In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions

In vitro "determination" of... | F1000Research "use strict";function _typeof(t){return(_typeof="function"==typeof Symbol&&"symbol"==typeof Symbol.iterator?function(t){return typeof t}:function(t){return t&&"function"==typeof Symbol&&t.constructor===Symbol&&t!==Symbol.prototype?"symbol":typeof t})(t)}!function(){var t=function(){var t,e,o=[],n=window,r=n;for(;r;){try{if(r.frames.__tcfapiLocator){t=r;break}}catch(t){}if(r===n.top)break;r=r.parent}t||(!function t(){var e=n.document,o=!!n.frames.__tcfapiLocator;if(!o)if(e.body){var r=e.createElement("iframe");r.style.cssText="display:none",r.name="__tcfapiLocator",e.body.appendChild(r)}else setTimeout(t,5);return!o}(),n.__tcfapi=function(){for(var t=arguments.length,n=new Array(t),r=0;r 3&&2===parseInt(n[1],10)&&"boolean"==typeof n[3]&&(e=n[3],"function"==typeof n[2]&&n[2]("set",!0)):"ping"===n[0]?"function"==typeof n[2]&&n[2]({gdprApplies:e,cmpLoaded:!1,cmpStatus:"stub"}):o.push(n)},n.addEventListener("message",(function(t){var e="string"==typeof t.data,o={};if(e)try{o=JSON.parse(t.data)}catch(t){}else o=t.data;var n="object"===_typeof(o)&&null!==o?o.__tcfapiCall:null;n&&window.__tcfapi(n.command,n.version,(function(o,r){var a={__tcfapiReturn:{returnValue:o,success:r,callId:n.callId}};t&&t.source&&t.source.postMessage&&t.source.postMessage(e?JSON.stringify(a):a,"*")}),n.parameter)}),!1))};"undefined"!=typeof module?module.exports=t:t()}(); dataLayer = dataLayer || []; // Standard GTM initialization - Google Consent Mode handles consent automatically (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start': new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0], j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src= 'https://www.googletagmanager.com/gtm.js?id='+i+dl+ '>m_auth=hzk0Vc3qFsQYhCrIoHz68A>m_preview=env-1>m_cookies_win=x';f.parentNode.insertBefore(j,f); })(window,document,'script','dataLayer','GTM-MWFK8L5J'); ;window.NREUM||(NREUM={});NREUM.init={distributed_tracing:{enabled:true},privacy:{cookies_enabled:true},ajax:{deny_list:["bam.nr-data.net"]}}; ;NREUM.loader_config={accountID:"438030",trustKey:"438030",agentID:"772317073",licenseKey:"97f8f67f26",applicationID:"772317073"} ;NREUM.info={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net",licenseKey:"97f8f67f26",applicationID:"772317073",sa:1} ;/*! For license information please see nr-loader-spa-1.236.0.min.js.LICENSE.txt */ (()=>{"use strict";var e,t,r={5763:(e,t,r)=>{r.d(t,{P_:()=>l,Mt:()=>g,C5:()=>s,DL:()=>v,OP:()=>T,lF:()=>D,Yu:()=>y,Dg:()=>h,CX:()=>c,GE:()=>b,sU:()=>_});var n=r(8632),i=r(9567);const o={beacon:n.ce.beacon,errorBeacon:n.ce.errorBeacon,licenseKey:void 0,applicationID:void 0,sa:void 0,queueTime:void 0,applicationTime:void 0,ttGuid:void 0,user:void 0,account:void 0,product:void 0,extra:void 0,jsAttributes:{},userAttributes:void 0,atts:void 0,transactionName:void 0,tNamePlain:void 0},a={};function s(e){if(!e)throw new Error("All info objects require an agent identifier!");if(!a[e])throw new Error("Info for ".concat(e," was never set"));return a[e]}function c(e,t){if(!e)throw new Error("All info objects require an agent identifier!");a[e]=(0,i.D)(t,o),(0,n.Qy)(e,a[e],"info")}var u=r(7056);const d=()=>{const e={blockSelector:"[data-nr-block]",maskInputOptions:{password:!0}};return{allow_bfcache:!0,privacy:{cookies_enabled:!0},ajax:{deny_list:void 0,enabled:!0,harvestTimeSeconds:10},distributed_tracing:{enabled:void 0,exclude_newrelic_header:void 0,cors_use_newrelic_header:void 0,cors_use_tracecontext_headers:void 0,allowed_origins:void 0},session:{domain:void 0,expiresMs:u.oD,inactiveMs:u.Hb},ssl:void 0,obfuscate:void 0,jserrors:{enabled:!0,harvestTimeSeconds:10},metrics:{enabled:!0},page_action:{enabled:!0,harvestTimeSeconds:30},page_view_event:{enabled:!0},page_view_timing:{enabled:!0,harvestTimeSeconds:30,long_task:!1},session_trace:{enabled:!0,harvestTimeSeconds:10},harvest:{tooManyRequestsDelay:60},session_replay:{enabled:!1,harvestTimeSeconds:60,sampleRate:.1,errorSampleRate:.1,maskTextSelector:"*",maskAllInputs:!0,get blockClass(){return"nr-block"},get ignoreClass(){return"nr-ignore"},get maskTextClass(){return"nr-mask"},get blockSelector(){return e.blockSelector},set blockSelector(t){e.blockSelector+=",".concat(t)},get maskInputOptions(){return e.maskInputOptions},set maskInputOptions(t){e.maskInputOptions={...t,password:!0}}},spa:{enabled:!0,harvestTimeSeconds:10}}},f={};function l(e){if(!e)throw new Error("All configuration objects require an agent identifier!");if(!f[e])throw new Error("Configuration for ".concat(e," was never set"));return f[e]}function h(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");f[e]=(0,i.D)(t,d()),(0,n.Qy)(e,f[e],"config")}function g(e,t){if(!e)throw new Error("All configuration objects require an agent identifier!");var r=l(e);if(r){for(var n=t.split("."),i=0;i {r.d(t,{D:()=>i});var n=r(50);function i(e,t){try{if(!e||"object"!=typeof e)return(0,n.Z)("Setting a Configurable requires an object as input");if(!t||"object"!=typeof t)return(0,n.Z)("Setting a Configurable requires a model to set its initial properties");const r=Object.create(Object.getPrototypeOf(t),Object.getOwnPropertyDescriptors(t)),o=0===Object.keys(r).length?e:r;for(let a in o)if(void 0!==e[a])try{"object"==typeof e[a]&&"object"==typeof t[a]?r[a]=i(e[a],t[a]):r[a]=e[a]}catch(e){(0,n.Z)("An error occurred while setting a property of a Configurable",e)}return r}catch(e){(0,n.Z)("An error occured while setting a Configurable",e)}}},6818:(e,t,r)=>{r.d(t,{Re:()=>i,gF:()=>o,q4:()=>n});const n="1.236.0",i="PROD",o="CDN"},385:(e,t,r)=>{r.d(t,{FN:()=>a,IF:()=>u,Nk:()=>f,Tt:()=>s,_A:()=>o,il:()=>n,pL:()=>c,v6:()=>i,w1:()=>d});const n="undefined"!=typeof window&&!!window.document,i="undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self.navigator instanceof WorkerNavigator||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis.navigator instanceof WorkerNavigator),o=n?window:"undefined"!=typeof WorkerGlobalScope&&("undefined"!=typeof self&&self instanceof WorkerGlobalScope&&self||"undefined"!=typeof globalThis&&globalThis instanceof WorkerGlobalScope&&globalThis),a=""+o?.location,s=/iPad|iPhone|iPod/.test(navigator.userAgent),c=s&&"undefined"==typeof SharedWorker,u=(()=>{const e=navigator.userAgent.match(/Firefox[/\s](\d+\.\d+)/);return Array.isArray(e)&&e.length>=2?+e[1]:0})(),d=Boolean(n&&window.document.documentMode),f=!!navigator.sendBeacon},1117:(e,t,r)=>{r.d(t,{w:()=>o});var n=r(50);const i={agentIdentifier:"",ee:void 0};class o{constructor(e){try{if("object"!=typeof e)return(0,n.Z)("shared context requires an object as input");this.sharedContext={},Object.assign(this.sharedContext,i),Object.entries(e).forEach((e=>{let[t,r]=e;Object.keys(i).includes(t)&&(this.sharedContext[t]=r)}))}catch(e){(0,n.Z)("An error occured while setting SharedContext",e)}}}},8e3:(e,t,r)=>{r.d(t,{L:()=>d,R:()=>c});var n=r(2177),i=r(1284),o=r(4322),a=r(3325);const s={};function c(e,t){const r={staged:!1,priority:a.p[t]||0};u(e),s[e].get(t)||s[e].set(t,r)}function u(e){e&&(s[e]||(s[e]=new Map))}function d(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:"",t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:"feature";if(u(e),!e||!s[e].get(t))return a(t);s[e].get(t).staged=!0;const r=[...s[e]];function a(t){const r=e?n.ee.get(e):n.ee,a=o.X.handlers;if(r.backlog&&a){var s=r.backlog[t],c=a[t];if(c){for(var u=0;s&&u {let[t,r]=e;return r.staged}))&&(r.sort(((e,t)=>e[1].priority-t[1].priority)),r.forEach((e=>{let[t]=e;a(t)})))}function f(e,t){var r=e[1];(0,i.D)(t[r],(function(t,r){var n=e[0];if(r[0]===n){var i=r[1],o=e[3],a=e[2];i.apply(o,a)}}))}},2177:(e,t,r)=>{r.d(t,{c:()=>f,ee:()=>u});var n=r(8632),i=r(2210),o=r(1284),a=r(5763),s="nr@context";let c=(0,n.fP)();var u;function d(){}function f(e){return(0,i.X)(e,s,l)}function l(){return new d}function h(){u.aborted=!0,u.backlog={}}c.ee?u=c.ee:(u=function e(t,r){var n={},c={},f={},g=!1;try{g=16===r.length&&(0,a.OP)(r).isolatedBacklog}catch(e){}var p={on:b,addEventListener:b,removeEventListener:y,emit:v,get:x,listeners:w,context:m,buffer:A,abort:h,aborted:!1,isBuffering:E,debugId:r,backlog:g?{}:t&&"object"==typeof t.backlog?t.backlog:{}};return p;function m(e){return e&&e instanceof d?e:e?(0,i.X)(e,s,l):l()}function v(e,r,n,i,o){if(!1!==o&&(o=!0),!u.aborted||i){t&&o&&t.emit(e,r,n);for(var a=m(n),s=w(e),d=s.length,f=0;fn,p:()=>i});var n=r(2177).ee.get("handle");function i(e,t,r,i,o){o?(o.buffer([e],i),o.emit(e,t,r)):(n.buffer([e],i),n.emit(e,t,r))}},4322:(e,t,r)=>{r.d(t,{X:()=>o});var n=r(5546);o.on=a;var i=o.handlers={};function o(e,t,r,o){a(o||n.E,i,e,t,r)}function a(e,t,r,i,o){o||(o="feature"),e||(e=n.E);var a=t[o]=t[o]||{};(a[r]=a[r]||[]).push([e,i])}},3239:(e,t,r)=>{r.d(t,{bP:()=>s,iz:()=>c,m$:()=>a});var n=r(385);let i=!1,o=!1;try{const e={get passive(){return i=!0,!1},get signal(){return o=!0,!1}};n._A.addEventListener("test",null,e),n._A.removeEventListener("test",null,e)}catch(e){}function a(e,t){return i||o?{capture:!!e,passive:i,signal:t}:!!e}function s(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;window.addEventListener(e,t,a(r,n))}function c(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2],n=arguments.length>3?arguments[3]:void 0;document.addEventListener(e,t,a(r,n))}},4402:(e,t,r)=>{r.d(t,{Ht:()=>u,M:()=>c,Rl:()=>a,ky:()=>s});var n=r(385);const i="xxxxxxxx-xxxx-4xxx-yxxx-xxxxxxxxxxxx";function o(e,t){return e?15&e[t]:16*Math.random()|0}function a(){const e=n._A?.crypto||n._A?.msCrypto;let t,r=0;return e&&e.getRandomValues&&(t=e.getRandomValues(new Uint8Array(31))),i.split("").map((e=>"x"===e?o(t,++r).toString(16):"y"===e?(3&o()|8).toString(16):e)).join("")}function s(e){const t=n._A?.crypto||n._A?.msCrypto;let r,i=0;t&&t.getRandomValues&&(r=t.getRandomValues(new Uint8Array(31)));const a=[];for(var s=0;s {r.d(t,{Bq:()=>n,Hb:()=>o,oD:()=>i});const n="NRBA",i=144e5,o=18e5},7894:(e,t,r)=>{function n(){return Math.round(performance.now())}r.d(t,{z:()=>n})},7243:(e,t,r)=>{r.d(t,{e:()=>o});var n=r(385),i={};function o(e){if(e in i)return i[e];if(0===(e||"").indexOf("data:"))return{protocol:"data"};let t;var r=n._A?.location,o={};if(n.il)t=document.createElement("a"),t.href=e;else try{t=new URL(e,r.href)}catch(e){return o}o.port=t.port;var a=t.href.split("://");!o.port&&a[1]&&(o.port=a[1].split("/")[0].split("@").pop().split(":")[1]),o.port&&"0"!==o.port||(o.port="https"===a[0]?"443":"80"),o.hostname=t.hostname||r.hostname,o.pathname=t.pathname,o.protocol=a[0],"/"!==o.pathname.charAt(0)&&(o.pathname="/"+o.pathname);var s=!t.protocol||":"===t.protocol||t.protocol===r.protocol,c=t.hostname===r.hostname&&t.port===r.port;return o.sameOrigin=s&&(!t.hostname||c),"/"===o.pathname&&(i[e]=o),o}},50:(e,t,r)=>{function n(e,t){"function"==typeof console.warn&&(console.warn("New Relic: ".concat(e)),t&&console.warn(t))}r.d(t,{Z:()=>n})},2587:(e,t,r)=>{r.d(t,{N:()=>c,T:()=>u});var n=r(2177),i=r(5546),o=r(8e3),a=r(3325);const s={stn:[a.D.sessionTrace],err:[a.D.jserrors,a.D.metrics],ins:[a.D.pageAction],spa:[a.D.spa],sr:[a.D.sessionReplay,a.D.sessionTrace]};function c(e,t){const r=n.ee.get(t);e&&"object"==typeof e&&(Object.entries(e).forEach((e=>{let[t,n]=e;void 0===u[t]&&(s[t]?s[t].forEach((e=>{n?(0,i.p)("feat-"+t,[],void 0,e,r):(0,i.p)("block-"+t,[],void 0,e,r),(0,i.p)("rumresp-"+t,[Boolean(n)],void 0,e,r)})):n&&(0,i.p)("feat-"+t,[],void 0,void 0,r),u[t]=Boolean(n))})),Object.keys(s).forEach((e=>{void 0===u[e]&&(s[e]?.forEach((t=>(0,i.p)("rumresp-"+e,[!1],void 0,t,r))),u[e]=!1)})),(0,o.L)(t,a.D.pageViewEvent))}const u={}},2210:(e,t,r)=>{r.d(t,{X:()=>i});var n=Object.prototype.hasOwnProperty;function i(e,t,r){if(n.call(e,t))return e[t];var i=r();if(Object.defineProperty&&Object.keys)try{return Object.defineProperty(e,t,{value:i,writable:!0,enumerable:!1}),i}catch(e){}return e[t]=i,i}},1284:(e,t,r)=>{r.d(t,{D:()=>n});const n=(e,t)=>Object.entries(e||{}).map((e=>{let[r,n]=e;return t(r,n)}))},4351:(e,t,r)=>{r.d(t,{P:()=>o});var n=r(2177);const i=()=>{const e=new WeakSet;return(t,r)=>{if("object"==typeof r&&null!==r){if(e.has(r))return;e.add(r)}return r}};function o(e){try{return JSON.stringify(e,i())}catch(e){try{n.ee.emit("internal-error",[e])}catch(e){}}}},3960:(e,t,r)=>{r.d(t,{K:()=>a,b:()=>o});var n=r(3239);function i(){return"undefined"==typeof document||"complete"===document.readyState}function o(e,t){if(i())return e();(0,n.bP)("load",e,t)}function a(e){if(i())return e();(0,n.iz)("DOMContentLoaded",e)}},8632:(e,t,r)=>{r.d(t,{EZ:()=>u,Qy:()=>c,ce:()=>o,fP:()=>a,gG:()=>d,mF:()=>s});var n=r(7894),i=r(385);const o={beacon:"bam.nr-data.net",errorBeacon:"bam.nr-data.net"};function a(){return i._A.NREUM||(i._A.NREUM={}),void 0===i._A.newrelic&&(i._A.newrelic=i._A.NREUM),i._A.NREUM}function s(){let e=a();return e.o||(e.o={ST:i._A.setTimeout,SI:i._A.setImmediate,CT:i._A.clearTimeout,XHR:i._A.XMLHttpRequest,REQ:i._A.Request,EV:i._A.Event,PR:i._A.Promise,MO:i._A.MutationObserver,FETCH:i._A.fetch}),e}function c(e,t,r){let i=a();const o=i.initializedAgents||{},s=o[e]||{};return Object.keys(s).length||(s.initializedAt={ms:(0,n.z)(),date:new Date}),i.initializedAgents={...o,[e]:{...s,[r]:t}},i}function u(e,t){a()[e]=t}function d(){return function(){let e=a();const t=e.info||{};e.info={beacon:o.beacon,errorBeacon:o.errorBeacon,...t}}(),function(){let e=a();const t=e.init||{};e.init={...t}}(),s(),function(){let e=a();const t=e.loader_config||{};e.loader_config={...t}}(),a()}},7956:(e,t,r)=>{r.d(t,{N:()=>i});var n=r(3239);function i(e){let t=arguments.length>1&&void 0!==arguments[1]&&arguments[1],r=arguments.length>2?arguments[2]:void 0,i=arguments.length>3?arguments[3]:void 0;return void(0,n.iz)("visibilitychange",(function(){if(t)return void("hidden"==document.visibilityState&&e());e(document.visibilityState)}),r,i)}},1214:(e,t,r)=>{r.d(t,{em:()=>v,u5:()=>N,QU:()=>S,_L:()=>I,Gm:()=>L,Lg:()=>M,gy:()=>U,BV:()=>Q,Kf:()=>ee});var n=r(2177);const i="nr@original";var o=Object.prototype.hasOwnProperty,a=!1;function s(e,t){return e||(e=n.ee),r.inPlace=function(e,t,n,i,o){n||(n="");var a,s,c,u="-"===n.charAt(0);for(c=0;c 2?n-2:0),o=2;o {r(A[T],e,w),r(E[T],e,w)})),r(l._A,"fetch",y),t.on(y+"end",(function(e,r){var n=this;if(r){var i=r.headers.get("content-length");null!==i&&(n.rxSize=i),t.emit(y+"done",[null,r],n)}else t.emit(y+"done",[e],n)})),t}const O={},j=["pushState","replaceState"];function S(e){const t=function(e){return(e||n.ee).get("history")}(e);return!l.il||O[t.debugId]++||(O[t.debugId]=1,s(t).inPlace(window.history,j,"-")),t}var P=r(3239);const C={},R=["appendChild","insertBefore","replaceChild"];function I(e){const t=function(e){return(e||n.ee).get("jsonp")}(e);if(!l.il||C[t.debugId])return t;C[t.debugId]=!0;var r=s(t),i=/[?&](?:callback|cb)=([^&#]+)/,o=/(.*)\.([^.]+)/,a=/^(\w+)(\.|$)(.*)$/;function c(e,t){var r=e.match(a),n=r[1],i=r[3];return i?c(i,t[n]):t[n]}return r.inPlace(Node.prototype,R,"dom-"),t.on("dom-start",(function(e){!function(e){if(!e||"string"!=typeof e.nodeName||"script"!==e.nodeName.toLowerCase())return;if("function"!=typeof e.addEventListener)return;var n=(a=e.src,s=a.match(i),s?s[1]:null);var a,s;if(!n)return;var u=function(e){var t=e.match(o);if(t&&t.length>=3)return{key:t[2],parent:c(t[1],window)};return{key:e,parent:window}}(n);if("function"!=typeof u.parent[u.key])return;var d={};function f(){t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}function l(){t.emit("jsonp-error",[],d),t.emit("jsonp-end",[],d),e.removeEventListener("load",f,(0,P.m$)(!1)),e.removeEventListener("error",l,(0,P.m$)(!1))}r.inPlace(u.parent,[u.key],"cb-",d),e.addEventListener("load",f,(0,P.m$)(!1)),e.addEventListener("error",l,(0,P.m$)(!1)),t.emit("new-jsonp",[e.src],d)}(e[0])})),t}var k=r(5763);const H={};function L(e){const t=function(e){return(e||n.ee).get("mutation")}(e);if(!l.il||H[t.debugId])return t;H[t.debugId]=!0;var r=s(t),i=k.Yu.MO;return i&&(window.MutationObserver=function(e){return this instanceof i?new i(r(e,"fn-")):i.apply(this,arguments)},MutationObserver.prototype=i.prototype),t}const z={};function M(e){const t=function(e){return(e||n.ee).get("promise")}(e);if(z[t.debugId])return t;z[t.debugId]=!0;var r=n.c,o=s(t),a=k.Yu.PR;return a&&function(){function e(r){var n=t.context(),i=o(r,"executor-",n,null,!1);const s=Reflect.construct(a,[i],e);return t.context(s).getCtx=function(){return n},s}l._A.Promise=e,Object.defineProperty(e,"name",{value:"Promise"}),e.toString=function(){return a.toString()},Object.setPrototypeOf(e,a),["all","race"].forEach((function(r){const n=a[r];e[r]=function(e){let i=!1;[...e||[]].forEach((e=>{this.resolve(e).then(a("all"===r),a(!1))}));const o=n.apply(this,arguments);return o;function a(e){return function(){t.emit("propagate",[null,!i],o,!1,!1),i=i||!e}}}})),["resolve","reject"].forEach((function(r){const n=a[r];e[r]=function(e){const r=n.apply(this,arguments);return e!==r&&t.emit("propagate",[e,!0],r,!1,!1),r}})),e.prototype=a.prototype;const n=a.prototype.then;a.prototype.then=function(){var e=this,i=r(e);i.promise=e;for(var a=arguments.length,s=new Array(a),c=0;c e())),t};function m(e,t){i.inPlace(t,["onreadystatechange"],"fn-",E)}function b(){var e=this,t=r.context(e);e.readyState>3&&!t.resolved&&(t.resolved=!0,r.emit("xhr-resolved",[],e)),i.inPlace(e,f,"fn-",E)}if(function(e,t){for(var r in e)t[r]=e[r]}(o,p),p.prototype=o.prototype,i.inPlace(p.prototype,J,"-xhr-",E),r.on("send-xhr-start",(function(e,t){m(e,t),function(e){h.push(e),a&&(y?y.then(A):u?u(A):(w=-w,x.data=w))}(t)})),r.on("open-xhr-start",m),a){var y=c&&c.resolve();if(!u&&!c){var w=1,x=document.createTextNode(w);new a(A).observe(x,{characterData:!0})}}else t.on("fn-end",(function(e){e[0]&&e[0].type===d||A()}));function A(){for(var e=0;e {r.d(t,{t:()=>n});const n=r(3325).D.ajax},6660:(e,t,r)=>{r.d(t,{A:()=>i,t:()=>n});const n=r(3325).D.jserrors,i="nr@seenError"},3081:(e,t,r)=>{r.d(t,{gF:()=>o,mY:()=>i,t9:()=>n,vz:()=>s,xS:()=>a});const n=r(3325).D.metrics,i="sm",o="cm",a="storeSupportabilityMetrics",s="storeEventMetrics"},4649:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageAction},7633:(e,t,r)=>{r.d(t,{Dz:()=>i,OJ:()=>a,qw:()=>o,t9:()=>n});const n=r(3325).D.pageViewEvent,i="firstbyte",o="domcontent",a="windowload"},9251:(e,t,r)=>{r.d(t,{t:()=>n});const n=r(3325).D.pageViewTiming},3614:(e,t,r)=>{r.d(t,{BST_RESOURCE:()=>i,END:()=>s,FEATURE_NAME:()=>n,FN_END:()=>u,FN_START:()=>c,PUSH_STATE:()=>d,RESOURCE:()=>o,START:()=>a});const n=r(3325).D.sessionTrace,i="bstResource",o="resource",a="-start",s="-end",c="fn"+a,u="fn"+s,d="pushState"},7836:(e,t,r)=>{r.d(t,{BODY:()=>A,CB_END:()=>E,CB_START:()=>u,END:()=>x,FEATURE_NAME:()=>i,FETCH:()=>_,FETCH_BODY:()=>v,FETCH_DONE:()=>m,FETCH_START:()=>p,FN_END:()=>c,FN_START:()=>s,INTERACTION:()=>l,INTERACTION_API:()=>d,INTERACTION_EVENTS:()=>o,JSONP_END:()=>b,JSONP_NODE:()=>g,JS_TIME:()=>T,MAX_TIMER_BUDGET:()=>a,REMAINING:()=>f,SPA_NODE:()=>h,START:()=>w,originalSetTimeout:()=>y});var n=r(5763);const i=r(3325).D.spa,o=["click","submit","keypress","keydown","keyup","change"],a=999,s="fn-start",c="fn-end",u="cb-start",d="api-ixn-",f="remaining",l="interaction",h="spaNode",g="jsonpNode",p="fetch-start",m="fetch-done",v="fetch-body-",b="jsonp-end",y=n.Yu.ST,w="-start",x="-end",A="-body",E="cb"+x,T="jsTime",_="fetch"},5938:(e,t,r)=>{r.d(t,{W:()=>o});var n=r(5763),i=r(2177);class o{constructor(e,t,r){this.agentIdentifier=e,this.aggregator=t,this.ee=i.ee.get(e,(0,n.OP)(this.agentIdentifier).isolatedBacklog),this.featureName=r,this.blocked=!1}}},9144:(e,t,r)=>{r.d(t,{j:()=>m});var n=r(3325),i=r(5763),o=r(5546),a=r(2177),s=r(7894),c=r(8e3),u=r(3960),d=r(385),f=r(50),l=r(3081),h=r(8632);function g(){const e=(0,h.gG)();["setErrorHandler","finished","addToTrace","inlineHit","addRelease","addPageAction","setCurrentRouteName","setPageViewName","setCustomAttribute","interaction","noticeError","setUserId"].forEach((t=>{e[t]=function(){for(var r=arguments.length,n=new Array(r),i=0;i 1?r-1:0),i=1;i {e.exposed&&e.api[t]&&o.push(e.api[t](...n))})),o.length>1?o:o[0]}(t,...n)}}))}var p=r(2587);function m(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:{},m=arguments.length>2?arguments[2]:void 0,v=arguments.length>3?arguments[3]:void 0,{init:b,info:y,loader_config:w,runtime:x={loaderType:m},exposed:A=!0}=t;const E=(0,h.gG)();y||(b=E.init,y=E.info,w=E.loader_config),(0,i.Dg)(e,b||{}),(0,i.GE)(e,w||{}),(0,i.sU)(e,x),y.jsAttributes??