Biomedical subjects

D L Park

Publications and source records attributed to D L Park.

At least 19 recordsLinked to original sources

PubMed2003

Response of Vibrio parahaemolyticus 03:K6 to a hot water/cold shock pasteurization process.

Vibrio vulnificus and V. parahaemolyticus are natural inhabitants of estuarine environments world wide. Pathogenic strains of these bacteria are often transmitted to humans through consumption of raw oysters, which flourish in the same estuaries. Previous studies reported the effective use of hot water pasteurization followed by cold shock to eliminate from raw oysters naturally and artificially incurred environmental strains of V. vulnificus and V. parahaemolyticus common to the Gulf of Mexico. The present study focused on the use of the same pasteurization method to reduce a highly process resistant Vibrio strain, V. parahaemolyticus O3:K6 to non-detectable levels. Oysters were artificially contaminated with 10(4) and 10(6) V. parahaemolyticus 03:K6 cfu g(-1) oyster meat. Contaminated oysters were pasteurized between 50 and 52 degrees C for up to 22 min. Samples of processed oysters were enumerated for V. parahaemolyticus O3:K6 at 2-min intervals beginning after the 'come-up time' to achieve an oyster internal temperature of at least 50 degrees C. The D value (D(52)deg C) was 1.3-1.6 min. V. parahaemolyticus O3:K6 proved more process resistant than non-pathogenic environmental strains found in Gulf of Mexico waters. A total processing time of at least 22 min at 52 degrees C was recommended to reduce this bacterium to non-detectable levels (< 3 g(-1) oyster meat).

Animals↗

PubMed2001

Partial chemical/structural elucidation of anti-mutagenic compounds from corn.

In this study, corn fractions obtained from an isolation process of anti-mutagenic factors in our previous research work (Burgos-Hernández et al., 2001), were subjected to several analyses for chemical/structural elucidation. The anti-mutagenic activity of these fractions was tested against aflatoxin B(1) (AFB(1)) and 1-methyl-3-nitro-1-nitrosoguanidine (MNNG), a mutagen that does not require bioactivation. Two concentrations of this agent in the corn fractions were tested for anti-mutagenicity in the Salmonella/microsomal mutagenicity assay, using tester strain TA100 with no metabolic activation. Corn fractions tested showed evidence of anti-mutagenic activity by producing a dose-response type of relationship between a constant amount of MNNG and several concentrations of tested corn fraction. Five different varieties of yellow corn were tested in order to determine if the anti-mutagenic factors were intrinsic to corn. Variety of the corn did not show an effect on the reduction of the mutagenic potential of AFB(1) suggesting that anti-mutagenic compounds are intrinsic to corn. Four corn fractions, previously obtained after the isolation process were analyzed by MALDI-MS and GC-MS. MALDI-MS showed the presence of two groups of molecules or molecular fragments. The molecular mass of one group ranged from 250 to 370 m/z, the other ranged from 540 to 640 m/z. GC-MS identified linoleic acid as one of the compounds responsible for the anti-mutagenic activity present in corn.

Aflatoxin B1↗

PubMed2001

Anti-mutagenic compounds from corn.

In previous studies with aflatoxin-contaminated corn an uncharacteristic response for AFB1 in the Salmonella/microsomal mutagenicity assay (Ames test) was observed and the presence of anti-aflatoxin factors in the corn was suggested. In the current study, corn was extracted and fractionated using thin layer chromatography (TLC) using different developing solvent systems and the Ames test was used to monitor for anti-mutagenic activity in the corn fractions. Both Salmonella tester strains TA98 and TA100 with metabolic activation (S9) were used. Several corn fractions, at different stages in the isolation and purification process, showed anti-mutagenic dose-responses when exposed to pure AFB1. Corti extracts were non-toxic to the tester strains and TLC fractions that showed the best anti-mutagenic dose-responses were selected for further partial characterization analyses.

Aflatoxin B1↗

PubMed2000

Central nervous system plasticity during hair cell loss and regeneration.

