1/27
2004


ISSUE:
101
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FDA to impose tighter restrictions on cattle feed
January 26, 2004

source from: http://www.meatami.com/
HHS Secretary Tommy G. Thompson today announced several new public health measures, to be implemented by the Food and Drug Administration (FDA) to "strengthen significantly" the agency's existing BSE firewalls, including banning the use of blood and blood products, so-called "plate waste" and poultry litter from the animal feed chain.
"Today's actions will make strong public health protections against BSE even stronger," Thompson told a national teleconference in Washington, D.C. "Although the current animal feed rule provides a strong barrier against the further spread of BSE, we must never be satisfied with the status quo where the health and safety of our animals and our population is at stake."FDA Commissioner Mark McClellan, PhD., added that the agency would also increase its inspections of feed mills and rendering plants.
"Today, we are bolstering our BSE firewalls to further protect the public from being exposed to any potentially risky materials from cattle," McClellan said. "FDA's vigorous inspection and enforcement program has helped us achieve a compliance rate of more than 99 percent with the feed ban rule, and we intend to increase our enforcement efforts to assure compliance with our enhanced regulations."
AMI reacted with a strong statement of support and noted the extraordinary nature of the new rules.
"It is our view that feed controls are an important line of defense in preventing BSE," said AMI President J. Patrick Boyle. ."With a greater than 99 percent level of compliance with existing feed restrictions (the highest level of compliance with any FDA rule), these new and aggressive actions will create an extraordinarily high level of assurance that U.S. cattle are protected from the risk of BSE."

Boyle added that FDAs actions, together with the additional precautions USDA announced on Dec. 30, 2003, should "reassure our international trading partners" that U.S. livestock are healthy and that U.S. beef products are safe."We therefore call upon our trading partners to act expeditiously to restore trade," Boyle said.To implement the new regulations, FDA will publish two interim final rules that will take effect immediately upon publication, although there will be an opportunity for public comment after publication.

The first interim final rule will ban the following materials from FDA-regulated human food, (including dietary supplements) and cosmetics, including:?Any material from "downer" cattle (animals that cannot walk).?Any material from "dead" cattle that die on the farm (before reaching the slaughter plant).
?Specified Risk Materials (SRMs), such as the brain, skull, eyes, and spinal cord of cattle 30 months or older, and a portion of the small intestine and tonsils from all cattle.
?The product known as "mechanically separated beef," which may contain SRMs, AMR, or meat obtained by Advanced Meat Recovery, may be used since USDA regulations do not allow the presence of SRMs in this product.
The second interim final rule will implement four specific changes to FDA's current animal feed regulations. The changes will:?Eliminate the feed rule exemption that allows mammalian blood and blood products to be fed to other ruminants, since recent scientific evidence suggests that blood can carry some BSE infectivity.?Ban the use of "poultry litter" (bedding, spilled feed, feathers and fecal matter) as a feed ingredient for ruminant animals, since poultry feed may contain ruminant-derived protein and spillage could occur, allowing collected litter with ruminant protein to be added to cattle feed.?Ban the use of plate waste (uneaten meat and other meat scraps collected from restaurants) as a feed ingredient for ruminants. Plate waste confounds FDA's ability to analyze ruminant feeds for the presence of prohibited proteins.
?Require requiring equipment, facilities or production lines to be dedicated to non-ruminant animal feeds if they use protein that is prohibited in ruminant feed.

McClellan also noted that FDA would step up its inspections of feed mills and renderers in 2004 and will conduct 2,800 inspections and work with state agencies to fund 3,100 contract inspections of feed mills and renderers that handle animal feed and feed ingredients. Through partnerships with states, FDA will also receive data on 700 additional inspections, for a total of 3,800 state contract and partnership inspections in 2004, including annual inspections of 100 percent of all known renderers and feed mills that process products containing materials prohibited in ruminant feed."With today's actions, FDA will be doing more than ever before to protect the public against BSE by eliminating additional potential sources of BSE exposure," McClellan concluded.Comprehensive information about FDA's BSE regulations and links to other related Web sites can be accessed at http://www.fda.gov

