In vivo screening of four phytochemicals/extracts and a fungal immunomodulatory protein against an Eimeria acervulina infection in broilers.

BACKGROUND: Besides the anticoccidial drug resistance problem, increasing consumer concerns about food safety and residues have propelled the quest for alternative prevention and control strategies amongst which phytotherapy has gained appeal due to a renewed interest in natural medicine. OBJECTIVE: The objective was in vivo screening of four phytochemicals/extracts and a fungal immunomodulatory protein (FIP) against an Eimeria acervulina infection in broilers. ANIMALS AND METHODS: Four phytochemicals/extracts (extract from Echinacea purpurea, betaine (Betain™), curcumin, carvacrol (two different doses)), and a recombinant FIP from Ganoderma lucidum cloned and expressed in Escherichia coli were investigated for their anticoccidial potential. The experiment was conducted in a battery cage trial with 54 cages of eight birds each. Broilers infected with E. acervulina (a low and high infection dose of 10(4) and 10(5) sporulated oocysts, respectively) and treated with the phytochemicals/extracts or the FIP were compared with broilers treated with the anticoccidial salinomycin sodium (Sacox®) and with an untreated uninfected and an untreated infected control group. Coccidiosis lesion scores, body weight gains and oocyst shedding were used as parameters. RESULTS: The results showed a coccidiosis infection dose effect on the mean coccidiosis lesion scores. The phytochemicals/extracts and the FIP failed to reduce coccidiosis lesion scores and oocyst shedding, while salinomycin efficiently controlled the E. acervulina infection and enabled significantly higher body weight gains. CONCLUSION: In conclusion, the selected phytochemicals/extracts and the FIP did not reduce the lesions of an experimentally induced E. acervulina infection.

In vivo screening of five phytochemicals/extracts and a fungal immunomodulatory protein against colibacillosis in broilers.

Five phytochemicals/extracts (an extract from Echinacea purpurea, a ß-glucan-rich extract from Shiitake, betaine [Betain™], curcumin from Curcuma longa [turmeric] powder, carvacrol and also a recombinant fungal immunomodulatory protein [FIP] from Ganoderma lucidum) cloned and expressed in Escherichia coli were investigated for their anticolibacillosis potential in three chicken experiments, which were conducted in floor pens. Birds that were inoculated with E. coli intratracheally were treated with the phytochemicals/extracts or the FIP and compared with doxycycline-medicated and non-medicated infected broilers. Non-medicated and non-infected birds were used as negative controls. Mortality, colibacillosis lesions and body weight gains were used as parameters. Considering the sum of dead birds and chickens with generalized colibacillosis per group, there was no significant difference between the positive control groups and birds treated with phytochemicals/extracts or the FIP. In contrast, doxycycline-treated birds showed significantly lower mortality and generalized colibacillosis. Moreover, none of the phytochemicals/extracts and the FIP improved recovery from colibacillosis lesions, while all doxycycline-treated broilers recovered completely. The negative control birds and doxycycline-treated groups consistently showed the highest weight gains. Pulsed-field gel electrophoresis of reisolates showed that they were genetically indistinguishable from the inoculation strain. In conclusion, none of the tested phytochemicals/extracts and the FIP significantly reduced the E. coli-induced mortality and generalized colibacillosis, and nor did they improve recovery from colibacillosis lesions.

Melanin biosynthesis pathway in Agaricus bisporus mushrooms.

With the full genome sequence of Agaricus bisporus available, it was possible to investigate the genes involved in the melanin biosynthesis pathway of button mushrooms. Based on different BLAST and alignments, genes were identified in the genome which are postulated to be involved in this pathway. Seven housekeeping genes were tested of which 18S rRNA was the only housekeeping gene that was stably expressed in various tissues of different developmental stages. Gene expression was determined for most gene homologs (26 genes) involved in the melanin pathway. Of the analysed genes, those encoding polyphenol oxidase (PPO), the PPO co-factor L-chain (unique for A. bisporus), and a putative transcription factor (photoregulator B) were among the highest expressed in skin tissue. An in depth look was taken at the clustering of several PPO genes and the PPO co-factor gene on chromosome 5, which showed that almost 25% of the protein encoding genes in this cluster have a conserved NACHT and WD40 domain or a P-loop nucleoside triphosphate hydrolase. This article will be the start for an in depth study of the melanin pathway and its role in quality losses of this economically important product.