Following cochlear ablation, auditory neurons in the central nervous system (CNS) undergo alterations in morphology and function, including neuronal cell death. The trigger for these CNS changes is the abrupt cessation of afferent input via eighth nerve fiber activity. Gentamicin can cause ototoxic damage to cochlear hair cells responsible for high frequency hearing, which seems likely to cause a frequency-specific loss of input into the CNS. In birds, these hair cells can regenerate, presumably restoring input into the CNS. This review summarizes current knowledge of how CNS auditory neurons respond to this transient, frequency-specific loss of cochlear function. A single systemic injection of a high dose of gentamicin results in the complete loss of high frequency hair cells by 5 days, followed by the regeneration of new hair cells. Both hair cell-specific functional measures and estimates of CNS afferent activity suggest that newly regenerated hair cells restore afferent input to brainstem auditory neurons. Frequency-specific neuronal cell death and shrinkage occur following gentamicin damage to hair cells, with an unexpected recovery of neuronal cell number at longer survival times. A newly-developed method for topical, unilateral gentamicin application will allow future studies to compare neuronal changes within a given animal.

Animals↗

PubMed2000

Low temperature pasteurization to reduce the risk of vibrio infections from raw shell-stock oysters.

Vibrio vulnificus and V. parahaemolyticus are natural inhabitants of estuarine environments and may be transmitted to humans by ingestion of raw oysters. This study focused on the use of low temperature pasteurization, to reduce these Vibrio spp. to nondetectable levels, thus reducing the risk of infection associated with raw oyster consumption. Artificially-inoculated V. vulnificus and V. parahaemolyticus and naturally-contaminated V. vulnificus in live oysters were pasteurized at 50 degrees C for up to 15 min. Samples of processed and unprocessed oysters were enumerated for V. vulnificus, V. parahaemolyticus, and aerobic spoilage bacteria for 0-14 days. Low temperature pasteurization was effective in reducing these pathogens from > 100,000 to non-detectable levels in less than 10 min of processing. Spoilage bacteria were reduced by 2-3 logs, thus increasing the shelf-life for up to 7 days beyond live unprocessed oysters. Vibrio vulnificus in control oysters was reduced by 10(2) during ice storage alone. Following pasteurization and during a temperature storage abuse study (24 h at 22 degrees C), V. vulnificus was not recovered. During this storage period spoilage bacteria exceeded 1 million/g oyster meat.

Animals↗

PubMed1999

Aquatic biotoxins: design and implementation of seafood safety monitoring programs.

Naturally occurring toxicants are usually odorless, tasteless, and generally undetectable by any simple chemical test. Various programs have been established that are effective in reducing risks associated with these toxicants in food. These programs include setting regulatory limits, monitoring susceptible commodities for toxin levels, and using decontamination procedures. Bioassays have been used traditionally to monitor suspect products. All traditional bioassays, however, have one common disadvantage, i.e., the lack of specificity for individual toxins. The lack of available reference standards for specific toxins has also hampered implementation of monitoring programs. Utilizing the knowledge gained with regulatory monitoring and decontamination programs for other toxins, e.g., aflatoxin, similar seafood safety programs can be developed for aquatic biotoxins that will reduce risks and hazards associated with the contaminant to practicable levels and help to preserve an adequate food supply. Research is needed in several areas identified in this article. International cooperation has an important role in achieving these essential elements. Global programs will help in the adequate management of risks associated with aquatic biotoxins. To have an effective monitoring program, it is necessary to define precisely the local needs for information in a short or long time range. It is necessary to have basic knowledge about the biological, chemical, and physical conditions as well as temporal and geographic variations within the region of interest (2). Regardless of the overall success of fish/shellfish toxin monitoring plans, emergencies will occur. Therefore, contingency plans should be developed so there will be no misunderstanding of what actions to take (148). In general, however, the structure of the program must be kept as simple as possible to facilitate fast and uncomplicated flow of information among the various organizations and individuals involved (2). Public health and safety requires the removal of any toxic shellfish from the market, within practicability, and closure of any suspect harvest area. It should be important to remember that economic value of the fish or shellfish resource is always secondary to public health and safety (148).