The FDA's fishy standards
January 24, 2004
The LA Times
http://www.latimes.com/news/printedition/opinion/la-ed-salmon24jan24,1,6501037.story?coll=la-news-comment
A study that found high levels of PCBs and other toxins in farm-raised salmon, according to this opinion piece, adds to the rising debate about the fish, which are artificially colored and far higher in fat than the wild variety ?with healthful Omega 3 fatty acids making up a lower proportion of those fats. Industry practices also pollute the ocean where the fish are raised in netted pens. Of even bigger concern to consumers, though, the report published in Science magazine this month demonstrated the need for the Food and Drug Administration to update its standards for such toxins in fish and all foods.
The story says that fed on pellets high in fish oils ?fat is where toxic residues tend to collect ?farmed salmon fatten up for market quickly. But if the salmon were caught by amateur anglers, they would fall under advisory guidelines set by the Environmental Protection Agency. With levels of dioxins, PCBs and pesticide residue at 10 times the level for wild salmon, the EPA would advise people not to eat that salmon more than once a month.
The FDA, however, is in charge of commercially produced food. Seeing its role as protecting the food industry as well as consumers, the FDA generally sets standards for toxins up to 40 times higher than the EPA does.
The FDA says: Eat all the farmed salmon you want. The EPA sticks to its guidelines. And the public is left clueless about whether it should still buy the fish, how much and how often.
The story goes on to say that a letter to the FDA last month urged the agency to overhaul its standards on contaminants, many of which date back a quarter of a century, when public eating patterns were different, toxins were harder to measure and less was known about their effects. Signed by dozens of scientists from such prestigious institutions as the Harvard School of Public Health, the letter's proposal merits attention.
The FDA should launch an expansive review that puts the priority on public health, not the food industry's profit margin.

USDA to update biotech regulations

Monday, January 26, 2004
IFT Daily News
http://www.ift.org/news_bin/news/news_home.shtml
1/26/2004-The USDA announced that it will update and strengthen its biotechnology regulations for the importation, interstate movement and environmental release of certain genetically engineered (GE) organisms. USDAs Animal and Plant Health Inspection Service (APHIS) will prepare an environmental impact statement evaluating its biotechnology regulations and several possible regulation changes, including the development of a multi-tiered, risk-based permitting system to replace the current permit/notification system, along with enhancements to the deregulation process to provide flexibility for long term monitoring. Any proposed changes to the regulations will be science and risk-based. APHIS welcomes comments and input from stakeholders and the public to assist in determining the scope of the EIS and any proposed regulations. This notice is scheduled for publication in the Jan. 23 Federal Register and is available for viewing at www.aphis.usda.gov/. APHIS documents published in the Federal Register and related information, including the names of organizations and individuals who have commented on APHIS dockets, are available on the Internet at http://www.aphis.usda.gov/ppd/rad/webrepor.html.

Worker says discovery of infected cow was 'a fluke'
Saturday, January 24, 2004 - Page updated at 12:00 A.M.

By Carol M. Ostrom
Seattle Times staff reporter

http://seattletimes.nwsource.com/html/localnews/2001842920_madcow24m.html

Dave Louthan says he remembers her well: an old dairy cow with specks of blood on her tail, spooky about going down the ramp into the slaughterhouse.
The government says the discovery that the Holstein had mad-cow disease proves its surveillance program, which focuses on "downer" or nonambulatory cows, works. After last month's discovery, the U.S. Department of Agriculture banned downer cows in the food supply.

But Louthan says it was "a fluke" that the Holstein, a cow he describes as "a good walker," was tested.

And even if it had been deemed a downer, under emergency rules enacted earlier this month it would have been sent to a rendering plant, where tests are not done at all.

Louthan, who killed the Holstein at Vern's Moses Lake Meat Co. on Dec. 9, says he has no doubt he remembers the right cow. "Every cow that comes in there, I kill. That kind of puts us in a relationship," said Louthan, who killed cows at Vern's for four years before he was laid off earlier this month after his bosses told him business had slowed. The plant manager, Tom Ellestad, has also confirmed that the cow was walking.

If he'd done what he should have, Louthan said, he would have taken the cow out of the truck and herded her around to a holding pen with other ambulatory animals.

But, he said, it was late in the day, the cow looked balky, and "I was cutting corners." So he shot a bolt through her head, scooped out a bit of brain, put it in a bag, labeled it with her number, and hung it on the wall with samples from others in the truckload. Later, he checked records to confirm that the "mad cow" was the cow he remembered, the balky Holstein from the Sunny Dene Ranch in Mabton, Yakima County.