={},d.v6&&(y.jsAttributes.isWorker=!0),(0,i.CX)(e,y),g();const T=function(e,t){t||(0,c.R)(e,"api");const h={};var g=a.ee.get(e),p=g.get("tracer"),m="api-",v=m+"ixn-";function b(t,r,n,o){const a=(0,i.C5)(e);return null===r?delete a.jsAttributes[t]:(0,i.CX)(e,{...a,jsAttributes:{...a.jsAttributes,[t]:r}}),x(m,n,!0,o||null===r?"session":void 0)(t,r)}function y(){}["setErrorHandler","finished","addToTrace","inlineHit","addRelease"].forEach((e=>h[e]=x(m,e,!0,"api"))),h.addPageAction=x(m,"addPageAction",!0,n.D.pageAction),h.setCurrentRouteName=x(m,"routeName",!0,n.D.spa),h.setPageViewName=function(t,r){if("string"==typeof t)return"/"!==t.charAt(0)&&(t="/"+t),(0,i.OP)(e).customTransaction=(r||"http://custom.transaction")+t,x(m,"setPageViewName",!0)()},h.setCustomAttribute=function(e,t){let r=arguments.length>2&&void 0!==arguments[2]&&arguments[2];if("string"==typeof e){if(["string","number"].includes(typeof t)||null===t)return b(e,t,"setCustomAttribute",r);(0,f.Z)("Failed to execute setCustomAttribute.\nNon-null value must be a string or number type, but a type of was provided."))}else(0,f.Z)("Failed to execute setCustomAttribute.\nName must be a string type, but a type of was provided."))},h.setUserId=function(e){if("string"==typeof e||null===e)return b("enduser.id",e,"setUserId",!0);(0,f.Z)("Failed to execute setUserId.\nNon-null value must be a string type, but a type of was provided."))},h.interaction=function(){return(new y).get()};var w=y.prototype={createTracer:function(e,t){var r={},i=this,a="function"==typeof t;return(0,o.p)(v+"tracer",[(0,s.z)(),e,r],i,n.D.spa,g),function(){if(p.emit((a?"":"no-")+"fn-start",[(0,s.z)(),i,a],r),a)try{return t.apply(this,arguments)}catch(e){throw p.emit("fn-err",[arguments,this,"string"==typeof e?new Error(e):e],r),e}finally{p.emit("fn-end",[(0,s.z)()],r)}}}};function x(e,t,r,i){return function(){return(0,o.p)(l.xS,["API/"+t+"/called"],void 0,n.D.metrics,g),i&&(0,o.p)(e+t,[(0,s.z)(),...arguments],r?null:this,i,g),r?void 0:this}}function A(){r.e(439).then(r.bind(r,7438)).then((t=>{let{setAPI:r}=t;r(e),(0,c.L)(e,"api")})).catch((()=>(0,f.Z)("Downloading runtime APIs failed...")))}return["actionText","setName","setAttribute","save","ignore","onEnd","getContext","end","get"].forEach((e=>{w[e]=x(v,e,void 0,n.D.spa)})),h.noticeError=function(e,t){"string"==typeof e&&(e=new Error(e)),(0,o.p)(l.xS,["API/noticeError/called"],void 0,n.D.metrics,g),(0,o.p)("err",[e,(0,s.z)(),!1,t],void 0,n.D.jserrors,g)},d.il?(0,u.b)((()=>A()),!0):A(),h}(e,v);return(0,h.Qy)(e,T,"api"),(0,h.Qy)(e,A,"exposed"),(0,h.EZ)("activatedFeatures",p.T),T}},3325:(e,t,r)=>{r.d(t,{D:()=>n,p:()=>i});const n={ajax:"ajax",jserrors:"jserrors",metrics:"metrics",pageAction:"page_action",pageViewEvent:"page_view_event",pageViewTiming:"page_view_timing",sessionReplay:"session_replay",sessionTrace:"session_trace",spa:"spa"},i={[n.pageViewEvent]:1,[n.pageViewTiming]:2,[n.metrics]:3,[n.jserrors]:4,[n.ajax]:5,[n.sessionTrace]:6,[n.pageAction]:7,[n.spa]:8,[n.sessionReplay]:9}}},n={};function i(e){var t=n[e];if(void 0!==t)return t.exports;var o=n[e]={exports:{}};return r[e](o,o.exports,i),o.exports}i.m=r,i.d=(e,t)=>{for(var r in t)i.o(t,r)&&!i.o(e,r)&&Object.defineProperty(e,r,{enumerable:!0,get:t[r]})},i.f={},i.e=e=>Promise.all(Object.keys(i.f).reduce(((t,r)=>(i.f[r](e,t),t)),[])),i.u=e=>(({78:"page_action-aggregate",147:"metrics-aggregate",242:"session-manager",317:"jserrors-aggregate",348:"page_view_timing-aggregate",412:"lazy-feature-loader",439:"async-api",538:"recorder",590:"session_replay-aggregate",675:"compressor",733:"session_trace-aggregate",786:"page_view_event-aggregate",873:"spa-aggregate",898:"ajax-aggregate"}[e]||e)+"."+{78:"ac76d497",147:"3dc53903",148:"1a20d5fe",242:"2a64278a",317:"49e41428",348:"bd6de33a",412:"2f55ce66",439:"30bd804e",538:"1b18459f",590:"cf0efb30",675:"ae9f91a8",733:"83105561",786:"06482edd",860:"03a8b7a5",873:"e6b09d52",898:"998ef92b"}[e]+"-1.236.0.min.js"),i.o=(e,t)=>Object.prototype.hasOwnProperty.call(e,t),e={},t="NRBA:",i.l=(r,n,o,a)=>{if(e[r])e[r].push(n);else{var s,c;if(void 0!==o)for(var u=document.getElementsByTagName("script"),d=0;d {s.onerror=s.onload=null,clearTimeout(h);var i=e[r];if(delete e[r],s.parentNode&&s.parentNode.removeChild(s),i&&i.forEach((e=>e(n))),t)return t(n)},h=setTimeout(l.bind(null,void 0,{type:"timeout",target:s}),12e4);s.onerror=l.bind(null,s.onerror),s.onload=l.bind(null,s.onload),c&&document.head.appendChild(s)}},i.r=e=>{"undefined"!=typeof Symbol&&Symbol.toStringTag&&Object.defineProperty(e,Symbol.toStringTag,{value:"Module"}),Object.defineProperty(e,"__esModule",{value:!0})},i.j=364,i.p="https://js-agent.newrelic.com/",(()=>{var e={364:0,953:0};i.f.j=(t,r)=>{var n=i.o(e,t)?e[t]:void 0;if(0!==n)if(n)r.push(n[2]);else{var o=new Promise(((r,i)=>n=e[t]=[r,i]));r.push(n[2]=o);var a=i.p+i.u(t),s=new Error;i.l(a,(r=>{if(i.o(e,t)&&(0!==(n=e[t])&&(e[t]=void 0),n)){var o=r&&("load"===r.type?"missing":r.type),a=r&&r.target&&r.target.src;s.message="Loading chunk "+t+" failed.\n("+o+": "+a+")",s.name="ChunkLoadError",s.type=o,s.request=a,n[1](s)}}),"chunk-"+t,t)}};var t=(t,r)=>{var n,o,[a,s,c]=r,u=0;if(a.some((t=>0!==e[t]))){for(n in s)i.o(s,n)&&(i.m[n]=s[n]);if(c)c(i)}for(t&&t(r);u {i.r(o);var e=i(3325),t=i(5763);const r=Object.values(e.D);function n(e){const n={};return r.forEach((r=>{n[r]=function(e,r){return!1!==(0,t.Mt)(r,"".concat(e,".enabled"))}(r,e)})),n}var a=i(9144);var s=i(5546),c=i(385),u=i(8e3),d=i(5938),f=i(3960),l=i(50);class h extends d.W{constructor(e,t,r){let n=!(arguments.length>3&&void 0!==arguments[3])||arguments[3];super(e,t,r),this.auto=n,this.abortHandler,this.featAggregate,this.onAggregateImported,n&&(0,u.R)(e,r)}importAggregator(){let e=arguments.length>0&&void 0!==arguments[0]?arguments[0]:{};if(this.featAggregate||!this.auto)return;const r=c.il&&!0===(0,t.Mt)(this.agentIdentifier,"privacy.cookies_enabled");let n;this.onAggregateImported=new Promise((e=>{n=e}));const o=async()=>{let t;try{if(r){const{setupAgentSession:e}=await Promise.all([i.e(860),i.e(242)]).then(i.bind(i,3228));t=e(this.agentIdentifier)}}catch(e){(0,l.Z)("A problem occurred when starting up session manager. This page will not start or extend any session.",e)}try{if(!this.shouldImportAgg(this.featureName,t))return void(0,u.L)(this.agentIdentifier,this.featureName);const{lazyFeatureLoader:r}=await i.e(412).then(i.bind(i,8582)),{Aggregate:o}=await r(this.featureName,"aggregate");this.featAggregate=new o(this.agentIdentifier,this.aggregator,e),n(!0)}catch(e){(0,l.Z)("Downloading and initializing ".concat(this.featureName," failed..."),e),this.abortHandler?.(),n(!1)}};c.il?(0,f.b)((()=>o()),!0):o()}shouldImportAgg(r,n){return r!==e.D.sessionReplay||!1!==(0,t.Mt)(this.agentIdentifier,"session_trace.enabled")&&(!!n?.isNew||!!n?.state.sessionReplay)}}var g=i(7633),p=i(7894);class m extends h{static featureName=g.t9;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];if(super(r,n,g.t9,i),("undefined"==typeof PerformanceNavigationTiming||c.Tt)&&"undefined"!=typeof PerformanceTiming){const n=(0,t.OP)(r);n[g.Dz]=Math.max(Date.now()-n.offset,0),(0,f.K)((()=>n[g.qw]=Math.max((0,p.z)()-n[g.Dz],0))),(0,f.b)((()=>{const t=(0,p.z)();n[g.OJ]=Math.max(t-n[g.Dz],0),(0,s.p)("timing",["load",t],void 0,e.D.pageViewTiming,this.ee)}))}this.importAggregator()}}var v=i(1117),b=i(1284);class y extends v.w{constructor(e){super(e),this.aggregatedData={}}store(e,t,r,n,i){var o=this.getBucket(e,t,r,i);return o.metrics=function(e,t){t||(t={count:0});return t.count+=1,(0,b.D)(e,(function(e,r){t[e]=w(r,t[e])})),t}(n,o.metrics),o}merge(e,t,r,n,i){var o=this.getBucket(e,t,n,i);if(o.metrics){var a=o.metrics;a.count+=r.count,(0,b.D)(r,(function(e,t){if("count"!==e){var n=a[e],i=r[e];i&&!i.c?a[e]=w(i.t,n):a[e]=function(e,t){if(!t)return e;t.c||(t=x(t.t));return t.min=Math.min(e.min,t.min),t.max=Math.max(e.max,t.max),t.t+=e.t,t.sos+=e.sos,t.c+=e.c,t}(i,a[e])}}))}else o.metrics=r}storeMetric(e,t,r,n){var i=this.getBucket(e,t,r);return i.stats=w(n,i.stats),i}getBucket(e,t,r,n){this.aggregatedData[e]||(this.aggregatedData[e]={});var i=this.aggregatedData[e][t];return i||(i=this.aggregatedData[e][t]={params:r||{}},n&&(i.custom=n)),i}get(e,t){return t?this.aggregatedData[e]&&this.aggregatedData[e][t]:this.aggregatedData[e]}take(e){for(var t={},r="",n=!1,i=0;i t.max&&(t.max=e),e 2&&void 0!==arguments[2])||arguments[2];super(e,r,j.t,n),c.il&&((0,t.OP)(e).initHidden=Boolean("hidden"===document.visibilityState),(0,N.N)((()=>(0,s.p)("docHidden",[(0,p.z)()],void 0,j.t,this.ee)),!0),(0,O.bP)("pagehide",(()=>(0,s.p)("winPagehide",[(0,p.z)()],void 0,j.t,this.ee))),this.importAggregator())}}var P=i(3081);class C extends h{static featureName=P.t9;constructor(e,t){let r=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(e,t,P.t9,r),this.importAggregator()}}var R,I=i(2210),k=i(1214),H=i(2177),L={};try{R=localStorage.getItem("__nr_flags").split(","),console&&"function"==typeof console.log&&(L.console=!0,-1!==R.indexOf("dev")&&(L.dev=!0),-1!==R.indexOf("nr_dev")&&(L.nrDev=!0))}catch(e){}function z(e){try{L.console&&z(e)}catch(e){}}L.nrDev&&H.ee.on("internal-error",(function(e){z(e.stack)})),L.dev&&H.ee.on("fn-err",(function(e,t,r){z(r.stack)})),L.dev&&(z("NR AGENT IN DEVELOPMENT MODE"),z("flags: "+(0,b.D)(L,(function(e,t){return e})).join(", ")));var M=i(6660);class B extends h{static featureName=M.t;constructor(r,n){let i=!(arguments.length>2&&void 0!==arguments[2])||arguments[2];super(r,n,M.t,i),this.skipNext=0;try{this.removeOnAbort=new AbortController}catch(e){}const o=this;o.ee.on("fn-start",(function(e,t,r){o.abortHandler&&(o.skipNext+=1)})),o.ee.on("fn-err",(function(t,r,n){o.abortHandler&&!n[M.A]&&((0,I.X)(n,M.A,(function(){return!0})),this.thrown=!0,(0,s.p)("err",[n,(0,p.z)()],void 0,e.D.jserrors,o.ee))})),o.ee.on("fn-end",(function(){o.abortHandler&&!this.thrown&&o.skipNext>0&&(o.skipNext-=1)})),o.ee.on("internal-error",(function(t){(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,o.ee)})),this.origOnerror=c._A.onerror,c._A.onerror=this.onerrorHandler.bind(this),c._A.addEventListener("unhandledrejection",(t=>{const r=function(e){let t="Unhandled Promise Rejection: ";if(e instanceof Error)try{return e.message=t+e.message,e}catch(t){return e}if(void 0===e)return new Error(t);try{return new Error(t+(0,D.P)(e))}catch(e){return new Error(t)}}(t.reason);(0,s.p)("err",[r,(0,p.z)(),!1,{unhandledPromiseRejection:1}],void 0,e.D.jserrors,this.ee)}),(0,O.m$)(!1,this.removeOnAbort?.signal)),(0,k.gy)(this.ee),(0,k.BV)(this.ee),(0,k.em)(this.ee),(0,t.OP)(r).xhrWrappable&&(0,k.Kf)(this.ee),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}onerrorHandler(t,r,n,i,o){"function"==typeof this.origOnerror&&this.origOnerror(...arguments);try{this.skipNext?this.skipNext-=1:(0,s.p)("err",[o||new F(t,r,n),(0,p.z)()],void 0,e.D.jserrors,this.ee)}catch(t){try{(0,s.p)("ierr",[t,(0,p.z)(),!0],void 0,e.D.jserrors,this.ee)}catch(e){}}return!1}}function F(e,t,r){this.message=e||"Uncaught error with no additional information",this.sourceURL=t,this.line=r}let U=1;const q="nr@id";function G(e){const t=typeof e;return!e||"object"!==t&&"function"!==t?-1:e===c._A?0:(0,I.X)(e,q,(function(){return U++}))}function V(e){if("string"==typeof e&&e.length)return e.length;if("object"==typeof e){if("undefined"!=typeof ArrayBuffer&&e instanceof ArrayBuffer&&e.byteLength)return e.byteLength;if("undefined"!=typeof Blob&&e instanceof Blob&&e.size)return e.size;if(!("undefined"!=typeof FormData&&e instanceof FormData))try{return(0,D.P)(e).length}catch(e){return}}}var X=i(7243);class W{constructor(e){this.agentIdentifier=e,this.generateTracePayload=this.generateTracePayload.bind(this),this.shouldGenerateTrace=this.shouldGenerateTrace.bind(this)}generateTracePayload(e){if(!this.shouldGenerateTrace(e))return null;var r=(0,t.DL)(this.agentIdentifier);if(!r)return null;var n=(r.accountID||"").toString()||null,i=(r.agentID||"").toString()||null,o=(r.trustKey||"").toString()||null;if(!n||!i)return null;var a=(0,_.M)(),s=(0,_.Ht)(),c=Date.now(),u={spanId:a,traceId:s,timestamp:c};return(e.sameOrigin||this.isAllowedOrigin(e)&&this.useTraceContextHeadersForCors())&&(u.traceContextParentHeader=this.generateTraceContextParentHeader(a,s),u.traceContextStateHeader=this.generateTraceContextStateHeader(a,c,n,i,o)),(e.sameOrigin&&!this.excludeNewrelicHeader()||!e.sameOrigin&&this.isAllowedOrigin(e)&&this.useNewrelicHeaderForCors())&&(u.newrelicHeader=this.generateTraceHeader(a,s,c,n,i,o)),u}generateTraceContextParentHeader(e,t){return"00-"+t+"-"+e+"-01"}generateTraceContextStateHeader(e,t,r,n,i){return i+"@nr=0-1-"+r+"-"+n+"-"+e+"----"+t}generateTraceHeader(e,t,r,n,i,o){if(!("function"==typeof c._A?.btoa))return null;var a={v:[0,1],d:{ty:"Browser",ac:n,ap:i,id:e,tr:t,ti:r}};return o&&n!==o&&(a.d.tk=o),btoa((0,D.P)(a))}shouldGenerateTrace(e){return this.isDtEnabled()&&this.isAllowedOrigin(e)}isAllowedOrigin(e){var r=!1,n={};if((0,t.Mt)(this.agentIdentifier,"distributed_tracing")&&(n=(0,t.P_)(this.agentIdentifier).distributed_tracing),e.sameOrigin)r=!0;else if(n.allowed_origins instanceof Array)for(var i=0;i 2&&void 0!==arguments[2])||arguments[2];super(r,n,Z.t,i),(0,t.OP)(r).xhrWrappable&&(this.dt=new W(r),this.handler=(e,t,r,n)=>(0,s.p)(e,t,r,n,this.ee),(0,k.u5)(this.ee),(0,k.Kf)(this.ee),function(r,n,i,o){function a(e){var t=this;t.totalCbs=0,t.called=0,t.cbTime=0,t.end=E,t.ended=!1,t.xhrGuids={},t.lastSize=null,t.loadCaptureCalled=!1,t.params=this.params||{},t.metrics=this.metrics||{},e.addEventListener("load",(function(r){_(t,e)}),(0,O.m$)(!1)),c.IF||e.addEventListener("progress",(function(e){t.lastSize=e.loaded}),(0,O.m$)(!1))}function s(e){this.params={method:e[0]},T(this,e[1]),this.metrics={}}function u(e,n){var i=(0,t.DL)(r);i.xpid&&this.sameOrigin&&n.setRequestHeader("X-NewRelic-ID",i.xpid);var a=o.generateTracePayload(this.parsedOrigin);if(a){var s=!1;a.newrelicHeader&&(n.setRequestHeader("newrelic",a.newrelicHeader),s=!0),a.traceContextParentHeader&&(n.setRequestHeader("traceparent",a.traceContextParentHeader),a.traceContextStateHeader&&n.setRequestHeader("tracestate",a.traceContextStateHeader),s=!0),s&&(this.dt=a)}}function d(e,t){var r=this.metrics,i=e[0],o=this;if(r&&i){var a=V(i);a&&(r.txSize=a)}this.startTime=(0,p.z)(),this.listener=function(e){try{"abort"!==e.type||o.loadCaptureCalled||(o.params.aborted=!0),("load"!==e.type||o.called===o.totalCbs&&(o.onloadCalled||"function"!=typeof t.onload)&&"function"==typeof o.end)&&o.end(t)}catch(e){try{n.emit("internal-error",[e])}catch(e){}}};for(var s=0;s 1?e[1]=i:e.push(i)}else e[0]&&e[0].headers&&s(e[0].headers,n)&&(this.dt=n);function s(e,t){var r=!1;return t.newrelicHeader&&(e.set("newrelic",t.newrelicHeader),r=!0),t.traceContextParentHeader&&(e.set("traceparent",t.traceContextParentHeader),t.traceContextStateHeader&&e.set("tracestate",t.traceContextStateHeader),r=!0),r}}function x(e,t){this.params={},this.metrics={},this.startTime=(0,p.z)(),this.dt=t,e.length>=1&&(this.target=e[0]),e.length>=2&&(this.opts=e[1]);var r,n=this.opts||{},i=this.target;"string"==typeof i?r=i:"object"==typeof i&&i instanceof Y?r=i.url:c._A?.URL&&"object"==typeof i&&i instanceof URL&&(r=i.href),T(this,r);var o=(""+(i&&i instanceof Y&&i.method||n.method||"GET")).toUpperCase();this.params.method=o,this.txSize=V(n.body)||0}function A(t,r){var n;this.endTime=(0,p.z)(),this.params||(this.params={}),this.params.status=r?r.status:0,"string"==typeof this.rxSize&&this.rxSize.length>0&&(n=+this.rxSize);var o={txSize:this.txSize,rxSize:n,duration:(0,p.z)()-this.startTime};i("xhr",[this.params,o,this.startTime,this.endTime,"fetch"],this,e.D.ajax)}function E(t){var r=this.params,n=this.metrics;if(!this.ended){this.ended=!0;for(var o=0;o 2&&void 0!==arguments[2])||arguments[2];super(e,t,we.t,r),this.importAggregator()}}new class{constructor(e){let t=arguments.length>1&&void 0!==arguments[1]?arguments[1]:(0,_.ky)(16);c._A?(this.agentIdentifier=t,this.sharedAggregator=new y({agentIdentifier:this.agentIdentifier}),this.features={},this.desiredFeatures=new Set(e.features||[]),this.desiredFeatures.add(m),Object.assign(this,(0,a.j)(this.agentIdentifier,e,e.loaderType||"agent")),this.start()):(0,l.Z)("Failed to initial the agent. Could not determine the runtime environment.")}get config(){return{info:(0,t.C5)(this.agentIdentifier),init:(0,t.P_)(this.agentIdentifier),loader_config:(0,t.DL)(this.agentIdentifier),runtime:(0,t.OP)(this.agentIdentifier)}}start(){const t="features";try{const r=n(this.agentIdentifier),i=[...this.desiredFeatures];i.sort(((t,r)=>e.p[t.featureName]-e.p[r.featureName])),i.forEach((t=>{if(r[t.featureName]||t.featureName===e.D.pageViewEvent){const n=function(t){switch(t){case e.D.ajax:return[e.D.jserrors];case e.D.sessionTrace:return[e.D.ajax,e.D.pageViewEvent];case e.D.sessionReplay:return[e.D.sessionTrace];case e.D.pageViewTiming:return[e.D.pageViewEvent];default:return[]}}(t.featureName);n.every((e=>r[e]))||(0,l.Z)("".concat(t.featureName," is enabled but one or more dependent features has been disabled (").concat((0,D.P)(n),"). This may cause unintended consequences or missing data...")),this.features[t.featureName]=new t(this.agentIdentifier,this.sharedAggregator)}})),(0,T.Qy)(this.agentIdentifier,this.features,t)}catch(e){(0,l.Z)("Failed to initialize all enabled instrument classes (agent aborted) -",e);for(const e in this.features)this.features[e].abortHandler?.