Environmental Monitoring↗

PubMed1999

Tonotopic changes in 2-deoxyglucose activity in chick cochlear nucleus during hair cell loss and regeneration.

Following cochlear ablation, auditory neurons in the central nervous system (CNS) undergo alterations in morphology and function, including neuronal cell death. The trigger for these CNS changes is the abrupt cessation of eighth nerve fiber activity. Gentamicin can cause ototoxic damage to cochlear hair cells responsible for high frequency hearing. In birds, these hair cells can regenerate. Therefore, gentamicin causes a partial, yet reversible insult to the ear. It is not known how this partial hair cell damage affects excitatory input to the cochlear nucleus. We examined chick cochlear nucleus activity during hair cell loss and regeneration by measuring 2-deoxyglucose (2DG) uptake. Normal animals showed a rostral to caudal gradient of 2DG activity, with higher activity in caudal regions. When hair cells are damaged (2, 5 days), 2DG uptake is decreased in cochlear nucleus. When hair cells regenerate (9, 16, 28 days), 2DG uptake returns to control levels. This decrease and subsequent return of activity only occurs in the rostral, high frequency region of the cochlear nucleus. No changes are seen in the caudal, low frequency region. These results suggest that changes in activity of cochlear nucleus occur at a similar time course to anatomical changes in the cochlea.

Acoustic Stimulation↗

PubMed1998

Evidence for loss and recovery of chick brainstem auditory neurons during gentamicin-induced cochlear damage and regeneration.

It is well documented that damage to the chick cochlea caused by acoustic overstimulation or ototoxic drugs is reversible. Second-order auditory neurons in nucleus magnocellularis (NM) are sensitive to changes in input from the cochlea. However, few experiments studying changes in NM during cochlear hair cell loss and regeneration have been reported. Chicks were given a single systemic dose of gentamicin, which results in maximal hair cell loss in the base of the cochlea after 5 days. Many new hair cells are present by 9 days. These new hair cells are mature but not completely recovered in organization by 70 days. We counted neurons in Nissl-stained sections of the brainstem within specific tonotopic regions of NM, comparing absolute cell number between gentamicin- and saline-treated animals at both short and long survival times. Our data suggest that neuronal number in rostral NM parallels hair cell number in the base of the cochlea. That is, after a single dose of gentamicin, we see a loss of both cochlear hair cells and NM neurons early, followed by a recovery of both cochlear hair cells and NM neurons later. These results suggest that neurons, like cochlear hair cells, can recover following gentamicin-induced damage.

Animals↗

PubMed1997

Risks associated with consumption of herbal teas.

Plants have been used for medicinal purposes for centuries. Health-oriented individuals are turning to herbal teas as alternatives to caffeinated beverages such as coffee, tea, and cocoa and for low-caloric supplements. The popularity of herbal tea consumption has increased significantly during the past two decades in the U.S. Hundreds of different teas made up of varied mixtures of roots, leaves, seeds, barks, or other parts of shrubs, vines, or trees are sold in health food stores. Although chemists have been characterizing toxic plant constituents for over 100 years, toxicological studies of herbal teas have been limited and, therefore, the safety of many of these products is unknown. Plants synthesize secondary metabolites that are not essential in the production of energy and whose role may be in the defense mechanisms as plant toxins to their interactions with other plants, herbivores, and parasites. Pyrrolizidine alkaloids (PAs) were among the first naturally occurring carcinogens identified in plant products, and their presence in herbal teas is a matter of public health significance. Some herbal tea mixtures and single-ingredient herbal teas have been analyzed for toxic/mutagenic potential by bioassay and chromatographic techniques. Numerous human and animal intoxications have been associated with naturally occurring components, including pyrrolizidine alkaloids, tannins, and safrole. Thus, the prevention of human exposure to carcinogens or mutagens present in herbal tea mixture extracts is crucial. Preparation of infusion drinks prepared from plants appears to concentrate biologically active compounds and is a major source of PA poisoning. The quantity and consumption over a long period of time is of major concern. It is recommended that widespread consumption of herbal infusions should be minimized until data on the levels and varieties of carcinogens, mutagens, and toxicants are made available.