If the Holstein had walked into the slaughterhouse, it probably would have been examined carefully, because it had apparent calving injuries and the veterinarian on site had tagged it for inspection.

But experts in the disease, formally known as bovine spongiform encephalopathy (BSE) say the inspector likely wouldn't have found anything, because the tiny protein particles implicated in the disease aren't visible to the naked eye.

Quite likely, the Holstein wouldn't have been tested for BSE, and no one would have known that the disease had been transported across the border in cows sent from Canada. Or that her meat, which was recalled Dec. 24, had found its way into the food chain.

The disease, which causes Swiss-cheese-like holes to form in a cow's brain, is fatal for humans who contract a related illness from eating infected meat.

Felicia Nestor, food-safety project director at the Government Accountability Project, a citizens' watchdog group, said investigation by her office confirms Louthan's account.

Even more troubling, said Nestor, is that the testing program is totally voluntary, and that the industry ?not inspectors ?chooses which animals are tested for BSE.

"Had this cow not given birth, and had a birthing injury, that cow would still be out in the field with BSE," Nestor said.

In another twist, emergency rules now in place would have let the Holstein fall through the surveillance net.

On Jan. 12, the U.S. Department of Agriculture (USDA) took emergency action banning all downer cows ?no matter what the cause of their inability to walk ?from the food supply.

Now, they're sent directly to rendering plants ?where, in this state, they're not being tested for the disease.

USDA spokesman Nolan Lemon said all resources in Washington are being used to track down the remaining cows that came with the Holstein from Canada.

"Right now, we're evaluating how our surveillance program will have to change," Lemon said. Setting up testing at rendering plants "is a possibility," he said.

If current rules had been in place when the Holstein came through, "we may not have caught that cow," said Frank Hendrix, a cattleman and Washington State University extension agent in Yakima.

But, Hendrix added, the USDA is well aware of the problem.

"I'm sure in the next couple of months, the government will get it straightened out, and start testing at rendering plants as well," he said.

Whether the infected Holstein was a downer is important, because the government's surveillance system has long concentrated on downers, deemed to be more likely to have BSE than cows that can walk.

"Downers weren't the only source of samples, but they were a high source of it," Lemon said. Out of a total of 20,277 tests last fiscal year, ending Sept. 30, 16,560 were "downers" and 3,090 were cows that had died before they arrived at the slaughterhouse.

According to USDA records, Vern's began testing in early October, gathering samples from 258 cows through December.

The veterinarian's notes from Dec. 9 at Vern's show the Holstein as "alert" but lying down on its sternum. Notes made after it was slaughtered show it had pelvic injuries, which indicate difficulties in calving, not BSE.

The department's surveillance was designed to catch the 1-in-a-million case that statisticians said would be a tip-off that there were perhaps 45 other cases in the U.S. adult cattle population of about 45 million, Lemon said.

Before the Holstein tested positive, the USDA had determined that the tests should increase to 40,000 a year to catch that 1-in-a-million case, he added.

Nestor, at the Government Accountability Project, said much of the voluntary sampling of livestock has been done at smaller facilities. "Thank goodness for us all that Vern's stepped up to the plate and was willing to take the responsibility to sample these animals," she said. "If they had not been willing to do this, then we would never have found this cow."

But, she added, such an event is financially traumatic for a plant owner, who is not protected from the economic fallout resulting from discovery of the disease. "There is no incentive for a plant to find BSE," she said.

Louthan, 44, said he's sorry to lose his job, because he enjoyed the work. "I did it because I liked to kill cows," he said. "I don't care if I'm hauling them, feeding them or killing them. As long as I'm around livestock, I'm happy. I'm a cowboy."

