Archive for category: Technical Reports

Isolation, screening and identification of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

Isolation, screening and identification of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

Original article doi: ©2015 by the Asian Pacific Journal of Tropical Biomedicine. All rights reserved.
Rosario Galarza-Seeber1, Juan David Latorre1, Xochitl Hernandez-Velasco2, Amanda Drake Wolfenden1, Lisa Renee Bielke1, Anita
Menconi1, Billy Marshall Hargis1, Guillermo Tellez1*

Isolation, screening and identification of Bacillus spp. As direct-fed microbial candidates for aflatoxin B1 biodegradation (PDF Download Available). Available from: https://www.researchgate.net/publication/282500849_Isolation_screening_and_identification_of_Bacillus_spp_As_direct-fed_microbial_candidates_for_aflatoxin_B1_biodegradation [accessed Aug 9, 2017].

Abstract
Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials (DFM) to biodegrade aflatoxin B1 (AFB1) by using an in vitro digestive model simulating in vivo conditions. Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 μg/mL of AFB1 in modified Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16S rRNA sequence analyzes for identification. Tolerance to acidic pH, osmotic concentrations of NaCl, bile salts were tested, and antimicrobial sensitivity profiles were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time (3:15 h), supernatants and digesta were collected for high-performance liquid chromatography fluorescence detection analysis by triplicate. Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of fluorescence and area of clearance around each colony in modified Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identified the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions (pH 2.0), tolerant to a high osmotic pressure (NaCl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No significant differences (P > 0.05) were observed in the concentrations of AFB1 in neither the supernatants nor digesta samples evaluated by high-performance liquid chromatography with fluorescence detection between positive control or DFM treated groups. Conclusions: In vitro digestion time was not enough to confirm biodegradation of AFB1. Further studies to evaluate the possible biodegradation effects of the Bacillus-DFM when continuously administered in experimentally contaminated feed with AFB1, are in progress.
For more please visit
https://www.researchgate.net/publication/282500849_Isolation_screening_and_identification_of_Bacillus_spp_As_direct-fed_microbial_candidates_for_aflatoxin_B1_biodegradation
SFFF Comment=The relevance of biological control in the search for the reduction of mycotoxins informed the republication of this excellent article
Aug 09 2017

Is this TRUE …..GLYPHOSATE has effect on AUTISM ???

MIT Researcher: Glyphosate Herbicide will Cause Half of All Children to Have Autism by 2025

 Half of All Children Will Be Autistic by 2025, Warns Senior Research Scientist at MIT

By Alliance For Natural Health
anh-usa.org

Why? Evidence points to glyphosate toxicity from the overuse of Monsanto’s Roundup herbicide on our food.

For over three decades, Stephanie Seneff, PhD, has researched biology and technology, over the years publishing over 170 scholarly peer-reviewed articles. In recent years she has concentrated on the relationship between nutrition and health, tackling such topics as Alzheimer’s, autism, and cardiovascular diseases, as well as the impact of nutritional deficiencies and environmental toxins on human health.

At a [recent] conference, in a special panel discussion about GMOs, she took the audience by surprise when she declared, “At today’s rate, by 2025, one in two children will be autistic.” She noted that the side effects of autism closely mimic those of glyphosate toxicity, and presented data showing a remarkably consistent correlation between the use of Roundup on crops (and the creation of Roundup-ready GMO crop seeds) with rising rates of autism. Children with autism have biomarkers indicative of excessive glyphosate, including zinc and iron deficiency, low serum sulfate, seizures, and mitochondrial disorder.

A fellow panelist reported that after Dr. Seneff’s presentation, “All of the 70 or so people in attendance were squirming, likely because they now had serious misgivings about serving their kids, or themselves, anything with corn or soy, which are nearly all genetically modified and thus tainted with Roundup and its glyphosate.”

