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class="content-sidebar-wrap"><main class="content"><article class="post-5687 post type-post status-publish format-standard has-post-thumbnail category-microbiology-testing tag-chocolate tag-cocoa tag-salmonella entry" aria-label="Cocoa: ‘Choc -a-block’ with Challenges for Food Microbiologists?"><header class="entry-header"><h1 class="entry-title">Cocoa: ‘Choc -a-block’ with Challenges for Food Microbiologists?</h1> <span class="post-info-author"> By <a href="https://www.thermofisher.com/blog/author/davidcrabtree/" title="Posts by David Crabtree" rel="author">David Crabtree</a> </span> <span class="post-info-date"> 09.10.2015 </span> </header><div class="entry-content"><p><img decoding="async" style="float: right;margin: 5px" alt="chocolate testing" src="https://www.thermofisher.com/blog/food/wp-content/uploads/sites/5/2015/09/chocolate.jpg" width="330" height="220" />If the average member of the public was asked the question, “how is chocolate made?”, <span style="color: #000000">I wouldn’t be surprised if the response was less than confident or even close to the right answer at all. </span>A mystery to some, the lengthy process of chocolate manufacturing is a highly refined procedure that ferments, dries, cleans, roasts, de-shells, winnows, grinds, cooks, presses and tempers the raw ingredient of cacao beans into a tasty treat that’s good to eat. While most could appreciate this extensive course of processing, few would anticipate the challenges presented to a microbiologist by cocoa and cocoa-containing products when testing for food borne pathogens.</p> <p>Cacao beans are subject to contamination with <a href="https://www.thermofisher.com/blog/food/controlling-salmonella-contamination/"><em>Salmonella</em></a> at the early stages of their processing where beans are removed from ripe pods by hand and left to ferment in the sun on large trays. The beans ferment for approximately 1 week, giving plenty of opportunity for contaminants to find their way into the crop during this time.</p> <p>The further processing of the beans includes drying and cooking steps that can kill bacteria which may be present post-fermentation. Studies have shown that, despite the efforts to kill any harmful bacteria present, drying and heating of cocoa beans may not be 100% efficient in killing <em>Salmonella</em> and very low numbers of injured cells can survive and persist in cocoa-containing products<sup>1</sup>.</p> <p>When screening for the presence of <em>Salmonella </em>in cocoa-containing products, microbiologists are faced with challenges that can make detection more difficult than many other foods. Using culture media to recover low numbers of highly injured cells can be difficult as the balance of nutrients has to be just right to encourage growth from such conditions. Traditional culture methods can be lengthy and involve more than just a single step of preparation for microbiologists to go through before conducting any tests so optimising these tests is essential to obtain a reliable result.</p> <p>Thermo Scientific<sup>TM</sup> <span></span><a href="http://www.thermoscientific.com/en/product/cocoa-sample-recovery-csr-broth.html" target="_blank"><span class="thread"><span>Cocoa Sample Recovery (</span>CSR) Broth</span></a> is a culture medium designed for the recovery of low numbers of injured cells of <em>Salmonella</em> from cocoa-containing products, in a one-step enrichment that is faster and simpler than traditional culture methods.</p> <p>Cocoa often contains polyphenols which are known to be toxic to bacteria, making recovery of injured cells difficult. The formulation of CSR Broth includes acid-free casein to reduce this bactericidal effect, providing a better environment from which to grow low numbers of stressed <em>Salmonella</em>. This is crucial in encouraging the target pathogen to grow to a level that can <span>be detected </span>(Limit of Detection, LOD).</p> <p>Not only can CSR Broth be used as a single-step enrichment medium for the detection of <em>Salmonella</em><span style="color: #000000"> in culture-based methods</span>, but it can also be used for more rapid PCR methods. Under traditional circumstances, PCR methods may struggle to perform well when used to test cocoa-containing products, due to the presence of polyphenols. Polyphenols are able to inhibit PCR reactions due to their ability to irreversibly bind covalently to free nucleic acids. This binding makes extracted DNA unavailable for amplification and halts the continuation of a PCR reaction, leading to potential false negative results or internal control failures.</p> <p>To overcome this issue, the <a href="http://www.thermoscientific.com/en/about-us/general-landing-page/suretect-real-time-pcr-system.html" target="_blank">Thermo Scientific™ SureTect™</a> Salmonella protocol for cocoa and cocoa-containing products uses an additional 1:5 dilution step post enrichment to remove these compounds without diluting the recovered Salmonella below the LOD, and without the use of lengthy centrifugation steps that may be used to remove inhibitors.