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This study aimed to investigate the physiochemical characterizations of the fermented product (FP) produced by the high-yield surfactin strain Bacillus subtilis LYS1 (LYS1), as well as its effects on growth performance, carcass traits, intestinal morphology, tibial bone characteristics, and clinical blood biochemistry in broilers. Accordingly, the optimal supplementation of FP for improving growth performance, intestinal villi development, and tibial bone strength in broilers was elucidated using a broken-line quadratic (BLQ) ascending model. Three hundred and sixty 0-day-old Ross 308 broiler chicks, with equal numbers of both sexes, were randomly assigned to dietary supplementation of 2.5% fish meal or 0, 1, 1.5, 2, or 2.5% FP. Each treatment had six replicates, and the experimental period was 5 wk. The LYS1 count, surfactin content, and surfactin composition of the FP were 9.1 log CFU/g, 11.23 mg/g, and C12 to C18, respectively. The FP-supplemented groups improved feed intake, weight gain (WG), and production efficiency factor at 0 to 5 weeks old (p < 0.05) compared with the 0% group. The villus height/crypt depth (V/C) in the jejunum and ileum of the FP-supplemented groups was higher than in the 0% group (p < 0.05). The tibiotarsal index, Ca, and P in the tibia showed a linear effect with increased FP supplementation (p < 0.05). Moreover, the tibiotarsus weight/length index (TWLI) showed a quadratic effect with increased FP supplementation (p < 0.05). The optimal supplementation of FP for WG, V/C in the jejunum and ileum, and TWLI was 1.8, 1.9, and 1.6%, respectively. In conclusion, dietary supplementation with 1 to 2.5% LYS1 FP in broilers can improve their growth performance and the development of intestinal villi. Moreover, 1.9% is the optimal supplementation of LYS1 FP in the diet, based on the fitting results obtained with the BLQ model.
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(1) Background: Agarwood is an aromatic resin produced by the host tree through an immunological response against biotic and abiotic stress. The aim was, first, to use the fungus Geotrichum candidum to stimulate compound changes in Aquilaria sinensis horizontally (color formation) and vertically (cutting layers) after injection with it. (2) Methods: Horizontal and vertical sections were collected and separated five months after injection with the fungal broth. Two grams of dry powder was mixed with 20 mL methanol for 3 h at room temperature, and the solution was vibrated in an ultrasonic cleaner bath at 40 °C for 1 h. After vacuum drying, a concentration of 10 mg/mL of the tested samples in methanol was prepared for reversed-phase high-performance liquid chromatography (RP-HPLC), gas chromatography/mass spectrometry (GC/MS), and thin-layer chromatography (TLC) analysis. (3) Results: The horizontal changes in the compounds and their concentrations were associated with color. Compared to the normal (N) group, G. candidum injection stimulated more compounds at RT 27-42 in the white (W) group, brown (BR) group, and black (B) group. Furthermore, a significant increase in fatty acids was observed in the W group, implying an early plant response after G. candidum injection. In the BR group, the compounds were more similar to commercial agarwood (Out group). In the B group, alkaloids were the main compounds. Vertical changes in the main compounds were not observed, although the compound level varied. A TLC analysis determined the main compounds in the BR group at 254 nm and in the B group at 365 nm. Higher fatty acid levels were found in L6 and L5 and were correlated with higher terpenoid and sesquiterpene levels, suggesting that these compounds were possibly the first stage of agarwood formation. A GC/MS analysis demonstrated that the main compound groups were almost identical to the BR parts. (4) Conclusions: The injection of G. candidum led A. sinensis to synthesize different phytochemicals horizontally, not vertically, in the BR group.
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Soy yogurt has been gaining popularity as a vegan food produced simply by soymilk fermentation with proper microbial manipulation. It is well known that soy containing rich isoflavones is beneficial for ameliorating hyperglycaemic disorders. Soy fermentation can improve the bioavailability of these precious nutrients. Lactiplantibacillus plantarum is one of the most abundant and frequently isolated species in soymilk manufacturing. Soy yogurts produced with efficient GABA (γ-aminobutyric acid)-producing L. plantarum and the deglycosylating activity of L. plantarum were functionally assessed in a STZ-induced hyperglycaemic mouse model. Hyperglycaemic mice were assigned into groups and treated with daily gavage of either dH2O, soymilk, soy yoghurts produced with high GABA-producing L. plantarum GA30 (LPGA30), low GABA-producing L. plantarum PV30 (LPPV30) or the soy yoghurts fortified with additional 30 mg g-1 GABA counterparts (GA + GABA and PV + GABA groups). Except the dH2O group, all soy yoghurt groups retained body weight with improved glucose homeostasis, glucose tolerance test results and renal tissue integrity, while the soymilk group shows partial benefits. Plasma GABA concentrations in the daily soy yoghurt-supplemented groups (LPGA30 and LPPV30) plateaued at 5 times higher than the average 0.5 µM in dH2O and soymilk groups, and their GABA-fortified soy yoghurt counterparts (GA + GABA and PV + GABA) groups were accountable for the restored plasma insulin levels. Gut microbiome analysis revealed dysbiosis in STZ-induced hyperglycemic mice of the dH2O group with breached out facultative anaerobic Proteobacteria over the normal phyla Firmicutes and Bacteroidetes. Restored gut microbiota with transitionally populated Actinobacteria was demonstrated in the LPGA30 group but not in the LPPV30 group. Soy yoghurts produced with efficient GABA-producing L. plantarum GA30 showed exceptional benefits in modulating gut microbiota with dominant genera of Enterococcus, Lactobacillus and Bifidobacterium, and the presence of some minor beneficial microbial communities including Akkermansia muciniphila, Butyricicoccus pullicaecorum, Corynebacterium spp. and Adlercreutzia spp. Efficient GABA-producing L. plantarum GA30 fermented soymilk to produce soy yoghurts that exhibit profound synergistic protections over rich soy isoflavones to restore pancreatic ß-cell functions for insulin production in STZ-induced hyperglycaemic mice. Additionally, the probiotic role of GABA-producing L. plantarum in re-establishing healthy gut microbiota in hyperglycaemic mice implies a possible symbiotic relationship, awaiting further exploration.