Single-longitudinal-mode optical parametric oscillator for spectroscopic applications.

We have developed a tunable, narrow-bandwidth nanosecond optical parametric oscillator system and applied it to spectroscopic studies. The system consists of a narrow-bandwidth grazing-incidence oscillator and a seeded power oscillator, generating Fourier-transform-limited 1.5-ns pulses (bandwidth <500 MHz) in the wavelength range 435 to 2000 nm with energy of 3.5 mJ at a pump energy of 22 mJ. Continuous scanning over 30 to 100 GHz (depending on wavelength) is demonstrated by recording of the resonance line of the Hg atom at 253.7 nm and a vibrational transition of the CO (2) molecule at 1528 nm.

Expression of the Fusarium resistance gene I-2 colocalizes with the site of fungal containment.

The tomato resistance gene I-2 is one of at least six members of a gene family that are expressed at low levels in the roots, stems and leaves of young tomato plants. Plants transformed with constructs containing a functional I-2 promoter fused to the beta-glucuronidase (GUS) reporter gene were used in detailed expression studies. Highest GUS activity was found in stems of young tomato plants. Histochemical analysis revealed that the I-2 promoter drives expression of the reporter gene in vascular tissue of fruits, leaves, stems and mature roots. In younger roots, expression was most abundant at the base of lateral root primordia. Microscopical analysis of young tomato plants revealed expression in tissue surrounding the xylem vessels. We show that in resistant plants, fungal growth into this region of the vascular tissue is prevented, suggesting a correlation with the I-2-mediated resistance response.

Foxy: an active family of short interspersed nuclear elements from Fusarium oxysporum.

A novel family of short interspersed nuclear elements (SINEs) has been identified in Fusarium oxysporum. This family has been called Foxy. The feature that makes Foxy unique among SINEs is the presence of 5' terminal tetranucleotide repeats. Both the number and the sequence of these repeats vary between individual members of the family. The genome of F. oxysporum f. sp. lycopersici contains at least 160 copies of Foxy. In a mutant obtained upon gamma irradiation of a wild-type isolate, 13 new Foxy insertions were identified. These observations, together with the occurrence of many Foxy-specific polymorphisms between isolates within one vegetative incompatibility group and the presence of Foxy-specific transcripts in the fungus, indicate that Foxy is currently active and may contribute to the genetic variability of F. oxysporum. Since we have not been able to detect Foxy sequences by PCR analyses in other fungi, this novel family of SINEs seems to be confined to Fusarium species.

Toxicological consequences of feeding PCB congeners to infant rhesus (Macaca mulatta) and cynomolgus (Macaca fascicularis) monkeys.