Dr. Seneff noted the ubiquity of glyphosate’s use. Because it is used on corn and soy, all soft drinks and candies sweetened with corn syrup and all chips and cereals that contain soy fillers have small amounts of glyphosate in them, as do our beef and poultry since cattle and chicken are fed GMO corn or soy. Wheat is often sprayed with Roundup just prior to being harvested, which means that all non-organic bread and wheat products would also be sources of glyphosate toxicity. The amount of glyphosate in each product may not be large, but the cumulative effect (especially with as much processed food as Americans eat) could be devastating. A recent study shows that pregnant women living near farms where pesticides are applied have a 60% increased risk of children having an autism spectrum disorder.

Other toxic substances may also be autism-inducing. You may recall our story on the CDC whistleblower who revealed the government’s deliberate concealment of the link between the MMR vaccine (for measles, mumps, and rubella) and a sharply increased risk of autism, particularly in African American boys. Other studies now show a link between children’s exposure to pesticides and autism. Children who live in homes with vinyl floors, which can emit phthalate chemicals, are more likely to have autism. Children whose mothers smoked were also twice as likely to have autism. Research now acknowledges that environmental contaminants such as PCBs, PBDEs, and mercury can alter brain neuron functioning even before a child is born.

This month, the USDA released a study finding that although there were detectable levels of pesticide residue in more than half of food tested by the agency, 99% of samples taken were found to be within levels the government deems safe, and 40% were found to have no detectable trace of pesticides at all. The USDA added, however, that due to “cost concerns,” it did not test for residues of glyphosate.Let’s repeat that:they never tested for the active ingredient in the most widely used herbicide in the world. “Cost concerns”? How absurd—unless they mean it will cost them too much in terms of the special relationship between the USDA and Monsanto. You may recall the revolving door between Monsanto and the federal government, with agency officials becoming high-paying executives—and vice versa! Money, power, prestige: it’s all there. Monsanto and the USDA love to scratch each others’ backs. Clearly this omission was purposeful.

In addition, as we have previously reported, the number of adverse reactions from vaccines can be correlated as well with autism, though Seneff says it doesn’t correlate quite as closely as with Roundup. The same correlations between applications of glyphosate and autism show up in deaths from senility.

Of course, autism is a complex problem with many potential causes. Dr. Seneff’s data, however, is particularly important considering how close the correlation is—and because it is coming from a scientist with impeccable credentials. Earlier this year, she spoke at the Autism One conference and presented many of the same facts; that presentation is available on YouTube.

Monsanto claims that Roundup is harmless to humans. Bacteria, fungi, algae, parasites, and plants use a seven-step metabolic route known as the shikimate pathway for the biosynthesis of aromatic amino acids; glyphosate inhibits this pathway, causing the plant to die, which is why it’s so effective as an herbicide. Monsanto says humans don’t have this shikimate pathway, so it’s perfectly safe.

Dr. Seneff points out, however, that our gut bacteria do have this pathway, and that’s crucial because these bacteria supply our body with crucial amino acids. Roundup thus kills beneficial gut bacteria, allowing pathogens to grow; interferes with the synthesis of amino acids including methionine, which leads to shortages in critical neurotransmitters and folate; chelates (removes) important minerals like iron, cobalt and manganese; and much more.

Even worse, she notes, additional chemicals in Roundup are untested because they’re classified as“inert,” yet according to a 2014 study in BioMed Research International, these chemicals are capable of amplifying the toxic effects of Roundup hundreds of times over.

Glyphosate is present in unusually high quantities in the breast milk of American mothers, at anywhere from 760 to 1,600 times the allowable limits in European drinking water. Urine testing shows Americans have ten times the glyphosate accumulation as Europeans.

“In my view, the situation is almost beyond repair,” Dr. Seneff said after her presentation. “We need to do something drastic.”