();const r=(0,T.fP)();return delete r.initializedAgents[this.agentIdentifier]?.api,delete r.initializedAgents[this.agentIdentifier]?.[t],delete this.sharedAggregator,r.ee?.abort(),delete r.ee?.get(this.agentIdentifier),!1}}}({features:[J,m,S,class extends h{static featureName=oe;constructor(t,r){if(super(t,r,oe,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;const n=this.ee;let i;(0,k.QU)(n),this.eventsEE=(0,k.em)(n),this.eventsEE.on(se,(function(e,t){this.bstStart=(0,p.z)()})),this.eventsEE.on(ae,(function(t,r){(0,s.p)("bst",[t[0],r,this.bstStart,(0,p.z)()],void 0,e.D.sessionTrace,n)})),n.on(ce+ne,(function(e){this.time=(0,p.z)(),this.startPath=location.pathname+location.hash})),n.on(ce+ie,(function(t){(0,s.p)("bstHist",[location.pathname+location.hash,this.startPath,this.time],void 0,e.D.sessionTrace,n)}));try{i=new PerformanceObserver((t=>{const r=t.getEntries();(0,s.p)(te,[r],void 0,e.D.sessionTrace,n)})),i.observe({type:re,buffered:!0})}catch(e){}this.importAggregator({resourceObserver:i})}},C,xe,B,class extends h{static featureName=de;constructor(e,r){if(super(e,r,de,!(arguments.length>2&&void 0!==arguments[2])||arguments[2]),!c.il)return;if(!(0,t.OP)(e).xhrWrappable)return;try{this.removeOnAbort=new AbortController}catch(e){}let n,i=0;const o=this.ee.get("tracer"),a=(0,k._L)(this.ee),s=(0,k.Lg)(this.ee),u=(0,k.BV)(this.ee),d=(0,k.Kf)(this.ee),f=this.ee.get("events"),l=(0,k.u5)(this.ee),h=(0,k.QU)(this.ee),g=(0,k.Gm)(this.ee);function m(e,t){h.emit("newURL",[""+window.location,t])}function v(){i++,n=window.location.hash,this[ve]=(0,p.z)()}function b(){i--,window.location.hash!==n&&m(0,!0);var e=(0,p.z)();this[pe]=~~this[pe]+e-this[ve],this[ye]=e}function y(e,t){e.on(t,(function(){this[t]=(0,p.z)()}))}this.ee.on(ve,v),s.on(be,v),a.on(be,v),this.ee.on(ye,b),s.on(ge,b),a.on(ge,b),this.ee.buffer([ve,ye,"xhr-resolved"],this.featureName),f.buffer([ve],this.featureName),u.buffer(["setTimeout"+le,"clearTimeout"+fe,ve],this.featureName),d.buffer([ve,"new-xhr","send-xhr"+fe],this.featureName),l.buffer([me+fe,me+"-done",me+he+fe,me+he+le],this.featureName),h.buffer(["newURL"],this.featureName),g.buffer([ve],this.featureName),s.buffer(["propagate",be,ge,"executor-err","resolve"+fe],this.featureName),o.buffer([ve,"no-"+ve],this.featureName),a.buffer(["new-jsonp","cb-start","jsonp-error","jsonp-end"],this.featureName),y(l,me+fe),y(l,me+"-done"),y(a,"new-jsonp"),y(a,"jsonp-end"),y(a,"cb-start"),h.on("pushState-end",m),h.on("replaceState-end",m),window.addEventListener("hashchange",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("load",m,(0,O.m$)(!0,this.removeOnAbort?.signal)),window.addEventListener("popstate",(function(){m(0,i>1)}),(0,O.m$)(!0,this.removeOnAbort?.signal)),this.abortHandler=this.#e,this.importAggregator()}#e(){this.removeOnAbort?.abort(),this.abortHandler=void 0}}],loaderType:"spa"})})(),window.NRBA=o})(); window.jQuery || document.write(' ') CKEDITOR_BASEPATH='https://f1000research.com/js/vendor/ckeditor/' window.reactTheme = 'research'; window.MathJax = { CommonHTML: { linebreaks: { automatic: true } }, 'HTML-CSS': { linebreaks: { automatic: true } }, SVG: { linebreaks: { automatic: true } }, AuthorInit: function() { MathJax.Hub.Register.MessageHook('End Process', function () { let timeout = false; // holder for timeout id const delay = 250; // delay after event is "complete" to run callback const reflowMath = function() { const dispFormulas = document.querySelectorAll('.disp-formula.panel'); if (!dispFormulas) { return; } for (const dispFormula of dispFormulas) { const child = dispFormula.querySelector('.MathJax_Preview').nextSibling.firstChild; const isMultiline = MathJax.Hub.getAllJax(dispFormula)[0].root.isMultiline; if (dispFormula.offsetWidth < child.offsetWidth || isMultiline) { MathJax.Hub.Queue(['Rerender', MathJax.Hub, dispFormula]); } } }; window.addEventListener('resize', function() { clearTimeout(timeout); // clear the timeout timeout = setTimeout(reflowMath, delay); // start timing for event "completion" }); }); }, }; if (window.location.hash == '#_=_'){ window.location = window.location.href.split('#')[0] } !function(f,b,e,v,n,t,s){if(f.fbq)return;n=f.fbq=function() {n.callMethod? n.callMethod.apply(n,arguments):n.queue.push(arguments)} ;if(!f._fbq)f._fbq=n; n.push=n;n.loaded=!0;n.version='2.0';n.queue=[];t=b.createElement(e);t.async=!0; t.src=v;s=b.getElementsByTagName(e)[0];s.parentNode.insertBefore(t,s)}(window, document,'script','https://connect.facebook.net/en_US/fbevents.js'); fbq('init', '1641728616063202'); fbq('track', "PixelInitialized", {}); (function(h,o,t,j,a,r){ h.hj=h.hj||function(){(h.hj.q=h.hj.q||[]).push(arguments)}; h._hjSettings={hjid:2318163,hjsv:6}; a=o.getElementsByTagName('head')[0]; r=o.createElement('script');r.async=1; r.src=t+h._hjSettings.hjid+j+h._hjSettings.hjsv; a.appendChild(r); })(window,document,'https://static.hotjar.com/c/hotjar-','.js?sv='); search file_upload Submit your research search menu close search Browse Gateways & Collections How to Publish Submit your Research My Submissions Article Guidelines Article Guidelines (New Versions) Open Data, Software and Code Guidelines Open Data and Accessible Source Materials Guidelines (HSS) Open Data, Software and Code Guidelines (PSE) Prepublication Checks Production Process Posters and Slides Guidelines Document Guidelines Article Processing Charges Peer Review Finding Article Reviewers About How it Works For Reviewers Our Advisors Policies Glossary FAQs For Developers Newsroom Contact My Research Submissions Content and Tracking Alerts My Details Sign In file_upload Submit your research { "@context": "https://schema.org", "@type": "ScholarlyArticle", "mainEntityOfPage": { "@type": "WebPage", "@id": "https://f1000research.com/articles/14-670" }, "headline": "In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in...", "datePublished": "2025-07-07T17:05:26", "dateModified": "2025-07-07T17:05:26", "author": [ { "@type": "Person", "name": "Tanes Sangsri" }, { "@type": "Person", "name": "Rathanin Seng" }, { "@type": "Person", "name": "Worrapot Pengpa" }, { "@type": "Person", "name": "Thanwa Wongsuk" }, { "@type": "Person", "name": "Patcharaporn Siwayaprahm" } ], "publisher": { "@type": "Organization", "name": "F1000Research", "logo": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 480, "width": 60 } }, "image": { "@type": "ImageObject", "url": "https://f1000research.com/img/AMP/F1000Research_image.png", "height": 1200, "width": 150 }, "description": " Backgrounds Lactiplantibacillus plantarum is a Gram-positive lactic acid bacterium known for its probiotic benefits, commonly found in fermented foods and mammalian gastrointestinal tracts. L. plantarum strain L47-2 was selected for its in vitro probiotic characteristics. Methods We investigated its antibacterial activity against pathogenic bacteria using the dual culture overlay method and assessed survival in a mimicked gastrointestinal tract by subjecting the bacteria to pH levels of 1.0–4.0 and bile salt concentrations of 0.1–0.5%. Adhesion to Caco-2 cells was also evaluated. Safety evaluations included phenotypic tests for antibiotic resistance and hemolytic activity, genotypic screening for virulence genes, and whole genome sequencing. Results After 4 hours, L47-2 survived well at pH 2.0 (57.0±3.6%) and 3.0 (66.3±2.5%), and maintained 35.0±3.0% survival in 0.4% bile salt. The L47-2 strain demonstrated antibacterial activity against gastrointestinal pathogens and exhibited safety as a probiotic strain. Notably, the L47-2 strain had high autoaggregation (76.3±3.2%) and coaggregation with specific pathogens ranging from 33.1 to 46.7%. Additionally, when compared to the assessed pathogens, the L47-2 strain showed the highest surface hydrophobicity, 68.4±0.4%. This strain exhibited potential adhesion to Caco-2 cells and inhibited the adhesion of all tested pathogens. It was most effective at inhibiting pathogenic bacterial strains rather than competing and displacing them. Pearson correlation analysis revealed significant positive relationships between autoaggregation and coaggregation (P=0.037), autoaggregation and adhesion to Caco-2 cells (P=0.020), and hydrophobicity and adhesion to Caco-2 cells (P=0.016). Conclusions The findings shed light on this strain’s probiotic potential and safety, as well as its already established functional capabilities, together with its potential applications as a biopreservative in the food industry and for the prevention and treatment of infectious diseases. " } { "@context": "http://schema.org", "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": "1", "item": { "@id": "https://f1000research.com/", "name": "Home" } }, { "@type": "ListItem", "position": "2", "item": { "@id": "https://f1000research.com/browse/articles", "name": "Browse" } }, { "@type": "ListItem", "position": "3", "item": { "@id": "https://f1000research.com/articles/14-670", "name": "In vitro "determination" of Lactiplantibacillus plantarum..." } } ] } Home Browse In vitro "determination" of Lactiplantibacillus plantarum... ALL Metrics - Views Downloads Get PDF Get XML Cite How to cite this article Sangsri T, Seng R, Pengpa W et al. In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.12688/f1000research.166421.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] Tanes Sangsri https://orcid.org/0000-0002-9937-2183 1 , Rathanin Seng 2 , Worrapot Pengpa https://orcid.org/0009-0007-9688-6107 3 , Thanwa Wongsuk 4 , Patcharaporn Siwayaprahm https://orcid.org/0009-0000-7714-3227 3 Tanes Sangsri https://orcid.org/0000-0002-9937-2183 1 , Rathanin Seng 2 , [...] Worrapot Pengpa https://orcid.org/0009-0007-9688-6107 3 , Thanwa Wongsuk 4 , Patcharaporn Siwayaprahm https://orcid.org/0009-0000-7714-3227 3 PUBLISHED 07 Jul 2025 Author details Author details 1 Department of Microbiology, Faculty of Medicine, Princess of Naradhiwas University, Narathiwat, 96000, Thailand 2 Department of Medical Science, Amnatcharoen Campus, Mahidol University, Amnatcharoen, 37000, Thailand 3 Department of Microbiology, Faculty of Science,, Kasetsart University, Bangkok, Bangkok, 10900, Thailand 4 Department of Clinical Pathology, Faculty of Medicine Vajira Hospital, Navamindradhiraj University, Bangkok, 10300, Thailand Tanes Sangsri Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Rathanin Seng Roles: Formal Analysis, Methodology, Writing – Original Draft Preparation, Writing – Review & Editing Worrapot Pengpa Roles: Investigation Thanwa Wongsuk Roles: Formal Analysis, Writing – Original Draft Preparation, Writing – Review & Editing Patcharaporn Siwayaprahm Roles: Conceptualization, Funding Acquisition, Methodology, Project Administration, Supervision, Validation, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS Abstract Backgrounds Lactiplantibacillus plantarum is a Gram-positive lactic acid bacterium known for its probiotic benefits, commonly found in fermented foods and mammalian gastrointestinal tracts. L. plantarum strain L47-2 was selected for its in vitro probiotic characteristics. Methods We investigated its antibacterial activity against pathogenic bacteria using the dual culture overlay method and assessed survival in a mimicked gastrointestinal tract by subjecting the bacteria to pH levels of 1.0–4.0 and bile salt concentrations of 0.1–0.5%. Adhesion to Caco-2 cells was also evaluated. Safety evaluations included phenotypic tests for antibiotic resistance and hemolytic activity, genotypic screening for virulence genes, and whole genome sequencing. Results After 4 hours, L47-2 survived well at pH 2.0 (57.0±3.6%) and 3.0 (66.3±2.5%), and maintained 35.0±3.0% survival in 0.4% bile salt. The L47-2 strain demonstrated antibacterial activity against gastrointestinal pathogens and exhibited safety as a probiotic strain. Notably, the L47-2 strain had high autoaggregation (76.3±3.2%) and coaggregation with specific pathogens ranging from 33.1 to 46.7%. Additionally, when compared to the assessed pathogens, the L47-2 strain showed the highest surface hydrophobicity, 68.4±0.4%. This strain exhibited potential adhesion to Caco-2 cells and inhibited the adhesion of all tested pathogens. It was most effective at inhibiting pathogenic bacterial strains rather than competing and displacing them. Pearson correlation analysis revealed significant positive relationships between autoaggregation and coaggregation ( P =0.037), autoaggregation and adhesion to Caco-2 cells ( P =0.020), and hydrophobicity and adhesion to Caco-2 cells ( P =0.016). Conclusions The findings shed light on this strain’s probiotic potential and safety, as well as its already established functional capabilities, together with its potential applications as a biopreservative in the food industry and for the prevention and treatment of infectious diseases. READ ALL READ LESS Keywords Lactiplantibacillus plantarum, antibacterial activity, lactic acid bacteria, anti-adhesion, whole genome sequencing Corresponding Author(s) Patcharaporn Siwayaprahm ( [email protected] ) Close Corresponding author: Patcharaporn Siwayaprahm Competing interests: No competing interests were disclosed. Grant information: This research was partially supported by the Undergraduate Research Matching Fund: URMF 2021, Faculty of Science, Kasetsart University and Microbiology Projects grant of the Department of Microbiology, Faculty of Science, Kasetsart University and Fundamental Fund (2024), Thailand Science Research and Innovation, Faculty of Medicine, Princess of Naradhiwas University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright: © 2025 Sangsri T et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. How to cite: Sangsri T, Seng R, Pengpa W et al. In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.12688/f1000research.166421.1 ) First published: 07 Jul 2025, 14 :670 ( https://doi.org/10.12688/f1000research.166421.1 ) Latest published: 07 Jul 2025, 14 :670 ( https://doi.org/10.12688/f1000research.166421.1 ) Introduction Lactic acid bacteria (LAB) are typical microbes in several fermented dairy products. Of these, Lactobacillus is a well-known genus that serves as probiotic and non-pathogenic bacteria in the intestine, inhibiting pathogenic bacteria and stimulating immune cell response. 1 – 3 LAB encompasses various genera, including the revised genus Lactobacillus , as well as Lactiplantibacillus , Limosilactobacillus , Lacticaseibacillus , Leuconostoc , Pediococcus , Streptococcus , and Weissella. 4 The potent probiotic qualities balance the microbial ecology in the intestine, preventing diarrheal diseases by defending against harmful intestinal bacteria. 5 , 6 Therefore, the probiotic bacterial properties should include being generally recognized as safe (GRAS), resistance to acidic and bile salt conditions, adherence to intestinal epithelial cell surfaces, antimicrobial activity against enteric bacterial pathogens, and no hemolytic activity or antibiotic resistance. 7 , 8 Recently, LAB have played a role as a bio-preservative in food products. LAB produces bacteriocins-antimicrobial peptides that inhibit Listeria monocytogenes and various strains of Salmonella. 9 , 10 These natural preservatives are becoming more important due to rising concerns over antibiotic resistance and consumer preference for foods with fewer chemical additives. 11 Studies have shown that LAB can effectively combat both Gram-positive and Gram-negative bacteria, suggesting that they have potential as safe alternatives in food preservation. 9 , 10 LAB also exhibit immunomodulatory effects, which can enhance human health by regulating immune responses. Research indicates that LAB-fermented products can activate innate immune responses and modulate adaptive immunity, making them beneficial for individuals with compromised immune systems. 12 Furthermore, LAB have been shown to inhibit viral infections, such as noroviruses and coronaviruses, through mechanisms that may involve competitive inhibition and modulation of the host’s immune response. 13 The therapeutic applications of LAB are expanding beyond traditional uses. Recent advancements include the development of genetically modified LAB strains that can serve as live delivery vectors for therapeutic agents. 11 This innovation is particularly relevant in the context of synthetic biology, where LAB are being engineered to produce bioactive compounds with medical applications. 11 Integration of artificial intelligence in this field is expected to enhance the efficiency and safety of these microbial systems, leading to novel therapeutic options. Lactiplantibacillus plantarum , previously known as Lactobacillus plantarum , is a prominent LAB species recognized for its diverse applications in food production and probiotic health benefits. 14 This Gram-positive, rod-shaped bacterium is commonly found in fermented foods, the gastrointestinal tracts of mammals, and various environmental settings, including plants and insects. 15 Its capability to adapt to different ecological niches is attributed to its robust metabolic versatility and genetic plasticity. L. plantarum cells typically measure 0.9–1.2 μm in width and 3–8 μm in length, often appearing singly, in pairs, or in short chains. 15 This species thrives across a broad range of temperatures (15 to 45°C) and pH levels (as low as 3.2), demonstrating significant acid and bile salt tolerance, which facilitates its survival in the human digestive system. 15 L. plantarum is also widely acknowledged for its probiotic potential. It exhibits several beneficial traits, including the capability to adhere to intestinal epithelial cells, produce antimicrobial compounds such as bacteriocins, and enhance gut health by modulating the microbiota composition. 16 – 18 The strain’s capacity to withstand low pH environments and bile salts further supports its role as a functional probiotic in various dietary applications. 17 , 18 In the food industry, L. plantarum serves as a starter culture for the fermentation of dairy products, vegetables, and meats. 19 It contributes to flavor development, texture improvement, and preservation by producing organic acids and bioactive compounds. 19 Additionally, its bacteriocin-producing strains are considered promising natural preservatives because of their capability to inhibit pathogenic bacteria. 