In a study designed to minimize interspecies extrapolation of toxicological data, nine rhesus (Macaca mulatta) and 15 cynomolgus (M. fascicularis) day-old infant monkeys were separated from their dams following parturition and hand-reared using a liquid non-human primate formulation. The infants were randomly divided into a control and a treated group which received a mixture of polychlorinated biphenyl (PCB) congeners analogous to those found in breast milk from Canadian women. The concentration of congeners in the dosing media resulted in each infant receiving a total of 7.5 microg PCB congeners/kg body weight/day. The congeners were added either to the liquid formulation or to corn oil and administered to the back of the monkey's mouth for 20 weeks. Monthly blood and adipose specimens were obtained during the dosing period and then periodically until the monkey was necropsied or taken off test (minimum of 66 weeks on test) for congener analysis. Parameters such as body weight, formula consumption, tooth eruption, somatic measurements, haematology and serum biochemistry were monitored throughout the study. In addition, a qualitative evaluation of the absorption and depletion of the various congeners was undertaken as was an immunological evaluation. For the monitored parameters, very few differences were found to be statistically significant. For the immunological parameters, the only statistically differences found were a reduction over time for immunoglobulins M and G antibodies to sheep red blood cells (cyno, P = 0.025; rhesus, P = 0.002) and a treatment-related reduction in the levels of the HLA-DR cell surface marker (mean percent, P = 0.016; absolute levels, P = 0.027). There were some qualitative differences regarding absorption and depletion rates for the various congeners, but it could not be definitely ascertained whether these differences were due to species differences or dosing mode. However, statistically significant differences were found for treatment (P = 0.0293) as well as for species and vehicle regarding the concentration of PCB in blood (species;--P = 0.0399; treatment--P = 0.0001) and adipose tissue (species--P = 0.0489; treatment--P = 0.0001).

Loss of Avirulence and Reduced Pathogenicity of a Gamma-Irradiated Mutant of Fusarium oxysporum f. sp. lycopersici.

ABSTRACT The tomato Fusarium resistance gene I-2 confers resistance to F. oxy-sporum f. sp. lycopersici race 2, which expresses the corresponding aviru-lence gene avrI-2. To elucidate the molecular basis of this gene-for-gene interaction, we initiated a search for the avrI-2 gene. Gamma irradiation mutagenesis, using (137)Cs, was performed to generate an avrI-2 mutant of F. oxysporum f. sp. lycopersici. To this end, a race 2 isolate was first transformed with a phleomycine resistance gene and a GUS marker gene in order to distinguish mutants from contaminating isolates. A total of 21,712 mutagenized colonies was tested for loss of avirulence on I-2-containing tomato seedlings. One mutant was selected that showed the expected loss of avirulence but, surprisingly, also showed reduced pathogenicity toward susceptible tomato plants. DNA analysis was subsequently used to visualize genomic changes in the mutant. Southern analysis on contour-clamped homogeneous electrophoretic field blots demonstrated a translocation of a 3.75-Mb chromosome in the mutant. Random amplified polymorphic DNA and amplified fragment length polymorphism analysis identified at least nine polymorphisms between the wild-type and mutant isolates. Most of these polymorphisms appeared as extra fragments in the mutant and contained repetitive DNA sequences.

Biological and Molecular Characterization of Fusarium oxysporum f. sp. lycopersici Divides Race 1 Isolates into Separate Virulence Groups.

ABSTRACT A collection of race 1 and race 2 isolates of Fusarium oxysporum f. sp. lycopersici was screened for vegetative compatibility and characterized by random amplified polymorphic DNA (RAPD) analysis to establish the identity and genetic diversity of the isolates. Comparison of RAPD profiles revealed two main groups that coincide with vegetative compatibility groups (VCGs). In addition, several single-member VCGs were identified that could not be grouped in one of the two main RAPD clusters. This suggests that F. oxysporum f. sp. lycopersici is a polyphyletic taxon. To assign avirulence genotypes to race 1 isolates, they were tested for their virulence on a small set of tomato lines (Lycopersicon esculentum), including line OT364. This line was selected because it shows resistance to race 2 isolates but, unlike most other race 2-resistant lines, susceptibility to race 1 isolates. To exclude the influence of other components than those related to the race-specific resistance response, we tested the virulence of race 1 isolates on a susceptible tomato that has become race 2 resistant by introduction of an I-2 transgene. The results show that both line OT364 and the transgenic line were significantly affected by four race 1 isolates, but not by seven other race 1 isolates nor by any race 2 isolates. This allowed a subdivision of race 1 isolates based on the presence or absence of an avirulence gene corresponding to the I-2 resistance gene. The data presented here support a gene-for-gene relationship for the interaction between F. oxysporum f. sp. lycopersici and its host tomato.

