Steam precooking, an effective pretreatment to reduce contamination with polycyclic aromatic hydrocarbons in traditionally smoked shrimp.

ABSTRACT: Traditionally smoked shrimps are highly contaminated with carcinogenic polycyclic aromatic hydrocarbons (PAHs). The effect of precooking methods such as boiling (7 and 15 min) and steaming (15 and 20 min) on shrimp smoking effectiveness was investigated through smoking duration and the change of moisture, protein content, and PAHs contamination. The consumer's perception of derived shrimps was highlighted by acceptability and Just-About-Right test using 7-point hedonic and 3-point scale respectively. Both precooking methods didn't affect the shrimp protein content while steaming reduced significantly moisture content. All the precooking methods reduced significantly the smoking duration with the lowest values for steaming. Smoking with or free from precooking increased significantly PAHs content of shrimp. Although, the PAHs trends showed similar pattern for both precooking methods, steaming during 20 min before smoking enables to have the shrimp with Benzo[a]pyrene (BaP) and sum of benzo[b]fluoranthene, benzo[a]anthracene, chrysene and BAP (PAH4) contents lower to the safety limit recommended by the European regulation. Irrespective of consumption forms, main sensory attributes of smoked steamed shrimps were scored as Just-About-Right by most consumers (65 to 88%). Steam precooking (20 min) is the better way for upgrading the shrimp safety with expected liking of consumers.

Corrigendum: Cell-Free Spent Media Obtained From Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3'-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium.

[This corrects the article DOI: 10.3389/fmicb.2016.01460.].

Assessment of the physicochemical characteristics, chemical and microbiological safety of two types of kilichi, a grilled meat produced in Niger.

Production of kilichi, a grilled meat of West Africa, is a common method of meat preservation in Niger. Thirty samples of condiments-coated kilichi and uncoated kilichi collected in Niger, were analyzed for microbiological contamination, as well as NaCl, protein and lipid contents, using standard methods. Contamination with Polycyclic Aromatic Hydrocarbons (PAHs) was also assessed using a HPLC-FLD technique. Highly significant differences (p < .001) were observed between coated kilichi and uncoated kilichi, for NaCl content (2.56% and 1.40%), for proteins (51% and 72%) and lipids (18% and 13%), respectively. Water activity was low in both kilichi, showing a potential microbial stability. Among the 15 European Union (EU) priority PAHs, 12 were detected in the samples. About 56.3% of coated kilichi samples exceeded the EU maximal limit for BaP, and 75% exceeded the EU maximal limit for the sum of 4 PAHs (PAH4). For uncoated kilichi, 28.6% of samples did not meet the standards for BaP and PAH4. About 6% of coated kilichi samples were not compliant with standards related to Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, and 31%, 50% for yeasts and fungi, respectively. Escherichia coli and Enterobacteria were below the detection limit in both kilichi, but Salmonella and Bacillus cereus were detected only in one coated kilichi. The noncompliant samples of uncoated kilichi were in the proportions varying between 7% -86% for S. aureus, C. perfringens, yeasts, and fungi. This study showed potential risks associated with the consumption of traditionally produced kilichi in Niger due to both PAHs and pathogen bacteria contamination.

Development of an analytical method to detect short-chain fatty acids by SPME-GC-MS in samples coming from an in vitro gastrointestinal model.

Short chain fatty acids (SCFA) are end-products of intestinal bacterial fermentation. The concentrations of fermentation metabolites are closely related to the microbial activity that occurs in various digestive compartments. The fermentation products may vary qualitatively and quantitatively, especially within the colon. The Simulator of the Human Intestinal Microbial Ecosystem (SHIME), an in vitro dynamic and multicompartment model of the human intestinal tract, can be adapted to mimic the piglet gastrointestinal tract. In this context, a quantitative method, based on solid phase microextraction gas chromatography coupled to mass spectrometry (SPME-GC-MS), was developed for the determination of seven short chain fatty acids, i.e. acetic, propionic, butyric, isobutyric, valeric, isovaleric and hexanoic acids, in samples coming from this experimental in vitro gastrointestinal model. The advantage of the SPME-GC-MS technique is that the seven compounds could be determined in a single run, after a simple and rapid sample treatment, without any other extraction than the automatic SPME. The developed method was validated in accordance to the European and US FDA guidelines and showed good specificity/selectivity. In addition, limits of detection and quantification ranged from 8 to 72 mg L-1 and from 16 to 144 mg L-1, respectively. Two internal quality control samples spiked at different concentrations were analyzed to assess the trueness of the developed method, which ranged between 97.7 and 122.4% of the expected value, for the seven compounds analyzed. The method was successfully applied to twenty samples coming from a gastrointestinal model, with different inocula. The developed method might be used as a general method for measuring SCFA in biological samples.