20 Due to its unique probiotic characteristics, L. plantarum has been recognized as a versatile and important LAB. It can tolerate acidic and bile environments and has antagonistic action against gut infections. 19 , 21 In our previous study, 22 L. plantarum strain L47-2 isolated from dairy effluent exhibited anti- Candida albicans activity and colonization, leading to its hypothesized probiotic use. The objective of this study was to evaluate the probiotic characteristics of L. plantarum strain L47-2 in relation to its safe probiotic use in gastrointestinal tract conditions. Methods Bacterial strains and growth conditions Lactiplantibacillus plantarum strain L47-2 was isolated from dairy effluent in our previous study. 22 The bacterium was kept at -80 °C in de Man Rogose Sharpe (MRS) broth (Difco Laboratories, Detroit) plus 20% (v/v) glycerol. The L. plantarum strain L47-2 was cultured in MRS medium under 5% CO 2 at 37°C, while Bacillus cereus BCC 6386, Salmonella Typhimurium DMST 562, Escherichia coli ATCC 8739, Enterococcus faecalis TISTR 379, Staphylococcus aureus ATCC 6538, and Listeria monocytogenes DMST 17303 were cultured in tryptic soy broth (Difco™, USA) at 37°C. Determining the probiotic properties of L. plantarum strain L47-2 Antibacterial activity assay L. plantarum strain L47-2 was investigated for its antibacterial activity against gastrointestinal pathogenic bacteria using the dual culture overlay method, as previously described. 23 The bacterial pathogens were B. cereus BCC 6386, E. coli ATCC 8739, S. aureus ATCC 6538, S. Typhimurium DMST 562, E. faecalis TISTR 379, and L. monocytogenes DMST 17303. First, 10 μL of an 18-h culture (1.0 × 10 5 CFU/mL) of the L47-2 strain was dropped onto the MRS agar plates and incubated in a 5-10% CO 2 atmosphere at 37°C for 48 h to allow colonies to develop. One mL of an 18-h culture in tryptic soy broth (TSB) (Difco™, USA) incubated at 37°C (1.0 × 10 8 CFU/mL) of each pathogenic bacterium was mixed well into 5 mL of soft-tryptic soy agar (0.75% agar) and poured into a plate. The plate was incubated under aerobic conditions at 37°C for 24 h. The diameter of the clear zone was equal to the inhibition zone. This experiment was performed in triplicate. Survival of L. plantarum strain L47-2 under low pH This assay was slightly modified from a previous study. 24 The L47-2 strain (1.0 × 10 8 CFU/mL) was inoculated into MRS broth with a pH of 2.0–5.0, adjusted with 1 N HCl. Unadjusted pH 6.5 MRS broth was used as a control. The cultures were incubated at 37°C for 0, 1, 2, 3, and 4 h. Colony-forming bacteria were subsequently enumerated on MRS agar. The percent survival was computed as the CFU/ml in MRS broth adjusted for pH at various times in MRS broth at pH 6.5. Survival of L. plantarum strain L47-2 under bile salt This assay was slightly modified from a previous study. 25 1.0 × 10 9 CFU/mL in PBS of the L47-2 strain was prepared, and 0.5 mL of the bacterial suspension was pipetted into 4.5 mL of MRS broth containing 0.1–3.0% (w/v) of oxgall (Sigma-Aldrich, USA) to a final cell density of 1.0 × 10 8 CFU/mL. MRS medium with no oxgall was used as a control. The inoculum was incubated at 37°C for 0, 1, 2, 3, and 4 h. Bacterial survival was enumerated as the CFU/ml representative of live bacteria. The percentage of survival was calculated by CFU/ml in MRS broth containing oxgall at a specific time/CFU/ml in MRS broth compared to a medium with no oxgall at the same time × 100. Cell surface hydrophobicity assay This assay was performed by measuring the adhesion of bacteria to xylene as in a previous study. 26 Three mL of each bacterial suspension (1.0 × 10 8 CFU/mL) in PBS was combined with 1 mL of xylene, and the mixture was incubated for 1 h without shaking. The optical density (OD) at 600 nm at the initial stage was spectrophotometrically measured (Thermo Scientific™, USA) as the absorbance (A 0 ). One mL of the aqueous phase was gently pipetted to measure OD for absorbance (A 1 ). Hydrophobicity was expressed as a percentage using the formula: (1- A 1 /A 0 ) × 100. Autoaggregation and coaggregation assays The same procedures as from the earlier research were followed for these assays. 27 For the autoaggregation assay, 4 mL of suspended bacterial cells (1.0×10 8 CFU/mL) in PBS were vortexed for 10 seconds and then allowed to stand at room temperature for 0, 1, 2, 3, 4, 5, 16, and 24 h. Absorbance (A) was measured at 600 nm after transferring 0.1 mL of the top suspension to 3.9 mL of sterile PBS in a separate tube. The formula used to calculate the percentage of autoaggregation was: 1-(A t /A 0 ) ×100, where A t is the absorbance at a specific time (1, 2, 4, 6, 8, 10, 20, and 24 h) and A 0 is the absorbance at time zero. For coaggregation, equal volumes (1 mL) of L. plantarum strain L47-2 and each cell suspension of pathogens, including L. monocytogenes DMST 17303, E. coli ATCC 8739, E. faecalis TISTR 379, S. Typhimurium DMST 562, S. aureus ATCC 6538, and B. cereus BCC 6386, were mixed in pairs by vortexing for 10 seconds. Coaggregation was measured after incubation at 0, 1, 2, 3, 4, 5, 16, and 24 h. After transferring 0.1 mL of the upper suspension into a 3.9 mL tube containing sterile PBS, the absorbance (A) at 600 nm was measured. At the same time, 4 mL of each bacterial suspension was used as a control. The percentage of coaggregation was computed as [(A L47-2 + A pathogen )/2 - (A mix )/(A L47-2 + A pathogen )/2 × 100]. 28 Adhesion and anti-adhesion of L. plantarum strain L47-2 against pathogens on Caco-2 cells The adherence capacity of L. plantarum strain L47-2 and gastrointestinal pathogenic bacteria to Caco-2 cells was performed as in a previous study, with modifications. 29 , 30 The Caco-2 cell line (ATCC ® Cat. No. HTB-37™) used in this study was obtained from the American Type Culture Collection (ATCC). Caco-2 cells were cultured in Dulbecco’s Modified Eagle Medium (Gibco BRL, USA) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (HyClone, USA) (complete DMEM; cDMEM) and incubated in a 5% CO 2 atmosphere at 37°C. 100 μL of a 1.5 × 10 4 cell suspension/well in a 96-well plate was prepared and incubated overnight at 37°C for the following assays. All assays were conducted in triplicate, and each experiment was performed independently as follows. Adhesion assay: Caco-2 cells were infected with each bacterium at a Multiplicity of Infection (MOI) of 10:1 (bacteria: cells) and incubated at 37°C under a 5% CO 2 atmosphere for 1 h. Non-adherent bacteria were gently washed out 5 times with sterile PBS (Gibco BRL, USA), and the attached bacteria were dislodged by incubating them with 0.1% Triton X-100 (Sigma-Aldrich, USA) for 5 min. All bacteria were counted as colony-forming units (CFU) by serial dilution and colony counting on MRS agar for the L47-2 strain and tryptic soy agar for each bacterial pathogen, respectively, after incubating them at 37°C for 24 h. Bacterial adhesion was computed from %Adhesion = [Adhered bacteria/Total of added bacteria] × 100. Inhibition assay: Caco-2 cells were first incubated with an MOI of 10:1 of the L47-2 strain for 1 h. Then, each bacterial pathogen at an MOI of 10:1 was subsequently added into the wells and further incubated for 1 h. Non-attached bacteria were removed by washing 5 times with sterile PBS. The bacteria were counted according to the adhesion assay protocol. Inhibition was computed using the formula, %Inhibition of adhesion = (1 - T 1 /T 2 ) × 100, where T 1 and T 2 are the percentages of adhesion by pathogens with or without the strain L47-2, respectively. Competition assay: Caco-2 cells were infected with an equal amount of the L47-2 strain and each pathogen at an MOI of 10:1 and incubated for 1 h. All unbound bacteria were removed, and the attached bacteria were dislodged and counted as performed in the adhesion assay. The percentages of competition were calculated from the adhesion of bacterial pathogens added together with the L47-2 strain relative to adhered bacteria without the L47-2 strain (control). Displacement assay : Caco-2 cells were infected with each pathogen and incubated for 1 h, after which the L47-2 strain was added and the cells were incubated for another 1 h. Bacterial enumeration was performed as described in the adhesion assay. The percentage of displacement was calculated from adhered bacterial pathogens with or without the L47-2 strain, as described above. Safety assessment of L. plantarum strain L47-2 Hemolytic activity assay A streak plate technique was used to determine hemolytic activity. L. plantarum strain L47-2 was streaked onto sheep blood agar (5% (v/v) of sheep blood) and incubated at 37°C for 2 days in a 5–10% CO 2 atmosphere. A zone of hemolysis was observed around the colonies. The β-hemolysis control strain was Staphylococcus aureus ATCC 6538. 7 Antibiotic susceptibility profile This method was performed as previously described. 31 An overnight culture of L. plantarum strain L47-2 was adjusted to a cell density of 1.0 × 10 8 CFU/mL in sterile PBS. Then, 0.1 mL of this adjusted culture was inoculated into 0.9 mL of MRS broth containing several antibiotics (Oxoid, USA), including ampicillin, vancomycin, chloramphenicol, gentamycin, streptomycin, kanamycin, and tetracycline. The antibiotics were in concentrations of 2, 4, 8, 16, 32, 64, 128, 256, 512, and 1024 μg/ml and were incubated at 37°C for 24 h. The CFU/mL count of the bacteria was enumerated by a plate count technique on MRS agar. A bacterial culture in MRS broth without antibiotics was used as a control. The minimal inhibitory concentration (MIC) was the lowest antibiotic concentration that fully suppressed observable growth compared to MRS broth with no antibiotic. The minimal bactericidal concentration (MBC) was determined by transferring an aliquot from each tube of the MIC test to be spread on an MRS agar plate and incubated at 37°C, under 5–10% CO 2 for 24 h. Molecular screening of virulence genes L. plantarum strain L47-2 was investigated for the presence of thirteen virulent genes as described in a previous study. 9 The DNA of the L47-2 strain was extracted with a DNeasy ® UltraClean ® Microbial Kit (Qiagen, Germany). The primer sets used for gene amplification included cylA , cylB , cylM , cylLS , cylLL , ccf , cob , cpd , efaAfm , efaAfs, esp, gelE, and agg as previously described. 32 E. faecalis TISTR 379 was used as a positive control. The PCR reaction was performed using a ThermoCycler (Bio-Rad, USA) in 0.2-ml PCR tubes containing 5.0 μL of 10× PCR buffer, 0.4 μL of 25 mM dNTPs, 0.2 μL of Taq DNA polymerase, 3.0 μL of 25 mM MgCl 2 , 1.0 μL each of 10 μM forward and reverse primers, 1 μL of 100 ng DNA template and DNase-free water to a 50 μL final volume. PCR amplifications were started at 94°C for 1 min, followed by 35 cycles of denaturation, annealing, and extension (at 94°C for 1 min, 55°C for 1 min, and 72°C for 2 min, respectively) and a final extension at 72°C for 7 min, before being stored at 4°C. The PCR products were electrophoresed at 100 V for 30 min and detected with a gel documentation system (Bio-Rad, USA). Whole genome sequencing analysis Genomic DNA was extracted from the L47-2 strain using a QIAamp DNA Mini Kit (Qiagen, Germany) and then subjected to 150-base-read library preparation and sequencing using the Illumina HiSeq PE150 system at Biomarker Technologies. The de novo assembly of short-read data was performed using SPAdes v3.13.1. 33 The number of contigs and N50 were assessed by QUAST v.5.0.2 ( https://github.com/ablab/quast ). Genome completeness and contamination were evaluated using CheckM v.1.2.2. 34 The number of contigs, N50, completeness, and contamination of the L47-2 strain assembled genome were 641 contigs, 254,624 bp, 99.38%, and 3.47%, respectively. Bacterial species were identified using the Type (Strain) Genome Server (TYGS) ( https://tygs.dsmz.de/ and https://pathogen.watch/ ). A CARD Resistance Gene Identifier v. 1.2.1 (cite: DOI: 10.1093/nar/gkac920) was used to detect antibiotic resistance genes in the L47-2 genome. Statistical analysis All assays were conducted in triplicate. Cell culture assays were performed in 3 independent experiments, with all results shown as mean ± standard deviation values. The Pearson correlation coefficient was used for the correlation analysis. A student’s t -test was used to compare samples in one group, and one-way ANOVA was employed for more than two groups. The GraphPad Prism software version 6.0 was used for statistical analysis. P < 0.05 was considered statistically significant. Results Antibacterial activity assay L. plantarum strain L47-2 significantly inhibited the growth of pathogenic bacteria. The inhibitory activity of this strain was ranked based on the size of inhibition zones against the pathogens listed in Table 1 . Table 1. Antibacterial activity of L. plantarum strain L47-2 by dual overlay method. Tested bacterial strains Inhibition zone diameter (mm) B. cereus BCC 6386 37.5 ± 0.50 S. aureus ATCC 6538 33.5 ± 1.5 E. coli ATCC 8739 32 ± 0.48 S. Typhimurium DMST 562 31.5 ± 0.15 L. monocytogenes DMST 17303 29 ± 0.24 E. faecalis TISTR 379 20 ± 0.55 Survival of L. plantarum strain L47-2 under acidic and bile salt conditions L. plantarum strain L47-2 was selected for its ability to survive under the acidic and bile salt conditions found in the gastrointestinal tract. The results showed that the L47-2 strain could survive in a pH range of 2.0–5.0 ( Figure 1A ). The total viable counts at pH values of 2.0 and 3.0 were 57.0 ± 3.6 and 66.3 ± 2.5%, respectively, after 4 h of exposure. The number of viable cells decreased gradually with lower pH values and longer times. Additionally, the L47-2 strain could survive at 0.1–0.4% concentrations of bile salt, showing rates of 58.3 ± 3.5, 47.6 ± 1.5, 44.0 ± 2.0, and 35.0 ± 3.0% ( Figure 1B ), indicating that the L47-2 strain could survive in a 0.4% bile salt concentration. These findings suggest that the L47-2 strain could survive in the small intestine. Figure 1. The survival rate of L. plantarum strain L47-2 in acidic and bile salt conditions. The L47-2 strain survived in MRS broth at 37°C in various pH ranges (A), or MRS broth containing oxgall at 0.1–0.5% (B), and at various incubation times. Cell surface hydrophobicity assay L. plantarum strain L47-2 had the highest hydrophobicity followed by E. coli ATCC 8739, L. monocytogenes DMST 17303 , S. Typhimurium DMST 562, S. aureus ATCC 6538, B. cereus BCC 6386, and E. faecalis TISTR 379, with values of 68.4 ± 0.4, 37.0 ± 1.9, 33.7 ± 1.8, 17.6 ± 0.5, 15.0 ± 2.7, 1.8 ± 0.8, and 1.7 ± 0.4%, respectively ( Figure 2 ). Figure 2. The percentage of bacterial cell surface hydrophobicity after incubation in xylene at 37°C for 1 h. Dashed lines represent the P- values of the ANOVA test. Autoaggregation and coaggregation assays Auto- and co-aggregation assays were used to determine the capabilities of L. plantarum strain L47-2 to aggregate and co-aggregate with individual pathogen strains. As a result, autoaggregation of individual bacterial strains was increased during incubation, rising significantly after 2 h at 37°C and then gradually until 24 h ( Figure 3A ). Interestingly, the L47-2 strain showed higher autoaggregation, 76.3 ± 3.2%, than all pathogen strains after 24 h, except for L. monocytogenes DMST 17303 and E. coli ATCC 8739. They showed values of 85.2 ± 3.3 and 82.8 ± 5.7%, respectively. B. cereus BCC 6386 exhibited the lowest autoaggregation, 50.3 ± 2.1%. Figure 3. Autoaggregation and coaggregation of L. plantarum strain L47-2 and individual tested pathogens. A) Shows autoaggregation of bacterial strains at 37°C and various incubation times. B) Shows coaggregation between the strain L47-2 and individual pathogen strains at 37°C and various incubation times. The data represent the mean ± standard deviation of tests performed in triplicate. Coaggregation was determined by the capability of the L47-2 strain to coaggregate with individual pathogen strains after 24 h of incubation at 37°C. The results, shown in Figure 3B , indicated that the highest percentage of coaggregation occurred between the L47-2 strain and L. monocytogenes DMST 17303 followed by E. coli ATCC 8739, with rates of 46.7 ± 2.4 and 41.2 ± 1.0%, respectively. The lowest coaggregation occurred between the L47-2 strain and B. cereus BCC 6386, 33.1 ± 3.2%, even though coaggregation increased considerably after 2 h. This result also suggests that the L47-2 strain could coaggregate with the other individual pathogens at levels of 33.1–46.7%. This was dependent on the specific pathogen strain and incubation time. These results also suggest a high percentage of autoaggregation linked to high coaggregation activities, which influenced the L47-2 strain to coaggregate with gastrointestinal bacterial pathogens. Adhesion and anti-adhesion of L. plantarum strain L47-2 against pathogens to Caco-2 cells Lactiplantibacillus plantarum L47-2 exhibited the highest adhesion (33.1 ± 3.9%), significantly greater than all tested pathogens ( Figure 4A ). Pathogenic bacteria such as B. cereus BCC 6386, E. coli ATCC 8739, S. aureus ATCC 6538, S. Typhimurium DMST 562, E. faecalis TISTR 379, and L. monocytogenes DMST 17303 showed lower adhesion, ranging between 15.6 ± 1 – 28.3 ± 0.8%. P -values indicated that L. plantarum L47-2 adheres significantly better than the pathogens. The L47-2 strain most effectively inhibited the adhesion of B. cereus BCC 6386 (up to ~80%) under the inhibition condition ( Figure 4B ). Inhibition is generally higher than competition or displacement across all pathogens ( Figure 4B ). The effectiveness of the L47-2 strain in reducing pathogen adhesion varied depending on the pathogen and the test conditions. This suggests its strong potential as a probiotic to prevent pathogen colonization. Figure 4. L. plantarum strain L47-2's adhesion and anti-adhesion to bacterial pathogens on Caco-2 cells. A) Shows bacterial adherence to Caco-2 cells, with the dotted line representing the P -values of the ANOVA test and the solid lines representing the P -values of t -tests. B) Shows the anti-adhesion of the L47-2 strain against pathogens, including inhibition, competition, and displacement. Individual scatter plots were displayed with mean ± standard deviation from three independent experiments, each carried out in triplicate. Pearson correlation coefficient analysis showed that autoaggregation and coaggregation between the L47-2 strain and each tested pathogen were positively correlated ( P = 0.037) ( Table 2 ). Auto- or co-aggregation and adhesion capabilities were positively correlated ( P = 0.020 and P = 0.080, respectively). Furthermore, this study also demonstrated that hydrophobicity and adherence to Caco-2 cells were significantly correlated ( P = 0.016). This finding indicates that the capabilities of the L47-2 strain to autoaggregate, coaggregate, and adhere were significantly correlated. Table 2. The Pearson correlation coefficients between the autoaggregation, coaggregation, hydrophobicity, and adhesion properties of all bacteria. Assay Autoaggregation Hydrophobicity Coaggregation Adhesion Autoaggregation - 0.676 (0.096) 0.837 (0.037) 0.832 (0.020) Hydrophobicity - - - 0.846 (0.016) Co-aggregation - 0.741 (0.092) - 0.758 (0.080) Safety assessments of L. plantarum strain L47-2 Hemolytic activity assay Hemolytic activity of L. plantarum strain L47-2 was tested to confirm the nonpathogenic character of this strain. The results revealed no hemolytic activity of the L47-2 strain with no inhibition zones surrounding colonies on sheep blood agar plates ( Figure 5 ). Figure 5. Determination of the hemolytic activity of L. plantarum strain L47-2 on sheep blood agar at 48 h. A) Shows no reaction (γ-hemolysis) of the L47-2 strain, and B) Shows a clear zone of β-hemolysis surrounding the S. aureus ATCC 6538 colony. Antibiotic susceptibility profile and the presence of virulence genes An antibiotic susceptibility test was applied, and virulence gene contents were determined for the L. plantarum L47-2 strain as a probiotic ( Tables 3 and 4 ). The results showed that the L47-2 strain was susceptible to ampicillin, tetracycline, chloramphenicol, and gentamycin, whereas it resisted vancomycin, streptomycin, and kanamycin ( Table 3 ). Antibiotic resistance was considered according to MIC EFSA breakpoints. 