Usporedba osjetljivosti Screening metoda na ostatke antibiotika - The four plate test (FPT), Screening test na ostatke antibiotika (STAR) i Premi(R) test na standarde sulfonamida

The sensitivity of three microbiological antibiotic residue screening methods, the FPT, the STAR and the Premi®Test, were compared for the detection of 10 different standards from the sulphonamide (SA) group. Phtalylsulphathiazole (PHT), sulphadimidine (SD), sulphaguanidine (SG), sulphachlorpyridazine (SCHP), sulphamerazine (SRZ), sulphamethoxazole (SMX), sulphanilamid (SAM), sulphanilic acid (SAC), sulphaquinoxaline (SQ) and sulphathiazole (STZ) were tested using the concentrations from 0.05 μg.ml-1 to 1 μg.ml-1. The detection sensitivity of the methods represented by minimum inhibiting concentration (MIC) was evaluated. The MIC of SA standards detected by FPT was 0.1 μg.ml-1 for SRZ, 0.2 μg.ml-1 for SMX, SQ and STZ, 0.3 μg.ml-1 for SCHP, and 1 μg.ml-1 for SD. No detection sensitivity was observed for PHT, SG, SAC and SAM standards. The MIC of SA standards detected by STAR method was 0.05 μg.ml-1 for SCHP, SMX, SQ and STZ, 0.1 μg.ml-1 for SRZ, and 0.3 μg.ml-1 for SD. No detection sensitivity was observed for PHT, SG, SAC and SAM standards. The MIC of SA standards detected by Premi®Test was 0.05 μg.ml-1 for SD, SCHP, SMX, SQ and STZ, 0.1 μg.ml-1 for SRZ and PHT, and 0.3 μg.ml-1 for SG, SAC and SAM. The MICs represent the detection limit of the methods (LOD). The results of examinations showed that Premi®Test is the most sensitive method to sulphonamides followed by STAR method and FPT. Premi®Test detected six SA at the level of the maximum residue limit (MRL) 0.1 μg.ml-1 set for SA group, STAR method detected five SA at the level of MRL, and FPT detected only one SA at the level of MRL.U radu je uspoređena osjetljivost mikrobioloških screening metoda u određivanju rezidua 10 različitih standarda sulfonamida. Korišteni su phtalylsulphathiazole (PHT), sulphadimidine (SD), sulphaguanidine (SG), sulphachlorpyridazine (SCHP), sulphamerazine (SRZ), sulphamethoxazole (SMX), sulphanilamid (SAM), sulphanilic acid (SAC), sulphaquinoxaline (SQ) i sulphathiazole (STZ) u koncentracijama od 0.05 μg.ml-1 do 1 μg.ml-1. Minimalna inhibicijska koncentracija standarda utvrđena Four Plate Testom bila je 0.1 μg.ml-1 za SRZ, 0.2 μg.ml-1 za SMX, SQ i STZ, 0.3 μg.ml-1 za SCHP, te 1 μg.ml-1 za SD. Minimalna inhibicijska koncentracija standarda utvrđena STAR metodom bila je 0.05 μg.ml-1 za SCHP, SMX, SQ i STZ, 0.1 μg.ml-1 za SRZ, i 0.3 μg.ml-1 za SD. Ni jednom od navedenih metoda nije zabilježena inhibicija za standarde PHT, SG, SAC i SAM. Upotrebom Premi®Testa minimalna inhibicijska koncentracija bila je 0.05 μg.ml-1 za SD, SCHP, SMX, SQ i STZ, 0.1 μg.ml-1 za SRZ i PHT, te 0.3 μg.ml-1 za SG, SAC i SAM. Minimalne inhibicijske koncentracije predstavljaju prag detekcije metode. Rezultati su pokazali da je najosjetljivija metoda za određivanje ostataka sulfonamida Premi®Test, potom STAR metoda i na kraju Four Plate Test. Premi®Testom utvrđeno je 6 sulfonamida na razini propisane najviše dopuštene količine (NDK) od 0,1 0.1μg.ml-1, STAR metodom njih 5, a Four Plate Testom samo 1 sulfonamid

Usporedba osjetljivosti Screening metoda na ostatke antibiotika - The four plate test (FPT), Screening test na ostatke antibiotika (STAR) i Premi(R) test na standarde sulfonamida

