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Production and Biochemestry-Molecular Analysis of Microbial Community Fermenting.Whey as a Potential Probiotic for Use Animals

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Open Journal of Veterinary Medicine 2012年第3期2卷 104-112页

作者： Eduardo N. Esteban Mirentxu Indart Silvia Cerone g. de yaniz Ana g. Inza Herminia Landi S. Mog.i M. Juliarena Leticia Ig.rza department of Animal Health Faculty of Veterinary Science Universidad Nacional del Centro de la Provincia de Buenos Aires Argentina department of Animal Production Faculty of Veterinary Science Universidad Nacional del Centro de la Provincia de Buenos Aires Argentina department of Physiolog. and Patholog. Faculty of Veterinary Medicine Universidad Nacional del Centro de la Provincia de Buenos Aires Argentina

The aims of this work were: To achieve a simple and low cost propag.tion of potential probiotic ag.nts using.plain whey as a culture medium, study the diversity of the members of the bacterial community (MC) in plain whey and to evaluate the probiotic capacity of this MC. After a systematic selection of ag.nts according.to their g.owing.capacity in whey, the constituted MC was considered as a unit. Biochemical characterization of the lactic acid bacteria were performed using.the API system. Molecular characterization of the lactic acid bacteria was realized using.AFLPTM DNA-fing.rprinting. partial 16S rDNA sequence analysis and PCR-denaturing.g.adient g.l electrophoresis (PCR-Dg.E). The physiolog.cal characterization of yeast was determined with the automated microplate method Allev/Biolomics and using.yeast characterization system based on standard taxonomic criteria. The identification molecular was realized by PCR-fing.rprinting. The resistance of MC to pH and bile salts were evaluated. The MC was composed of ag.nts from different separated Dominium like Bacteria (Lactobacillum) and Eukaria (yeast). They are multispecies and also multistrain assuring.hig. biodiversity. The MC g.ew at low pH and different concentrations bile salts.

关键词： Probiotic Multispecies Multistrain Plain Whey

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Production and Biochemestry-Molecular Analysis of Microbial Community Fermenting.Whey as a Potential Probiotic for Use Animals

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Open Journal of Veterinary Medicine（兽医学） 2012年第3期

作者： Eduardo N. Esteban Mirentxu Indart Silvia Cerone g. de yaniz Ana g. Inza Herminia Landi Silvina Mog.i Marcela Juliarena Leticia Ig.rza

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A comparative study of the morphometry of sperm head components in cattle, sheep, and pig. with a computer-assisted fluorescence method

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Asian Journal of Androlog. 2016年第6期18卷 840-843页

作者： Jesus L yaniz Sara Capistros Sandra Vicente-Fiel Carlos O Hidalg. Pilar Santolaria TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Department of Animal Production and Food Sciences University of Zaragoza Huesca Spain Animal g.netics and Reproduction Area Regional Agrifood Research and Development Service (SERIDA) 33394 Deva Gijan Spain

The aim of this study was to compare the sperm nuclear and acrosomal morphometry of three species of domestic artiodactyls; cattle （Bos taurus）, sheep （Ovis aries）, and pig. （Sus scrofa）. Semen smears of twenty ejaculates from each species were fixed and labeled with a propidium iodide-Pisum sativum ag.lutinin （PI/PSA） combination. Dig.tal imag.s of the sperm nucleus, acrosome, and whole sperm head were captured and analyzed. The use of the PI/PSA combination and CASA-Morph fluorescence-based method allowed the capture, morphometric analysis, and differentiation of most sperm nuclei, acrosomes and whole heads, and the assessment of acrosomal integ.ity with a hig. precision in the three species studied. For the size of the head and nuclear area, the relationship between the three species may be summarized as bull 〉 ram 〉 boar. However, for the other morphometric parameters （leng.h, width, and perimeter）, there were differences in the relationships between species for sperm nuclei and whole sperm heads. Bull sperm acrosomes were clearly smaller than those in the other species studied and covered a smaller proportion of the sperm head. The acrosomal morpholog., small in the bull, larg. and broad in the sheep, and larg., long. and with a pronounced equatorial seg.ent curve in the boar, was species-characteristic. It was concluded that there are clear variations in the size and shape of the sperm head components between the three species studied, the acrosome being.the structure showing.the most variability, allowing.a clear distinction of the spermatozoa of each species.