Dissection of the fusarium I2 gene cluster in tomato reveals six homologs and one active gene copy.

The I2 locus in tomato confers resistance to race 2 of the soil-borne fungus Fusarium oxysporum f sp lycopersici. The selective restriction fragment amplification (AFLP) positional cloning strategy was used to identify I2 in the tomato genome. A yeast artificial chromosome (YAC) clone covering approximately 750 kb encompassing the I2 locus was isolated, and the AFLP technique was used to derive tightly linked AFLP markers from this YAC clone. Genetic complementation analysis in transgenic R1 plants using a set of overlapping cosmids covering the I2 locus revealed three cosmids giving full resistance to F. o. lycopersici race 2. These cosmids shared a 7-kb DNA fragment containing an open reading frame encoding a protein with similarity to the nucleotide binding site leucine-rich repeat family of resistance genes. At the I2 locus, we identified six additional homologs that included the recently identified I2C-1 and I2C-2 genes. However, cosmids containing the I2C-1 or I2C-2 gene could not confer resistance to plants, indicating that these members are not the functional resistance genes. Alignments between the various members of the I2 gene family revealed two significant variable regions within the leucine-rich repeat region. They consisted of deletions or duplications of one or more leucine-rich repeats. We propose that one or both of these leucine-rich repeats are involved in Fusarium wilt resistance with I2 specificity.

Postmortem tissue levels of polychlorinated biphenyls in female rhesus monkeys after more than six years of daily dosing with Aroclor 1254 and in their non-dosed offspring.

Polychlorinated biphenyl (PCBs) analyses were made on prenecropsy blood samples and postmortem adipose, liver, kidney, and brain tissues from female rhesus monkeys fed a daily dose of 0, 5, 20, 40, or 80 micrograms Aroclor 1254/kg body weight for approximately 6 years. During this time, the females were bred with non-dosed males. All resulting offspring were nursed for 22 weeks and fed no additional PCBs until they were necropsied at approximately 120 weeks after birth. PCBs were also measured in necropsied infant tissues to determine PCB levels due to intake of PCB-contaminated milk from the dosed dams, in addition to in utero exposure. Polychlorinated biphenyl levels in all tissues of the adult monkeys increased with their dosage. The highest PCB levels were found in adipose tissue and the lowest levels were found in the brain. Polychlorinated biphenyl residues in the cortex of the kidney were lower than in the medulla, while in the brain no appreciable differences were observed between the occipital and frontal lobes. Necropsy tissues of infants from dosed dams contained more PCBs than those nursed by controls, but less than tissues from stillborn infants. Although no differences were observed between PCB tissue levels from monkeys having offspring and those having no offspring, those having a stillborn infant had higher PCB levels in their tissues than those with a viable infant. Similarly, monkeys that were euthanized because of poor health had higher PCB levels in their tissues than those necropsied at the conclusion of the study and showed a dramatic shift from tetra- and hexachlorobiphenyls to penta- and heptachlorobiphenyls in their tissues. The PCB distribution pattern in tissues from a dosed mother/infant pair differed considerably. A larger percentage of heptachlorobiphenyls was found in the infant than in its dam. The adipose/blood PCB ratio in the adult monkeys remained remarkably constant.

Female rhesus monkeys dosed with Aroclor 1254: analysis of polychlorinated biphenyl congeners in dam's milk and in the blood of dams and their offspring before, during, and after gestation.

Specific polychlorinated biphenyl (PCB) congeners were measured before, during, and after gestation in the blood of rhesus monkeys, as well as in their milk and in the blood of their infants during lactation, as part of a long-term feeding study to evaluate the toxicology of Aroclor 1254 on pre- and postnatal development of infant monkeys. During gestation a considerable shift from the higher to lower chlorinated biphenyls in the blood was observed in both dosed and nondosed animals. The contribution of penta- and hexachlorobiphenyls in the milk slightly increased with higher dosage. In addition, the percentages of 2,2'3,4,5'-,2,2',4,5,5'-, and 2,3,3',4',6-pentachlorobiphenyls were remarkably lower in the milk of dosed dams than in the originally ingested Aroclor 1254. PCB congener levels in infant blood increased during the lactation period but immediately decreased upon weaning. The lower chlorinated biphenyls virtually disappeared from infant blood after 16 weeks of nursing. Some correlations were observed between PCB congener levels in mother and infant and the congener ratios calculated.