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Diabetes Mellitus Experimental , Microbioma Gastrointestinal , Hiperglicemia , Insulinas , Isoflavonas , Probióticos , Animais , Camundongos , Estreptozocina , Iogurte , Hiperglicemia/terapia , Diabetes Mellitus Experimental/terapia , Ácido gama-Aminobutírico , Camundongos Obesos , FermentaçãoRESUMO
Soy isoflavones possess antioxidative, anti-inflammatory, anti-diabetic and phytoestrogenic properties. Soybean residue contains a fair amount of nutrients such as glycosylated isoflavones, minerals and dietary fibers, and is a substantial waste product produced from soymilk and tofu manufacturing. A solid-state fermentation of soybean residue by Rhizopus oligosporus or co-inoculated with Lactiplantibacillus plantarum improves the availability of isoflavones and GABA content which is attributed to ameliorated hyperglycemic symptoms in STZ-induced hyperglycemic mice. The effortless solid-state fermentation with present microbial manipulation supports an anti-hyperglycemia value-added application of soybean residue for functional food development. Background: Due to an awareness of the food crisis and with a rapidly rising prevalence of diabetes, recycling the substantial fibrous soybean residue disposed from soy industries has received consideration. Methods: Lactiplantibacillus plantarum was previously screened for active glutamate decarboxylase, and ß-glucosidase activities were adopted for the fermenting of soybean residue using a traditional tempeh solid-state fermenting process with fungal Rhizopus oligosporus. Fermented soybean residue was chemically analyzed and functionally assessed in in vitro and in vivo hyperglycemic conditions. Results: A 48 h longer solid-state fermentation of the soybean residue co-inoculated with R. oligosporus and L. plantarum showed improved contents of isoflavone aglycones and GABA which were attributed to augmented antioxidative capacity, lowered ROS level, improved blood biochemistry, and better blood glucose homeostasis in STZ-induced hyperglycemic mice. Conclusion: The advantages of a food industrial effortless fermentation process, and a health nutritional endorsing anti-hyperglycemic value-added property offer a practical alternative in recycled soybean residue.
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Streptococcus agalactiae (GBS) can infect newborns, pregnant women and immunocompromised or elderly people. This study aimed to investigate differences in three pilus genes and virulence genes pavA, cfb, rib and scpB and changes in predominant serotypes III, V and VI from 2008 to 2012. The susceptibilities to penicillin, ceftriaxone, azithromycin, erythromycin, clindamycin, levofloxacin and moxifloxacin of 145 GBS strains of serotype III, V and VI strains from 2008 and 2012 were determined using disc diffusion method. PCR identification of ST-17, the pilus genes and virulence genes; multilocus sequence typing (MLST); and conserved domain and phylogenetic analysis of scpB-1 and scpB-2 proteins were performed. A dramatic number reduction was observed in serotype V, not III and V, from 2008 to 2012. The rate of resistance to azithromycin, clindamycin and erythromycin was the highest in serotype V. ST-17 was only found in serotype III with pilus genes PI-1+PI-2b. The major pilus genotype was PI-1+PI-2a. Serotype V without the rib gene was reduced in number between two studied years. Compared to scpB-1, scpB-2 had a 128-bp deletion in a PA C5a-like peptidase domain and putative integrin-binding motif RGD. In conclusion, reduction in serotype V may be due to presence of scpB-2 or lack of genes scpB and rib.
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Staphylococcus aureus is a major human pathogen that produces various virulence factors which promote the binding of bacteria to tissues and medical devices such as vascular access devices, thereby developing a wide range of invasive infections. Vascular access serves as an entry site for S. aureus and elevates the risk of infection in the hemodialysis population. Nevertheless, the distribution of virulence genes in Staphylococcus spp. associated with vascular access infections (VAIs) has not been studied previously. In this study, we determined the relationship between the molecular characteristics and virulence profiles of S. aureus isolates obtained from VAIs. We collected isolates from patients with VAIs between August 2017 and December 2020 and further analyzed the molecular characteristics, antimicrobial resistance profiles, and virulence gene distribution in the isolates. Overall, 15 sequence types (STs), including a new ST (ST6892) and 19 spa types, were identified among the 56 isolates. Of the 53 S. aureus isolates, ST8, ST239, ST45, and ST59 were the predominant STs, whereas ST2250 was the only ST in 3 S. argenteus isolates. ST45-SCCmecIV-t026 (abbreviated as ST45-IV-t026), ST59-V-t437, and ST8-IV-t008 were the predominant clones that belonged to agr type I. All isolates harbored clfB and eno, whereas all S. aureus isolates harbored clfA. In addition, 10 Panton-Valentine leucocidin-positive isolates belonged to ST8 and ST59, with ST8-IV-t008 and ST59-V-t437 being the predominant clones. In brief, the distribution of virulence genes associated with STs may assist in the spread of molecular types of Staphylococcus spp.