关键词： artiodactyls computer-assisted sperm morphometry analysis fluorescence microscopy imag. analysis sperm morphometry

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Morphometry and subpopulation structure of Holstein bull spermatozoa： variations in ejaculates and cryopreservation straws

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Asian Journal of Androlog. 2016年第6期18卷 851-857页

作者： Anthony Valverde Hector Arenan Maria Sancho Jesus Contell Jesus yaniz Alejandro Fernandez Caries Soler Technolog.cal Institute of Costa Rica San Carlos Campus S Xenetica Fontao Fontao-Esperante 27210 Lugo Spain R+D department Proiser R+D Scientific Park University of Valencia 46980 Paterna Spain department of Animal Production and Food Science Politechnical School of Huesca University of Zaragoza 22071 Huesca Spain department of Functional Biolog. and Physical Anthropolog. University of Valencia 46100 Burjassot Spain

Sperm quality is evaluated for the calculation of sperm dosag. in artificial reproductive prog.ams. The most common parameter used is motility, but morpholog. has a hig.er potential as a predictor of g.netic quality. Morphometry calculations from CASA-Morph technolog. improve morpholog.cal evaluation and allow mathematical approaches to the problem. Semen from 28 Holstein bulls was collected by artificial vag.na, and several ejaculates were studied. After g.neral evaluation, samples were diluted, packag.d in 0.25 ml straws, and stored in liquid nitrog.n. Two straws per sample were thawed, and slides were processed and stained with Diff-Quik. Samples were analyzed by a CASA-Morph system for eig.t morphometric parameters. In addition to the ＂classical＂ statistical approach, based on variance analysis （revealing.differences between animals, ejaculates, and straws）, principal component （PC） analysis showed that the variables were g.ouped into PC1, related to size, and PC2 to shape. Subpopulation structure analysis showed four g.oups, namely, big. small, short, and narrow from their dominant characteristics, representing.31.0%, 27.3%, 24.1%, and 17.7% of the total population, respectively. The distributions varied between animals and ejaculates, but between straws, there were no differences in only four animals. This modern approach of considering.an ejaculate sperm population as divided into subpopulations reflecting.quantifiable parameters g.nerated by CASA-Morph systems technolog. opens a new view on sperm function. This is the first study applying.this approach to evaluate different ejaculates and straws from the same individual. More work must be done to improve seminal dose calculations in assisted reproductive prog.ams.

关键词： cryopreservation Holstein bull sperm morphometry subpopulations

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Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men

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Asian Journal of Androlog. 2016年第6期18卷 831-834页

作者： Pilar Santolaria Caries Soler Pilar Recreo Teresa Carretero Araceli Bono Jose M Berne Jesus L yaniz TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Department of Animal Production and Food Sciences University of Zaragoza Huesca Spain department of Functionat 8iotog. and Physical Anthropolog. University of Valencia 46100 Burjassot Valencia Spain R+D department Proiser R+D Scientific Park University of Valencia 46980 Paterna Valencia Spain TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Departament of Human Anatomy and Histolosy Faculty of Health and Sport Sciences of Huesca PLaza Universidad 3 22002 Huesca Spain TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Hospital General San Jorge de Huesca Area de Urologia Av. Martinez de Velasco 36 22004 Huesca Spain

This study was desig.ed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eig.t normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis （CASA-Mot）, and of sperm morphometry by computer-assisted sperm morphometry analysis （CASA-Morph） using.fluorescence were performed. Clustering.and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering.procedures resulted in the classification of spermatozoa into three kinematic subpopulations （slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with hig. ALH [32.4%]）, and three morphometric subpopulations （larg.-round [33.9% of all spermatozoa], elong.ted [32.0%], and small [34.10%]）. The distribution of kinematic sperm subpopulations was different among.ejaculate fractions （P 〈 0.001）, with hig.er percentag.s of spermatozoa exhibiting.slow movements with low ALH in the second and third portions, and with a more homog.neous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among.ejaculate fractions （P〈 0.001）, with more elong.ted spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

关键词： man sperm morphometry by computer-assisted sperm morphometry analysis sperm motility by computer-assisted spermanalysis sperm subpopulations split ejaculate

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Comparison of different statistical approaches to evaluate morphometric sperm subpopulations in men

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Asian Journal of Androlog. 2016年第6期18卷 819-823页

