis of B. ovis infection in semen, urine, preputial wash and tissue samples from infected rams.

To evaluate percutaneous mitral balloon valvuloplasty (PMBV) in surgical high risk patients.Twenty (12%) patients out of 172 submitted to a PMBV were considered high surgical risk cases; 17 (85%) were women and mean age was 43 (18-69). Thirteen (65%) were in acute pulmonary edema (3 were pregnant, 2 had previous cerebrovascular event, 1 had pulmonary thromboembolism and other had chronic renal failure), and 7 (35%) were in functional class (CF) III (2 had coronary artery disease, 2 severe obesity, 1 pulmonary neoplasia, 1 cardiac cachexia and one with previous cerebrovascular event). All patients underwent PMBV through transseptal technique. Double balloon was used in 10 (50%) patients; Inoue balloon in 5 (25%), mono-foil in 3 (15%) and bi-foil in 2 (10%).Seventeen (85%) obtained success and 2 (10%) had clinical improvement. The hemodynamic results pre versus post-PMBV showed: left atrium pressure (LAP) mmHg 28.2 +/- 10.0 x 15.2 +/- 9.2 (n = 20) (p0.001), mitral medium gradient (G) mmHg 21.2 +/- 10.7 x 10.7 +/- 6.7 (n = 18) (p0.001), mitral valve area (MVA) cm2 0.73 +/- 0.3 x 1.73 +/- 0.6 (n = 11) (p0.001), pulmonary artery pressure mmHg 52.0 +/- 18.2 x 40.1 +/- 14.7 (n = 18) (p0.001) and cardiac index L/min/m2 2.1 +/- 0.4 x 2.5 +/- 0.6 (n = 14) (p0.001). Comparative echocardiography results pre, post and 6 months after PMBV showed: MVA 1.06 +/- 0.39 x 1.92 +/- 0.51 x 1.65 +/- 0.5 and G 13.8 +/- 4.7 x 7.3 +/- 3.6 x 7.3 +/- 4.4. There were two immediate deaths (pulmonary thromboembolism and multiple organs failure) and other after hospital discharge. The complications were: 2 (10%) patients with neurological complications, 1 (5%) atrial septal defect and other right atrium perforation (5%).PMBV offers an alternative to surgical treatment in high risk surgical patients and the results are maintained in the 6 month follow-up.

Minas artisanal cheese is made from endogenous starter cultures, including lactic acid bacteria (LAB). Some LAB may possess probiotic potential. Thus, this study aimed to evaluate the in vitro probiotic properties of lactobacilli isolated from Minas artisanal cheeses produced in Minas Gerais. Ten samples of lactobacilli, formerly isolated from those cheeses, were submitted to the following assays: antimicrobial susceptibility, tolerance to artificial gastric juice and biliary salts, production of hydrogen peroxide and antagonism against pathogenic and non-pathogenic micro-organisms. Only L. plantarum (C0) was sensitive to all tested antimicrobials, while the other LAB samples were resistant to at least one drug. Six samples were tolerant to artificial gastric juice, and L. brevis (A6) even grew in that medium. Three samples were tolerant to biliary salts. Only L. brevis (E35) produced hydrogen peroxide. Difference (P< 0.05) was observed among the means of inhibition haloes of lactobacilli against Enterococcus faecalis ATCC 19433 and Lactobacillus plantarum C24 in spot-on-the-lawn assay. All samples of lactobacilli inhibited Escherichia coli ATCC 25922, Salmonella enterica var. Typhimurium ATCC 14028 in co-culture antagonism test (P< 0.0001). Most lactobacilli samples showed in vitro probiotic potential. From the tested samples, L. brevis (A6) presented the best results considering all in vitro probiotic tests. RESUMO O queijo minas artesanal é produzido por culturas starters endógenas, incluindo bactérias ácido-láticas (BAL). Algumas BAL podem possuir potencial probiótico. Com isso, este estudo teve como objetivo avaliar as propriedades probióticas in vitro de lactobacilos isolados de queijo minas artesanal produzido no estado de Minas Gerais. Dez amostras de lactobacilos, previamente isoladas desses queijos, foram submetidas aos seguintes testes: susceptibilidade aos antimicrobianos, tolerância ao suco gástrico artificial e aos sais biliares, produção de peróxido de hidrogênio e antagonismo contra micro-organismos patogênicos e não patogênicos. Apenas L. plantarum (C0) foi sensível a todos os antimicrobianos testados, enquanto as outras amostras de BAL foram resistentes a, pelo menos, uma droga testada. Seis amostras foram tolerantes ao suco gástrico artificial, e L. brevis (A6) apresentou crescimento nesse meio. Três amostras foram tolerantes aos sais biliares. Apenas L. brevis (E35) produziu peróxido de hidrogênio. Diferença (P