Beverages↗

PubMed1997

Human disease associated with Clostridium perfringens enterotoxin.

Clostridium perfringens continues to be a common cause of food-borne disease. Characteristics of this organism that contribute to its ability to cause food-borne illness include the formation of heat-resistant spores that survive normal cooking/heating temperatures, a rapid growth rate in warm food, and the production of enterotoxin (CPE) in the human gut. Time and temperature abuse associated with food preparation contributes to the majority of outbreaks of C. perfringens food-borne disease. CPE-induced diarrhea has been reported in the absence of a defined food vehicle. These cases have been typically associated with the elderly and following a course of antibiotic therapy. The incidence of CPE-induced diarrhea may be expected to increase with the growing population of immunocompromised (disease-, treatment-, or age-induced) individuals. Clostridium perfringens has been implicated as a possible contributor to the development of SIDS in susceptible individuals. Specifically, it has been hypothesized that CPE acts as a triggering agent, initiating the events associated with the development of SIDS. Continued refinement of both immunoassays and molecular methods for toxin and gene detection, respectively, will facilitate their eventual availability as commercial kits, providing rapid and simplified methods for the detection of C. perfringens isolates that produce or have the capacity to produce CPE as well as other toxins associated with this organism.

Clostridium Infections↗

PubMed1997

Anti-aflatoxin mutagenic factors in corn.

Extracts, isolated through sequential fractionation and partition procedures described previously (Martinez et al. 1994) from aflatoxin-free corn and aflatoxin-contaminated corn with and without ammonia treatment, were investigated for mutagenic potential using the Ames test (TA 100 tester strain). 2-Aminofluorene (2-AF) and pure aflatoxin B1 (AFB1) were used as positive controls. Although TA100 showed mutagenic response to pure AFB1 at a dose of ca 10 ng/plate, all isolates tested from ammonia-treated aflatoxin-contaminated corn containing 7500 ng AFB1/g did not exhibit positive results in the Ames test. Additionally, isolates from non-ammonia-treated aflatoxin-contaminated corn failed to give positive mutagenic potentials. These results indicate that differences between the mutagenic potentials of pure aflatoxins and of aflatoxins in naturally-contaminated corn exist. CH2Cl2 extracts (the fractions containing aflatoxins) obtained from aflatoxin-contaminated corn with and without ammonia treatment were applied to preparative thin layer chromatography (TLC) in an effort to separate aflatoxins and/or ammonia/aflatoxin reaction products from the "unknown interfering materials' existing in the corn matrix. Each of the fractions separated by TLC was tested by the Ames test with S9 activation and none of them gave a mutagenic response to TA100. CH2Cl2 extracts in dimethylsulphoxide (DMSO) obtained from non-ammonia-treated aflatoxin-free corn were spiked with pure AFB1 and tested by TA100 with S9 activation. Again, no positive responses were observed. These findings provide further evidence of "unknown interfering materials' in corn which may bind with aflatoxin and/or can be extracted by CH2Cl2 together with aflatoxin, and, therefore, block the mutagenic activity of aflatoxin in the Ames test. Those materials were not separated from the aflatoxins by the TLC technique used in the present study. Possible reasons and further studies required to evaluate this phenomenon are discussed.

Aflatoxins↗

PubMed1997

Effect of leak location on measured respirator fit.