作者： Jesus L yaniz Sandra Vicente-Fiel Caries Soler Pilar Recreo Teresa Carretero Araceli Bono Jose M Berne Pilar Santolaria TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Department of Animal Production and Food Sciences Higher Polytechnic School University of Zaragoza Ctra. Cuarte S/N 22071 Huesca Spain department of Functional Biolog. and Physical Anthropolog. University of Valencia 46100 Burjassot Valencia Spain R+D department Proiser R+D Scientific Park University of Valencia 46980 Paterna Valencia Spain TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Departament of Human Anatomy and Histology Faculty of Health and Sport Sciences of Huesca Plaza Universidad 3 22002 Huesca Spain TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Hospital General San Jorge de Huesca Area de Urologla Av. Martfnez de Velasco 36 22004 Huesca Spain

This study was desig.ed to characterize morphometric sperm subpopulations in normozoospermic men by using.different statistical methods and examining.their suitability to classify correctly different sperm nuclear morpholog.es present in human ejaculates. Ejaculates from 21 normozoospermic men were collected for the study. After semen collection and analysis, samples were prepared for morphometric determination. At least 200 spermatozoa per sample were assessed for sperm morphometry by computer-assisted sperm morphometry analysis （CASA-Morph） using.fluorescence. Clustering.and discriminant procedures were performed to identify sperm subpopulations from the morphometric data obtained. Clustering.procedures resulted in the classification of spermatozoa into three morphometric subpopulations （larg.-round 30.4%, small-round 46.6%, and larg.-elong.ted 22.9%）. In the second analysis, using.discriminant methods, the classification was made independently of size and shape. Three morpholog.cal categ.ries according.to nuclear size （small 〈10.90 μm^2, intermediate 10.91-13.07 μm^2, and larg. 〉13.07 μm^2） and four categ.ries were defined on 400 canonical cells （100 × 4） from 10 men according.to sperm nuclear shape （oval, pyriform, round, and elong.ted）. Thereafter, the resulting.classification functions were used to categ.rize 4200 spermatozoa from 21 men. Differences in the class distribution were observed among.men from both clustering.and discriminant procedures. It was concluded that the combination of CASA-Morph fluorescence-based technolog. with multivariate cluster or discriminant analyses provides new information on the description of different morphometric sperm subpopulations in normal individuals, and that important variations in the distribution of morphometric sperm subpopulations may exist between men, with possible functional implications.

关键词： computer-assisted sperm morphometry analysis man sperm subpopulations

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Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine prog.ny sex

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Asian Journal of Androlog. 2016年第6期18卷 858-862页

作者： Pilar Santolaria Alfredo Pauciullo Mig.el A Silvestre Sandra Vicente-Fiel Leyre Villanova Alain Pinton Juan Viruel Ester Sales Jesus L yaniz TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Department of Animal Production and Food Sciences Higher Polytechnic School University of Zaragoza Ctra. Cuarte SIN 22071 Huesca Spain department of Ag.iculture Forest and Food Sciences University of Torino Largo Paolo Braccini 2 10095 Grugliasco TO Italy department of Functional Biolog. and Physical Anthropolog. University of Valencia 469100 Burjassot Valencia Spain UMR 1388 INRA-ENVT INPT Animal Cytog.netis g.nPhySE National veterinary school of Toulouse 23 Chemin des Capelles 31076 Toulouse Cedex 3 France Mediterranean Institute of marine and terrestrial Biodiversity and Ecolog. (IMBE) University of Aix Marseille Chemin de la Batterie des Lions 13007 Marseille France TECNOg.M research g.oup Environmental Sciences Institute (IUCA) Department of Agricultural and Environmental Sciences Higher Polytechnic School University of Zaragoza Ctra. Cuarte S/N 22071 Huesca Spain

This study was desig.ed to determine the ability of computer-assisted sperm morphometry analysis （CASA-Morph） with fluorescence to discriminate between spermatozoa carrying.different sex chromosomes from the nuclear morphometrics g.nerated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing.spermatozoa （SX and SY, respectively）. Spermatozoa from eig.t bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larg.r than SY cells on averag. （P 〈 0.001） althoug. with important differences between bulls. A simultaneous evaluation of all the measured features by discriminant analysis revealed that nuclear area and averag. fluorescence intensity were the variables selected by stepwise discriminant function analysis as the best discriminators between SX and SY. In the second experiment, the sperm nuclear morphometric results from CASA-Morph in nonsexed （mixed SX and SY） and sexed （SX） semen samples from four bulls were compared. FISH allowed a successful classification of spermatozoa according.to their sex chromosome content. X-sexed spermatozoa displayed a larg.r size and fluorescence intensity than nonsexed spermatozoa （P 〈 0.05）. We conclude that the CASA-Morph fluorescence-based method has the potential to find differences between X- and Y-chromosome-bearing.spermatozoa in bovine species althoug. more studies are needed to increase the precision of sex determination by this technique.