35 Furthermore, the L47-2 strain was negative for all virulence genes examined, suggesting that it can be safely utilized as a probiotic strain ( Table 4 ). Table 3. Antibiotic susceptibility profile of L. plantarum strain L47-2. Antibiotics Antibiotic sensitivity MIC (μg/ml) MBC (μg/ml) Strain L47-2 Ampicillin 1,024 Resistant Chloramphenicol <2 64 Sensitive Gentamycin 4 16 Sensitive Streptomycin 64 128 Resistant Kanamycin 128 512 Resistant Tetracycline <2 8 Sensitive Table 4. The presence of virulence genes in L. plantarum strain L47-2. Bacteria Genes cylA cylB cylM cylLS cylLL ccf cob cpd efaAfm efaAfs esp gelE agg E. faecalis TISTR 379 - - - + + + - + + + - + - L. plantarum strain L47-2 - - - - - - - - - - - - - Genomic characteristics of the L47-2 strain The assembly of the raw reads produced bacterial chromosomes, each measuring approximately the same size as those previously documented for sequenced L. plantarum isolates, from 3 to 3.6 Mbp. 17 The genome-based taxonomy was identified by the Type (Strain) Genome Server (TYGS) ( https://tygs.dsmz.de/ and https://pathogen.watch/ ). The type strain of L47-2 was Lactiplantibacillus plantarum. The ANI value for L. plantarum L47-2 and L. plantarum ATCC 14917 was calculated, yielding an ANI value between the two strains of 98.8548%, indicating their strong relationship ( Figure 6 ). We detected vancomycin resistance genes ( vanH and vanY ), which is consistent with the MIC results. Figure 6. The ANI values for L. plantarum L47-2 (query) and L. plantarum ATCC 14917 (reference). Each red line segment represents a reciprocal mapping of the analyzed genome, highlighting the regions that are evolutionarily conserved. Discussion This study’s findings reveal that L. plantarum strain L47-2 has the potential for probiotic use due to its anti-pathogenic bacterial capabilities, survivability in acidic and bile salt conditions, favorable adhesion and anti-adhesion, lack of hemolysis, antibiotic susceptibility profile, and negative results for virulence genes. These factors confirm its safety for host consumption and promising potential for probiotic use. Antimicrobial activity is a crucial criterion for selecting probiotics as they can inhibit gastrointestinal pathogens. In this study, the L47-2 strain exhibited antagonistic activity against all tested pathogens, in agreement with previous findings. 7 , 26 , 27 , 36 – 42 Furthermore, data from our previous work and other studies showed that LAB could inhibit fungal growth, particularly opportunistic infections in AIDS patients. 22 , 43 , 44 L. plantarum is classified into the LAB group as it produces secondary metabolites, including lactic acid, and can affect the vital cell functions of other microorganisms. 45 Several studies have reported that the antagonistic activity of LAB was dependent on bacteriocin or bacteriocin-like substances, organic acids, hydrogen peroxide, ethanol, 1,3-propanediol, short-chain fatty acids, carbon dioxide, lactic acid, acetic acid, and phenyllactic acid. 31 , 46 – 48 LAB gains the advantageous capability to prevent infections with these antimicrobial compounds. According to our recent findings and previous work, 22 the L47-2 strain has antibacterial and antifungal activities. Probiotics must survive in acidic and bile salt environments, which is a requirement for their use. The L47-2 strain survived following exposure to pH 2.0 for 4 h, suggesting it may be a good probiotic candidate. These findings are consistent with previous reports 7 , 24 , 49 which reported that Lactobacillus spp. could survive under acidic and bile salt conditions for 4 h or more, indicating survival while passing through the stomach and reaching the entry of the small intestine, where the pH is 2.0-3.0 and the concentration of bile salt is 0.3%. 50 , 51 The capability of strain L47-2 to survive under bile salt conditions was due to modifications of carbohydrate and glycosidase activity, 52 production of exopolysaccharide, 53 , 54 the configuration of proteins and fatty acids on the cell membrane, 55 and enhanced adhesion to host mucus membranes. 56 , 57 In this study, the L47-2 strain exhibited the highest percentage of cell surface hydrophobicity, followed by E. coli ATCC 8739 and L. monocytogenes DMST 17303, which is related to the autoaggregation results. The L47-2 strain could coaggregate with each tested pathogen, depending on the bacterial-specific strain and incubation time. Surprisingly, the highest coaggregation activity (46.7 ± 2.4%) was observed between the L47-2 strain and L. monocytogenes DMST 17303, followed by E. coli ATCC 8739 (41.2 ± 1.0%) due to the high autoaggregation activity of these bacteria. The results of this study concur with the premise that autoaggregation and coaggregation capacities are correlated, as reported in previous studies. 31 , 42 , 58 , 59 The coaggregation capability of the L47-2 strain beneficially interferes with gastrointestinal pathogen colonization of host intestinal epithelial cells. Additionally, it could inhibit bacterial pathogens by producing inhibitory substances. 30 , 60 The coaggregation results of our work agree with previous studies, which illustrated that coaggregation is strain-specific (probiotic and pathogen) and incubation time dependent. 28 , 58 , 61 In the current study, the probiotic bacteria’s colonization of the gastrointestinal tract was shown by an adhesion assay. However, this capability is also characteristic of gastrointestinal pathogenic bacteria. The current study suggests that the L47-2 strain has the most significant adhesion capability at the surface of Caco-2 cells compared to the tested pathogens. Bacterial adhesion is a complicated mechanism where the bacterial cell membrane communicates with the contacting surfaces. 62 However, all bacterial pathogens could adhere to the surfaces of Caco-2 cells, indicating their capability to penetrate the human intestinal mucosa. Factors affecting the Lactobacilli’s adhesion to Caco-2 cells were their proteinaceous components, carbohydrate moieties, and surface proteins. 63 – 66 The obtained results in this study show that the L47-2 strain could protect Caco-2 cells from adhesion by all tested pathogenic bacteria, with inhibition as the most effective mechanism rather than competition and displacement. This suggests the potential use of the L47-2 strain in preventing bacterial infection in the gastrointestinal tract. The correlation coefficient revealed that autoaggregation is significantly correlated with coaggregation ( P = 0.037) and adhesion to Caco-2 cells ( P = 0.020). This implies that autoaggregation and adhesion capabilities are linked to bacterial attachment to intestinal cells. 67 , 68 Hydrophobic contacts are powerful non-covalent interactions and are regarded as critical factors in facilitating microbial adherence to host epithelial cells. 58 , 59 As expected, higher hydrophobicity was found to be associated with increased autoaggregation, indicating high adhesion to the cell surface as described in a previous study of L. plantarum strains. 69 Nonetheless, bacterial adhesion to the host cell surface depends on multifactor interactions and other factors, for instance, exopolysaccharides, S-layer proteins, mucus-binding proteins, lipoteichoic acid, and mannose-specific adhesions, which contribute to adhesion to host epithelial cells. 30 , 70 The safety of the L47-2 strain for probiotic use was determined using antibiotic susceptibility profiles, hemolysis, as well as the presence of virulence genes. In this study, the L47-2 strain was susceptible to ampicillin, tetracycline, gentamycin, and chloramphenicol, which concurs with previous research. 48 , 54 Kwon et al. (2021) reported that Lactiplantibacillus plantarum Q180 is sensitive to gentamycin, tetracycline, ampicillin, clindamycin, erythromycin, kanamycin, and chloramphenicol. 71 However, an intrinsic resistance of Lactobacillus sp. to vancomycin has also been reported. 72 The L47-2 strain has none of the 13 virulence genes according to PCR results, indicating its safety for probiotic use. The agg gene encodes an aggregation protein for adherence to eukaryotic cells; gelE encodes an extracellular metalloendopeptidase that involves hydrolysis of hemoglobin, collagen, gelatin, and other bioactive substances; and the esp is a cell wall-associated protein connected with immunological evasion. 73 efaAfs and efaAfm are cell wall adhesion molecules. 74 ccf , cob , and cpd encode for chemotactic attraction to human leukocytes and enable conjugation. 9 The cylLL , cylLS , cylLM , cylB , and cylA genes are cytolysin (hemolysin-bacteriocin) precursors that express the formation of active cytolysin, which is highly poisonous to human and bacterial cells. 73 Further research into in vivo anti-adhesion and immunomodulatory effects of the L47-2 strain would be beneficial in advancing our understanding of this strain’s potential as a promising probiotic for humans. The commercialization prospects for new probiotics, particularly next-generation probiotics (NGPs), are promising due to several key factors influencing the market dynamics. This growth is driven by rising consumer awareness regarding gut health, increased demand for functional foods, and a shift towards preventive healthcare. Next-generation probiotics are designed not only for traditional dietary uses but also for therapeutic applications. They show promise in treating various chronic ailments and are being tailored for personalized therapies. 75 This innovation aligns with trends in the healthcare sector focusing on individualized medicine, which could enhance their market acceptance and commercial viability. Despite the positive outlook, there are challenges to commercialization of NGPs, including strain selection, survivability, and regulatory hurdles. 75 Overall, Lactiplantibacillus plantarum represents a highly adaptable and beneficial microorganism with significant implications for food science and human health. Its extensive use in fermentation processes and probiotic formulations underscores its importance in both traditional and modern food systems. Further research into its genomic diversity and functional properties reveals new potential applications for this versatile bacterium. 16 – 19 These findings show that L. plantarum L47-2 may be deemed safe as a probiotic strain. More trials are necessary to determine the acute and subacute toxicity, immunotoxicity, embryotoxicity, and other properties of these probiotic bacteria in preclinical investigations for medicinal applications. Conclusions These findings indicate that L. plantarum strain L47-2 has potential for probiotic use due to its relevant antimicrobial activities, survival in acidic and bile salt environments, as well as its anti-adhesion to the host cells of all pathogens studied, and the absence of harmful characteristics. This study conducted a safety assessment of L47-2 utilizing phenotypic and genomic analyses. Moreover, the L47-2 strain exhibited an antipathogenic effect against six pathogenic bacteria. Based on our thorough safety evaluation, we have determined that the L. plantarum strain L47-2 can be used as a probiotic food supplement. However, additional comprehensive functional studies are needed to clarify the health benefits associated with this strain. Data availability The raw sequence data of L. plantarum strain L47-2 has been deposited in the European Nucleotide Archive (ENA) under the accession number PRJNA1220260 ( http://identifiers.org/ena.embl:PRJNA1220260 ). The sample accession number is SAMN46715021 ( http://identifiers.org/ena.embl:SAMN46715021 ). Acknowledgments We are grateful to the staff at the Department of Microbiology, Faculty of Science, Kasetsart University. References 1. Choi JH, Moon CM, Shin TS, et al. : Lactobacillus paracasei -derived extracellular vesicles attenuate the intestinal inflammatory response by augmenting the endoplasmic reticulum stress pathway. Exp. Mol. Med. 2020; 52 (3): 423–437. PubMed Abstract | Publisher Full Text | Free Full Text 2. Cristofori F, Dargenio VN, Dargenio C, et al. : Anti-Inflammatory and Immunomodulatory Effects of Probiotics in Gut Inflammation: A Door to the Body. Front. Immunol. 2021; 12 : 578386. PubMed Abstract | Publisher Full Text | Free Full Text 3. Taha-Abdelaziz K, Astill J, Kulkarni RR, et al. : In vitro assessment of immunomodulatory and anti- Campylobacter activities of probiotic lactobacilli. Sci. Rep. 2019; 9 (1): 17903. PubMed Abstract | Publisher Full Text | Free Full Text 4. Zheng J, Wittouck S, Salvetti E, et al. : A taxonomic note on the genus Lactobacillus : Description of 23 novel genera, emended description of the genus Lactobacillus Beijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae. Int. J. Syst. Evol. Microbiol. 2020; 70 (4): 2782–2858. PubMed Abstract | Publisher Full Text 5. Kim JA, Bayo J, Cha J, et al. : Investigating the probiotic characteristics of four microbial strains with potential application in feed industry. PLoS One. 2019; 14 (6): e0218922. PubMed Abstract | Publisher Full Text | Free Full Text 6. Sniffen JC, McFarland LV, Evans CT, et al. : Choosing an appropriate probiotic product for your patient: An evidence-based practical guide. PLoS One. 2018; 13 (12): e0209205. PubMed Abstract | Publisher Full Text | Free Full Text 7. Xu Y, Tian Y, Cao Y, et al. : Probiotic Properties of Lactobacillus paracasei subsp. paracasei L1 and Its Growth Performance-Promotion in Chicken by Improving the Intestinal Microflora. Front. Physiol. 2019; 10 : 937. PubMed Abstract | Publisher Full Text | Free Full Text 8. Yadav R, Puniya AK, Shukla P: Probiotic Properties of Lactobacillus plantarum RYPR1 from an Indigenous Fermented Beverage Raabadi. Front. Microbiol. 2016; 7 : 1683. PubMed Abstract 9. Gomez NC, Ramiro JM, Quecan BX, et al. : Use of Potential Probiotic Lactic Acid Bacteria (LAB) Biofilms for the Control of Listeria monocytogenes , Salmonella Typhimurium, and Escherichia coli O157:H7 Biofilms Formation. Front. Microbiol. 2016; 7 : 863. 10. Sharma H, Fidan H, Ozogul F, et al. : Recent development in the preservation effect of lactic acid bacteria and essential oils on chicken and seafood products. Front. Microbiol. 2022; 13 : 1092248. PubMed Abstract | Publisher Full Text | Free Full Text 11. Peter SB, Qiao Z, Godspower HN, et al. : Biotechnological Innovations and Therapeutic Application of Pediococcus and Lactic Acid Bacteria: The Next-Generation Microorganism. Front. Bioeng. Biotechnol. 2021; 9 : 802031. 12. Mathur H, Beresford TP, Cotter PD: Health Benefits of Lactic Acid Bacteria (LAB) Fermentates. Nutrients. 2020; 12 (6). PubMed Abstract | Publisher Full Text | Free Full Text 13. Farahmandi F, Parhizgar P, Mozafari Komesh Tape P, et al. : Implications and Mechanisms of Antiviral Effects of Lactic Acid Bacteria: A Systematic Review. Int. J. Microbiol. 2023; 2023 : 9298363. 14. Gizachew S, Engidawork E: Genomic Characterization of Lactiplantibacillus plantarum Strains: Potential Probiotics from Ethiopian Traditional Fermented Cottage Cheese. Genes (Basel). 2024; 15 (11). PubMed Abstract | Publisher Full Text | Free Full Text 15. Arasu MV, Al-Dhabi NA, Ilavenil S, et al. : In vitro importance of probiotic Lactobacillus plantarum related to medical field. Saudi J. Biol. Sci. 2016; 23 (1): S6–S10. PubMed Abstract | Publisher Full Text | Free Full Text 16. Alan Y, Keskin AO, Sonmez M: Probiotic and functional characterization of newly isolated Lactiplantibacillus plantarum strains from human breast milk and proliferative inhibition potential of metabolites. Enzym. Microb. Technol. 2025; 182 : 110545. PubMed Abstract | Publisher Full Text 17. Surve S, Shinde DB, Kulkarni R: Isolation, characterization and comparative genomics of potentially probiotic Lactiplantibacillus plantarum strains from Indian foods. Sci. Rep. 2022; 12 (1): 1940. PubMed Abstract | Publisher Full Text | Free Full Text 18. Zhang S, Wang T, Zhang D, et al. : Probiotic characterization of Lactiplantibacillus plantarum HOM3204 and its restoration effect on antibiotic-induced dysbiosis in mice. Lett. Appl. Microbiol. 2022; 74 (6): 949–958. PubMed Abstract | Publisher Full Text | Free Full Text 19. Yilmaz B, Bangar SP, Echegaray N, et al. : The Impacts of Lactiplantibacillus plantarum on the Functional Properties of Fermented Foods: A Review of Current Knowledge. Microorganisms. 2022; 10 (4). PubMed Abstract | Publisher Full Text | Free Full Text 20. Barbosa J, Albano H, Silva B, et al. : Characterization of a Lactiplantibacillus plantarum R23 Isolated from Arugula by Whole-Genome Sequencing and Its Bacteriocin Production Ability. Int. J. Environ. Res. Public Health. 2021; 18 (11). PubMed Abstract | Publisher Full Text | Free Full Text 21. Fidanza M, Panigrahi P, Kollmann TR: Lactiplantibacillus plantarum -Nomad and Ideal Probiotic. Front. Microbiol. 