17 research outputs found

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    <p>Antibody responses and protection of offspring when mothers were immunized via the IN route and their offspring via the IN route<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157041#t004fn001" target="_blank"><sup>a</sup></a>.</p

    Rapid and Accurate Quantification of Viable <i>Lactobacillus</i> Cells in Infant Formula by Flow Cytometry Combined with Propidium Monoazide and Signal-Enhanced Fluorescence In Situ Hybridization

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    Lactobacillus is an important member of the probiotic bacterial family for regulating human intestinal microflora and preserving its normalcy, and it has been widely used in infant formula. An appropriate and feasible method to quantify viable Lactobacilli cells is urgently required to evaluate the quality of probiotic-fortified infant formula. This study presents a rapid and accurate method to count viable Lactobacilli cells in infant formula using flow cytometry (FCM). First, Lactobacillus cells were specifically and rapidly stained by oligonucleotide probes based on a signal-enhanced fluorescence in situ hybridization (SEFISH) technique. A DNA-binding fluorescent probe, propidium monoazide (PMA), was then used to accurately recognize viable Lactobacillus cells. The entire process of this newly developed PMA-SEFISH-FCM method was accomplished within 2.5 h, which included pretreatment, dual staining, and FCM analysis; thus, this method showed considerably shorter time-to-results than other rapid methods. This method also demonstrated a good linear correlation (R2 = 0.9994) with the traditional plate-based method with a bacterial recovery rate of 91.24%. To the best of our knowledge, the present study is the first report of FCM combined with PMA and FISH for the specific detection of viable bacterial cells

    Body weight changes in the offspring within 21 days after the lethal virus challenge.

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    <p>A). Both mothers and offspring were immunized with the inactivated vaccine through the IP route. B). Mothers were immunized through the IP route and offspring through the IN route. C). Both mothers and offspring were immunized through the IN route. D). Mothers were immunized through the IN route and offspring through the IP route. Data points represent the means ± SD of each group of mice. “MDA- control” indicates that the offspring in this group had no MDAs and were unimmunized; “1μg MDA+” indicates that the offspring in this group had MDAs and were immunized with 1 μg of the vaccine.</p

    Antibody titers in mother mice after immunization and in their offspring before immunization <sup>a</sup>.

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    <p>Antibody titers in mother mice after immunization and in their offspring before immunization <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157041#t001fn001" target="_blank"><sup>a</sup></a>.</p

    Antibody responses and protection of offspring when mothers were immunized via the IP route and their offspring via the IN route<sup>a</sup>.

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    <p>Antibody responses and protection of offspring when mothers were immunized via the IP route and their offspring via the IN route<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157041#t003fn001" target="_blank"><sup>a</sup></a>.</p

    Antibody responses and protection of offspring when both mothers and offspring were immunized via the IP route<sup>a</sup>.

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    <p>Antibody responses and protection of offspring when both mothers and offspring were immunized via the IP route<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0157041#t002fn001" target="_blank"><sup>a</sup></a>.</p

    Survival rates of offspring over time after lethal virus challenge.

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    <p>A). Both mothers and offspring were immunized with the inactivated vaccine through the IP route. B). Mothers were immunized through the IP route and offspring through the IN route. C). Both mothers and offspring were immunized through the IN route. D). Mothers were immunized through the IN route and offspring through the IP route. Data points represent the means ± SD of each group of mice. “MDA- control” indicates that the offspring in this group had no MDAs and were unimmunized; “1μg MDA+” indicates that the offspring in this group had MDAs and were immunized with 1 μg of the vaccine. <sup>a</sup>Significant differences (<i>p</i> < 0.05) compared with the MDA- control determined by the Log Rank test.</p

    Genotype Diversity of H9N2 Viruses Isolated from Wild Birds and Chickens in Hunan Province, China

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    <div><p>Three H9N2 avian influenza viruses were isolated from the Dongting Lake wetland, among which one was from fresh egret feces, the other two were from chicken cloacal swabs in poultry markets. Phylogenetic analyses suggested that eight genes of the egret-derived H9N2 virus might come from Korean-like or American-like lineages. The two poultry-derived H9N2 viruses were reassortants between the CK/BJ/94-like and G1-like viruses. Except the PB1 genes (90.6%), the nucleotide sequence of other internal genes of the two viruses exhibited high homology (>95%). In addition, they also exhibited high homology (96–98.3%) with some genes of the H7N9 virus that caused an epidemic in China in 2013. Nucleotide sequence of the poultry-derived and egret-derived H9N2 viruses shared low homology. Infection studies showed that the egret-derived H9N2 virus was non-pathogenic to both mice and chickens, and the virus was unable to infect chickens even through 8 passages continuously in the lung. On the other hand, the chickens infected by poultry-derived viruses showed obvious clinical symptoms and even died; the infected mice showed no noticeable clinical symptoms and weight loss, but viruses could be detected in their lungs. In conclusion, for the egret-derived H9N2 virus, it would take a long adaptation process to achieve cross-species transmission in poultry and mammals. H9N2 viruses isolated at different times from the same host species in the same geographical region presented different evolutionary status, and virus isolated from different hosts in the same geographical region exhibited genetic diversity. Therefore, it is important to continue the H9N2 virus surveillance for understanding their evolutionary trends so as to provide guidance for disease control and prevention.</p></div

    Molecular characterizations of HA, NA, PB2, PB1, PA, NP, M and NS at representative sites.

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    a<p>E1: A/Egret/Hunan/1/2012; C1: A/Chicken/Hunan/1/2012; C12: A/Chicken/Hunan/12/2011.</p><p>-: There was no deletion. +: There was deletion.</p

    Homology (%) of nucleotide sequences of eight genes of virus from chickens and relevant sequences of birds or H7N9.

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    a<p>E1: A/Egret/Hunan/1/2012; C1: A/Chicken/Hunan/1/2012; C12: A/Chicken/Hunan/12/2011;</p><p>BJ16:A/Brambling/Beijing/16/2012(H9N2); S69: A/Pigeon/Shanghai/S1069/2013(H7N9).</p><p>The identity was not done.</p
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