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The National Bovine Tuberculosis (bTB) Eradication Program for dairy cattle has been operating in Taiwan for many years and has allowed the prevalence of bTB to decrease gradually. However, 29% of intradermal tuberculin test (ITT)-positive dairy cows were later found to be TB negative based on necropsy, histopathological examination, and mycobacterial isolation results. Studies in Taiwan have indicated that Mycobacterium avium subsp. paratuberculosis (MAP) may lead to false-positive ITT. Due to the high prevalence (over 90%) of paratuberculosis (PTB) serum antibody among Taiwan's farms, comparative ITT (CITT) has been recommended to differentiate between bTB and PTB infections. In this study, we used ITT, CITT, and enzyme-linked immunosorbent assay (ELISA) to evaluate the prevalence of bTB from 2012 to 2018. We also used pathological and bacterial examination from ITT-positive dairy cows to evaluate CITT's diagnostic ability and adjust its cutoff point accordingly. After careful selection, 36 cows (including 31 cows from 11 ITT-positive farms and 5 from 2 ITT-negative farms) were examined by CITT. The cutoff point was adjusted using a receiver operating characteristic (ROC) analysis. Overall, our results identified the ITT-positive prevalence in Taiwan as 0.03-0.22%, and PTB-positive prevalence as 54.55-73.53%. The results of sensitivity, specificity, kappa, and ROC analyses have identified the optimal cutoff point for the CITT in Taiwan as ≥ 3 mm. At this cutoff point value, the sensitivity and specificity were 62.5% and 96.43%, respectively. Our findings can be used to reduce the false-positive response rate caused by PTB cross-reaction and accelerate the eradication of bTB in Taiwan.
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Doenças dos Bovinos , Mycobacterium bovis , Paratuberculose , Tuberculose Bovina , Animais , Bovinos , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Paratuberculose/diagnóstico , Paratuberculose/epidemiologia , Sensibilidade e Especificidade , Taiwan/epidemiologia , Tuberculina , Teste Tuberculínico/veterinária , Tuberculose Bovina/diagnóstico , Tuberculose Bovina/epidemiologiaRESUMO
BACKGROUND: Nontyphoid Salmonella and Shigella can cause gastroenteritis in humans. Ceftriaxone (CRO) has been used to treat their infection, however, development of CRO resistance are often associated with plasmid-mediated blaCMY. Here, we investigated the presence of plasmid-mediated ISEcp-1 tnpA-blaCMY-2-blc-sugE and the role of these genes in regulation of CRO susceptibility in different hosts. METHODS: 194 strains of Salmonella serovars and Shigella were tested for CRO susceptibility. Non-susceptibility strains were examined for plasmid-mediated ISEcp-1 tnpA-blaCMY-2-blc-sugE by PCR amplification, Southern blot, and DNA sequencing. The plasmid profiles were determined by HindIII-digested restriction fragment length polymorphism (RFLP). Four recombinant plasmids with different genes from ISEcp-1 tnpA-blaCMY-2-blc-sugE were constructed and then were transferred into Escherichia coli and different Salmonella serovars to evaluate the CRO susceptibility. RESULTS: Among 20 CRO-nonsusceptible isolates of Salmonella Choleraesuis (5), S. Typhimurium (4), S. Mons (1), S. Stanley (2) and Shigella sonnei (8) with plasmid-mediated blaCMY-2, 19 isolates carried the ISEcp-1 tnpA-blaCMY-2-blc-sugE and only one isolate with tnpA-blaCMY-2. Transformation of these plasmids into E. coli pir116 produced multidrug resistance. Furthermore, PCR-RFLP analysis determined 5 different plasmid profiles and identical RFLP pattern between S. Typhimurium and S. sonnei. Transformation of the recombinant plasmids into E. coli and different Salmonella serovars resulted in phenotypes ranging from susceptible to resistant (especially inducible resistance) to CRO that were dependent on the genes, and host. CONCLUSION: The CRO susceptibility associated with the ISEcp-1 tnpA-blaCMY-2-blc-sugE element is regulated positively by ISEcp-1 tnpA and SugE and negatively regulated by Blc and unknown species-dependent host factor(s).