2021; 12 : 712236. PubMed Abstract | Publisher Full Text | Free Full Text 22. Sungsri T, Lertcanawanichakul M, Siwayaprahm P: Isolation and Selection of Anti- Candida albicans Metabolites Producing Lactic Acid Bacteria from Various Sources. KKU Res. J. 2012; 17 (4): 630–638. 23. Magnusson J, Schnurer J: Lactobacillus coryniformis subsp. coryniformis strain Si3 produces a broad-spectrum proteinaceous antifungal compound. Appl. Environ. Microbiol. 2001; 67 (1): 1–5. PubMed Abstract | Publisher Full Text | Free Full Text 24. Anandharaj M, Sivasankari B, Santhanakaruppu R, et al. : Determining the probiotic potential of cholesterol-reducing Lactobacillus and Weissella strains isolated from gherkins (fermented cucumber) and south Indian fermented koozh. Res. Microbiol. 2015; 166 (5): 428–439. PubMed Abstract | Publisher Full Text 25. de Albuquerque TMR , Garcia EF, de Oliveira AA , et al. : In Vitro Characterization of Lactobacillus Strains Isolated from Fruit Processing By-Products as Potential Probiotics. Probiotics Antimicrob. Proteins. 2018; 10 (4): 704–716. PubMed Abstract | Publisher Full Text 26. Rokana N, Singh BP, Thakur N, et al. : Screening of cell surface properties of potential probiotic lactobacilli isolated from human milk. J. Dairy Res. 2018; 85 (3): 347–354. PubMed Abstract | Publisher Full Text 27. Zommiti M, Connil N, Hamida JB, et al. : Probiotic Characteristics of Lactobacillus curvatus DN317, a Strain Isolated from Chicken Ceca. Probiotics Antimicrob. Proteins. 2017; 9 (4): 415–424. PubMed Abstract | Publisher Full Text 28. Collado MC, Meriluoto J, Salminen S: Adhesion and aggregation properties of probiotic and pathogen strains. Eur. Food Res. Technol. 2008; 226 (5): 1065–1073. Publisher Full Text 29. Sangsri T, Saiprom N, Tubsuwan A, et al. : Tetraspanins are involved in Burkholderia pseudomallei -induced cell-to-cell fusion of phagocytic and non-phagocytic cells. Sci. Rep. 2020; 10 (1): 17972. PubMed Abstract | Publisher Full Text | Free Full Text 30. Garcia-Ruiz A, Gonzalez de Llano D, Esteban-Fernandez A, et al. : Assessment of probiotic properties in lactic acid bacteria isolated from wine. Food Microbiol. 2014; 44 : 220–225. PubMed Abstract | Publisher Full Text 31. Feng YY, Qiao L, Liu R, et al. : Potential probiotic properties of lactic acid bacteria isolated from the intestinal mucosa of healthy piglets. Ann. Microbiol. 2017; 67 (3): 239–253. Publisher Full Text 32. Eaton TJ, Gasson MJ: Molecular screening of Enterococcus virulence determinants and potential for genetic exchange between food and medical isolates. Appl. Environ. Microbiol. 2001; 67 (4): 1628–1635. PubMed Abstract | Publisher Full Text | Free Full Text 33. Prjibelski A, Antipov D, Meleshko D, et al. : Using SPAdes De Novo Assembler. Curr. Protoc. Bioinformatics. 2020; 70 (1): e102. Publisher Full Text 34. Parks DH, Imelfort M, Skennerton CT, et al. : CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Res. 2015; 25 (7): 1043–1055. PubMed Abstract | Publisher Full Text | Free Full Text 35. EFSA: Guidance on the assessment of bacterial susceptibility to antimicrobials of human and veterinary importance. EFSA J. 2012; 10 (6): 2740. 36. Giri SS, Sukumaran V, Sen SS, et al. : Use of a Potential Probiotic, Lactobacillus casei L4, in the Preparation of Fermented Coconut Water Beverage. Front. Microbiol. 2018; 9 : 1976. PubMed Abstract | Publisher Full Text | Free Full Text 37. Fernandez MF, Boris S, Barbes C: Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract. J. Appl. Microbiol. 2003; 94 (3): 449–455. Publisher Full Text 38. Fuochi V, Cardile V, Petronio Petronio G, et al. : Biological properties and production of bacteriocinslike-inhibitory substances by Lactobacillus sp. strains from human vagina. J. Appl. Microbiol. 2018; 126 : 1541–1550. 39. Jena PK, Trivedi D, Thakore K, et al. : Isolation and characterization of probiotic properties of Lactobacilli isolated from rat fecal microbiota. Microbiol. Immunol. 2013; 57 (6): 407–416. PubMed Abstract | Publisher Full Text 40. Fossi BT, Ndjouenkeu R: Probiotic potential of thermotolerant lactic acid bacteria isolated from Gari a cassava-based African fermented food. J. Appl. Biol. Biotechnol. 2017; 5 (4): 001–005. 41. Bernbom N, Licht TR, Saadbye P, et al. : Lactobacillus plantarum inhibits growth of Listeria monocytogenes in an in vitro continuous flow gut model, but promotes invasion of L. monocytogenes in the gut of gnotobiotic rats. Int. J. Food Microbiol. 2006; 108 (1): 10–14. PubMed Abstract | Publisher Full Text 42. Abdelazez A, Abdelmotaal H, Evivie SE, et al. : Screening Potential Probiotic Characteristics of Lactobacillus brevis Strains in vitro and Intervention Effect on Type I Diabetes In Vivo. Biomed. Res. Int. 2018; 2018 : 7356173. 43. Ambe NF, Longdoh NA, Tebid P, et al. : The prevalence, risk factors and antifungal sensitivity pattern of oral candidiasis in HIV/AIDS patients in Kumba District Hospital, South West Region, Cameroon. Pan. Afr. Med. J. 2020; 36 : 23. Publisher Full Text 44. Kang CH, Han SH, Kim Y, et al. : In Vitro Probiotic Properties of Lactobacillus salivarius MG242 Isolated from Human Vagina. Probiotics Antimicrob. Proteins. 2018; 10 (2): 343–349. PubMed Abstract | Publisher Full Text 45. Kivanc M, Yilmaz M, Cakir E: Isolation and identification of lactic acid bacteria from boza, and their microbial activity against several reporter strains. Turk. J. Biol. 2011; 35 (3): 313–324. 46. Greifova G, Majekova H, Greif G, et al. : Analysis of antimicrobial and immunomodulatory substances produced by heterofermentative Lactobacillus reuteri. Folia Microbiol. (Praha). 2017; 62 (6): 515–524. PubMed Abstract | Publisher Full Text 47. Arslan S, Durak AN, Erbas M, et al. : Determination of microbiological and chemical properties of probiotic boza and its consumer acceptability. J. Am. Coll. Nutr. 2015; 34 (1): 56–64. PubMed Abstract | Publisher Full Text 48. Mulaw G, Sisay Tessema T, Muleta D, et al. : Corrigendum to “In Vitro Evaluation of Probiotic Properties of Lactic Acid Bacteria Isolated from Some Traditionally Fermented Ethiopian Food Products”. Int. J. Microbiol. 2020; 2020 : 1. Publisher Full Text 49. Dowarah R, Verma AK, Agarwal N, et al. : Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status in weaned piglets. PLoS One. 2018; 13 (3): e0192978. PubMed Abstract | Publisher Full Text | Free Full Text 50. du Toit M , Franz CM, Dicks LM, et al. : Characterisation and selection of probiotic lactobacilli for a preliminary minipig feeding trial and their effect on serum cholesterol levels, faeces pH and faeces moisture content. Int. J. Food Microbiol. 1998; 40 (1-2): 93–104. PubMed Abstract | Publisher Full Text 51. Wang J, Ji HF, Zhang DY, et al. : Assessment of probiotic properties of Lactobacillus plantarum ZLP001 isolated from gastrointestinal tract of weaning pigs. Afr. J. Biotechnol. 2011; 10 (54): 11303–11308. 52. Taheri HR, Moravej H, Tabandeh F, et al. : Screening of lactic acid bacteria toward their selection as a source of chicken probiotic. Poult. Sci. 2009; 88 (8): 1586–1593. PubMed Abstract | Publisher Full Text 53. Petsuriyawong B, Khunajakr N: Screening of probiotic lactic acid bacteria from piglet feces. Kasetsart J. 2011; 45 : 245–253. 54. Shazali N, Foo HL, Loh TC, et al. : Prevalence of antibiotic resistance in lactic acid bacteria isolated from the faeces of broiler chicken in Malaysia. Gut Pathog. 2014; 6 (1): 1. PubMed Abstract | Publisher Full Text | Free Full Text 55. Fairbrother JM, Nadeau E, Gyles CL: Escherichia coli in postweaning diarrhea in pigs: an update on bacterial types, pathogenesis, and prevention strategies. Anim. Health Res. Rev. 2005; 6 (1): 17–39. PubMed Abstract | Publisher Full Text 56. Cho IJ, Lee NK, Hahm YT: Characterization of Lactobacillus spp. isolated from the feces of breast-feeding piglets. J. Biosci. Bioeng. 2009; 108 (3): 194–198. PubMed Abstract | Publisher Full Text 57. Venkatesan SM, Narayan GS, Ramachandran AK, et al. : The effect of two bleaching agents on the phosphate concentration of the enamel evaluated by Raman spectroscopy: An ex vivo study. Contemp. Clin. Dent. 2012; 3 (Suppl 2): S172–S176. PubMed Abstract | Publisher Full Text 58. Abushelaibi A, Al-Mahadin S, El-Tarabily K, et al. : Characterization of potential probiotic lactic acid bacteria isolated from camel milk. Lwt-Food Sci. Technol. 2017; 79 : 316–325. Publisher Full Text 59. Somashekaraiah R, Shruthi B, Deepthi BV, et al. : Probiotic Properties of Lactic Acid Bacteria Isolated From Neera: A Naturally Fermenting Coconut Palm Nectar. Front. Microbiol. 2019; 10 : 1382. PubMed Abstract | Publisher Full Text | Free Full Text 60. Kaewnopparat S, Dangmanee N, Kaewnopparat N, et al. : In vitro probiotic properties of Lactobacillus fermentum SK5 isolated from vagina of a healthy woman. Anaerobe. 2013; 22 : 6–13. PubMed Abstract | Publisher Full Text 61. Ben Taheur F, Kouidhi B, Fdhila K, et al. : Anti-bacterial and anti-biofilm activity of probiotic bacteria against oral pathogens. Microb. Pathog. 2016; 97 : 213–220. PubMed Abstract | Publisher Full Text 62. Solieri L, Bianchi A, Mottolese G, et al. : Tailoring the probiotic potential of non-starter Lactobacillus strains from ripened Parmigiano Reggiano cheese by in vitro screening and principal component analysis. Food Microbiol. 2014; 38 : 240–249. PubMed Abstract | Publisher Full Text 63. Dimitrov Z, Gotova I, Chorbadjiyska E: In vitro characterization of the adhesive factors of selected probiotics to Caco-2 epithelium cell line. Biotechnol. Biotechnol. Equip. 2014; 28 (6): 1079–1083. PubMed Abstract | Publisher Full Text | Free Full Text 64. Greene JD, Klaenhammer TR: Factors involved in adherence of lactobacilli to human Caco-2 cells. Appl. Environ. Microbiol. 1994; 60 (12): 4487–4494. PubMed Abstract | Publisher Full Text | Free Full Text 65. Lim SM, Ahn DH: Factors affecting adhesion of lactic acid bacteria to Caco-2 cells and inhibitory effect on infection of Salmonella typhimurium. J. Microbiol. Biotechnol. 2012; 22 (12): 1731–1739. PubMed Abstract | Publisher Full Text 66. Polak-Berecka M, Wasko A, Paduch R, et al. : The effect of cell surface components on adhesion ability of Lactobacillus rhamnosus. Antonie Van Leeuwenhoek. 2014; 106 (4): 751–762. PubMed Abstract | Publisher Full Text | Free Full Text 67. Kos B, Suskovic J, Vukovic S, et al. : Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J. Appl. Microbiol. 2003; 94 (6): 981–987. PubMed Abstract | Publisher Full Text 68. Hojjati M, Behabahani BA, Falah F: Aggregation, adherence, anti-adhesion and antagonistic activity properties relating to surface charge of probiotic Lactobacillus brevis gp104 against Staphylococcus aureus. Microb. Pathog. 2020; 147 : 104420. PubMed Abstract | Publisher Full Text 69. Sun Y, Zhang S, Li H, et al. : Assessments of Probiotic Potentials of Lactiplantibacillus plantarum Strains Isolated From Chinese Traditional Fermented Food: Phenotypic and Genomic Analysis. Front. Microbiol. 2022; 13 : 895132. PubMed Abstract | Publisher Full Text | Free Full Text 70. Garcia-Cayuela T, Korany AM, Bustos I, et al. : Adhesion abilities of dairy Lactobacillus plantarum strains showing an aggregation phenotype. Food Res. Int. 2014; 57 : 44–50. Publisher Full Text 71. Kwon YJ, Chun BH, Jung HS, et al. : Safety Assessment of Lactiplantibacillus (formerly Lactobacillus ) plantarum Q180. J. Microbiol. Biotechnol. 2021; 31 (10): 1420–1429. PubMed Abstract | Publisher Full Text | Free Full Text 72. Gueimonde M, Sanchez B, de los Reyes-Gavilán CG , et al. : Antibiotic resistance in probiotic bacteria. Front. Microbiol. 2013; 4 : 202. 73. AlKalbani NS, Turner MS, Ayyash MM: Isolation, identification, and potential probiotic characterization of isolated lactic acid bacteria and in vitro investigation of the cytotoxicity, antioxidant, and antidiabetic activities in fermented sausage. Microb. Cell Factories. 2019; 18 (1): 188. PubMed Abstract | Publisher Full Text | Free Full Text 74. Popovic N, Dinic M, Tolinacki M, et al. : New Insight into Biofilm Formation Ability, the Presence of Virulence Genes and Probiotic Potential of Enterococcus sp. Dairy Isolates. Front Microbiol. 2018; 9 : 78. PubMed Abstract | Publisher Full Text | Free Full Text 75. Abouelela ME, Helmy YA: Next-Generation Probiotics as Novel Therapeutics for Improving Human Health: Current Trends and Future Perspectives. Microorganisms. 2024; 12 (3). PubMed Abstract | Publisher Full Text | Free Full Text Comments on this article Comments (0) Version 1 VERSION 1 PUBLISHED 07 Jul 2025 ADD YOUR COMMENT Comment Author details Author details 1 Department of Microbiology, Faculty of Medicine, Princess of Naradhiwas University, Narathiwat, 96000, Thailand 2 Department of Medical Science, Amnatcharoen Campus, Mahidol University, Amnatcharoen, 37000, Thailand 3 Department of Microbiology, Faculty of Science,, Kasetsart University, Bangkok, Bangkok, 10900, Thailand 4 Department of Clinical Pathology, Faculty of Medicine Vajira Hospital, Navamindradhiraj University, Bangkok, 10300, Thailand Tanes Sangsri Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Rathanin Seng Roles: Formal Analysis, Methodology, Writing – Original Draft Preparation, Writing – Review & Editing Worrapot Pengpa Roles: Investigation Thanwa Wongsuk Roles: Formal Analysis, Writing – Original Draft Preparation, Writing – Review & Editing Patcharaporn Siwayaprahm Roles: Conceptualization, Funding Acquisition, Methodology, Project Administration, Supervision, Validation, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information This research was partially supported by the Undergraduate Research Matching Fund: URMF 2021, Faculty of Science, Kasetsart University and Microbiology Projects grant of the Department of Microbiology, Faculty of Science, Kasetsart University and Fundamental Fund (2024), Thailand Science Research and Innovation, Faculty of Medicine, Princess of Naradhiwas University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Article Versions (1) version 1 Published: 07 Jul 2025, 14:670 https://doi.org/10.12688/f1000research.166421.1 Copyright © 2025 Sangsri T et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Sangsri T, Seng R, Pengpa W et al. In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.12688/f1000research.166421.1 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 1 VERSION 1 PUBLISHED 07 Jul 2025 Views 0 Cite How to cite this report: Kingkaew E. Reviewer Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r402055 ) The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-402055 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 01 Oct 2025 Engkarat Kingkaew , King Mongkut’s Institute of Technology Ladkrabang, Bangkok, Thailand Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.183407.r402055 1. Please make the statistically analysis for showing the significant between factors/time ? Introduction part: Opening scope Broaden first sentence: LAB occur in many fermented foods (vegetables, ... Continue reading READ ALL 1. Please make the statistically analysis for showing the significant between factors/time ? Introduction part: Opening scope Broaden first sentence: LAB occur in many fermented foods (vegetables, meats, etc.), not only dairy products. Grammar & wording “serves as probiotic and non-pathogenic bacteria” → revise to “serves as a probiotic and contains non-pathogenic species.” “LAB produces bacteriocins-antimicrobial peptides” → revise to “LAB produce bacteriocins—antimicrobial peptides…” “has antagonistic action against gut infections” → revise to “exhibits antagonistic activity against gut pathogens.” Redundancy Acid and bile tolerance, bacteriocin production, and probiotic traits are repeated multiple times; condense to avoid overlap. Paragraph structure The second paragraph is overloaded (bio-preservation, immunomodulation, antiviral activity, genetic engineering, AI) → split into separate focused sections. Speculative or unclear content The mention of artificial intelligence lacks context and clear link to the study’s probiotic focus; clarify or remove. Provide justification or reference for the statement about AI improving LAB safety and efficiency. Scientific precision When listing LAB genera, briefly note the reason for taxonomic revision (phylogenetic reclassification). In the probiotic criteria list, consider adding “ability to confer a health benefit to the host” to align with FAO/WHO definitions. Morphological details Cell size (0.9–1.2 µm × 3–8 µm) may be unnecessary unless tied to functional relevance; consider shortening. Reference placement Avoid repeating identical citations (e.g., “9,10” multiple times in one sentence); merge or distribute logically. Research objective Place the study aim (“to evaluate the probiotic characteristics of L. plantarum strain L47-2…”) in a separate concluding paragraph with a clearer link to the preceding background Methods: Opening & Growth Conditions Correct spelling: “ de Man Rogose Sharpe (MRS) ” → “ de Man, Rogosa and Sharpe (MRS) ” . Standardize brand/supplier format (e.g., “Difco Laboratories, Detroit” vs. “Difco™, USA”). Use consistent units and spacing (e.g., 37 °C, CFU/mL, μL). General Style & Structure Excessive repetition of “ L. plantarum strain L47-2”; after first mention in each section, shorten to “ L47-2 ” or “ the strain ” . Break long sections (e.g., Adhesion and anti-adhesion , Autoaggregation and coaggregation ) into smaller subheadings or paragraphs for clarity. Ensure all Latin names ( Lactiplantibacillus plantarum , Escherichia coli , etc.) are italicized consistently. Antibacterial Activity Assay Clarify how i nhibition zones were measured (e.g., ruler, caliper, digital imaging) and specify units (mm). Indicate how triplicate results were averaged and analyzed. Low pH and Bile Salt Survival State whether pH was monitored/maintained during incubation (acid drift can affect results). Clarify whether survival counts were log-transformed before statistical analysis. Cell Surface Hydrophobicity Confirm the formula (1 − A1/A0) × 100 includes blank correction for OD measurements. Autoaggregation & Coaggregation Unify time points: text lists 0,1,2,3,4,5,16,24 h but formula mentions 1,2,4,6,8,10,20,24 h. Clarify if coaggregation assessment was only OD-based or also verified microscopically. Adhesion / Anti-adhesion (Caco-2) Define Multiplicity of Infection (MOI) calculation and how bacterial counts were confirmed. Add details on washing stringency (PBS volume per wash). Indicate whether viability staining was used to distinguish live vs. dead attached cells. Safety Assessment Hemolysis: specify blood agar base used and criteria for α-, β-, or γ-hemolysis interpretation. Antibiotic susceptibility: Indicate whether EUCAST/EFSA or CLSI guidelines were used for MIC/MBC interpretation. Clarify if broth pH was adjusted after antibiotic addition, as acidity can affect LAB growth. Molecular Screening of Virulence Genes Confirm if 55 °C annealing temperature is optimal for all primer sets. Provide positive and negative control strains for each gene, not just overall. Whole Genome Sequencing Include total genome size and GC content for completeness. State if raw reads/assemblies were deposited in a public database (e.g., NCBI SRA) and provide accession numbers. Define identity and coverage cut-offs used by CARD for antibiotic resistance gene detection. Statistical Analysis Correct phrase: “ A student’s t-test” → “Student’s t-test. ” Specify normality testing (e.g., Shapiro–Wilk) prior to t-test/ANOVA. Indicate if post-hoc multiple-comparison corrections (e.g., Tukey’s) were applied after ANOVA. Results: 1. Please show the data for confirming of the identification (such as ANI, AAI, dDDH) 2. Please attached the picture for showing the antibacterial activity of strain L47-2 and why the author use "dual culture overlay method" ? 