</p> <p><strong>To read more, follow these links to technical posters evaluating the performance of CSR Broth as an enrichment medium for rapid culture and rapid PCR methods:</strong></p> <ul> <li><a href="http://www.thermoscientific.com/content/dam/tfs/SDG/MBD/MBD%20Documents/Application%20&%20Technical%20Notes/Microbiology/Evaluation-Rapid-Culture-Method-Detection-Salmonella-Cocoa-Samples-LT2179-EN.pdf" target="_blank">Evaluation of a New Rapid Culture Method for Detection of <em>Salmonella</em> spp. in Cocoa-Containing Samples</a></li> <li><a href="http://www.thermoscientific.com/content/dam/tfs/SDG/MBD/MBD%20Documents/Application%20&%20Technical%20Notes/Microbiology/Evaluation-Enrichment-Broth-Cocoa-Samples-SureTect-Salmonella-PCR-Assay-LT2180-EN.pdf" target="_blank">Evaluation of a New Enrichment Broth for the Preparation of Cocoa-Containing Samples Preceding the Thermo Scientific SureTect Salmonella species PCR Assay</a></li> </ul> <p>Learn more about <a href="http://www.thermoscientific.com/en/product/cocoa-sample-recovery-csr-broth.html" target="_blank"><span>Cocoa Sample Recovery Broth</span></a> or <a href="http://www.thermoscientific.com/en/community/microbiology-testing.html" target="_blank">visit our food and beverage community</a> for further discussion on food safety testing</p> <p><strong>References</strong></p> <p><span>da Silva do Nascimento,  <em>et al.</em>, 2012. <a href="http://www.ncbi.nlm.nih.gov/pubmed/23107501" target="_blank">Inactiviation of <em>Salmonella</em> during cocoa roasting and chocolate conching</a>. International Journal of Food Microbiology. 159 (2012), 225-229</span></p> <aside class="mashsb-container mashsb-main "><div class="mashsb_above_buttons">Share this article </div><div class="mashsb-box"><div class="mashsb-buttons"><a class="mashicon-facebook mash-large mashsb-noshadow" href="https://www.facebook.com/sharer.php?u=https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Ffood%2Fcocoa-choc-a-block-with-challenges-for-food-microbiologists%2F" target="_top" rel="nofollow"><span class="icon"></span><span class="text">Facebook</span></a><a class="mashicon-linkedin mash-large mashsb-noshadow" href="https://www.linkedin.com/shareArticle?trk=Cocoa%3A+%E2%80%98Choc+-a-block%E2%80%99+with+Challenges+for+Food+Microbiologists%3F&url=https%3A%2F%2Fwww.thermofisher.com%2Fblog%2Ffood%2Fcocoa-choc-a-block-with-challenges-for-food-microbiologists%2F" target="_top" rel="nofollow"><span class="icon"></span><span class="text">Linkedin</span></a><div class="onoffswitch2 mash-large mashsb-noshadow" style="display:none"></div></div> </div> <div style="clear:both"></div><div class="mashsb-toggle-container"></div></aside> <div class="jp-relatedposts"> <div class="related-post related-1"> <a href="https://www.thermofisher.com/blog/food/the-important-of-portion-control-on-thanksgiving-for-food-manufacturers/"><img width="310" height="274" src="https://www.thermofisher.com/blog/food/wp-content/uploads/sites/5/2024/11/iStock-471063125-scaled-e1732575698271-310x274.jpg" class="attachment-grid size-grid wp-post-image" alt="Cranberry Sauce / Jellied Cranberries" decoding="async" /></a> <span class="grid-content"> <h4 class="entry-title"><a href="https://www.thermofisher.com/blog/food/the-important-of-portion-control-on-thanksgiving-for-food-manufacturers/">The Importance of Portion Control on Thanksgiving — For Food Manufacturers</a></h4> <p>It’s that time of year when holidays are approaching a... </p> <a href="https://www.thermofisher.com/blog/food/the-important-of-portion-control-on-thanksgiving-for-food-manufacturers/" class="read-more">Read More <i class="fa fa-icon fa-chevron-right"></i></a> </span> </div> <div class="related-post related-2"> <a href="https://www.thermofisher.com/blog/food/quantifying-caffeine-in-steeped-tea-using-uv-vis/"><img width="310" height="274" src="https://www.thermofisher.com/blog/food/wp-content/uploads/sites/5/2024/10/iStock-1336864023_steepingtea-310x274.jpg" class="attachment-grid size-grid wp-post-image" alt="Steeping herbal pyramid tea bag in a glass cup" decoding="async" loading="lazy" /></a> <span class="grid-content"> <h4 class="entry-title"><a href="https://www.thermofisher.com/blog/food/quantifying-caffeine-in-steeped-tea-using-uv-vis/">Quantifying Caffeine in Steeped Tea Using UV-Vis</a></h4> <p>In the realm of food science, precise quantification of key ... </p> <a href="https://www.thermofisher.com/blog/food/quantifying-caffeine-in-steeped-tea-using-uv-vis/" class="read-more">Read More <i class="fa fa-icon fa-chevron-right"></i></a> </span> </div> <div class="related-post related-3"> <a href="https://www.thermofisher.com/blog/food/dairy-production-in-india/"><img width="310" height="274" src="https://www.thermofisher.com/blog/food/wp-content/uploads/sites/5/2023/10/iStock-1297005108_milkpour-310x274.jpg" class="attachment-grid size-grid wp-post-image" alt="Close-up of raw milk being poured into container with dairy cows in background" decoding="async" loading="lazy" /></a> <span class="grid-content"> <h4 class="entry-title"><a href="https://www.thermofisher.com/blog/food/dairy-production-in-india/">Dairy Production in India</a></h4> <p>The cow is a revered animal in throughout much of India. 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} $.each(elems, function(index, elem) { var inElementsArray = $.inArray(elem, options.elements); var inCacheArray = $.inArray(elem, cache); if (inElementsArray != -1) { options.elements.splice(inElementsArray, 1); } if (inCacheArray != -1) { cache.splice(inCacheArray, 1); } }); }; /* * Throttle function borrowed from: * Underscore.js 1.5.2 * http://underscorejs.org * (c) 2009-2013 Jeremy Ashkenas, DocumentCloud and Investigative Reporters & Editors * Underscore may be freely distributed under the MIT license. */ function throttle(func, wait) { var context, args, result; var timeout = null; var previous = 0; var later = function() { previous = new Date; timeout = null; result = func.apply(context, args); }; return function() { var now = new Date; if (!previous) previous = now; var remaining = wait - (now - previous); context = this; args = arguments; if (remaining <= 0) { clearTimeout(timeout); timeout = null; previous = now; result = func.apply(context, args); } else if (!timeout) { timeout = setTimeout(later, remaining); 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