Current Food Safety Informaiton
01/27. Testing All US Cattle For Mad Cow Not Needed-USDA
01/27. Inquiry At Boardman Dairy Ends; Test Results On 20 Cows Awai
01/27. Ban On Meat From 'Downers' Grows Rules Affect Canned Food, Cosmetics
01/27. 'For the consumer'; Despite controversy swirling around food
01/27. USDA produce quality inspection training program
01/27. Serving tripe instead of science
01/27. Task force report recommends creation of international centr
01/27. DeHaven: BSE investigation winding down
01/27. Quarantines ended for five herds in BSE investigation
01/27. BSE: Trade Talks Continue
01/27. Case for Kosher
01/27. Are EU consumers at risk?
01/27. UK supermarkets to stay GM-free
01/27. FDA to impose tighter restrictions on cattle feed
01/27. USDA narrows focus of BSE investigation; report due 'in matt
01/27. Have an action plan to cope with allergies
01/27. U.S. Works to Ease Mad-Cow Trade Fears
01/27. South Korea to maintain ban on US beef
01/27. Newspaper Tests Show Traces of Listeria in Some Scottish Smo
01/27. Irradiated beef Congress encourages schools to use the meat,
01/27. Administration's mad cow response irks both sides
01/27. Mad cow's quiet crisis: The disease has not inspired predict
01/27. Perils of eating produce
01/27. Rules Issued on Animal Feed and Use of Disabled Cattle
01/27. World appeal to contain bird flu
01/27. U.S. Nears End of Mad Cow Investigation
01/27. Health Agencies Discuss Mercury Levels In Fish
01/27. Food Businesses Advised To Use Properly Cooked Or Pasteurise
01/27. Who is minding the USA's food store?

01/26. Legislature Might Have To Wait On Congress For Mad Cow Actio
01/26. Cow From Suspect Herd Traced To Moxee
01/26. Beef Industry Unveils Post - Mad Cow Ad
01/26. The FDA's fishy standards
01/26. The wages of fear
01/26. Illegal eggs sold across Toronto
01/26. McGuinty government unveils Canada's first Emergency Medical
01/26. Research to determine safety of farm-raised fish
01/26. City wins court challenge of DineSafe - Toronto's restaurant
01/26. USDA to update biotech regulations
01/26. U.S., Japanese officials meet but reach no new agreements on
01/26. Another view: Japanese perspectives on U.S. beef ban
01/26. EU GM food to be identified

01/25. Physicists to provide solution to BSE?
01/25. GM free in the Garden of Eden
01/25. EU GM food to be identified
01/25. OIE re-emphasizes that beef is 'safe to trade' under group's
01/25. Last Chance to Enter 2003/2004 Oxoid Food Awards
01/25. Food Safety Summit
01/25. Mom urges school to protect allergic kids
01/25. Schools, Peanuts And The Law
01/25. Worker says discovery of infected cow was 'a fluke'

01/24. Health jitters shortening China's menus
01/24. Mad Cow Probe Spreads to Seventh Facility
01/24. UI economist says BSE worries are not over
01/24. Mad Cow Creates New Animal-Disposal Woes
01/24. BSE fears spread to Canadian Forces in Kabul
01/24. Mad-Cow Worries May Force Change in Rules for Feed
01/24. Nation develops mobile food safety monitoring facilities
01/24. Mad cow crisis has been a no-win situation for Bush administ

Current Recall Information

Current USDA/FDA NEWS
U.S. Codex Office "What's New" Page: Updated January 27, 2003
USDA BSE Update
Transmissible Spongiform Encephalopathies Advisory Com
mittee; Notice of Meeting
Food Safety Mobile Game
Verification Instructions For The Interim Final Rule Regarding Specified Risk Materials In Cattle
Awareness Meeting Regarding New Regulations that Prohibit Non-Ambulatory Disabled Cattle
U.S. Codex Office "What's New" Page: Updated January 20, 2003
FSIS to Conduct Teaching Workshops on BSE

FSIS Constituent Update/Alert: Updated January 16, 2004
Substances Prohibited From Use in Animal Food or Feed, Animal Proteins Prohibited
Questions and Answers For FSIS Notice 4-04 Regarding FSIS¡¯s BSE Regulations
Substances Affirmed as Generally Recognized as Safe: Menhaden Oil
Interim Guidance For Non-Ambulatory Disabled Cattle and Age Determination

Current Outbreaks
01/27. Source of E. coli at Fort Bend fair under dispute
01/26. Kansas Cryptosporidium Worse Than Thought
01/25. Woman dies after coming down with food poisoning
01/25. MB visits food poisoning victim
01/22. Hepatitis A - Russia (Buryatiya)
01/22. Princess cruises ship reports outbreak of intestinal illness
01/22. Cholera kills 10 in Mozambique

01/21. Botulism, fish-related 2003 - Norway, Germany
01/21. Stomach bug hits Open
01/20. Trichinellosis - Turkey