Occurrence of Acetamiprid Residues in Water Reservoirs in the Cotton Basin of Northern Benin.

An Ultra performance liquid chromatography (UPLC) coupled to UV detection method was developed to determine acetamiprid residues in water reservoirs of northern Benin, close to cotton fields. The quantification limit of this method was 0.2 µg L-1 acetamiprid in water, its precision ranged between 8% and 22%, and its trueness between 99% and 117% (for concentrations ranging from 0.2 to 5.0 µg L-1). Acetamiprid residues were determined in water samples collected in four reservoirs from northern Benin during the phytosanitary treatment period of cotton. The minimum and maximum concentrations of acetamiprid residues in water were 0.2 and 7.7 µg L-1, respectively. These levels do not represent any risk for human consumption of this water, but indicate a regular use of acetamiprid, possibly together with other pesticides which could be more harmful for both humans and aquatic species.

Cell-Free Spent Media Obtained from Bifidobacterium bifidum and Bifidobacterium crudilactis Grown in Media Supplemented with 3'-Sialyllactose Modulate Virulence Gene Expression in Escherichia coli O157:H7 and Salmonella Typhimurium.

Complex oligosaccharides from human milk (HMO) possess an antimicrobial activity and can promote the growth of bifidobacteria such as Bifidobacterium bifidum and Bifidobacterium longum subsp. infantis. In addition, fermentation of carbohydrates by bifidobacteria can result in the production of metabolites presenting an antivirulence effect on several pathogenic bacteria. Whey is rich in complex bovine milk oligosaccharides (BMO) structurally similar to HMO and B. crudilactis, a species of bovine origin, is able to metabolize some of those complex carbohydrates. This study focused on the ability of B. bifidum and B. crudilactis to grow in a culture medium supplemented in 3'-sialyllactose (3'SL) as the main source of carbon, a major BMO encountered in cow milk. Next, the effects of cell-free spent media (CFSM) were tested against virulence expression of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium. Both strains were able to grow in presence of 3'SL, but B. crudilactis showed the best growth (7.92 ± 0.3 log cfu/ml) compared to B. bifidum (6.84 ± 0.9 log cfu/ml). Then, CFSM were tested for their effects on virulence gene expression by ler and hilA promoter activity of luminescent mutants of E. coli and S. Typhimurium, respectively, and on wild type strains of E. coli O157:H7 and S. Typhimurium using RT-qPCR. All CFSM resulted in significant under expression of the ler and hilA genes for the luminescent mutants and ler (ratios of -15.4 and -8.1 respectively) and qseA (ratios of -2.1 and -3.1) for the wild type strain of E. coli O157:H7. The 3'SL, a major BMO, combined with some bifidobacteria strains of bovine or human origin could therefore be an interesting synbiotic to maintain or restore the intestinal health of young children. These effects observed in vitro will be further investigated regarding the overall phenotype of pathogenic agents and the exact nature of the active molecules.

Guiana Dolphins (Sotalia guianensis) and DR-CALUX for Screening Coastal Brazilian Environments for Dioxins and Related Compounds.

Guiana dolphin is the top predator of highest toxicological concern in Brazil and many studies on levels of persistent, bioaccumulative, and toxicant (PBT) pollutants have been performed on the species. However, due to high costs of the analyses, only one investigation comprised the determination of dioxins and related compounds (DRCs) in Guiana dolphin tissues. The dioxin responsive-chemically activated luciferase gene expression (DR-CALUX(®)) cell bioassay was used in the present study for the analyses of hepatic samples from 28 male Guiana dolphins in order to screen estuarine environments for DRCs, comprising three regions (Northeastern, Southeastern, and Southern) and four states [Paraná (PR), Rio de Janeiro (RJ), Espírito Santo (ES), and Ceará (CE)] of Brazil. High bioanalytical equivalent (BEQ) concentrations [dioxins (pg BEQ/g lipid)] were found, varying from 1.94 to 15.6 pg BEQ/g. A significant negative correlation between BEQ concentrations and total length was found in Guiana dolphins from Brazil (all analysed dolphins). This pattern also was verified for RJ state, pointing to (1) chemically induced developmental disruption or to (2) increasing efficiency of the detoxifying activity with the growth of the animal. Comparison was performed with literature data and significantly higher BEQ levels were found in Brazilian Guiana dolphins than in those reported for North Sea harbour porpoises. Higher levels were found in Southeastern (the most PBT-contaminated area of the country) than in Southern region. However, it is not possible to affirm that Guiana dolphins are more contaminated by DRCs in SE than in S region, because individuals were lengthier in S than in SE region. Our results seem to have mirrored dolphin exposure to PCBs in Brazil according to the literature. Further studies are required for investigating the hypotheses 1 and 2 mentioned above.