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Antibacterianos/farmacologia , Ceftriaxona/farmacologia , Farmacorresistência Bacteriana/genética , Regulação Bacteriana da Expressão Gênica , Plasmídeos/genética , Salmonella/genética , Shigella/genética , beta-Lactamases/genética , China/epidemiologia , DNA Bacteriano/genética , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/microbiologia , Humanos , Salmonella/efeitos dos fármacos , Infecções por Salmonella/epidemiologia , Infecções por Salmonella/microbiologia , Shigella/efeitos dos fármacosRESUMO
Burkholderia cepacia complex (BCC) is a group of closely related bacteria with widespread environmental distribution. BCC bacteria are opportunistic pathogens that cause nosocomial infections in patients, especially cystic fibrosis (CF). Multilocus sequence typing (MLST) is used nowadays to differentiate species within the BCC complex. This study collected 41 BCC isolates from vascular access infections (VAIs) and other clinical infections between 2014 and 2020. We preliminarily identified bacterial isolates using standard biochemical procedures and further conducted recA gene sequencing and MLST for species identification. We determined genetic diversity indices using bioinformatics software. We studied 14 isolates retrieved from patients with VAIs and observed that Burkholderia cepacia was the predominant bacterial species, and B. contaminans followed by B. cenocepacia were mainly retrieved from patients with other infections. According to MLST data, we identified that all B. contaminans isolates belonged to ST102, while a wide variety of sequence types (STs) were found in B. cenocepacia isolates. In summary, the high diversity and easy transmission of BCC increase BCC infections, which provides insights into their potential clinical effects in non-CF infections.
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Toxoplasma gondii and Neospora caninum are intracellular protozoan parasites that cause reproductive disorders in ruminants and humans. Information on the risk factors of T. gondii and N. caninum infections in goats is very limited in Taiwan. The aim of the study was to investigate the epidemiology and identify the risk factors of these two infections in goats. A total of 630 caprine sera were collected from 42 dairy goat farms and the owners were interviewed by a structured questionnaire. The apparent seroprevalences of T. gondii in farm- and individual- levels were respectively 88.1% and 32.22%, while those of N. caninum were 19.05% and 2.54%, respectively. Toxoplasma gondii B1 gene was identified in 7 feed samples and 8 from the water samples whereas N. caninum was not found. Wooden flooring was the main risk factor for T. gondii infection while the frequency of visits by staff to other farms and the breed of goat were risk factors for N. caninum. The improvement of flooring materials or thorough cleaning, periodic disinfection and maintenance of dryness on the floor are highly recommended for the prevention of T. gondii infection in farmed goats. In addition, unnecessary visits to other farms should be limited to prevent the spread of N. caninum. These factors should be highlighted for the prevention of T. gondii and N. caninum in goats, particularly when raised in intensive housing system with flooring on height.
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Coccidiose/veterinária , Doenças das Cabras/parasitologia , Neospora/imunologia , Toxoplasma/imunologia , Toxoplasmose Animal/epidemiologia , Análise de Variância , Ração Animal/parasitologia , Animais , Anticorpos Antiprotozoários/sangue , Coccidiose/epidemiologia , Coccidiose/parasitologia , Coccidiose/prevenção & controle , Água Potável/parasitologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/prevenção & controle , Cabras , Imunoglobulina G/sangue , Neospora/genética , Neospora/isolamento & purificação , Razão de Chances , Reação em Cadeia da Polimerase/veterinária , Fatores de Risco , Estudos Soroepidemiológicos , Inquéritos e Questionários , Taiwan/epidemiologia , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Toxoplasmose Animal/prevenção & controleRESUMO
BACKGROUND: Extraintestinal pathogenic E. coli (ExPEC) is a common gram-negative organism causing various infections, including urinary tract infections (UTIs), bacteremia, and neonatal meningitis. The cjrABC-senB gene cluster of E. coli contributes to ExPEC virulence in the mouse model of UTIs. Consistently, the distribution of cjrABC-senB is epidemiologically associated with human UTIs caused by E. coli. cjrABC-senB, which has previously been proposed to encode an iron uptake system, may facilitate ExPEC survival in the iron availability-restricted urinary tract. Given that the bloodstream is also an iron limited environment to invading bacteria, the pathogenic role of cjrABC-senB in ExPEC bacteremia, however, remains to be investigated. METHODS: The ability of ExPEC RS218 strains with and without cjrABC-senB to survive in the mouse bloodstream and human serum was evaluated. Subsequently, the role of this gene cluster in the ExPEC interaction with the complement system was evaluated. Finally, the distribution of cjrABC-senB in human clinical E. coli isolates was determined by PCR. The frequency of cjrABC-senB in bacteremia isolates that were not associated with UTIs (non-UTI bacteremia isolates) was compared with that in UTI-associated isolates and fecal isolates. RESULTS: Expression of cjrABC-senB attenuated the survival of RS218 in the mouse bloodstream and human serum. The cjrABC-senB-harboring strains triggered enhanced classical- and alternative-complement pathway activation and became more vulnerable to complement-mediated killing in serum. cjrA was identified as the major gene responsible for the attenuated serum survival. Expressing cjrABC-senB and cjrA increased bacterial susceptibility to detergent and induced periplasmic protein leakage, suggesting that the expression of these genes compromises the integrity of the outer membrane of ExPEC. In addition, the frequency of cjrABC-senB in non-UTI bacteremia isolates was significantly lower than that in UTI-associated isolates, while the frequencies in non-UTI bacteremia isolates and fecal isolates showed no significant difference. Consistently, this epidemiological investigation suggests that cjrABC-senB does not contribute to E. coli bacteremia in humans. CONCLUSION: The contribution of cjrABC-senB to the pathogenesis of ExPEC is niche dependent and contradictory because the genes facilitate ExPEC UTIs but hinder bacteremia. The contradictory niche-dependent characteristic may benefit the development of novel strategies against E. coli-caused infections.