 

Source=http://healthimpactnews.com/2014/mit-researcher-glyphosate-herbicide-will-cause-half-of-all-children-to-have-autism-by-2025/Source

 

Aug 6, 2017

 

SFFF Comments=We welcome your comments on this report  urgently

Some answers to food production problems

Scientists seek answers to food production problems

 Some answers have been provided as interventions to some food production issues. Wish you happy reading

Introduction

Many factors can compromise the quality and/or safety of a food product. But one of the most common scenarios that faces a manufacturer is an unexpected problem that comes with a change of supplier.

The irony is that everything seems fine when the ingredient is received. It meets its specification (and this is similar to the previous supplier’s), yet nonetheless, the finished product ends up being far from satisfactory. The problem might be one of flavour, appearance, performance or smell, but whatever it is, the challenge is to find out what has changed as a consequence of changing the supplier, and to take steps to put things right.

 

Analytical approaches to problem solving

Some problems stand out such as a bad smell, the wrong colour, failure of performance, or presence of foreign bodies. Other problems may not be so obvious e.g. microbial spoilage, adulteration, inauthentic ingredients.

However, whether obvious or not, every problem must be investigated. There are a range of analytical techniques to help find out what has gone wrong and to inform what needs to be done to correct it.

Few manufacturers will have all of these techniques available on-site, so are likely to need to outsource some or all of the investigative work to a specialist laboratory. It makes a lot of sense to find a laboratory to partner with well in advance of a problem occurring, so that they are better able to assist when a problem arises!

 

Potential problems:

Appearance/foreign body

Blemishes and discolouration may be the result of chemical contamination or perhaps microbial spoilage; their investigation will likely involve the chemical techniques referred to in the discussion of taints (below). Other cosmetic/aesthetic problems such as sediments or a visible separation of emulsions, will be investigated by the physical techniques applicable to performance problems (also below).

Foreign bodies are a separate case. They can be broadly classified as extrinsic or intrinsic: The former come from external sources either by deliberate or accidental means. Intrinsic foreign bodies include ingredients such as a leaf or stalk, or an ingredient in an unusual/unexpected state.

A broad spectrum of technologies are routinely used in foreign body investigations, but simple light microscopy is often the starting point. It can be used to determine features that are typical or characteristic of the likely source, thereby directing scientists to the sophisticated methods that will provide confirmation.

Different techniques are required for different types of contaminant. For example, a scanning electron microscope fitted with an energy dispersive X-ray (EDX) detector is useful for identifying the elemental composition of items such as metal fragments. Plastic fragments, which may look like glass, require confirmation using a technique known as Fourier transform – infra red spectroscopy (FT-IR).

 

Taste/smell

An inappropriate taste or smell (or discoloration) in a product is usually due to the unexpected/unwanted presence of any number of chemicals. Or there may be an absence of key ingredients (e.g. bitterness blockers), or failure of a taste-masking system (e.g. encapsulation of oils), that might otherwise be expected to keep the unwanted flavours in check.

Taints can come from direct contamination, packaging or the airspace in a bottle/jar. The unwanted chemical may come from microbial spoilage, or perhaps from reaction of ingredients within the product.

Isolating and identifying a tainting chemical is often a challenge because certain taints can be very potent at very low concentrations (e.g. 20 parts per trillion). It depends on the product they are found in but chemicals like halo-anisoles and benzene are especially potent. Whilst certain groups or families of chemicals give rise to specific flavours/odours that an experienced chemist will recognise, there are thousands of potential tainting chemicals in widespread use, so there is never one obvious candidate.

A taint or off-flavour can originate at any point in the lifetime of a product, from production of the raw materials through to eventual consumption. Hence it may be necessary to trace the source of a taint back to any point in the supply chain.

Investigations will usually involve an initial assessment by human senses. If the offending chemical is believed to be organic (i.e. carbon-based) there will follow a chemical extraction procedure, separation via chromatography (either liquid or gas) and identification of the chemical using mass spectrometry or possibly nuclear magnetic resonance (NMR) spectrometry. A metallic contaminant will be detected using inductively coupled plasma (ICP) mass spectrometry or ICP-optimal emission spectroscopy.