Toxicological consequences of aroclor 1254 ingestion by female rhesus (Macaca mulatta) monkeys. Part 2. Reproduction and infant findings.

A group of 80 menstruating rhesus (Macaca mulatta) monkeys were randomly allocated to four similar test rooms (20 monkeys/room) and then randomly allocated within each room to one of five dose groups (four females/dose group/room). Each day, the monkeys self-ingested capsules containing doses of 0, 5, 20, 40 or 80 micrograms Aroclor 1254/kg body weight. After 25 months of continuous dosing, approximately 90% of the treated females had attained a qualitative pharmacokinetic steady state with respect to the concentration of polychlorinated biphenyl (PCB) in their adipose tissue. Commencing on test month 37, each female was paired with an untreated male until either an impregnation occurred or the 29-month breeding phase of the study was completed. The females continued to receive their daily test dose during mating and gestation. To preclude an infant ingesting the mother's dosing capsule, dosing of the dam was discontinued when a nursing infant was approximately 7 wk old. Treatment was restarted when the infant was weaned at 22 wk of age. At parturition, and every 4 wk until weaning, milk and blood samples were obtained from the dam and a blood sample was obtained from the infant for PCB analysis. When the infant was 20 wk old, immunological testing was initiated and an adipose sample was obtained from the infant and dam for PCB analysis. Subsequently, further adipose and blood samples were obtained from the infant and blood specimens were obtained from the dam for PCB analysis. Concurrently, each infant was subjected to anthropometric measurements and detailed clinical examinations until it was approximately 122 wk old. At 122 wk some of the control and all of the treated infants were killed humanely and autopsied. A statistical analysis of the reproduction data provided evidence for a significant decreasing dose-related trend in conception rates and a significant increasing dose-related trend in foetal mortality. Several comparisons between impregnated and non-impregnated females did not implicate 'age' as a confounding factor regarding these results. The major findings with the infants involved some immunological test differences and mild clinical manifestations of PCB ingestion.

The elimination and estimated half-lives of specific polychlorinated biphenyl congeners from the blood of female monkeys after discontinuation of daily dosing with Aroclor 1254.

The levels of thirty polychlorinated biphenyl congeners in the blood of female rhesus monkeys, previously dosed with Aroclor 1254 for over six years, were monitored every two weeks during the first year and monthly during the subsequent two years after dosing was discontinued. Both blood lipid and polychlorinated biphenyl congener levels generally declined during this post dosing period. The percent distribution of the PCB congeners during the post dosing period remained relatively constant with more than half of all polychlorinated biphenyls consisting of the mono-orthochlorine substituted biphenyls. The contribution of the mono-orthochlorine substituted biphenyls was significantly different for one out of three monkeys in two of the three dose groups, during the post dosing period. Half-life, estimations for nine of the congeners ranged from 0.3-7.6 years.

Determination of polychlorinated biphenyls in postpartum blood, adipose tissue, and milk from female rhesus monkeys and their offspring after prolonged dosing with Aroclor 1254.

Analytical and quality control procedures are described for the determination of polychlorinated biphenyls in blood, adipose tissue, and milk from dosed female monkeys and their offspring, as part of a study to measure the toxicological effect of Aroclor 1254 on the pre- and postnatal development of fetus and infant, respectively. Recoveries of polychlorinated biphenyls from fortified blood, fat, and milk of monkeys ranged from 81 to 96%, whereas recoveries from fortified corn oil, used to evaluate routine analysis, ranged from 94 to 108%. The coefficient of variation for triplicate analyses of lipids and polychlorinated biphenyls in blood or adipose tissue or both was less than 10%. Polychlorinated biphenyl levels in blood, milk, and fat rose with increasing dosage. After weaning, when the infants were no longer exposed to polychlorinated biphenyls, their blood levels declined rapidly and approached maternal levels within 40-50 weeks. Approximately 100 weeks after weaning, polychlorinated biphenyl levels in adipose tissue of infants from treated dams reached the background levels of those in the control group.

