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Next-generation sequencing has revolutionized our ability to investigate the microbiota composition of diverse and complex environments. However, a number of factors can affect the accuracy of microbial community assessment, such as the DNA extraction method, the hypervariable region of 16S rRNA gene targeted, or the PCR primers used for amplification. The aim of this study was to assess the influence of commercially available DNA extraction kits and different primer pairs to provide a non-biased vision of the composition of bacterial communities present in olive xylem sap. For that purpose, branches from "Picual" and "Arbequina" olive cultivars were used for xylem sap extraction using a Scholander chamber device. The DNA extraction protocol significantly affected xylem sap bacterial community assessment. That resulted in significant differences in alpha (Richness) and beta diversity (UniFrac distances) metrics among DNA extraction protocols, with the 12 DNA extraction kits evaluated being clustered in four groups behaving differently. Although the core number of taxa detected by all DNA extraction kits included four phyla, seven classes, 12 orders, 16 or 21 families, and 12 or 14 genera when using the Greengenes or Silva database for taxonomic assignment, respectively, some taxa, particularly those identified at low frequency, were detected by some DNA extraction kits only. The most accurate depiction of a bacterial mock community artificially inoculated on sap samples was generated when using the PowerPlant DNA extraction kit, the combination of 799F/1193R primers amplifying the hypervariable V5–V7 region, and the Silva 132 database for taxonomic assignment. The DESeq2 analysis displayed significant differences among genera abundance between the different PCR primer pairs tested. Thus, Enterobacter, Granulicatella, Prevotella, and Brevibacterium presented a significant higher abundance in all PCR protocols when compared with primer pair 799F/1193R, while the opposite was true for Pseudomonas and Pectobacterium. The methodological approach followed in this study can be useful to optimize plant-associated microbiome analysis, especially when exploring new plant niches. Some of the DNA extraction kits and PCR primers selected in this study will contribute to better characterize bacterial communities inhabiting the xylem sap of olives or other woody crop species. ; This study was funded by the project AGL2016-75606-R (Programa Estatal de I+D Orientado a los Retos de la Sociedad from Spanish Government, the Spanish State Research Agency, and FEDER-EU) and Project XF-ACTORS (grant 727987) from the European Union's Horizon 2020 Framework Research Programme. MA-M is a recipient of a research fellowship BES-2017-082361 from the Spanish Ministry of Economy and Competitiveness.

JJA is funded by a National Institute of Health Research Academic Clinical Fellowship and has received an Action Medical Research training fellowship. TC is funded by a Crohn's in Childhood research association fellowship. CMC received a PhD studentship from SULSA Spirit industrial studentship. The NGS analysis was made possible by the award of a grant from the Source Bioscience 110th year anniversary promotion to CMC. The Rowett Institute receives funding from the Scottish Government (RESAS). ; Peer reviewed ; Publisher PDF

The widely accepted dogma of intrauterine sterility and initial colonization of the newborn during birth has been blurred by recent observations of microbial presence in meconium, placenta, and amniotic fluid. Given the importance of a maternal-derived in utero infant seeding, it is crucial to exclude potential environmental or procedural contaminations and to assess fetal colonization before parturition. To this end, we analyzed sterilely collected intestinal tissues, placenta, and amniotic fluid from rodent fetuses and tissues from autoptic human fetuses. Total bacterial DNA was extracted from collected samples and analyzed by Next Generation Sequencing (NGS) techniques using hypervariable 16S ribosomal RNA (rRNA) regions (V3-V4). Colonizing microbes were visualized in situ, using labeled probes targeting 16S ribosomal DNA by fluorescent in situ hybridization. The NGS analysis showed the presence of pioneer microbes in both rat and human intestines as well as in rodent placentas and amniotic fluids. Microbial communities showed fetus- and dam-dependent clustering, confirming the high interindividual variability of commensal microbiota even in the antenatal period. Fluorescent in situ hybridization analysis confirmed the microbes' presence in the lumen of the developing gut. These findings suggest a possible antenatal colonization of the developing mammalian gut.