Current New Methods
01/27. Making ready-to-eat meals safer
01/27. BRIGHT IDEA: Water, debugged
01/27. VeriPrime Inc. Certifies Abbott Laboratories for Rapid BSE T
01/26. Mad cow live test on way
01/26. Safety water marks
01/26. Essential oils also fight foodborne bacteria
01/26. Warnex E.coli 0157 Test Validated
01/25. Collecting environmental samples is simple with AssureSwab¢ç
01/25. New Range of Rapid Listeria Tests for Food Processing Enviro
01/24. Alicyclobacillus, the Beverage Industry and the BioSys
01/24. SEQUENOM Collaborates With HPA to Apply MassARRAY¢â Technolog
01/24. Bacteriophages may foil E.coli

Kansas Cryptosporidium Worse Than Thought
Yahoo! News Mon, Jan 26, 2004

http://news.yahoo.com/LAWRENCE, Kan. - An outbreak of infection by a diarrhea-inducing parasite might have been more than six times worse than previously reported, a new federal report shows. The report from the federal Centers for Disease Control, made public Friday by the Kansas Department of Health and Environment, shows that the cryptosporidiosis outbreak last summer in Lawrence might have infected 600 people. The report also suggests that the disease's spread could have been slowed if health authorities had alerted the community sooner. "The speed of diffusion of an outbreak like the one (in Lawrence) requires prompt and swift actions," it said. But Douglas County and state authorities said they took the correct action during the outbreak based on what they knew at the time. "Keep in mind that cryptosporidium is a fairly new disease," said KDHE spokeswoman Sharon Watson. "It's fairly easy to look back and see how things may have been done differently once you have all the information." She added: "There is no standard in place that says, 'If you have a cryptosporidiosis case in the community, you do A, B, C.'" The first cases in Lawrence were diagnosed in July. Among the initial victims was at least one member of the University of Kansas swim team. The Lawrence-Douglas County Health Department came under fierce criticism because it didn't alert the broader public about the parasite until late August, when authorities closed swimming pools after 11 cases had been confirmed. The parasite is easily spread in pools and at day-care centers. Eventually, 96 cases were confirmed in northeast Kansas ?89 in Douglas County and the vast majority of those in Lawrence. More than 600 other probable, but unconfirmed, cases were identified by community surveys. Only the confirmed cases were reported during the outbreak. The source of the outbreak was not determined, CDC officials wrote, but the genotype of the parasite found in stool samples is found only in human-to-human contact, not the animal-to-human contact that often starts a cryptosporidium outbreak. CDC's investigators suggested that in future cryptosporidium cases, the community be alerted immediately when multiple, apparently unrelated cases are reported or when diagnosis is confirmed of cryptosporidiosis in any patient who has been in a swimming pool during the infection period. Either condition would have resulted in a public announcement weeks earlier than late August had the protocols been in place. "We were working with limited data at the time, making decisions as data came in," Watson said. "Not everything pointed to a specific source at the time."Authorities said they waited to alert the public because they weren't sure the cases were related. Lawrence-Douglas County Health Department Director Kay Kent said Friday her department followed traditional methods in waiting to make the announcement.

"I think the (CDC) report basically does say that there might be some difference in the future than what might be traditionally done," Kent said. "This is the first time the CDC has put that in writing, and I think that will be extremely helpful in the future."

And she pointed out that the Health Department followed many other recommendations in the CDC report during the outbreak. Among them were "hyperchlorination" of pools linked to cryptosporidium, increased public education to discourage swimmers from entering pools until two weeks after a bout of diarrhea, and waiting three weeks after the last laboratory-confirmed case before declaring the outbreak over.

Making ready-to-eat meals safer

http://www.foodproductiondaily.com/news/news-NG.asp?id=49400

- 27/01/2004 - New aqueous ozone technology, designed to make food safer by eliminating Listeria, has been accepted by the US Department of Agriculture’s food safety inspection service (FSIS). Manufacturer BOC claims that the anti-microbial process is proven to kill Listeria.

The product is being targeted at makers of ready-to-eat (RTE) foods, since these foods eliminate the final, in-home cooking step that can kill any Listeria organisms that may remain on the food product. For these foods, Listeria organisms must be controlled in the food production environment to ensure consumer safety.
A recent risk assessment conducted by the FSIS, in conjunction with the Food and Drug Administration, ranks certain RTE meat and poultry products as having a very high potential for contamination. This is partly because the L. monocytogenes organism is capable of growing at refrigerated storage temperatures during the extended shelf life of the RTE meat and poultry products.