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Bacteriemia/microbiologia , Ativação do Complemento , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Escherichia coli Extraintestinal Patogênica/fisiologia , Genes Bacterianos , Família Multigênica , Animais , Escherichia coli Extraintestinal Patogênica/genética , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Salmonella infection is most common in patients with infected aortic aneurysm, especially in Asia. When the aortic wall is heavily atherosclerotic, the intima is vulnerable to invasion by Salmonella, leading to the development of infected aortic aneurysm. By using THP-1 macrophage-derived foam cells to mimic atherosclerosis, we investigated the role of three Salmonella enterica serotypes - Typhimurium, Enteritidis, and Choleraesuis - in foam cell autophagy and inflammasome formation. Herein, we provide possible pathogenesis of Salmonella-associated infected aortic aneurysms. Three S. enterica serotypes with or without virulence plasmid were studied. Through Western blotting, we investigated cell autophagy induction and inflammasome formation in Salmonella-infected THP-1 macrophage-derived foam cells, detected CD36 expression after Salmonella infection through flow cytometry, and measured interleukin (IL)-1ß, IL-12, and interferon (IFN)-α levels through enzyme-linked immunosorbent assay. At 0.5 h after infection, plasmid-bearing S. Enteritidis OU7130 induced the highest foam cell autophagy - significantly higher than that induced by plasmid-less OU7067. However, plasmid-bearing S. Choleraesuis induced less foam cell autophagy than did its plasmid-less strain. In foam cells, plasmid-less Salmonella infection (particularly S. Choleraesuis OU7266 infection) led to higher CD36 expression than did plasmid-bearing strains infection. OU7130 and OU7266 infection induced the highest IL-1ß secretion. OU7067-infected foam cells secreted the highest IL-12p35 level. Plasmid-bearing S. Typhimurium OU5045 induced a higher IFN-α level than did other Salmonella serotypes. Salmonella serotypes are correlated with foam cell autophagy and IL-1ß secretion. Salmonella may affect the course of foam cells formation, or even aortic aneurysm, through autophagy.Salmonella infection is most common in patients with infected aortic aneurysm, especially in Asia. When the aortic wall is heavily atherosclerotic, the intima is vulnerable to invasion by Salmonella, leading to the development of infected aortic aneurysm. By using THP-1 macrophage-derived foam cells to mimic atherosclerosis, we investigated the role of three Salmonella enterica serotypes Typhimurium, Enteritidis, and Choleraesuis in foam cell autophagy and inflammasome formation. Herein, we provide possible pathogenesis of Salmonella-associated infected aortic aneurysms. Three S. enterica serotypes with or without virulence plasmid were studied. Through Western blotting, we investigated cell autophagy induction and inflammasome formation in Salmonella-infected THP-1 macrophage-derived foam cells, detected CD36 expression after Salmonella infection through flow cytometry, and measured interleukin (IL)-1ß, IL-12, and interferon (IFN)-α levels through enzyme-linked immunosorbent assay. At 0.5 h after infection, plasmid-bearing S. Enteritidis OU7130 induced the highest foam cell autophagy significantly higher than that induced by plasmid-less OU7067. However, plasmid-bearing S. Choleraesuis induced less foam cell autophagy than did its plasmid-less strain. In foam cells, plasmid-less Salmonella infection (particularly S. Choleraesuis OU7266 infection) led to higher CD36 expression than did plasmid-bearing strains infection. OU7130 and OU7266 infection induced the highest IL-1ß secretion. OU7067-infected foam cells secreted the highest IL-12p35 level. Plasmid-bearing S. Typhimurium OU5045 induced a higher IFN-α level than did other Salmonella serotypes. Salmonella serotypes are correlated with foam cell autophagy and IL-1ß secretion. Salmonella may affect the course of foam cells formation, or even aortic aneurysm, through autophagy.
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Aneurisma Aórtico/microbiologia , Infecções por Salmonella/microbiologia , Salmonella typhimurium/patogenicidade , Animais , Aneurisma Aórtico/genética , Aneurisma Aórtico/imunologia , Linhagem Celular , Humanos , Interferon-alfa/genética , Interferon-alfa/imunologia , Interleucina-12/genética , Interleucina-12/imunologia , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Antígeno Ki-1/genética , Antígeno Ki-1/imunologia , Macrófagos/imunologia , Macrófagos/microbiologia , Monócitos/imunologia , Monócitos/microbiologia , Plasmídeos/genética , Plasmídeos/metabolismo , Infecções por Salmonella/genética , Infecções por Salmonella/imunologia , Salmonella typhimurium/genética , Salmonella typhimurium/fisiologia , Sorogrupo , VirulênciaRESUMO
S. Choleraesuis (Choleraesuis) and S. Typhimurium (Typhimurium) cause salmonellosis in pigs and humans. The effects of vaccine strains pSV-less Typhimurium OU5048 and Choleraesuis OU7266 and SPI-2-mutant Choleraesuis SC2284 on the immune responses of pigs against Typhimurium, Choleraesuis, and S. Enteritidis (Enteritidis) with or without the virulence plasmid (pSV) were determined. After oral vaccination of three vaccine groups and challenge with Choleraesuis CN36, the level of Salmonella-specific IgG in sera and the bactericidal effects and superoxide generation of peripheral blood mononuclear cells (PBMCs) and polymorphonuclear leukocytes (PMNs) against the above strains were determined using ELISA and NBT assay, respectively. Among three vaccine strains tested, OU7266 stimulated the highest Salmonella-specific IgG levels. Complement inactivation increased IgG concentration, while E. coli absorption reduced IgG levels. The pSV-containing strains were less resistant to serum killing than the pSV-less strains, and Enteritidis exhibited the lowest resistance to serum killing. Serovars tested, vaccine strains, and timeline periods postvaccination and challenge were important factors affecting superoxide production. The two Choleraesuis vaccine strains stimulated greater levels of superoxide from PMNs and PBMCs than the Typhimurium strains. The PMNs and PBMCs in challenged and vaccinated pigs reduced more superoxide than those in challenged hosts. In vaccinated hosts, pSV-less Salmonella strains triggered lower levels of PMN/PBMC-generated superoxide upon challenge than strains with pSV against Enteritidis and Choleraesuis. Overall, Choleraesuis OU7266 may be better than the other vaccine strains in generating the greatest IgG levels, serum bactericidal activity and superoxide levels. The pSV likely influences the immune responses.