This simple description belies the complexity that might be involved. As noted above, taints can be present in ppt concentrations. There is huge skill involved in extracting the taint, at detectable levels, free of other chemicals that might interfere with the analysis, and (in the case of volatile odours) without simply losing the chemical to the atmosphere before it can be identified.

 

Performance

Problems with the flow of liquids and semi-solids can be investigated using rheological instruments. These instruments apply different shearing forces which can mimic how materials might flow against different surfaces and internally. Such measurements will inform manufacturers of potential processing problems (e.g. when pumping) as well as problems with the behaviour of finished products. The resistance to flow by powders within hoppers, for example, can lead to clogging, or an accumulation of out-of-date material. The propensity for powders to behave in a disadvantageous manner can be predicted via a combination of powder rheology (Shear cell), particle sizing and microscopy.

Performance issues often come down to a problem with particle sizes or shape, as with the instability of an emulsion, or the gritty mouth feel of a product. This can be investigated using laser diffraction instruments, often in combination with microscopy techniques. The size and shape of particles can also be measured using static image analysis, which is a useful alternative to manual microscopy.

It is sometimes possible to observe how ingredients are distributed within a product by using X-ray tomography techniques along with the scanning electron microscope. This can give useful information about why a product performs badly. Even simple light microscopy, used with staining procedures can identify some of the microstructural features that are giving rise to performance problems.

The modern laboratory also has access to a range of instruments that can load products with crushing or stretching forces to investigate structural weaknesses (or strength).

 

Customer complaint/illness

Customers may attribute illness to a particular food item that shows no other signs of being at fault. Clearly, a manufacturer will want to investigate a complaint of this kind, with the possibility of allergen or pathogen contamination being a particular cause for concern.

Microbiological testing can address the latter, however allergen testing can be more complicated. As such, much depends on the nature of the complaint and the likelihood, or otherwise, that specific allergens might have come into contact with the product or its ingredients/packaging.

Tests for allergens usually rely on enzyme-linked immunosorbent assay (ELISA) techniques directed at specific proteins, or DNA techniques that can detect trace amounts of DNA associated with allergenic ingredients. In both cases, but especially with ELISA, it is important to be aware of the potential for interferences inherent to biological assays that can lead to false positive or false negative results. A laboratory must have robust protocols in place to reduce the risk of any false results.

Authenticity/purity

High profile scandals (horsemeat in beef, melamine in milk) remind us that the food supply chain is extremely complex and global, and tracing the origin of ingredients is near impossible. Moreover, not every player in the supply chain is necessarily honest or legitimate.

Whether there has been a specific incident, notification of a wider industry-concern or merely a desire on behalf of the manufacturer to protect their interests (perhaps with a new supplier), there are analytical approaches that will assist in determining the authenticity of a particular supply. However, this is very much dependent on the ingredient in question and authenticity is not always easy to prove.

Testing for authenticity in the case of meat and fish is relatively routine, using DNA methods that can target gene sequences found in one species but not in another. However the quantification of cross contamination in meat against the level set by the FSA still requires specialist knowledge and testing.

Other authenticity issues are more complex. For example, olive oil has well defined acceptable ranges for a variety of naturally occurring compounds giving the analyst a set of parameters that can be measured to assess authenticity.

Other techniques such as isotope ratio analysis have become established but these depend on building a large database of samples, at considerable cost. So new approaches such as non-targeted screening (NTS) are now being applied. Rather than looking for specific chemical compounds, analysts are applying chemo-metric or food-omic approaches to identify differences between samples and a dataset of known authentic ingredients. This ‘probabilistic’ approach to testing is useful for identifying ‘outliers’, allowing analysts to focus on the suspect samples more easily, and to devote time to more careful scrutiny of these.

Where adulteration is suspected, new methods may need to be developed as a matter of urgency to address the specific problem. In the melamine in milk example, a nitrogen-rich chemical had been added to milk to fool a test that judged quality by nitrogen content. That particular test had no means of detecting the source of nitrogen, and an entirely different way of testing milk was needed for this fraud to come to light.