"Meat and poultry processors can incorporate antimicrobial ingredients such as salts of organic acids to control L.monocytogenes growth,?said James Marsden, regent's distinguished professor at Kansas State University (KSU).

“However, with RTE products, it is also necessary to incorporate a lethality step in the production process that will reduce the levels of this pathogen and leave surviving cells injured."

While surface heat can be used to achieve the lethality required for surface L. monocytogenes contamination, it can result in undesirable changes in product quality and the capital investment costs can be restrictive. BOC claims that the new aqueous ozone technology is effective in helping processors achieve the desired lethality for surface L. monocytogenes contamination, at a lower cost and without negatively impacting the food product.

"Producers of RTE products can now have confidence that there is a proven effective, accepted and economical means of killing Listeria on food products and food contact surfaces," said BOC food safety markets business manager Mark DiMaggio.

BOC submitted its proprietary aqueous ozone technology to KSU for testing and validation. BOC then submitted the KSU results to the USDA as evidence that the technology will reduce surface contamination of L. monocytogenes and reduce the risk of this pathogen in the RTE products.

BOC provides a range of products such as ozone and UV light pathogen intervention systems, chilling and freezing technologies and modified atmosphere packaging. The group employs more than 44,500 people and had annual sales of some $7 billion in 2003.

Alicyclobacillus, the Beverage Industry and the BioSys

http://www.rapidmicrobiology.com/news/1206h2.php

Positive and Negative Vials
the vial on the left contains Alicyclobacillus whilst the one on the right is negative.
In 1982 a spore-forming bacillus was isolated from spoiled apple juice that could grow at pH values as low as 2.5 (Cerny et al., 1984). A new genus, Alicyclobacillus, was created for this and similar organisms based on their fatty acid composition and 16S rDNA sequences (Wisotzkey et al., 1992). Alicyclobacillus is a non-pathogenic bacterial spore-former that causes sporadic spoilage of various shelf-stable juice products.

This spore-forming soil microorganism has been a cause of spoilage problems for the fruit juice industry. Spoilage has been observed in apple, pear, orange, peach and white grape juices. It has also caused problems with juice blends, fruit juice containing drinks, tomato juice and canned tomatoes. In apple juice, the spoilage manifests itself as an off-flavor and off-odor in shelf stable products. This spoilage problem has proved to be expensive in the United States and Europe, resulting in consumer complaints and product withdrawals.


Alicyclobacillus are sometimes called acidophilic thermophilic bacteria (ATB). As the name implies, ATB grow well in acidic environments, surviving at pH levels as low as 2.5. The bacteria also grow well at elevated temperatures. The spores can survive the pasteurization treatment given to most shelf stable, glass packaged fruit juices. The heat treatment activates the spores to begin growth. Most contamination problems show up several days after bottling. The organisms tend to be resistant to Sorbate and SO2.

Research has shown that the off-odor defect in pasteurized apple juice is due to guaiacol, produced by ATB contamination from vanillin in the juice. The odor of guaiacol is quite distinctive and a clear sign that there is a spoilage problem due to ATB. Guaiacol detection in finished juice products is accomplished by methylene chloride extraction, concentration and analysis by GC-MS.

Studies show that Alicyclobacillus grows well in apple juice, tomato juice, white grape juice and various blends of juices (Splittstoesser, et al., 1994; Splittstoesser et al., 1998). The bacteria have been isolated from citrus processing lines and are potential spoilage agents of beverages containing citrus juices (Wisse and Parish, 1998).

Some studies have used Potato Dextrose Agar (PDA), pH 3.5 as a selective medium for Alicyclobacillus. The juice or other sample is heated at 60°C for 60 minutes prior to culturing to destroy non-heat resistant organisms such as yeasts and to activate dormant spores. The plates are incubated 5 to 7 days at 43°C. Media used by other investigators include K Media (Walls and Chuyate, 1998) and ALI Agar (Wisse and Parish, 1998). Because only low populations of the spores may be present, detection often requires culturing large volumes of juice. When the juice is filterable, several hundred milliliters may be passed through 0.45 um membrane filters. ATB can be detected in spoiled products by the pour plate method or membrane filtration method, with growth on K Agar medium. Concentrates are analyzed for Alicyclobacillus spores by treating the sample to a preliminary heat shock, followed by plating on K Agar medium. Heat resistance studies have shown that Alicyclobacillus spores can survive the usual hot fill processes that are given to commercial juices. The data also shows that increasing the Brix makes the spores more resistant to heat. The spores are less resistant as the pH is decreased and the type of organic acid can be important (Pontius et al., 1998).