A significant difference in leak detection as a function of leak location was observed during a study assessing how well current models of quantitative fit-test systems detect leakage. Known sources of leakage (matched hypodermic needles) were introduced at three fixed locations into factory-probed half-mask and full-face respirators mounted on a headform-breathing machine system. The leak locations were the bridge of the nose, the cheek, and the chin. Baseline leakage into each respirator was determined by conducting a fit-test with all fixed leak sources capped. Fit tests were repeated with each individual source uncapped. Study objectives included determining (1) how well each system measured the leakage, and (2) whether leak location had any effect on leak measurement. An ambient aerosol fit-test system (Portacount Plus) and a controlled negative pressure (CNP) fit-test system (FitTester 3000) were used. The ambient aerosol system detected an overall average of 37.2% of the known leakage, with a coefficient of variation of 44.7%. An analysis of variance showed significant differences in aerosol system measurements of leakage as a function of leak location and mask type (p < 0.001). A different pattern of aerosol leak detection as a function of leak location was observed between half-mask and full-face respirators, which appears to be related to differences in in-mask airflow dynamics. The CNP system detected an overall average of 97.9% of the known leakage through the same hypodermic needles, with a coefficient of variation of 4.3%. CNP system results were not affected by leak location (p > 0.43) or mask type (p > 0.32).

Aerosols↗

PubMed1996

Reduction of risks associated with fumonisin contamination in corn.

Fumonisins, produced by Fusarium moniliforme, have been recognized as an important group of chemicals which cause health risks in domestic animals and humans. Decontamination procedures for fumonisin B1 (FB1) were evaluated to determine chemical modification and reduction in toxic/carcinogenic potentials. Ammoniation, a procedure used for decontamination of aflatoxins, yielded a 79% reduction in FB1 levels in naturally contaminated corn. Authentic FB1 and FB1-contaminated corn were exposed to alternative treatments containing various combinations of Ca(OH)2, NaHCO3, and H2O2 simulating a modified nixtamalization procedure. Treatments also included NH4Cl alone or in combination with H2O2 or horseradish peroxidase. The brine shrimp assay (Artemia spp.) was used to monitor toxicity of reaction products and the Salmonella/microsomal mutagenicity assay, using tester strains TA-100 and TA-102, was used to evaluate mutagenicity. Treatments of FB1-contaminated corn simulating modified nixtamalization (Ca(OH)2 alone or with Na-HCO3 + H2O2) gave 100% reduction of FB1 and reduced brine shrimp toxicity by ca. 40%. The positive mutagenic potential (without S-9) for extracts of corn naturally contaminated with FB1 was eliminated following exposure to modified nixtamalization. Reaction products formed when pure FB1 was treated with Ca(OH)2 and H2O2/NaHCO3 were inhibitory to Bacillus cereus, B. subtilis, and B. megaterium. No inhibitory potential was evident for contaminated corn extracts following the chemical treatments.

Animals↗

PubMed1995

Immunochemical detection methods for Salmonella spp., Escherichia coli O157:H7, and Listeria monocytogenes in foods.

Immunochemical assays should prove to be a significant improvement over standard culture methods for the detection of foodborne pathogens. These techniques take advantage of the specificity and sensitivity of the antibody/antigen reaction for analyte detection. ELISA is the most useful form of immunochemical method for the detection of foodborne pathogens in the food-processing setting, based on their simplicity and ability to analyze large numbers of samples at a time. Concerns with immunochemical techniques include problems with cross-reactivity and difficulties obtaining species specific assays. Also, most immunochemical methods continue to require an enrichment technique; however, subsequent identification is quite rapid when compared with standard culture methods, therefore allowing for the quicker release of negative products.

Culture Media↗

PubMed1995

Ciguatera poisoning after ingestion of imported jellyfish: diagnostic application of serum immunoassay.