 

Conclusion

It is impossible to give a definitive list of problems and solutions, even in a very broad sense. Each problem needs to be assessed and investigated on its own merits.

It is also impossible for every manufacturer to equip itself with the resources and expertise that would be needed to investigate every potential problem. Rather every manufacturer should seek to partner with an expert laboratory that understands its processes, and can work with them to highlight potential weaknesses, and understand vulnerabilities.

At least this approach will ensure that when a problem does occur, whatever it happens to be, there is help at hand to identify what has gone wrong, and recommendations on what needs to be done to put it right.

First published in 2016  by

Reading Scientific Services Limited

 

Source=http://www.foodqualitynews.com/Product-Categories/Audit-and-Certification/Scientists-seek-answers-to-food-production-problems

Aug 1 2017

MOBILE APP TO TEST FOR AFLATOXIN

The app was developed by Mobile Assay, which specializes in mobile lab innovations. The test, more formally known as an immunoassay test, involves analysing a colour-changing test strip for the presence of a particular substance, in this case Aflatoxin. The test strip is photographed using the smartphone’s camera and the app calculates the pixel density of the coloured strip to determine how much Aflatoxin is present within a certain threshold. Current available lab tests are priced at $15 each plus the cost of transporting samples from rural areas to those equipped with testing facilities. This is out of reach for many in the Global South. Some turn to cheaper tests, which return only a positive or negative result and don’t provide any information about whether or not the level of contamination poses a health threat or is fit for consumption. This test is set to cost about $2-3 and provides such detailed information. What’s more, it is immediately uploaded to an online database tracking Aflatoxin outbreaks.

feb, 2017

source=http://innovatedevelopment.org/2014/03/12/mobile-rapid-diagnostic-testing-helping-farmers-protect-crops-people-in-east-africa

 

Affordable aflatoxin test kit for farmers launched in Malawi

A new technology that can detect aflatoxin on location has been launched by an international agricultural research organisation. The test kit, which was launched by Dr Wilkson Makumba, director, department of Agricultural Research Services (DARS) in Lilongwe, Malawi requires limited technical knowledge or training and can be used on location.

“The new test is simple to perform and can detect contamination at levels of 10 parts per billion in less than 15 minutes .

This exciting advancement combined with a mobile extraction kit is a simple non-laboratory based kit that can be used directly by non-technical people such as farmers, agro-dealers and food processors and will be ready in two months at a cost of Sh200.

“This kit can be used by traders to check for contamination before concluding a sale,” he said. The rapid detection is useful for public health authorities to help identify suspected samples in cases of an outbreak of aflatoxin poisoning.

The compact, portable device is based on the lateral flow immunoassay test (popularly known as the strip test like that used to detect glucose in human blood). If aflatoxin is present in the sample, then one pink line appears on the strip, whereas if the sample doesn’t have any aflatoxin, two pink lines will appear.

“The device will contribute to manage and reduce the entry of aflatoxins in the food value chains, improve diagnosis for local and export trade and support the food processing industry to maintain low exposure levels in food products in our local markets as well as for export markets,” said Dr Anitha Seetha, a scientist in Malawi.

According to Food and Agriculture Organisation, 25 per cent of all crops—including groundnut, maize, sorghum, pearl millet—in the world are affected by aflatoxins. In 2010, about 20,000 people died globally from aflatoxin poisoning and an equal number fell ill, says WHO.

By Milliam Murigi

source=http://www.mediamaxnetwork.co.ke/people-daily/243884/243884/

EU PROLONGS GLYPHOSATE LICENCE BY 18 MONTHS

A new study has found that low-dose and long-term exposure to the  weed killer Roundup causes liver disease in female rats.

However, the company,Monsanto has reacted, saying that the study used “flawed data” from 2012.