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Imunoglobulina G/sangue , Salmonelose Animal/prevenção & controle , Vacinas contra Salmonella/uso terapêutico , Doenças dos Suínos/prevenção & controle , Animais , Anticorpos Antibacterianos/sangue , Proteínas do Sistema Complemento/imunologia , Escherichia coli/imunologia , Escherichia coli/metabolismo , Feminino , Leucócitos Mononucleares/imunologia , Neutrófilos/imunologia , Espécies Reativas de Oxigênio/imunologia , Salmonella , Salmonelose Animal/imunologia , Vacinas contra Salmonella/imunologia , Salmonella typhimurium , Ensaios de Anticorpos Bactericidas Séricos , Suínos , Vacinação/veterinária , Vacinas Atenuadas/imunologia , Vacinas Atenuadas/uso terapêuticoRESUMO
Salmonella Typhimurium and S. Stanley are the most prevalent serogroup B serovars to infect humans in Taiwan. The aim was to determine possible factors to influence the prevalence between S. Typhimurium and S. Stanley. Genotypes were determined by pulsed field gel electrophoresis (PFGE) analysis and the intracellular survival, phagocytosis, reactive oxygen species (ROS) production of human monocyte THP-1 cell and tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6), and IL-1ßexpression in peripheral blood CD14+ cells after infection were analyzed. 182 S. Stanley was clonal disseminated with main pulsotypes 2 from 2004 to 2007. Overall S. Typhimurium evolved more genotypes, while S. Stanley conserved in genotypes. Human blood CD14+ monocytes expressed TNF-α, IL-6 and IL-1ß differently among serovars and bacterial conditions (live vs. killed). Live S. Stanley and S. Typhimurium suppressed the TNF-α and IL-6 expression compared to killed bacteria. However, live S. Typhimurium stimulated more IL-1ß expression than the killed bacteria, but S. Stanley expressed similar IL-1ß levels in both conditions. Furthermore, S. Stanley and S. Typhimurium differed in intracellular survival in the THP-1 cells, an early decrease for S. Stanley, not for S. Typhimurium. Additionally, higher reactive oxygen species (ROS) production in THP-1 cells was found agsinst S. Stanley infection, not found in S. Typhimurium. However, some isolates of S. Stanley could recover from early loss to become more in the monocytes than S. Typhimurium. Difference in phagocytized number, intracellular survival, ROS production and IL-1ß expression may contribute to prevalence different between two serovars.
Assuntos
Monócitos/imunologia , Fagocitose/imunologia , Infecções por Salmonella/imunologia , Salmonella typhimurium/imunologia , beta-Lactamases/genética , Linhagem Celular Tumoral , Eletroforese em Gel de Campo Pulsado , Humanos , Interleucina-1beta/biossíntese , Interleucina-6/biossíntese , Espécies Reativas de Oxigênio/metabolismo , Infecções por Salmonella/epidemiologia , Infecções por Salmonella/microbiologia , Salmonella typhimurium/genética , Salmonella typhimurium/isolamento & purificação , Células THP-1 , Taiwan , Fator de Necrose Tumoral alfa/biossínteseRESUMO
Hemodialysis patients are particularly vulnerable to Staphylococcus aureus infection, with the vascular access serving as the site of entry for this formidable pathogen. Patients with arteriovenous grafts (AVGs) and tunneled-cuffed catheters (TCCs) are at elevated risk of S. aureus infection. In this study, we investigated the correlation between the clinical characteristics of S. aureus vascular access infection (VAI), molecular profiles, and the biofilm formation abilities of clinical isolates of S. aureus. We collected samples of methicillin-resistant S. aureus (MRSA), methicillin-sensitive S. aureus (MSSA), and methicillin-sensitive S. argenteus (MSSAg) from patients with S. aureus VAI and patients with other infections. The molecular profiles of the clinical isolates were determined using disk diffusion testing and molecular typing. The biofilm formation ability was determined by microtiter plate assay. In total, 63 S. aureus and 10 S. argenteus isolates were identified: 40 MRSA, 23 MSSA, and ten MSSAg. MRSA was highly prevalent (77.8%) in TCC isolates and was multidrug resistant. Of the 40 MRSA isolates, ST239-SCCmec III was the predominant clone. SCCmec type IV was the predominant type (35%) in isolates from AVGs, while SCCmec type III was prevalent in TCC infection and showed significantly higher biofilm formation ability than types IV and V. In dialysis VAI by S. aureus, patients with TCC were more often infected with MRSA than patients with AVG, and MRSA in TCC-VAI was predominantly SCCmec type III, which had the strongest drug resistance and biofilm formation ability.
