Survey of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and non-ortho-polychlorinated biphenyls in U.S. meat and poultry, 2007-2008: effect of new toxic equivalency factors on toxic equivalency levels, patterns, and temporal trends.

A statistically based survey of dioxins and dioxin-like compounds in domestic meat and poultry was conducted by the U.S. Department of Agriculture (USDA) from September 2007 to September 2008. Seventeen toxic polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and four non-ortho-polychlorinated biphenyls (no-PCBs) were measured in 510 beef (steer/heifer), market hog, young turkey, and young chicken samples. The results of the survey showed the sum of PCDD/F and no-PCB toxic equivalencies (sum-TEQs) ranging from not detected to 4.5 pg/g of lipid. Mean sum-TEQ levels for beef, turkey, chicken, and pork were 0.66, 0.61, 0.17, and 0.16 pg/g of lipid, respectively. To compare the new survey data with data from previous USDA surveys in the mid-1990s and 2002-2003, TEQs from all data sets were calculated using the most recent 2005 toxic equivalency factors (TEFs). The results of the recalculation on the older survey data was a small increase (4-13%) in mean TEQs for the mid-1990s data, which initially used pre-1994 TEFs, and a small decrease (2-4%) for the 2002-2003 data, which initially used 1998 TEFs. A comparison of the three surveys indicates declining TEQ trends in all slaughter classes over the 10 year period; however, the congener patterns remain relatively constant between 2002 and 2008, indicating similar animal exposures to dioxins and dioxin-like compounds during these time periods. Several samples from the 2008 survey with the highest TEQ values are undergoing follow-up investigations to determine possible sources that may be contributing to these levels.

Cadmium and lead residue control in a hazard analysis and critical control point (HACCP) environment.

In 2003-2004, the U.S. Department of Agriculture Food Safety and Inspection Service (FSIS) conducted an exploratory assessment to determine the occurrence and levels of cadmium and lead in randomly collected samples of kidney, liver, and muscle tissues of mature chickens, boars/stags, dairy cows, and heifers. The data generated in the study were qualitatively compared to data that FSIS gathered in a 1985-1986 study in order to identify trends in the levels of cadmium and lead in meat and poultry products. The exploratory assessment was necessary to verify that Hazard Analysis and Critical Control Point plans and efforts to control exposure to these heavy metals are effective and result in products that meet U.S. export requirements. A comparison of data from the two FSIS studies suggests that the incidence and levels of cadmium and lead in different slaughter classes have remained stable since the first study was conducted in 1985-1986. This study was conducted to fulfill FSIS mandate to ensure that meat, poultry, and egg products entering commerce in the United States are free of adulterants, including elevated levels of environmental contaminants such as cadmium and lead.

Clavulanic acid inactivation of SHV-1 and the inhibitor-resistant S130G SHV-1 beta-lactamase. Insights into the mechanism of inhibition.

Clavulanic acid is a potent mechanism-based inhibitor of TEM-1 and SHV-1beta-lactamases, enzymes that confer resistance to beta-lactams in many gram-negative pathogens. This compound has enjoyed widespread clinical use as part of beta-lactam beta-lactamase inhibitor therapy directed against penicillin-resistant pathogens. Unfortunately, the emergence of clavulanic acid-resistant variants of TEM-1 and SHV-1 beta-lactamase significantly compromise the efficacy of this combination. A single amino acid change at Ambler position Ser130 (Ser --> Gly) results in resistance to inactivation by clavulanate in the SHV-1 and TEM-1beta-lactamases. Herein, we investigated the inactivation of SHV-1 and the inhibitor-resistant S130G variant beta-lactamases by clavulanate. Using liquid chromatography electrospray ionization mass spectrometry, we detected multiple modified proteins when SHV-1 beta-lactamase is inactivated by clavulanate. Matrix-assisted laser desorption ionization-time of flight mass spectrometry was used to study tryptic digests of SHV-1 and S130Gbeta-lactamases (+/- inactivation with clavulanate) and identified peptides modified at the active site Ser70. Ultraviolet (UV) difference spectral studies comparing SHV-1 and S130Gbeta-lactamases inactivated by clavulanate showed that the formation of reaction intermediates with absorption maxima at 227 and 280 nm are diminished and delayed when S130Gbeta-lactamase is inactivated. We conclude that the clavulanic acid inhibition of the S130G beta-lactamase must follow a branch of the normal inactivation pathway. These findings highlight the importance of understanding the intermediates formed in the inactivation process of inhibitor-resistant beta-lactamases and suggest how strategic chemical design can lead to novel ways to inhibit beta-lactamases.

Tazobactam inactivation of SHV-1 and the inhibitor-resistant Ser130 -->Gly SHV-1 beta-lactamase: insights into the mechanism of inhibition.

The increasing number of bacteria resistant to combinations of beta-lactam and beta-lactamase inhibitors is creating great difficulties in the treatment of serious hospital-acquired infections. Understanding the mechanisms and structural basis for the inactivation of these inhibitor-resistant beta-lactamases provides a rationale for the design of novel compounds. In the present work, SHV-1 and the Ser(130) --> Gly inhibitor-resistant variant of SHV-1 beta-lactamase were inactivated with tazobactam, a potent class A beta-lactamase inhibitor. Apoenzymes and inhibited beta-lactamases were analyzed by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI/MS), digested with trypsin, and the products resolved using LC-ESI/MS and matrix-assisted laser desorption ionization-time of flight mass spectrometry. The mass increases observed for SHV-1 and Ser(130) --> Gly (+ Delta 88 Da and + Delta 70 Da, respectively) suggest that fragmentation of tazobactam readily occurs in the inhibitor-resistant variant to yield an inactive beta-lactamase. These two mass increments are consistent with the formation of an aldehyde (+ Delta 70 Da) and a hydrated aldehyde (+ Delta 88 Da) as stable products of inhibition. Our results reveal that the Ser --> Gly substitution at amino acid position 130 is not essential for enzyme inactivation. By examining the inhibitor-resistant Ser(130) --> Gly beta-lactamase, our data are the first to show that tazobactam undergoes fragmentation while still attached to the active site Ser(70) in this enzyme. After acylation of tazobactam by Ser(130) --> Gly, inactivation proceeds independent of any additional covalent interactions.