3. Can you discuss the about antimicrobial compound which related to your antimicrobial activity and analysis through genome sequences and shows the location of genes 4. acidic and bile salt conditions < please statistically analyzed and showed p-value 5. Why you compare the hydrophobicity to pathogens / indicators strains ? Possibly, please compare to the other probiotic strains or make the more comprehensive and reasonable to clarify the results 6. What is meaning of dahsed lines ? 7. Why did you use "pathogen.watch" ? please indicate the reason in the method section 8. According to the adhesion ability of L47-2, please make the discussion about the ability and probiotic-associated genes Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests: No competing interests were disclosed. Reviewer Expertise: Probiotics, Food microbiology, Whole-genome sequenceing I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Kingkaew E. Reviewer Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r402055 ) The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-402055 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Kulkarni R. Reviewer Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r399880 ) The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-399880 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 01 Sep 2025 Ram Kulkarni , Symbiosis School of Biological Sciences, Symbiosis International (Deemed University), Pune, Maharashtra, India Not Approved VIEWS 0 https://doi.org/10.5256/f1000research.183407.r399880 The manuscript by Sangsri et al. evaluates a few phenotypic characteristics of L. plantarum L47-2 and reports its whole genome sequence. I consider the paper unsuitable for publication in F1000Research for the following reasons. The phenotypic data ... Continue reading READ ALL The manuscript by Sangsri et al. evaluates a few phenotypic characteristics of L. plantarum L47-2 and reports its whole genome sequence. I consider the paper unsuitable for publication in F1000Research for the following reasons. The phenotypic data and genome sequence presented do not provide new insights that advance the characterization of lactobacilli as probiotics. The experimental design, results, and discussion are often superficial and also carry flaws in a few places. Numerous previous studies have already explored the characterization of L. plantarum strains and established phenotype–genotype correlations. The Introduction contains considerable redundancy and a lack of clarity in places. For instance, the statement regarding the integration of artificial intelligence at the end of the second paragraph is irrelevant to this work. There is also redundant content on the antimicrobial activity of LAB / L. plantarum and the stress tolerance of L. plantarum in the second and third paragraphs. There is ambiguity regarding the source and precise identity of the strain L. plantarum L47-2 used in this study. The previously cited source (Sungsri et al. 2012) does not report isolation of any L. plantarum strains. Moreover, the strain referred to as L47-2 was previously identified as L. paracasei. Table 2: Correlation analysis between two variables is not meaningful when the data includes six bacteria for co-aggregation and seven for other properties. The conclusion, “This finding indicates that the capabilities of the L47-2 strain to autoaggregate, coaggregate, and adhere were significantly correlated,” is incorrect, as correlation analysis requires data from multiple strains. After conducting ANOVA to determine differences among group means of adhesion (Fig. 4A), a post hoc test rather than multiple t-tests should be used to identify which bacteria differ significantly from L47-2. A post hoc test should also be applied to Fig. 2. Please clarify why statistical analysis was performed only for select data, and not for other results presented in Table 1, Figures 1, 3, and 4B. The results of the genomic characterization are insufficiently detailed. Did the authors search only for vancomycin resistance genes? The evidence for the presence of vanH and vanY genes should be provided. It is also important to note that vancomycin resistance in L. plantarum is mostly intrinsic and typically related to variations in the D-alanine D-alanine ligase involved in cell wall biosynthesis. When whole genome sequencing has been performed, evaluating virulence genes solely by PCR is unnecessary, and such assessment could be conducted bioinformatically using WGS data. The Discussion lacks insight into possible mechanisms underlying key observations and comparison of the observations with the previously published literature in addition to being confusing at some places. For example, in the third paragraph, the statement regarding the reasons for the survival of L47-2 under bile salt conditions, along with the accompanying references, incorrectly suggests that the cited studies are on L47-2, which is not the case. Why authors find it “surprising” to see the highest coaggregation activity between the L47-2 strain and L. monocytogenes DMST 17303 is not clear. Furthermore, the statement that "the coaggregation capability of the L47-2 strain beneficially interferes with gastrointestinal pathogen colonization" is unsupported as there are no in vivo studies in this manuscript. Additional comments: Title: The use of the word "determination" in quotation marks in the title is potentially confusing and is not standard in scientific writing. Other words such as assessment or characterization could be used without quotation marks. Methodology: Please clarify whether L. plantarum was grown under 5% CO2, as stated in the first paragraph, or under 5–20% CO2, as stated in the second. The explanation of acid and bile salt tolerance calculations is confusing and contains errors; this section should be revised for accuracy and clarity. In the section on adhesion and anti-adhesion assays involving L. plantarum L47-2 and pathogens, the specific pathogens tested are not clearly specified. Additionally, in the inhibition, competition, and displacement assays, it is unclear which medium was used for counting pathogen CFUs. Was a selective medium employed to prevent the growth of L. plantarum ? Note that MIC stands for "minimum inhibitory concentration," not "minimal inhibitory concentration," and the same applies to MBC. Fig. 1: The variables shown on the x-axis should be grouped in a consistent manner for both panels A and B to facilitate comparison. For example, subgroups in both panels could represent varying concentrations of the stressor or, alternatively, varying incubation times. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? No Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? No Are all the source data underlying the results available to ensure full reproducibility? No source data required Are the conclusions drawn adequately supported by the results? No Competing Interests: No competing interests were disclosed. Reviewer Expertise: Lactic acid bacteria I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Kulkarni R. Reviewer Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r399880 ) The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-399880 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Comments on this article Comments (0) Version 1 VERSION 1 PUBLISHED 07 Jul 2025 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 Version 1 07 Jul 25 read read Ram Kulkarni , Symbiosis International (Deemed University), Pune, India Engkarat Kingkaew , King Mongkut’s Institute of Technology Ladkrabang, Bangkok, Thailand Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Kingkaew E. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 01 Oct 2025 | for Version 1 Engkarat Kingkaew , King Mongkut’s Institute of Technology Ladkrabang, Bangkok, Thailand 0 Views copyright © 2025 Kingkaew E. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions 1. Please make the statistically analysis for showing the significant between factors/time ? Introduction part: Opening scope Broaden first sentence: LAB occur in many fermented foods (vegetables, meats, etc.), not only dairy products. Grammar & wording “serves as probiotic and non-pathogenic bacteria” → revise to “serves as a probiotic and contains non-pathogenic species.” “LAB produces bacteriocins-antimicrobial peptides” → revise to “LAB produce bacteriocins—antimicrobial peptides…” “has antagonistic action against gut infections” → revise to “exhibits antagonistic activity against gut pathogens.” Redundancy Acid and bile tolerance, bacteriocin production, and probiotic traits are repeated multiple times; condense to avoid overlap. Paragraph structure The second paragraph is overloaded (bio-preservation, immunomodulation, antiviral activity, genetic engineering, AI) → split into separate focused sections. Speculative or unclear content The mention of artificial intelligence lacks context and clear link to the study’s probiotic focus; clarify or remove. Provide justification or reference for the statement about AI improving LAB safety and efficiency. Scientific precision When listing LAB genera, briefly note the reason for taxonomic revision (phylogenetic reclassification). In the probiotic criteria list, consider adding “ability to confer a health benefit to the host” to align with FAO/WHO definitions. Morphological details Cell size (0.9–1.2 µm × 3–8 µm) may be unnecessary unless tied to functional relevance; consider shortening. Reference placement Avoid repeating identical citations (e.g., “9,10” multiple times in one sentence); merge or distribute logically. Research objective Place the study aim (“to evaluate the probiotic characteristics of L. plantarum strain L47-2…”) in a separate concluding paragraph with a clearer link to the preceding background Methods: Opening & Growth Conditions Correct spelling: “ de Man Rogose Sharpe (MRS) ” → “ de Man, Rogosa and Sharpe (MRS) ” . Standardize brand/supplier format (e.g., “Difco Laboratories, Detroit” vs. “Difco™, USA”). Use consistent units and spacing (e.g., 37 °C, CFU/mL, μL). General Style & Structure Excessive repetition of “ L. plantarum strain L47-2”; after first mention in each section, shorten to “ L47-2 ” or “ the strain ” . Break long sections (e.g., Adhesion and anti-adhesion , Autoaggregation and coaggregation ) into smaller subheadings or paragraphs for clarity. Ensure all Latin names ( Lactiplantibacillus plantarum , Escherichia coli , etc.) are italicized consistently. Antibacterial Activity Assay Clarify how i nhibition zones were measured (e.g., ruler, caliper, digital imaging) and specify units (mm). Indicate how triplicate results were averaged and analyzed. Low pH and Bile Salt Survival State whether pH was monitored/maintained during incubation (acid drift can affect results). Clarify whether survival counts were log-transformed before statistical analysis. Cell Surface Hydrophobicity Confirm the formula (1 − A1/A0) × 100 includes blank correction for OD measurements. Autoaggregation & Coaggregation Unify time points: text lists 0,1,2,3,4,5,16,24 h but formula mentions 1,2,4,6,8,10,20,24 h. Clarify if coaggregation assessment was only OD-based or also verified microscopically. Adhesion / Anti-adhesion (Caco-2) Define Multiplicity of Infection (MOI) calculation and how bacterial counts were confirmed. Add details on washing stringency (PBS volume per wash). Indicate whether viability staining was used to distinguish live vs. dead attached cells. Safety Assessment Hemolysis: specify blood agar base used and criteria for α-, β-, or γ-hemolysis interpretation. Antibiotic susceptibility: Indicate whether EUCAST/EFSA or CLSI guidelines were used for MIC/MBC interpretation. Clarify if broth pH was adjusted after antibiotic addition, as acidity can affect LAB growth. Molecular Screening of Virulence Genes Confirm if 55 °C annealing temperature is optimal for all primer sets. Provide positive and negative control strains for each gene, not just overall. Whole Genome Sequencing Include total genome size and GC content for completeness. State if raw reads/assemblies were deposited in a public database (e.g., NCBI SRA) and provide accession numbers. Define identity and coverage cut-offs used by CARD for antibiotic resistance gene detection. Statistical Analysis Correct phrase: “ A student’s t-test” → “Student’s t-test. ” Specify normality testing (e.g., Shapiro–Wilk) prior to t-test/ANOVA. Indicate if post-hoc multiple-comparison corrections (e.g., Tukey’s) were applied after ANOVA. Results: 1. Please show the data for confirming of the identification (such as ANI, AAI, dDDH) 2. Please attached the picture for showing the antibacterial activity of strain L47-2 and why the author use "dual culture overlay method" ? 3. Can you discuss the about antimicrobial compound which related to your antimicrobial activity and analysis through genome sequences and shows the location of genes 4. acidic and bile salt conditions < please statistically analyzed and showed p-value 5. Why you compare the hydrophobicity to pathogens / indicators strains ? Possibly, please compare to the other probiotic strains or make the more comprehensive and reasonable to clarify the results 6. What is meaning of dahsed lines ? 7. Why did you use "pathogen.watch" ? please indicate the reason in the method section 8. According to the adhesion ability of L47-2, please make the discussion about the ability and probiotic-associated genes Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Probiotics, Food microbiology, Whole-genome sequenceing I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Kingkaew E. Peer Review Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r402055) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-402055 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Kulkarni R. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 01 Sep 2025 | for Version 1 Ram Kulkarni , Symbiosis School of Biological Sciences, Symbiosis International (Deemed University), Pune, Maharashtra, India 0 Views copyright © 2025 Kulkarni R. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Not Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The manuscript by Sangsri et al. evaluates a few phenotypic characteristics of L. plantarum L47-2 and reports its whole genome sequence. I consider the paper unsuitable for publication in F1000Research for the following reasons. The phenotypic data and genome sequence presented do not provide new insights that advance the characterization of lactobacilli as probiotics. The experimental design, results, and discussion are often superficial and also carry flaws in a few places. Numerous previous studies have already explored the characterization of L. plantarum strains and established phenotype–genotype correlations. The Introduction contains considerable redundancy and a lack of clarity in places. For instance, the statement regarding the integration of artificial intelligence at the end of the second paragraph is irrelevant to this work. There is also redundant content on the antimicrobial activity of LAB / L. plantarum and the stress tolerance of L. plantarum in the second and third paragraphs. There is ambiguity regarding the source and precise identity of the strain L. plantarum L47-2 used in this study. The previously cited source (Sungsri et al. 2012) does not report isolation of any L. plantarum strains. Moreover, the strain referred to as L47-2 was previously identified as L. paracasei. Table 2: Correlation analysis between two variables is not meaningful when the data includes six bacteria for co-aggregation and seven for other properties. The conclusion, “This finding indicates that the capabilities of the L47-2 strain to autoaggregate, coaggregate, and adhere were significantly correlated,” is incorrect, as correlation analysis requires data from multiple strains. After conducting ANOVA to determine differences among group means of adhesion (Fig. 4A), a post hoc test rather than multiple t-tests should be used to identify which bacteria differ significantly from L47-2. A post hoc test should also be applied to Fig. 2. Please clarify why statistical analysis was performed only for select data, and not for other results presented in Table 1, Figures 1, 3, and 4B. The results of the genomic characterization are insufficiently detailed. Did the authors search only for vancomycin resistance genes? The evidence for the presence of vanH and vanY genes should be provided. It is also important to note that vancomycin resistance in L. plantarum is mostly intrinsic and typically related to variations in the D-alanine D-alanine ligase involved in cell wall biosynthesis. When whole genome sequencing has been performed, evaluating virulence genes solely by PCR is unnecessary, and such assessment could be conducted bioinformatically using WGS data. The Discussion lacks insight into possible mechanisms underlying key observations and comparison of the observations with the previously published literature in addition to being confusing at some places. For example, in the third paragraph, the statement regarding the reasons for the survival of L47-2 under bile salt conditions, along with the accompanying references, incorrectly suggests that the cited studies are on L47-2, which is not the case. Why authors find it “surprising” to see the highest coaggregation activity between the L47-2 strain and L. monocytogenes DMST 17303 is not clear. Furthermore, the statement that "the coaggregation capability of the L47-2 strain beneficially interferes with gastrointestinal pathogen colonization" is unsupported as there are no in vivo studies in this manuscript. Additional comments: Title: The use of the word "determination" in quotation marks in the title is potentially confusing and is not standard in scientific writing. Other words such as assessment or characterization could be used without quotation marks. Methodology: Please clarify whether L. plantarum was grown under 5% CO2, as stated in the first paragraph, or under 5–20% CO2, as stated in the second. The explanation of acid and bile salt tolerance calculations is confusing and contains errors; this section should be revised for accuracy and clarity. In the section on adhesion and anti-adhesion assays involving L. plantarum L47-2 and pathogens, the specific pathogens tested are not clearly specified. Additionally, in the inhibition, competition, and displacement assays, it is unclear which medium was used for counting pathogen CFUs. Was a selective medium employed to prevent the growth of L. plantarum ? Note that MIC stands for "minimum inhibitory concentration," not "minimal inhibitory concentration," and the same applies to MBC. Fig. 1: The variables shown on the x-axis should be grouped in a consistent manner for both panels A and B to facilitate comparison. For example, subgroups in both panels could represent varying concentrations of the stressor or, alternatively, varying incubation times. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? No Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? No Are all the source data underlying the results available to ensure full reproducibility? No source data required Are the conclusions drawn adequately supported by the results? No Competing Interests No competing interests were disclosed. Reviewer Expertise Lactic acid bacteria I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. reply Respond to this report Responses (0) Kulkarni R. Peer Review Report For: In vitro "determination" of Lactiplantibacillus plantarum strain L47-2 for safe probiotic use in gastrointestinal tract conditions [version 1; peer review: 1 approved with reservations, 1 not approved] . F1000Research 2025, 14 :670 ( https://doi.org/10.5256/f1000research.183407.r399880) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/14-670/v1#referee-response-399880 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions Adjust parameters to alter display View on desktop for interactive features Includes Interactive Elements View on desktop for interactive features Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Stay Updated Sign up for content alerts and receive a weekly or monthly email with all newly published articles Register with F1000Research Already registered? Sign in Not now, thanks close PLEASE NOTE If you are an AUTHOR of this article, please check that you signed in with the account associated with this article otherwise we cannot automatically identify your role as an author and your comment will be labelled as a “User Comment”. If you are a REVIEWER of this article, please check that you have signed in with the account associated with this article and then go to your account to submit your report, please do not post your review here. If you do not have access to your original account, please contact us . All commenters must hold a formal affiliation as per our Policies . The information that you give us will be displayed next to your comment. User comments must be in English, comprehensible and relevant to the article under discussion. We reserve the right to remove any comments that we consider to be inappropriate, offensive or otherwise in breach of the User Comment Terms and Conditions . Commenters must not use a comment for personal attacks. When criticisms of the article are based on unpublished data, the data should be made available. I accept the User Comment Terms and Conditions Please confirm that you accept the User Comment Terms and Conditions. Affiliation ✕ refresh Please enter your institution. Note: To add your institution or organisation, start typing the name and then select the correct name from the list. Where applicable, the name will appear in both the original language and in English. Do not paste in the name. If the name does not appear in the drop-down list, we will display the information you have entered. ✕ refresh Country/Region * USA UK Canada China France Germany Afghanistan Aland Islands Albania Algeria American Samoa Andorra Angola Anguilla Antarctica Antigua and Barbuda Argentina Armenia Aruba Australia Austria Azerbaijan Bahamas Bahrain Bangladesh Barbados Belarus Belgium Belize Benin Bermuda Bhutan Bolivia Bosnia and Herzegovina Botswana Bouvet Island Brazil British Indian Ocean Territory British Virgin Islands Brunei Bulgaria Burkina Faso Burundi Cambodia Cameroon Canada Cape Verde Cayman Islands Central African Republic Chad Chile China Christmas Island Cocos (Keeling) Islands Colombia Comoros Congo Cook Islands Costa Rica Cote d'Ivoire Croatia Cuba Cyprus Czech Republic Democratic Republic of the Congo Denmark Djibouti Dominica Dominican Republic Ecuador Egypt El Salvador Equatorial Guinea Eritrea Estonia Ethiopia Falkland Islands Faroe Islands Federated States of Micronesia Fiji Finland France French Guiana French Polynesia French Southern Territories Gabon Georgia Germany Ghana Gibraltar Greece Greenland Grenada Guadeloupe Guam Guatemala Guernsey Guinea Guinea-Bissau Guyana Haiti Heard Island and Mcdonald Islands Holy See (Vatican City State) Honduras Hong Kong Hungary Iceland India Indonesia Iran Iraq Ireland Israel Italy Jamaica Japan Jersey Jordan Kazakhstan Kenya Kiribati Kosovo (Serbia and Montenegro) Kuwait Kyrgyzstan Lao People's Democratic Republic Latvia Lebanon Lesotho Liberia Libya Liechtenstein Lithuania Luxembourg Macao Madagascar Malawi Malaysia Maldives Mali Malta Marshall Islands Martinique Mauritania Mauritius Mayotte Mexico Minor Outlying Islands of the United States Moldova Monaco Mongolia Montenegro Montserrat Morocco Mozambique Myanmar Namibia Nauru Nepal Netherlands Antilles New Caledonia New Zealand Nicaragua Niger Nigeria Niue Norfolk Island North Korea North Macedonia Northern Mariana Islands Norway Oman Pakistan Palau Palestinian Territory Panama Papua New Guinea Paraguay Peru Philippines Pitcairn Poland Portugal Puerto Rico Qatar Reunion Romania Russian Federation Rwanda Saint Helena Saint Kitts and Nevis Saint Lucia Saint Pierre and Miquelon Saint Vincent and the Grenadines Samoa San Marino Sao Tome and Principe Saudi Arabia Senegal Serbia Seychelles Sierra Leone Singapore Slovakia Slovenia Solomon Islands Somalia South Africa South Georgia and the South Sandwich Is South Korea South Sudan Spain Sri Lanka Sudan Suriname Svalbard and Jan Mayen Swaziland Sweden Switzerland Syria Taiwan Tajikistan Tanzania Thailand The Gambia The Netherlands Timor-Leste Togo Tokelau Tonga Trinidad and Tobago Tunisia Turkey Turkmenistan Turks and Caicos Islands Tuvalu UK USA Uganda Ukraine United Arab Emirates United States Virgin Islands Uruguay Uzbekistan Vanuatu Venezuela Vietnam Wallis and Futuna West Bank and Gaza Strip Western Sahara Yemen Zambia Zimbabwe Please select your country/region. You must enter a comment. Competing Interests Please disclose any competing interests that might be construed to influence your judgment of the article's or peer review report's validity or importance. Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. Consider the following examples, but note that this is not an exhaustive list: Examples of 'Non-Financial Competing Interests' Within the past 4 years, you have held joint grants, published or collaborated with any of the authors of the selected paper. You have a close personal relationship (e.g. parent, spouse, sibling, or domestic partner) with any of the authors. You are a close professional associate of any of the authors (e.g. scientific mentor, recent student). You work at the same institute as any of the authors. You hope/expect to benefit (e.g. favour or employment) as a result of your submission. You are an Editor for the journal in which the article is published. Examples of 'Financial Competing Interests' You expect to receive, or in the past 4 years have received, any of the following from any commercial organisation that may gain financially from your submission: a salary, fees, funding, reimbursements. You expect to receive, or in the past 4 years have received, shared grant support or other funding with any of the authors. You hold, or are currently applying for, any patents or significant stocks/shares relating to the subject matter of the paper you are commenting on. Please state your competing interests The comment has been saved. An error has occurred. Please try again. Cancel Post var lTitle = "In vitro \"determination\" of Lactiplantibacillus...".replace("'", ''); var linkedInUrl = "http://www.linkedin.com/shareArticle?url=https://f1000research.com/articles/14-670/v1" + "&title=" + encodeURIComponent(lTitle) + "&summary=" + encodeURIComponent('Read the article by '); var deliciousUrl = "https://del.icio.us/post?url=https://f1000research.com/articles/14-670/v1&title=" + encodeURIComponent(lTitle); var redditUrl = "http://reddit.com/submit?url=https://f1000research.com/articles/14-670/v1" + "&title=" + encodeURIComponent(lTitle); linkedInUrl += encodeURIComponent('Sangsri T et al.'); var offsetTop = /chrome/i.test( navigator.userAgent ) ? 4 : -10; var addthis_config = { ui_offset_top: offsetTop, services_compact : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_expanded : "facebook,twitter,www.linkedin.com,www.mendeley.com,reddit.com", services_custom : [ { name: "LinkedIn", url: linkedInUrl, icon:"/img/icon/at_linkedin.svg" }, { name: "Mendeley", url: "http://www.mendeley.com/import/?url=https://f1000research.com/articles/14-670/v1/mendeley", icon:"/img/icon/at_mendeley.svg" }, { name: "Reddit", url: redditUrl, icon:"/img/icon/at_reddit.svg" }, ] }; var addthis_share = { url: "https://f1000research.com/articles/14-670", templates : { twitter : "In vitro \"determination\" of Lactiplantibacillus plantarum strain.... Sangsri T et al., published by " + "@F1000Research" + ", https://f1000research.com/articles/14-670/v1" } }; if (typeof(addthis) != "undefined"){ addthis.addEventListener('addthis.ready', checkCount); addthis.addEventListener('addthis.menu.share', checkCount); } $(".f1r-shares-twitter").attr("href", "https://twitter.com/intent/tweet?text=" + addthis_share.templates.twitter); $(".f1r-shares-facebook").attr("href", "https://www.facebook.com/sharer/sharer.php?u=" + addthis_share.url); $(".f1r-shares-linkedin").attr("href", addthis_config.services_custom[0].url); $(".f1r-shares-reddit").attr("href", addthis_config.services_custom[2].url); $(".f1r-shares-mendelay").attr("href", addthis_config.services_custom[1].url); function checkCount(){ setTimeout(function(){ $(".addthis_button_expanded").each(function(){ var count = $(this).text(); if (count !== "" && count != "0") $(this).removeClass("is-hidden"); else $(this).addClass("is-hidden"); }); }, 1000); } close How to cite this report {{reportCitation}} Cancel Copy Citation Details $(function(){R.ui.buttonDropdowns('.dropdown-for-downloads');}); $(function(){R.ui.toolbarDropdowns('.toolbar-dropdown-for-downloads');}); $.get("/articles/acj/166421/183407") new F1000.Clipboard(); new F1000.ThesaurusTermsDisplay("articles", "article", "183407"); $(document).ready(function() { $( "#frame1" ).on('load', function() { var mydiv = $(this).contents().find("div"); var h = mydiv.height(); console.log(h) }); var tooltipLivingFigure = jQuery(".interactive-living-figure-label .icon-more-info"), titleLivingFigure = tooltipLivingFigure.attr("title"); tooltipLivingFigure.simpletip({ fixed: true, position: ["-115", "30"], baseClass: 'small-tooltip', content:titleLivingFigure + " " }); tooltipLivingFigure.removeAttr("title"); $("body").on("click", ".cite-living-figure", function(e) { e.preventDefault(); var ref = $(this).attr("data-ref"); $(this).closest(".living-figure-list-container").find("#" + ref).fadeIn(200); }); $("body").on("click", ".close-cite-living-figure", function(e) { e.preventDefault(); $(this).closest(".popup-window-wrapper").fadeOut(200); }); $(document).on("mouseup", function(e) { var metricsContainer = $(".article-metrics-popover-wrapper"); if (!metricsContainer.is(e.target) && metricsContainer.has(e.target).length === 0) { $(".article-metrics-close-button").click(); } }); var articleId = $('#articleId').val(); if($("#main-article-count-box").attachArticleMetrics) { $("#main-article-count-box").attachArticleMetrics(articleId, { articleMetricsView: true }); } }); var figshareWidget = $(".new_figshare_widget"); if (figshareWidget.length > 0) { window.figshare.load("f1000", function(Widget) { // Select a tag/tags defined in your page. In this tag we will place the widget. _.map(figshareWidget, function(el){ var widget = new Widget({ articleId: $(el).attr("figshare_articleId") //height:300 // this is the height of the viewer part. [Default: 550] }); widget.initialize(); // initialize the widget widget.mount(el); // mount it in a tag that's on your page // this will save the widget on the global scope for later use from // your JS scripts. This line is optional. //window.widget = widget; }); }); } close Error Close Add Reset F1000.MICROSERVICES.AFFILIATION = ''; $(document).ready(function () { $('.js-affiliations-form').each((index, form) => { new AffiliationForm({ formId: form.id, institutionErrorSelector: '.comment-enter-institution', departmentErrorSelector: '.comment-enter-department', placeSelector: '.js-add-comment-place', stateSelector: '.js-add-comment-state', zipCodeSelector: '.js-add-comment-zipcode', countrySelector: '.js-add-comment-country', countryErrorSelector: '.comment-enter-country', }); }); }); $(document).ready(function () { var reportIds = { "399878": 0, "402054": 0, "405638": 0, "402055": 18, "399879": 0, "405639": 0, "397700": 0, "402052": 0, "405636": 0, "402053": 0, "405637": 0, "397698": 0, "402050": 0, "405634": 0, "397699": 0, "402051": 0, "405635": 0, "397696": 0, "402048": 0, "397697": 0, "402049": 0, "405633": 0, "399886": 0, "399887": 0, "399884": 0, "399885": 0, "399882": 0, "405642": 0, "399883": 0, "405640": 0, "399880": 17, "399881": 0, "405641": 0, "408238": 0, "413615": 0, "408239": 0, "413612": 0, "408236": 0, "408237": 0, "408234": 0, "408235": 0, "408233": 0, "413618": 0, "408242": 0, "413619": 0, "413616": 0, "408240": 0, "413617": 0, "408241": 0, "410983": 0, "410990": 0, "410988": 0, "410989": 0, "410986": 0, "410987": 0, "410984": 0, "410985": 0, "397694": 0, "402046": 0, "397695": 0, "402047": 0, "397692": 0, "397693": 0, "397691": 0, }; $(".referee-response-container,.js-referee-report").each(function(index, el) { var reportId = $(el).attr("data-reportid"), reportCount = reportIds[reportId] || 0; $(el).find(".comments-count-container,.js-referee-report-views").html(reportCount); }); var uuidInput = $("#article_uuid"), oldUUId = uuidInput.val(), newUUId = "e204b8d3-7a48-410a-8037-9980a484d2e1"; uuidInput.val(newUUId); $("a[href*='article_uuid=']").each(function(index, el) { var newHref = $(el).attr("href").replace(oldUUId, newUUId); $(el).attr("href", newHref); }); }); An innovative open access publishing platform offering rapid publication and open peer review, whilst supporting data deposition and sharing. Browse Gateways Collections How it Works Contact For Developers Cookie Notice Privacy Notice RSS Submit Your Research Follow us © 2012-2026 F1000 Research Ltd. ISSN 2046-1402 | Legal | Partner of Research4Life • CrossRef • ORCID • FAIRSharing R.templateTests.simpleTemplate = R.template(' $text $text $text $text $text '); R.templateTests.runTests(); var F1000platform = new F1000.Platform({ name: "f1000research", displayName: "F1000Research", hostName: "f1000research.com", id: "1", editorialEmail: "[email protected]", infoEmail: "[email protected]", usePmcStats: true }); $(function(){R.ui.dropdowns('.dropdown-for-authors, .dropdown-for-about, .dropdown-for-myresearch');}); // $(function(){R.ui.dropdowns('.dropdown-for-referees');}); $(document).ready(function () { if ($(".cookie-warning").is(":visible")) { $(".sticky").css("margin-bottom", "35px"); $(".devices").addClass("devices-and-cookie-warning"); } $(".cookie-warning .close-button").click(function (e) { $(".devices").removeClass("devices-and-cookie-warning"); $(".sticky").css("margin-bottom", "0"); }); $("#tweeter-feed .tweet-message").each(function (i, message) { var self = $(message); self.html(linkify(self.html())); }); $(".partner").on("mouseenter mouseleave", function() { $(this).find(".gray-scale, .colour").toggleClass("is-hidden"); }); }); Sign In Remember me Forgotten your password? Sign In Cancel Email or password not correct. Please try again Please wait... $(function(){ // Note: All the setup needs to run against a name attribute and *not* the id due the clonish // nature of facebox... $("a[id=googleSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("GOOGLE"); $("form[id=oAuthForm]").submit(); }); $("a[id=facebookSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("FACEBOOK"); $("form[id=oAuthForm]").submit(); }); $("a[id=orcidSignInButton]").click(function(event){ event.preventDefault(); $("input[id=oAuthSystem]").val("ORCID"); $("form[id=oAuthForm]").submit(); }); }); If you've forgotten your password, please enter your email address below and we'll send you instructions on how to reset your password. The email address should be the one you originally registered with F1000. Email address not valid, please try again You registered with F1000 via Google, so we cannot reset your password. To sign in, please click here . If you still need help with your Google account password, please click here . You registered with F1000 via Facebook, so we cannot reset your password. To sign in, please click here . If you still need help with your Facebook account password, please click here . Code not correct, please try again Reset password Cancel Email us for further assistance. Server error, please try again. If your email address is registered with us, we will email you instructions to reset your password. If you think you should have received this email but it has not arrived, please check your spam filters and/or contact for further assistance. Please wait... Register $(document).ready(function () { signIn.createSignInAsRow($("#sign-in-form-gfb-popup")); $(".target-field").each(function () { var uris = $(this).val().split("/"); if (uris.pop() === "login") { $(this).val(uris.toString().replace(",","/")); } }); });