Assuntos
Infecções Relacionadas a Cateter/microbiologia , Diálise Renal/efeitos adversos , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificação , Idoso , Biofilmes/crescimento & desenvolvimento , Testes Diagnósticos de Rotina , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Farmacorresistência Bacteriana Múltipla , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Tipagem Molecular , Staphylococcus , Staphylococcus aureus/classificação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/genética , Taiwan , Centros de Atenção TerciáriaRESUMO
Salmonella is a common foodborne and zoonotic pathogen. Only a few serovars carry a virulence plasmid (pSV), which enhances the pathogenicity of the host. Here, we investigated the pathogenicity roles of the pSVs among wild-type, plasmid-less, and complemented S. Typhimurium, S. Enteritidis S. Choleraesuis in invasion, phagocytosis, and intracellular bacterial survival in human THP-1â¯cells and cell death patterns by flow cytometry and difference in cell death patterns between pig and human S. Choleraesuis isolates with large pSCVs. Virulence plasmid (pSTV) led to slightly increasing cellular apoptosis for S. Typhimurium; virulence plasmid (pSEV) enhanced apoptosis and necrosis significantly for S. Enteritidis; and pSCV reduced apoptosis significantly for S. Choleraesuis. After complementation, pSTV increased the intracellular survival of pSCV-less Choleraesuis and the cytotoxicity against human THP-1â¯cells. Using the Cytochalasin D to differentiate the invasion of S. Choleraaesuis and phagocytosis of THP-1â¯cells determined that pSCV were responsible for invasion and phagocytosis at 0â¯h and inhibited intracellular replication in THP-1â¯cells, and pSTV were responsible for invasion and increased intracellular survival for S. Choleraesuis in THP-1â¯cells. The human isolates with large pSCV induced more cellular apoptosis and necrosis than the pig isolates. In conclusion, human S. Choleraesuis isolates carrying large pSCVs were more adapted to human THP-1â¯cells for more cell death than pig isolates with large pSCV. The role of pSVs in invasion, phagocytosis, intracellular survival and apoptosis differed among hosted serovars.
Assuntos
Morte Celular , Interações Hospedeiro-Patógeno , Plasmídeos/genética , Salmonella enterica/genética , Salmonella enteritidis/genética , Salmonella typhimurium/genética , Fatores de Virulência/genética , Animais , Apoptose , Citocalasina D/farmacologia , Replicação do DNA , Genes Bacterianos , Humanos , Camundongos , Viabilidade Microbiana , Necrose , Células RAW 264.7 , Salmonelose Animal/microbiologia , Salmonella enterica/patogenicidade , Salmonella enteritidis/patogenicidade , Salmonella typhimurium/patogenicidade , Sorogrupo , Suínos , Células THP-1 , VirulênciaRESUMO
Natural compounds have been candidates for anticancer medicine over the last 20 years. During the process of isolating seed oil from Calophyllum inophyllum L., yellow and green pigments containing multiple compounds with an aromatic structure were identified. High-performance liquid chromatography and nuclear magnetic resonance analysis of these pigments revealed that the compounds present were identical, but the concentration of the compounds was different. Treatment with the pigments was able to induce the death of DLD-1 human colon cancer cells and increase the percentage of the cells in the sub-G1 and sub-G2/M phases in a dose-dependent manner. Additionally, the pigments were able to exhibit cytotoxic activity on A549 and H1975 human non-small cell lung cancer (NSCLC) cell lines at 24 h, with half-maximal inhibitory concentrations (IC50) values of 0.1206 and 0.0676%, respectively for green pigments, and 0.0434 and 0.0501%, respectively for yellow pigments. Furthermore, a decrease in IC50 value was associated with an increase in the duration of treatment. However, a sharp decrease in IC50 value of the yellow pigment was observed for H1975 cells at 48 h and for A549 cells at 72 h compared with no change in IC50 value for the green pigment with time, suggesting that the pigments function and induce cell death differently in the two cell lines. An investigation was performed into the synergistic effect of the green pigment and gefitinib (Iressa®, ZD1839), which is a selective epidermal growth factor receptor-tyrosine kinase inhibitor to block growth factor-mediated cell proliferation. The combination of the green pigment and gefitinib resulted in an enhancement of the decrease in viability of A549 and H1975 cells compared with treatment with gefitinib alone, which suggested that treatment with the green pigments was able to enhance the sensitivity of NSCLC cells to gefitinib. In conclusion, these pigments may be considered for development as anti-colon cancer agents.