The Roundup herbicide contains controversial ingredient glyphosate, which is now one of the world’s most popular weed killers. The use of glyphosate has triggered strong reactions in the EU amid fears the product is carcinogenic.

In late June, the European Commission decided to extend the authorisation licence for glyphosate by 18 months, until the European Chemicals Agency (ECHA) gives an opinion on the substance. The ECHA’s opinion is expected in June or September this year.

Commission prolongs glyphosate licence by 18 months

The European Commission has decided to extend the licence for glyphosate by 18 months, after member states failed to achieve a qualified majority in favour or against the executive’s proposal.

EurActiv.com

 

Liver disease

A new study published this week (9 January) in Scientific Reports found that low levels of exposure to the weed killer Roundup over an extended period causes liver disease in rats.

The study focused on Roundup and not glyphosate alone and stated that it was not possible “to attribute the toxicity of the whole agricultural herbicide formulation to a given component”.

“Future studies involving the administration of glyphosate alone would shed light on this issue,” the study concluded.

Female rats were administered with an extremely low dose of Roundup weed killer over a two-year period and found to suffer from non-alcoholic fatty liver disease (NAFLD). The dose selected was below what people are commonly exposed to in the everyday environment and 75,000 times below what is permitted by EU regulators.

For a two-year period, female rats were administered a dose of Roundup, approximately 75,000 times below what is permitted by EU regulators for human exposure, and were found to suffer from non-alcoholic fatty liver disease.

Researchers pointed out that it was the first time that a causative link between consumption of Roundup at a “real world” environmental dose and a serious disease condition was identified.

Dr. Michael Antoniou, an author of the study, stressed that the findings were “worrying” and called EU policy-makers to reconsider the safety evaluation of glyphosate-based herbicides.

According to Antoniou, fatty liver disease occurs in at least one in five of the general population, and in the vast majority of people with type 2 diabetes.

source=euractiv.com

Mycotoxins and urine testing

 

Of recent, a lot of research activities have been on regarding diagnosis of mycotoxicosis in man or livestock. Detecting the particular mycotoxin eg aflatoxin, fumonisin  may be  the expected alarm that could send victims on an endless spending errands focussing on chemotherapy . Blood and urine are regular candidates of such test

However, the  National Institute for Occupational Safety and Health (NIOSH), a part of the Centers for Disease Control and Prevention (CDC),noted that these test results can lead to “misinformation and fear in the workplace; incorrect diagnoses; unnecessary, inappropriate and potentially harmful medical interventions; and unnecessary or inappropriate environmental and occupational evaluations.”

Also the US  FDA has not approved any urine test for mycotoxins. The CDC warns that such tests have not been approved for diagnostic purposes, meaning that you can’t trust them to tell you if anything is wrong with you or what to do about it.

Be guided, seek advice from professionals

Source:
“Use of Unvalidated Urine Mycotoxin Tests for the Clinical Diagnosis of Illness — United States, 2014.” Centers for Disease Control and Prevention, Morbidity and Mortality Weekly Report, February 20, 2015, http://www.cdc.gov/mmwr/preview/mmwrhtml/mm6406a7.htm

 

Mycotoxins, livestock and death

Mycotoxins in animal fodder, particularly silage, can decrease the performance of farm animals and potentially kill them

So beware of what you feed your animals. Seek appropriate guide from professionals

SPECIAL ANNOUNCEMENT

 

Do you have news on production, processing and packaging of food and feed, devoid of contaminants? Do you have information on farmer –friendly interventions that will guarantee healthy consumption? .Are you current on information regarding effects of chemicals/ contaminants, organic food  and Genetic modification in  agriculture and nutrition? Can you promote entrepreneurship through the supply of low-income equipment in Agriculture and drive women and the youth along ?

 

Please send your information to sfoodfeedf@yahoo.com

AFLATOXIN and CANCER

About 40% of all cases of liver cancer in Africa are traceable to AFLATOXIN  !!!!!…..  (EC)

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