Ciguatera fish poisoning is an important public health problem wherever humans consume tropical and subtropical fish. It accounts for over half of fish-related poisonings in the United States but is uncommonly diagnosed and underreported. Produced by dinoflagellates, ciguatoxin accumulates up the food chain in herbivorous and carnivorous fishes. Cnidaria jellyfish and related invertebrates) have not previously been associated with direct ciguatera intoxication in humans. We report the first case of ciguatera fish poisoning associated with cnidarian ingestion. A 12-year-old Tongan female presented to our Emergency Department with mid-abdominal pain, nausea, change in mental status, and new-onset movement disorder after ingestion of jellyfish imported from American Samoa. Clinical diagnosis was confirmed by strongly positive serum identification of ciguatoxin and related polyether toxins (including okadaic acid) with a rapid extraction method (REM) and highly reliable solid-phase immunobead assay (S-PIA) performed by the Food Toxicology Research Group, University of Arizona. Ciguatera pathophysiology, clinical presentation, differential diagnosis (including consideration of palytoxin poisoning), and treatment are briefly reviewed. We emphasize the growing incidence of ciguatera fish poisoning outside "high-risk" areas. In regions with immigrant populations, privately imported exotic fish may be toxin vectors. Marine species other than carnivorous fish are now suspect in human ciguatera intoxication. Reliable tests can aid in premarket fish testing, diagnosis, and follow-up of ciguatera fish poisoning. The global prevalence of marine toxins demands fishermen, consumers, and physicians maintain a high index of suspicion for ciguatera fish poisoning.

Animals↗

PubMed1994

Evolution of methods for assessing ciguatera toxins in fish.

Ciguatera toxins are odorless, tasteless, and generally undetectable by any simple chemical test; therefore, bioassays have traditionally been used to monitor suspect fish. Many native tests for toxicity in fish have been examined, including the discoloration of silver coins or copper wire or the repulsion of flies and ants, but all of these were rejected as invalid. Oral feeding of fish to cats or mongoose is a simple and relatively sensitive assay, but cats often regurgitate part of the meal. Feeding tests described above have been used in screening fish for toxicity, but they are cumbersome and nonquantitative. A mouse bioassay was developed; unfortunately, this procedure requires purification fish extracts, since mice are not very sensitive to ciguatoxin. An alternative to the use of mice is the mosquito bioassay, which was recently used to obtain a dose-response relationship between ingested ciguatoxin and clinical symptoms in man. This assay correlates reasonably well with cat and mouse bioassay. Many other bioassays have been developed using chicken, brine shrimp, and the guinea pig atrium. All traditional bioassays have one common disadvantage, the lack of specificity for individual toxins. Recent studies have also focused on the development of chemical methods, i.e., TLC and HPLC, for the detection and quantification of ciguatera-related toxins (okadaic acid and ciguatoxin and its analogs). Alternative assays based on immunochemical technology have been developed and show the greatest promise for use in seafood safety monitoring programs. For earlier methods (RIA and ELISA formats), liver extracts from ciguatoxic eels were used to prepare ciguatoxin antisera. The assay has been further adapted to a solid-phase immunobead assay (S-PIA), which has a very high potential for use as a marketplace screening tool to separate ciguateric fish. This assay can also be used to monitor reefs for ciguatera potential. Historically, attempts to validate methods used to measure ciguatera toxicity have been plagued with a lack of specificity and reference standards. These restrictions have been resolved, and a study is under way to evaluate the precision and accuracy of a low-cost, simplified field and laboratory S-PIA kit (Ciguatect, Hawaii Chemtect International, Pasadena, California, 91109), through an international interlaboratory trial using the AOAC/IUPAC validation mechanism.

Animals↗

PubMed1994

Antimicrobials in shrimp aquaculture in the United States: regulatory status and safety concerns.