Microbial endophytes are well known to protect host plants against pathogens, thus representing a promising strategy for the control of xylem-colonizing pathogens. To date, the vast majority of microbial communities inhabiting the olive xylem are unknown; therefore, this work pursues the characterization of the xylem-limited microbiome and determines whether the culture isolation medium, olive genotype, and the plant material used to analyze it can have an effect on the bacterial populations retrieved. Macerated xylem tissue and xylem sap extracted with the Scholander chamber from olive branches obtained from two cultivated and a wild olive genotypes were analyzed using culture-dependent and -independent approaches. In the culture-dependent approach using four solid culture media, a total of 261 bacterial isolates were identified after performing Sanger sequencing of 16S rRNA. Culturable bacteria clustered into 34 genera, with some effect of culture media for bacterial isolation. The cultivated bacteria belonged to four phyla and the most abundant genera included Frigoribacterium (18.8%), Methylobacterium (16.4%), and Sphingomonas (14.6%). On the other hand, in the culture-independent approach conducted using Illumina MiSeq 16S rRNA amplicon sequencing [next-generation sequencing (NGS)] of the xylem extracts, we identified a total of 48 operational taxonomic units (OTUs) belonging to five phyla, being Sphingomonas (30.1%), Hymenobacter (24.1%) and Methylobacterium (22.4%) the most representative genera (>76% of reads). In addition, the results indicated significant differences in the bacterial communities detected in the xylem sap depending on the genotype of the olive tree studied and, to a minor extent, on the type of sap extraction method used. Among the total genera identified using NGS, 14 (41.2%) were recovered in the culture collection, whereas 20 (58.8%) in the culture collection were not captured by the NGS approach. Some of the xylem-inhabiting bacteria isolated are known biocontrol agents of plant pathogens, whereas for others little information is known and are first reported for olive. Consequently, the potential role of these bacteria in conferring olive tree protection against xylem pathogens should be explored in future research. ; This study was funded by project AGL2016-75606-R (Programa Estatal de I+D Orientado a los Retos de la Sociedad from the Spanish Government and the Spanish State Research Agency and FEDER-EU) and Project XF-ACTORS (grant 727987) from the European Union's Horizon 2020 Framework Research Programme. MA-M is a recipient of a research fellowship BES-2017-082361 from the Spanish Ministry of Economy and Competitiveness.

Issue 11.5 of the Review for Religious, 1952. ; A. M. D.G. ' ' Review for Religious SEPTEMBER i5, 1952 Congress of Religious . The Editors Towards Continual Prayer . Pa.I DeJaegher Cases o~ Illegitimacy . Joseph F. Gallen ' "~ . Pius XII and Our Lady . JohnA. Hardon "Meaning" of A.M.D.G . Walter ~J. Ong Q, uesfions and Answers Book No÷ices VOLUME XI NUMBER 5 REVIEW FOR. RELIGIOUS VOLUME XI SEPTEMBER, 1952 NUMBER 5 CONTENTS CONGRESS OF RELIGIOUS~The Editors . 225 TOWARDS CONTINUAL PRAYER--Paul De Jaegher, S.J . 231 OUR CONTRIBUTORS . ¯ . 241 PRACTICAL AND PASTORAL C/(SES ON ILLEGITIMACY~ Joseph F. Gallen, S.J . " . 242 COMMUNION CARD FOR HOSPITALS . 248 COMPLETE TEXT OF SPONSA CHRIST1 IN ENGLISH . 248 LEGISLATION OF SPON,$A CHRI,~TI . 248 POPE PIUS XII AND OUR LADY---3ohn A. Hardon. S.J . 249 "A.M.D.G.": DEDICATION OR DIRECTIVE~Waher J. Ong, S.J. 257 TEN-YEAR INDEX--LIMITED SUPPLY . 264 QUESTIONS AND ANSWERSm 21. Moral Obligation of Voting . 265 22. Boundary of Novitiate . 269 23. On Reading Rodriguez . 269 24. Change in the Habit . 270. 25. Applying Indulgences to Souls in Purgatory . 270 26. Honoring'Bequests for Masses . . . '. . 270 27. Unrealizable Desires for Sanctification . 271 BOOK ANNOUNCEMENTS . 272 BOOK NOTICES . 274 REVIEW FOR RELIGIOUS, September, 1952. Vol. XL No. 4. Published bi-monthly: January, March, May, July, September, and November at the Colleg.e Press, 606 Harrison Street, Topeka, Kansas, by Sty Mary's College, St. Marys, Kansas, with ecclesiastical approbation. Entered as second class matteb ,January 15, 1942, at the Post Office, Topeka, Kansas, under the a~t of March 3, 1879. Editorial Board: Jerome Breunig, S.J.; Augustine G. Ellard, S.J.; Adam C. Ellis, S.J.; Gerald Kelly, S.3.; Francis N. Korth, S.3. Copyright, 1952, by Adam C. Ellis, S.J. Permission is hereby granted for quota- ~ions of reasonable le.ngth; provided due credit be given this review and the author. Subscription price: 3 dollars a year; 50 cents a copy. Printed in U. S. A. Before writing to us, please consult notice on inside back cover. Congress ot: Religious The Editors ~"HE first National Congress of Religious of'the United States w.as .