Polychlorinated biphenyl levels in the tissues of exposed and nonexposed humans.

Polychlorinated biphenyls (PCBs) are synthetic chemicals, manufactured in volume from about 1929 to the 1970s. Environmental contamination by PCBs has been documented in various substances, including human tissue. PCBs have been measured in human tissue by a variety of analytical methods. PCB levels have been reported as an approximation of total PCB content expressed in terms of a commercial mixture, by identification and quantification of chromatographic peaks, or by qualitative and quantitative characterization of specific congeners. Until recently, the coplanar mono-ortho- and di-ortho substituted PCBs, which are especially toxic and present in significant concentration in humans from industrial countries, had not been measured in human tissues. Examples of various types of commonly used analyses are presented in general population subjects and in persons who experienced special exposure. In this paper, the usefulness of PCB blood determinations following potential exposure is demonstrated, and their application in health studies is illustrated from a number of case studies. Coplanar PCB, mono-ortho-substituted and di-ortho-substituted PCB levels in human blood are presented and compared with polychlorinated dioxin (PCDD) and polychlorinated dibenzofuran (PCDF) levels in the U.S. population. Dioxin toxic equivalents for the two groups of chemicals are calculated and compared. It is found that mono-ortho-substituted and, to a lesser extent, coplanar PCBs, contribute substantially to dioxin toxic equivalents (TEq) in blood from U.S. adults. Because of substantial PCB contribution to dioxin toxic equivalents, total dioxinlike toxicity can only be determined if dioxins, dibenzofurans, and dioxinlike PCBs are measured.(ABSTRACT TRUNCATED AT 400 WORDS)

Temporal changes in some chlorinated hydrocarbon residue levels of Canadian breast milk and infant exposure.

Between 1967 and 1986, five national surveys were carried out to determine organochlorine residues in Canadian breast milk. In these surveys the number of samples, their geographic sampling distribution and fat content changed over the years. In addition, the number of samples obtained from younger mothers decreased, while those from older mothers (31-40 years) increased. Total DDTs showed the most consistent decline of all declining residues, while PCBs only decreased after 1982. Levels of oxychlordane and trans-nonachlor remained constant. Ratios of p,p'-DDE/p,p'-DDT gradually increased from 3.1 to 10.0 over the same period of time. With few exceptions, regional trends in residue levels followed those observed for the whole country. Although the number of reported chemicals increased since 1967, this was not necessarily reflected in the total accumulated residues in breast milk. By 1986, the daily intakes of all compounds at 8 weeks of nursing were well below the established acceptable daily intakes.

Toxicological consequences of Aroclor 1254 ingestion by female rhesus (Macaca mulatta) monkeys. Part 1B. Prebreeding phase: clinical and analytical laboratory findings.