The consumption of seafood, especially shrimp, increases yearly in the U.S. The U.S. is the second largest importer of shrimp in the world, consuming more than 11% of the total world production. Aquaculture is becoming an increasingly important source of the world's shrimp, currently accounting for approximately 30% of the world's supply. Unfortunately, in this era of international trade deficits, U.S. production of aquacultured shrimp is insignificant (< 0.1%) compared with world production. As shrimp aquaculture expands in the U.S., so does the use of intensive farming techniques. Shrimp aquaculture is like any other animal husbandry industry in that shrimp are subject to disease, especially under intensive farming methods. In penaeid shrimp, the primary diseases associated with mortalities are usually viral or bacterial. The majority of bacterial infections in penaeid shrimp are attributable to Vibrio species, with mortalities ranging from insignificant to 100%. However, the rapid growth of this industry has outpaced efforts by researchers, pharmaceutical companies, and federal regulatory agencies to provide approved therapeutants for shrimp disease management. Approval of drugs and their surveillance for compliance with regulations applicable to seafoods, including aquacultured goods, is the responsibility of the FDA. There are three general areas of concern regarding human health when chemotherapeutants are used in aquaculture: (1) residues of drugs in fish destined for human consumption; (2) development of drug resistance in human pathogenic bacteria; and (3) direct toxic effects to humans from handling of drugs. Currently, there are no antibacterials approved for shrimp aquaculture in the U.S. One of the major obstacles in the development and approval of new drugs for aquaculture is the cost of conducting the required studies. The high cost to pharmaceutical companies discourages investment in shrimp chemotherapeutant research, since the current U.S. market for such products is small. Unfortunately, the U.S. shrimp aquaculture industry will remain small without legal availability of chemotherapeutants. Oxytetracycline (OTC) and Romet-30 are two antibacterials currently approved in the U.S. for catfish and salmonid aquaculture. Shrimp aquaculture facilities outside of the U.S. routinely use these drugs, as well as others, in the treatment of bacterial disease outbreaks. Much of the work required for OTC approval by the FDA for penaeid shrimp has been completed.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1994

Effect of ammoniation of aflatoxin B1-contaminated cottonseed feedstock on the aflatoxin M1 content of cows' milk and hepatocarcinogenicity in the trout bioassay.

The effectiveness of ammonia in inactivating aflatoxins in contaminated cottonseed was investigated. Two aflatoxin-contaminated cottonseed lots were treated separately using an atmospheric pressure, ambient temperature ammoniation procedure (APAT) or a high pressure, high temperature ammoniation procedure (HPHT), and incorporated into dairy cow rations. Isocalorific diets containing 25% defatted, dried milk from cows fed aflatoxin-contaminated cottonseed without or with APAT or HPHT treatment, or an aflatoxin-free human grade commercial milk powder, were then fed for 12 months to rainbow trout (Oncorhynchus mykiss). Aflatoxin M1 (AFM1) concentrations in milk powders without and with seed treatment were: APAT, 85 and < 0.05 microgram/kg; HPHT, 32 and < 0.05 microgram/kg. In the APAT experiment, trout consuming the diet containing milk from cows fed the aflatoxin-contaminated cottonseed had a 42% incidence of hepatic tumours; APAT cottonseed treatment reduced this to 2.5%. Positive controls were included to demonstrate trout responsiveness. AFB1 fed continuously for 12 months at 4 micrograms/kg resulted in a 34% tumour incidence, whereas positive controls fed 20 micrograms AFB1/kg, 80 micrograms AFM1/kg, or 800 micrograms AFM1/kg for 2 wk and killed 9 months later had a 37, 5.7 and 50% incidence of tumours, respectively. These data demonstrate that APAT ammonia treatment of aflatoxin-contaminated dairy cattle cottonseed feedstock abolished the detectable transfer of AFM1 or AFB1 into milk powder, and greatly reduced the carcinogenic risk posed by any carry-over of aflatoxins or their derivatives into milk. In addition, the results confirm AFM1 to be a lower level hepatocarcinogen in comparison with AFB1 in the trout carcinogenicity assay. In the separate HPHT experiment, no tumours were observed in the livers of trout fed diets containing milk from either the ammonia-treated or untreated source, or the control diet containing 8 micrograms AFM1/kg. Positive controls fed 64 micrograms AFB1/kg for 2 wk exhibited a 29% tumour incidence 12 months later. Thus in this experiment, neither AFM1 at 8 micrograms/kg nor any HPHT-derived aflatoxin derivatives that might have been carried over into milk, represented a detectably carcinogenic hazard to trout.

Aflatoxin B1↗