Development of an LC-MS/MS analytical method for the simultaneous measurement of aldehydes from polyunsaturated fatty acids degradation in animal feed.

Knowing that polyunsaturated fatty acids can lead to the formation of potentially toxic aldehydes as secondary oxidation products, an analytical method using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) detection was developed to measure the concentration of eight aldehydes in animal feed: malondialdehyde (MDA), 4-hydroxy-2-nonenal (4-HNE), 4-hydroxy-2-hexenal (4-HHE), crotonaldehyde (CRT), benzaldehyde (BNZ), hexanal (HXL), 2,4-nonadienal, and 2,4-decadienal. The developed method was validated according to the criteria and procedure described in international standards. The evaluated parameters were specificity/selectivity, recovery, precision, accuracy, uncertainty, limits of detection and quantification, using the concept of accuracy profiles. These parameters were determined during experiments conducted over three different days with ground Kellogg's® Corn Flakes® cereals as model matrix for animal feed and spiked at different levels of concentration. Malondialdehyde, 4-HHE, 4-HNE, crotonaldehyde, benzaldehyde, and hexanal can be analyzed in the same run in animal feed with a very good accuracy, with recovery rates ranging from 86 to 109% for a working range going from 0.16 to 12.50 mg/kg. The analysis of 2,4-nonadienal and 2,4-decadienal can also be performed but in a limited range of concentration and with a limited degree of accuracy. Their recovery rates ranged between 54 and 114% and coefficient of variation for the intermediate precision between 11 and 25% for these two compounds. Copyright © 2016 John Wiley & Sons, Ltd.

Estrogenic evaluation and organochlorine identification in blubber of North Sea harbour porpoise (Phocoena phocoena) stranded on the North Sea coast.

Thirteen individual organochlorine compounds at 3 concentrations (80, 400, and 2000 ng/mL culture medium), as well as mixtures, were assayed for the estrogen receptor (ER) activation or inhibition, using a luciferase reporter gene assay (RGA). None of the PCB 138, 153, or 180 or their mixture induced a response in the RGA. o,p'-DDT was the most potent xenoestrogen from the DDT group, inducing a response already at 80 ng/mL. From the HCH and HCB group, only ß-HCH (at 400 and 2000 ng/mL) and δ-HCH (at 2000 ng/mL) displayed estrogenic activities. These 13 organochlorines were determined by GC-MS in 12 samples of North Sea harbor porpoise blubber. The PCBs were the main contaminants. Within each group, PCB 153 (6.0 × 10(2)~4.2 × 10(4) µg/kg), p,p'-DDE (5.1 × 10(2)~8.6 × 10(3) µg/kg), and HCB (7.6 × 10(1)~1.5 × 10(3) µg/kg) were the compounds found in highest concentrations. The hormonal activity of the porpoise blubber samples was also assayed in RGA, where two samples showed estrogenic activity, seven samples showed antiestrogenic activity, and one sample showed both estrogenic and antiestrogenic activity. Our results suggest that the 13 POPs measured by GC-MS in the samples cannot explain alone the estrogenicity of the extracts.

Effect of storage and cooking on the fatty acid profile of omega-3 enriched eggs and pork meat marketed in Belgium.

The fatty acids (FA) profile was determined in n-3 enriched (Columbus™) Belgian eggs and pork in order to evaluate to what extent the n-3 fatty acids, which are very sensitive to oxidation, are resistant to storage or cooking. In standard eggs or pork, no change of the fatty acid profile was observed after storage or cooking without culinary fat, as well as in Columbus™ eggs and pork after storage. Some cooking processes (eggs in custard and meat in oven) induced a slight significant loss of n-3 fatty acids in Columbus™ eggs or pork (11.1% in fat from eggs cooked in custard vs. 15.3% in raw Columbus™ eggs and 11.0% in fat from oven cooked meat vs. 11.6% in raw Columbus™ meat). As expected, when Columbus™ pork is cooked with culinary fat, its fatty acid profile is modified according to the nature of the fat used.

Monitoring Antibiotic Use and Residue in Freshwater Aquaculture for Domestic Use in Vietnam.

Vietnam is an important producer of aquaculture products, and aquatic products are essential to the Vietnamese diet. However, Vietnam also has very little enforced regulation pertaining to antibiotic usage in domestic aquaculture, which raises concerns for antibiotic resistance in pathogenic bacteria. In this study, analysis was conducted on the presence of antibiotic residues in domestically sold fish and shrimp raised in freshwater farms in Vietnam, and an assessment of farmers' knowledge of proper antibiotics usage was performed. The results indicated that a quarter of tested aquaculture products were antibiotic screening test positive, and there is a general lack of knowledge about the purpose and proper usage of antibiotics by aquaculture producers. Farmers' decision-making processes about antimicrobial use are influenced by biased sources of information, such as drug manufacturers and sellers, and by financial incentives.