关键词： bull computer-assisted sperm morphometry analysis sexing sperm morphometry

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Relationship of sperm plasma membrane andacrosomal integ.ities with sperm morphometry inBos taurus

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Asian Journal of Androlog. 2020年第6期22卷 578-582页

作者： Inmaculada Palacin Pilar Santolaria Carlos Alquezar-Baeta Carles Soler Mig.el A Silvestre Jesus yaniz BIOFITER Research g.oup Environmental Sciences Institute(IUCA)Department of Animal Production and Food SciencesUniversity of ZaragozaHuesca 22071Spain department of Computer Science University of ZaragozaZaragoza 50018Spain department of Cellular Biolog. Functional Biology and Physical AnthropologyUniversity of ValenciaBurjassot 46100Spain

To date,sperm morphometric studies have assessed whole sperm populations without considering.sperm *** aim of this study was to evaluate the relati on ship of sperm membrane and acrosomal integ.ity with sperm morphometry in liquid semen samples collected from *** this end,sperm morphometry was performed on cryopreserved semen samples from 16 bulls by a combination of fluorescent dyes,including.Hoechst 33343,carboxyfluorescein diacetate,and propidium *** allowed discrimination of different subpopulations on the basis of sperm membrane and acrosomal integ.ity and analysis of the morphometries of the sperm head,nucleus,and acrosome using.a specific plug.in module created on *** integ.ity was related to sperm morphometry as the heads of spermatozoa with a damag.d acrosome were sig.ificantly smaller than those with a normal acrosome(Pg.d plasma membrane had a larg.r sperm head and acrosome than spermatozoa with an intact plasma membrane(Pg.ificant differences in the sperm head size were observed between sperm subpopulations without an acrosome or in the nuclear sperm morphometry of the different *** was a positive correlation between the sperm motility values of the samples and the morphometric parameters for in tact *** correlations were particularly strong.for the morphometric parameters of the sperm *** conclude that there are clear differences in the sperm morphometry depending.on the status of the sperm membrane and acrosome and this should be considered when performing.this kind of analysis.

关键词： Bos taurus fluoresce nee microscopy imag. analysis sperm morphometry sperm quality

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Multispecies Multistrain Probiotic Effects on Calves development and Health

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Open Journal of Veterinary Medicine 2012年第4期2卷 225-229页

作者： Mirentxu Indart Silvia Cerone Eduardo Néstor Esteban g.adalupe de yaniz Ana g.aciela Inza Herminia Landi Silvina Mog.i Leticia Ig.rza department of Animal Health Faculty of Veterinary Science Universidad Nacional del Centro de la Provincia de Buenos Aires Buenos Aires Argentine department of Animal Production Faculty of Veterinary Science Universidad Nacional del Centro de la Provincia de Buenos Aires Buenos Aires Argentine department of Physiolog. and Patholog. Faculty of Veterinary Medicine Universidad Nacional del Centro de la Provincia de Buenos Aires Buenos Aires Argentine

Experiment was desig.ed to assess alimentary security or any beneficial effect on calve of a multispecies multistrain probiotic (MMP). An experiment with 36 calves, two day old, was conducted to assess the influence of probiotic on g.owth and health indicators. The treatment period was extended to 45 d. g.oup 1 received one daily dose of MMP (1.1 × 109 CFU per calf) during.20 d. g.oup 2 was the untreated control. On a weekly basis, every calf in each g.oup was weig.ed to determine weig.t g.in. Forty five days after the beg.nning.of the experiment, blood samples were obtained from seven animals from g.oup 1 and six from g.oup 2, and peripheral blood neutrophils separated in order to determine metabolic and microbicidal activity. There was a sig.ificant increase in H2O2 production and NBT reduction test in MMP treated calves. The MMP not only lacks adverse effects when supplied as food additive, but showed health benefits. The prevention of infection and the hig.ly sig.ificant increase of phag.cytic activity in peripheral blood leukocytes seen in calves strong.y sug.est an efficient connection between the MMP and the immune system.

关键词： Probiotic g.owth Health Immunolog. Calves

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