| held at "th~ University of Notre Dame, August 9 to 12, 1952. Three members of our editorial staff attended as delegates. At the various sessions we tried to note points that would be bf special interest to our readers: and, since it was impossible for us tobe per-sonally present at ail the different sectional, meetings, we asked many friends t~ make similar n6tes. ,The present report is made up from these notes "jottings" might be a better word. .Regarding our report, let us make two observations. First, it is not intended to be a complete a&ount of the Congress. Official Proceedings of the Congress will be published soon and will give this complete accountl ' Secondly, even as an incomplete account, our re-port is not adequate; it represents merely what a comparatively tiny number of de'legates considered point.s of spedal interest. It would .help us greatly if other delegates who read this rePort would Suppl~- ¯ ment (or~ correct, if need be) this material by sending us communi-cations containing their own impressions: And it would also ,help if any r.eaders, whether delegates or not, w, ould send their observations on the points recorded her~. This "request for commtinications r~eeds emp,hasis. The Congress opened up for discussion many vital points concerning our religious life; but because of lack of time it could do,no more than start the discussion. Readers' of this Review would do a great service to the ~ellgious life in this country if they would continue this discussion. by the frank and constructive expression of theii own observations-on these ~;ital points. Purposes of Congress " The Congress was summonedby the Sacred Congre~gfition of Religious, as a mean~ of intensifying and. strengthe.ning the religious life in the United States, of givii~g religious of all institutes an op-portunity to exchange ideas and particularly to discuss the problems ~ertaining to the adjustment of the religious life to ~onditions, pie- ¯ vailing in our land without compromising the principles on which the religious life is based. We believe that the. first purpose of.the Cbngress the intensi- ' ,225 THE EDITOP~ Review f6r, Retigious lying ~nd strengtt~ening of. our own. r~eligious life--~as. easily .the central point of the meetings andthat' ~ good start was m~de towards its accomplishment.~ C~rtainly all of us were~ inspire~d with-the de'sire of becoming better religious arid of making our" institutes more.effec-tive means in the great work of the Church. As c~n'e Sisterwho helped us p~epare?these notes put it:. "Probabl~ among the fi, nest outcomes of the First National .Congress of Religio,us of the Ufiited States will be a'faller.realization'on the.part ofeach community, whether large or ~all, of iis.actual, p~iticipation and impqrtance.in i~he Mystical -Body. of ~hrist; a deeper feeling of p~rspnal love for our Holy Father' and an appreciation Of his interest in our spiritual and temporal well- .,being; an~under.standila~g of the ,eagerness,. of the members 6f the Sacred Congregation of Religiou~ to serve our need~s and tO assist us to b~come holy, saintly' religious; and finally, a cloker bond among th( religious communities'of the United States, with e~ich ~roup cor~: scious of its imp6rtance to the good of the whole 'and, ready to assist in furthering the 'apostolic endeavors of every, other institute:" ¯ The other purposeF-~-the ekchange of ideas and the consideration of nece~sary adjustment~--also received their due attention" at both the scheduled sessiofis and the informal meetings of smaller groupL The on.e flaw in the plaorfing of the Congress, if there was 0ily fl~w,. was that there were so many.papers that the.re was not sufficient time for discussion°fr0m the floi3r. Nevertheless, a fine beginning was made, and w,e hope that what was begun at Notre Dame can be car- ¯ tied on much further in' the discussi6ns in this R~view and in smaller meetings that can be arranged from time ~o time. . Organization .The Sacred Congregation of." Rhligioussent. f~ur representatives to theCongress: .The Most Reverend Atcadio LarraonL C.MIF., Secretary of the Congregation: the Very Reverend Giuseppe\Gi.am- ~pietro, S.J., the assistant to Father Larraona i~ the organization of regional "meetings of religious; the .Very Reverend Elio Gambari, S.M.M'., .whose specihl duty is-to handlethe affairs of refigious :in the United States; and the ~i~y Re;gerend Edward 'L. Heston, C.S.C:, the Secretary G~ner~l for, the Congress in the United States. ¯ The Congress was divided into two sections, for religious n~dn and religious women respective.ly. The Very-P(everend John ~J. Cavanaugh, C.S.C., who was President of the University of Notre .Dame atthe time the C6ngress was.plhnned, wasHonora.ry~Chair- 226 ~September, 1952 , (~ONGRESS OF RELI,.GIOUS manfor both groups. Executiye Chairman for Religious Men was. the "Very Reverend FranCis J.Connell, C.SS.R.; and for Religious W~men, the_Reverend Mother Mary. Gerald Barry,. O.P., the Supe-rior, General 6f the.Domlnican Sisters of Adrian; Michigan. Co- - ordinato.r of the. Congress'was the Re~'e~end Alfred F: .Mendez. C.S.C. The work done by Fathers Connell and Mendez and Mother " Mary Geriild in preparing for th~ Congress "was little shdrt of miraculous. They had only al~out three months tb make their pre-parati6ns, yet every detail, both Of the preparations and Of the'actual carrying out of the"Congre.ss,: was S~l~erbfy:planned and executed. As one small sample.migh~ l~e instanced the plans for daily Masses: there, were seven hur~dred Masses.each day; yet there was not the 'slightest -c~nfusion or difficulty in getting, iri ~he Masses. ~ ¯ Other members of the Committee for the sectlon of Rehgmus ¯Men were: theVery Rev~:end Godf.rey Diekmann, O.S.B.; the Very Reverend Philip F. Mulhern, O.P.: the,.Very Reverend'Thomas .