RESUMO
Arachidin-1 [trans-4-(3-methyl-1-butenyl)-3,5,3',4'-tetrahydroxystilbene] is a polyphenol produced by peanut kernels during germination. The aim of the present study was to investigate the mechanism underlying the anti-inflammatory effect of arachidin-1 in endothelial cells (ECs). The results of cell adhesion and western blotting assays demonstrated that arachidin-1 attenuated tumor necrosis factor (TNF)-α-induced monocyte/EC adhesion and intercellular adhesion molecule-1 (ICAM-1) expression. Arachidin-1 was demonstrated to exert its inhibitory effects by the attenuation of TNF-α-induced nuclear factor-κB (NF-κB) nuclear translocation and inhibitor of κB-α (IκBα) degradation. Furthermore, arachidin-1 upregulated nuclear factor-E2-related factor-2 (Nrf-2), a known mediator of phase II enzyme expression, and increased the transcriptional activity of antioxidant response element. Transfection of ECs with Nrf-2 siRNA blocked the inhibitory effect of arachidin-1 on ICAM-1 expression, NF-κB nuclear translocation and IκBα degradation. In addition, arachidin-1 induced the expression of the phase II enzymes thioredoxin-1, thioredoxin reductase-1, heme oxygenase-1, glutamyl-cysteine synthetase and glutathione S-transferase. Following arachidin-1 pretreatment, the H2O2-induced generation of reactive oxygen species was reduced. Therefore, the present results indicate that arachidin-1 suppresses TNF-α-induced inflammation in ECs through the upregulation of Nrf-2-related phase II enzyme expression.
Assuntos
Inflamação/tratamento farmacológico , Molécula 1 de Adesão Intercelular/genética , Fator 2 Relacionado a NF-E2/genética , Estilbenos/administração & dosagem , Fator de Necrose Tumoral alfa/genética , Transporte Ativo do Núcleo Celular/efeitos dos fármacos , Arachis/química , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana , Humanos , Peróxido de Hidrogênio/toxicidade , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/patologia , Desintoxicação Metabólica Fase II/genética , Inibidor de NF-kappaB alfa/genética , NF-kappa B/genética , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Estilbenos/química , TransfecçãoRESUMO
This protocol describes the development of a colloidal gold immunochromatographic test strip based on the sandwich format that can be used to differentiate the myoglobin (Mb) of cetaceans from that of seals and other animals. The strip provides rapid and on-the-spot screening for cetacean meat, thereby restraining its illegal trade and consumption. Two monoclonal antibodies (mAbs) with reactivity toward the Mb of cetaceans were developed. The amino acid sequences of Mb antigenic reactive regions from various animals were analyzed in order to design two synthetic peptides (a general peptide and a specific peptide) and thereafter raise the mAbs (subclass IgG1). The mAbs were selected from hybridomas screened by indirect ELISA, western blot and dot blot. CGF5H9 was specific to the Mbs of rabbits, dogs, pigs, cows, goats, and cetaceans while it showed weak to no affinity to the Mbs of chickens, tuna and seals. CSF1H13 can bind seals and cetaceans with strong affinity but showed no affinity to other animals. Cetacean samples from four families (Balaenopteridae, Delphinidae, Phocoenidae and Kogiidae) were used in this study, and the results indicated that these two mAbs have broad binding ability to Mbs from different cetaceans. These mAbs were applied on a sandwich-type colloidal gold immunochromatographic test strip. CGF5H9, which recognizes many species, was colloid gold-labeled and used as the detection antibody. CSF1H13, which was coated on the test zone, detected the presence of cetacean and seal Mbs. Muscle samples from tuna, chicken, seal, five species of terrestrial mammals and 15 species of cetaceans were tested in triplicate. All cetacean samples showed positive results and all the other samples showed negative results.
Assuntos
Anticorpos Monoclonais/química , Cetáceos , Cromatografia de Afinidade/métodos , Coloide de Ouro/química , Mioglobina/análise , Animais , Técnicas de Química Sintética , Sensibilidade e Especificidade , Especificidade da EspécieRESUMO
BACKGROUND: Streptococcus agalactiae (GBS) is a common pathogen to infect newborn, woman, the elderly, and immuno-compromised human and fish. 37 fish isolates and 554 human isolates of the GBS in 2007-2012 were investigated in serotypes, antibiotic susceptibility, genetic difference and pathogenicity to tilapia. RESULTS: PCR serotyping determined serotype Ia for all fish GBS isolates and only in 3.2 % (3-4.2 %) human isolates. For fish isolates, all consisted a plasmid less than 6 kb and belonged to ST7 type, which includes mainly pulsotypes I and Ia, with a difference in a deletion at the largest DNA fragment. These fish isolates were susceptible to all antimicrobials tested in 2007 and increased in non-susceptibility to penicillin, and resistance to clindamycin and ceftriaxone in 2011. Differing in pulsotype and lacking plasmid from fish isolates, human serotype Ia isolates were separated into eight pulsotypes II-IX. Main clone ST23 included pulsotypes II and IIa (50 %) and ST483 consisted of pulsotype III. Human serotype Ia isolates were all susceptible to ceftriaxone and penicillin and few were resistant to erythromycin, azithromycin, clindamycin, levofloxacin and moxifloxacine with the resistant rate of 20 % or less. Using tilapia to analyze the pathogenesis, fish isolates could cause more severe symptoms, including hemorrhage of the pectoral fin, hemorrhage of the gill, and viscous black and common scites, and mortality (>95 % for pulsotype I) than the human isolates (<30 %); however, the fish pulostype Ia isolate 912 with deletion caused less symptoms and the lowest mortality (<50 %) than pulsotype I isolates. CONCLUSION: Genetic, pathogenic, and antimicrobial differences demonstrate diverse origin of human and fish serotype Ia isolates. The pulsotype Ia of fish serotype Ia isolates may be used as vaccine strains to prevent the GBS infection in fish.