Effect of fuel and kiln type on the polycyclic aromatic hydrocarbon (PAH) levels in smoked shrimp, a Beninese food condiment.

Smoked shrimp is a food condiment widely used in Beninese local cooking practices. A previous study revealed that this product is highly contaminated with polycyclic aromatic hydrocarbons (PAH). The present study explored possibilities to reduce PAH levels in shrimp smoked using cottage industry smoking techniques with barrel and chorkor kilns, by replacing wood by charcoal from Acacia auriculiformis and Mangifera indica, as fuels. Results showed that only shrimp smoked using acacia charcoal in a chorkor kiln had PAH levels (benzo[a]pyrene = 5 µg kg(-1) and sum of benzo[a]pyrene, chrysene, benzo[a]anthracene and benzo[b]fluoranthene = 28 µg kg(-1)) in accordance with a European standard of 5 and 30 µg kg(-1), respectively, and suitable physicochemical characteristics for good storage (moisture content = 11.9% ± 1.5%; water activity = 0.46 ± 0.03). However, further investigations still needs to be done to reduce the duration of product contact with combustion gasses in order to reduce the PAH content of smoked shrimp to safer levels, largely below standards.

Understanding the gap between the estrogenicity of an effluent and its real impact into the wild.

To study the reliability between in vitro and in vivo data collected downstream 2 sewage treatment plants (STP) as well as from bleached kraft mill industry (BKME), 5 rivers (3 impacted and 2 references) were investigated in the Walloon region (southern of Belgium). For the in vitro part of the work, water samples were collected to measure the estrogenicity of the 'out' effluent compared to reference sample point by MCF-7 assay. Results indicated significant estrogenicity of effluents from STP and BKME and a weak estrogenicity in reference sites. However, estradiol equivalents (EEQ) estimated into rivers were probably too low to impact wild population. Chemical analysis of 13 compounds of interest indicated that extraction procedure used in this study gave low recoveries of estrogen-like xenobiotics, leading to probably under-estimated MCF-7 responses. Surprisingly, a full scan mode has revealed an unexpected compound in the sample of BKME which was: 7-isopropyl-1,1,4a-trimethyl-1,2,3,4a,9,10,10a-octahydrophenanthrene, a product of pulp mill manufacture. In parallel to in vitro, in vivo assessment of estrogenic impact of effluent was followed on the gudgeon (Gobio gobio). Samples were achieved during 2 different periods of the reproductive cycle, resting period (RP) and pre-spawning period (pSP). Unspecific physiological parameters to estrogenic exposure (gonadosomatic index and systematic testis cell counting) displayed no significant differences related to endocrine disruption of the reproductive tract, only differences were correlated with the reproductive state of fish (RP versus pSP). Concerning the potent biomarker of estrogen exposure, vitellogenin (vtg), only basal induction was revealed but not related to estrogenic exposure. Nevertheless, vtg over-expression was found for male fish presenting a feminization of the reproductive tract captured downstream the STP station of Wégnez in the Vesdre River. Intersexuality, another indicator of the estrogenicity impact in fish, was observed in every site. Actually, ovotestis was systematically formed by protoplasmic oocyte observed in low percentage in every group analysed (impacted and references). Moreover, in fish captured in Wégnez, oocyte diameter was significantly higher compared to the other groups. In this study, only moderate to none impact in population of gudgeon was noticed. Moreover, in this case no discrepancy between in vitro and vivo was viewed although both approaches revealed gaps in monitoring effluent incidence into the environment. We should remain careful in the interpretation when only partial approaches are used in order to characterize impact in the aquatic milieu.

Analysis of EU priority polycyclic aromatic hydrocarbons in food supplements using high performance liquid chromatography coupled to an ultraviolet, diode array or fluorescence detector.

High performance liquid chromatography coupled to an ultraviolet, diode array or fluorescence detector (HPLC/UV-FLD) has been used to set up a method to detect the 15(+1) EU priority polycyclic aromatic hydrocarbons (PAHs) in food supplements covering the categories of dried plants and plant extracts excluding oily products. A mini validation was performed and the following parameters have been determined: limit of detection, limit of quantification, precision, recovery and linearity. They were in close agreement with quality criteria described in the Commission Regulation (EC) No 333/2007 concerning the PAH benzo[a]pyrene in foodstuffs, except the not fluorescent cyclopenta[c,d]pyrene for which the UV detection leads to a higher limit of detection. Analysis of twenty commercial food supplements covering mainly the class of dried plants was performed to evaluate their PAHs contamination levels and to test the applicability of the method to various plant matrices. Fifty percent of analyzed samples showed concentration exceeding 2 microgkg(-1) for one or more PAHs.