~. Plas'smann, O.F.M.: the Ve~y Reverend Adam C. Ellis, "S.J.: the Very Reverend Basil Frison, C.M.F.; Venerable Brother Alexis Vic-.- tor, F.S~C.; Venerable Brother Ephrern O'Dwyer, C.S.C.; Vener-able Brother William, ~.F.X. O~her members of the committee .for Religious Women were: the Re~,erend .Mother M. Catherine Sullivan, D.C.: the Reverend'. Mother-M. Rose Elizabeth, C.S.C." the Reverend Mother M. li~n, C.S.J.; the Reverend 'Mother M. Joan 0t: Arc Cronin, O.S.U. the Reverend. Mot'her Mari~Helene,. S.P.: and Sister M. Madeleva.;" Spiritual Ideals In one wa.y'or anothe'r many. of the discussions at the C~ngr~ss centered on the clarification of the spiritual ideals common to reli-gious. ins~itfltes and on the .means. of¯ attaining these ideals., Particu-larly stressed was.~he fact that religious.need a deep pers'orial devo~ tion to Christ. Basic to such a devotion is the knowledge of.Christ; iand the young r~llg~ous must be.helped ~o get th~s knowledge, partly through Well-planned reading, and~mostly through prayer--for it is ~i knowledge.of the heart, and it is given by the Holy Ghost to ~hose " who humbly and perse~'erifigly seek it. ¯The fervent-reception of. Holy Communion-is a great help to.the atta!ning of this interior -knowledge and devotion. Incidentally, in. the men:s discussion 6f this tiepin, it was poin~e~, out that here, as i~ Other aspec~so,of their religious training, youri~ religious are inspired.by the goqd example ¯ 227 THE EDITOR,S " Review/:or Religigus of thei~ elders-and are proportib.nate!y harmed by thelack of such example. Several discussions also emphasized the need of a sense of per, sor~al responsibility. For. instance, one danger oK the religious life, with its many exercises in common, is .what migl~t be called "~herd-spirituality": one goes to the exercise automatically and takes.part with a sort, of detached numbness, ,as though ¯partially anesthetized. The chief wa~ to counteract this is the constant striving on the part of the individual to make the exercls~ personal. Also, some religious who lead an active aposl~olate, especially in small houses, .are fre-quently unable to have common exercises. They can lose the rell-gious spirit completely Unless through their own personal efforts they try to form a plan for making their various spiritual exercises in private. " The same idea of personal responsibility, under the formality of per_sonal initiative, was prominent in the men's discussion of religious obedience. It was pointed out,that apostolic initiative is n6t stifled by obedience, though it must often be controlled for the common good, as well as for the good of the individual. The rel, igious who always waits to be told what to do. is by no. means the model of perfect obedience, and the superior who requires this of his subjects is by no,means the perf.ect superior. , In a paper t, bat all will read With interest and profit, the Very Reverend Giles Staab, O.F.M.C~'p., reduced the moral qualifications of candida.tes to the religious life to the)four virtues of generosity, docility, prudence,~and loyalty. The generous candidate will¯have the r~quisite piety, the fight intention,' the chastity, and the zeal. The docile candidate will be obedient and thus further the Work of the i.nstitute. The prudent candidate will have good jiadgment and emotional control. And ~he candidat~ imbued with a spirit Of loya[t~t.will, be ready to subordinate his own interests to thqse of the community and will, as a natural consequence, b~e a, gobd communi@ man: a religious withsocial 'balanch, cgurtesy, and considerateness." Conte~ptative Life The .Right Reverend Abbot M. James Fox, O.C.S.O., gave' an interesting and informative talk about the contemplative life in gen-eral and the Trappists' life inparticular. He said that there is.a great hunger for thec0ntemplative life in modern America, . and he illustrated this statement.by quotin~ excerpts from man'y letters'that h'e has received from applicants to the' Trappi~ts. The Trappists,., 228 " ' v Sep~ei'hber~ 1952.- " CONGRESS OF RELIGIOUS he said, have about ~700' novices in their varigus" houses ~throu.ghmit the world';" ,approximately half:of these novices ,are, in the' United State~.'In less than ten years the" Trappist monasteries in 'this country have increased from three' to ten.," °, "'- .~, Why the attraction to "contemplative orders?.+. At one of Sisters' sessions it was suggested that yout.h are attracted to the co.n= templative life because they,feel that in this life they can