Using the BioSys Instrument to Detect Alicyclobacillus

The ViV (vial-in-vial) system consists of two containers, an internal vial and an external vial. Organisms grow in the liquid medium in the internal vial and generate carbon-dioxide gas from their metabolic activity. Since the internal vial is placed unsealed into the external vial containing dye indicator, the generated gas is released from the internal vial to fill the headspace of the external vial. The generated gas travels to the surface of the indicator and interacts with its chemical compounds. The generated carbon dioxide interacts with the KOH compound to lower its pH. Thymolphthalein dye changes from purple at pH=9 (or higher) to colorless at lower pH values. The BioSys instrument records the time taken for an identified change in the absorbance. This will occur when the CO2 that is released by growing organisms is absorbed by the KOH in the external vial, resulting in a pH change and a concurrent change in dye color.

Method
Several strains of Alicyclobacillus were tested in modified K Broth developed for the BioSys system. For each strain, 0.1ml of a suspension containing 100-1,000 cfu/ml was introduced into the modified K Broth in the internal vial. The internal vials were inserted into the external vials containing dye indicator. The cap was sealed tightly and the vial inserted into the BioSys instrument. The vials were monitored for 36 hours.

Algorithm Parameters
The following temperature and algorithm parameters were used in the detection of Alicyclobacillus:

Vial Threshold Skip Shuteye Duration Temperature
ViV 10 1 25 36 43°C

Curves Obtained

A typical curve and clear detection time in the BioSys instrument was observed for the presence of Alicyclobacillus in all cases. Typical curves obtained are shown below:

Key: Culture Detection Time (hrs) cfu/ml
Alicyclobacillus sp. 231-471-1 32.1 5.8 x 101
Alicyclobacillus sp. 231-476-6 27.5 2.74 x 102
Alicyclobacillus sp. K-AJ 30.3 3.4 x 101

Positive and Negative Vials
The figure at the top of the page, shows a vial with Alicyclobacillus in the left and a vial with a sample that did not contain the organisms on the right.

Conclusion:
After heat shock, product samples can be inoculated or membrane filters inserted into the BioSys ViV inner vial containing modified K Broth Medium. The inner vial is inserted into the outer vial, which is tightly capped, placed in the BioSys instrument and monitored for 36 hours in the system. If the sample contains the target organism, the computerized system will detect the presence of the organism and will alert the user in "real time" of such occurrence. Samples that do not detect within 36 hours are negative and do not contain Alicyclobacillus.

References

Cerny, G., W. Hennlich, K. Poralla. 1984. Spoilage of fruit juice by bacilli: isolation and characterization of the spoilage organism. Z. Lebensm. Unters. Forsch. 179:224-227.

Pontius, A. J., J. E. Rushing, and P. M. Foegeding. 1998. Heat resistance of Alicyclobacillus acidoterrestris spores as affected by various pH values and organic acids. J. Food Protection 61:41-46.

Splittstoesser, D. F. 1993. Recent developments in the microbiology of fruit juices, p. 59-61. In D. L. Downing (ed), Juice Technology Workshop, Report 67, NYS Agric. Expt. Station, Geneva, NY

Splittstoesser, D. F., J. J. Churey, and C. Y. Lee. 1994. Growth characteristics of aciduric sporeforming bacteria isolated from fruit juices. J. Food Protection 57:1080.

Splittstoesser, D. F., and J. J. Churey. 1996. Unique spoilage organisms of musts and wines, p 36-41. In T. Toland and K. C. Fugelsang (ed), Wine spoilage microbiology Conference. California State University, Fresno.

Splittstoesser, D. F., C. Y. Lee and J. J. Churey. 1998. Control of Alicyclobacillus in the juice industry. Dairy Food and Environ. Sanitation 18:585-587.

Walls, I. And R. Chuyate, 1998. Alicyclobacillus - Historical perspective and preliminary characterization study. Dairy Food and Environ. Sanitation 18:499-503.

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