A group of 80 menstruating rhesus (Macaca mulatta) monkeys, with an average estimated age of 11.1 +/- 4.1 yr SD were first randomly allocated to four similar test rooms (20 monkeys/room), and then randomly allocated to one of five dose groups (four females/dose group/room). Each day, the monkeys self-ingested capsules containing doses of 0, 5, 20, 40 or 80 micrograms Aroclor 1254/kg body weight. After 25 months of daily dosing, approximately 90% of the treated females attained a qualitative pharmacokinetic steady state with respect to the concentration of polychlorinated biphenyl (PCB) in their adipose tissue. Subsequently, oestrogen and progesterone concentrations in serum were determined for one complete oestrous cycle and various immunological tests were conducted, while the monkeys continued to receive their daily dose of PCB. During the prebreeding phase of the study, blood for clinical and analytical monitoring including haematology, serum biochemistry, serum hydrocortisone, serum proteins (alpha 1, alpha 2, beta and gamma-globulins), serum immunoglobulins (A, G and M) and thyroid variables (thyroxine/triiodothyronine (T3) uptake ratio, percentage T3 uptake and free thyroxine index), were obtained monthly, as were specimens to ascertain the concentration of PCB in the blood, adipose tissue and faeces. Major findings among treated monkeys included the following: changes in haematology (decreased erythrocyte count, haematocrit, reticulocyte count, and mean platelet volume), serum biochemistry (decreased cholesterol and total bilirubin), immunotoxicity (decreased antibody production to sheep red blood cells and alterations in the percentage of T helper and T suppressor cells) and pathology (the number of regions of sebaceous gland lobules per unit of histological length was significantly reduced). These effects were observed at PCB doses lower than those previously reported for non-human primates.

Levels of chlorinated hydrocarbon residues in Canadian human breast milk and their relationship to some characteristics of the donors.

A total of 412 breast milk samples from women in all provinces of Canada were analysed for polychlorinated biphenyls, eight chlorinated benzenes, 2,3-dichloronaphthalene, Mirex, alpha, beta, gamma and delta hexachlorocyclohexane, alpha and gamma chlordane, oxychlordane, transnonachlor, p,p'-DDT and some analogues, heptachlor epoxide, dieldrin and octachlorostyrene. No delta-hexachlorocyclohexane, heptachlor or aldrin were found in any of the samples, while median levels of the 1,2,4- and 1,3,5-trichlorobenzene, 1,2,3,4- and 1,2,3,5-tetrachlorobenzenes, gamma chlordane, o,p'-DDT and octachlorostyrene were all less than the minimum detectable level (MDL). All other compounds were present at median levels ranging from < 0.1 to 7.2 ng/g whole milk, but did not occur in all samples. Di to tetrachlorobenzenes, except 1,2,4-trichlorobenzene, 2,3-dichloronaphthalene and alpha-chlordane were observed in < 50% of the samples. From 1967 to 1986 a steady decline of hexachlorobenzene, gamma-hexachlorocyclohexane, DDTs, heptachlor epoxide and dieldrin was observed in Canadian breast milk. In addition, a decrease in some chlorinated benzenes, gamma-hexachlorocyclohexane and PCBs was also observed between 1982 and 1986. Levels of oxychlordane and trans-nonachlor remained constant. Canadian breast milk contamination appears to be one of the lowest among the industrialized nations. Regional and provincial differences in residue levels appeared minimal, although more often samples from Quebec and British Columbia had higher levels of some contaminants, such as oxychlordane, than samples from other provinces. No relationship was found between maternal age and residue levels, but some contaminants, such as PCBs, were significantly higher in the milk of mothers who breastfed their first child as compared to multiparous mothers. Breast milk residue levels were not related to fish consumption.

Assessment of human exposure to chemical contaminants in foods.

One of the most important factors in assessing risk to human health from potentially harmful chemicals in foods is the availability of good data on the exposure of the public to such substances. The means of acquiring these data generally involves monitoring programmes using appropriate sampling procedures and reliable analytical methods for measuring the compounds of concern in a variety of substrates. Two approaches are generally employed: a biological monitoring programme which measures substances in human fluids and tissues, and a food analysis monitoring programme, preferably a total diet study, wherein food is prepared for consumption prior to analysis. The choice of approach to use and chemicals to monitor depend on the situation within a particular country. The analysis of food has the advantage of short term impact since problems can be identified relatively quickly and control measures established. Biological monitoring on the other hand tends to indicate both accumulated and current exposure from all sources, including air, water and food. In Canada both approaches have been used for a number of years with major surveys of human milk and adipose tissue, and the total diet study, being conducted approximately every five years. Details of these programmes together with some of the pertinent findings are presented.