The sensitivity of three microbiological antibiotic residue screening methods, the FPT, the STAR and the Premi®Test, were compared for the detection of 10 different standards from the sulphonamide (SA) group. Phtalylsulphathiazole (PHT), sulphadimidine (SD), sulphaguanidine (SG), sulphachlorpyridazine (SCHP), sulphamerazine (SRZ), sulphamethoxazole (SMX), sulphanilamid (SAM), sulphanilic acid (SAC), sulphaquinoxaline (SQ) and sulphathiazole (STZ) were tested using the concentrations from 0.05 μg.ml-1 to 1 μg.ml-1. The detection sensitivity of the methods represented by minimum inhibiting concentration (MIC) was evaluated. The MIC of SA standards detected by FPT was 0.1 μg.ml-1 for SRZ, 0.2 μg.ml-1 for SMX, SQ and STZ, 0.3 μg.ml-1 for SCHP, and 1 μg.ml-1 for SD. No detection sensitivity was observed for PHT, SG, SAC and SAM standards. The MIC of SA standards detected by STAR method was 0.05 μg.ml-1 for SCHP, SMX, SQ and STZ, 0.1 μg.ml-1 for SRZ, and 0.3 μg.ml-1 for SD. No detection sensitivity was observed for PHT, SG, SAC and SAM standards. The MIC of SA standards detected by Premi®Test was 0.05 μg.ml-1 for SD, SCHP, SMX, SQ and STZ, 0.1 μg.ml-1 for SRZ and PHT, and 0.3 μg.ml-1 for SG, SAC and SAM. The MICs represent the detection limit of the methods (LOD). The results of examinations showed that Premi®Test is the most sensitive method to sulphonamides followed by STAR method and FPT. Premi®Test detected six SA at the level of the maximum residue limit (MRL) 0.1 μg.ml-1 set for SA group, STAR method detected five SA at the level of MRL, and FPT detected only one SA at the level of MRL.U radu je uspoređena osjetljivost mikrobioloških screening metoda u određivanju rezidua 10 različitih standarda sulfonamida. Korišteni su phtalylsulphathiazole (PHT), sulphadimidine (SD), sulphaguanidine (SG), sulphachlorpyridazine (SCHP), sulphamerazine (SRZ), sulphamethoxazole (SMX), sulphanilamid (SAM), sulphanilic acid (SAC), sulphaquinoxaline (SQ) i sulphathiazole (STZ) u koncentracijama od 0.05 μg.ml-1 do 1 μg.ml-1. Minimalna inhibicijska koncentracija standarda utvrđena Four Plate Testom bila je 0.1 μg.ml-1 za SRZ, 0.2 μg.ml-1 za SMX, SQ i STZ, 0.3 μg.ml-1 za SCHP, te 1 μg.ml-1 za SD. Minimalna inhibicijska koncentracija standarda utvrđena STAR metodom bila je 0.05 μg.ml-1 za SCHP, SMX, SQ i STZ, 0.1 μg.ml-1 za SRZ, i 0.3 μg.ml-1 za SD. Ni jednom od navedenih metoda nije zabilježena inhibicija za standarde PHT, SG, SAC i SAM. Upotrebom Premi®Testa minimalna inhibicijska koncentracija bila je 0.05 μg.ml-1 za SD, SCHP, SMX, SQ i STZ, 0.1 μg.ml-1 za SRZ i PHT, te 0.3 μg.ml-1 za SG, SAC i SAM. Minimalne inhibicijske koncentracije predstavljaju prag detekcije metode. Rezultati su pokazali da je najosjetljivija metoda za određivanje ostataka sulfonamida Premi®Test, potom STAR metoda i na kraju Four Plate Test. Premi®Testom utvrđeno je 6 sulfonamida na razini propisane najviše dopuštene količine (NDK) od 0,1 0.1μg.ml-1, STAR metodom njih 5, a Four Plate Testom samo 1 sulfonamid

Verification for the presence of inhibitory substances in poultry meat after the consumption of the feed mixture supplemented with fermented feed

The European Union has an obligation to ensure that feed produced in the European Union is safe for animals and also humans by ensuring food of animal origin is safe and wholesome. An increasing demand for safe, wholesome and nutritious animal products has led to the search for alternative substances in animal feed. Fermented feed has gained a lot of popularity in many animal diets today. They meet the demand for animal nutrition due to the formation of target substances with the desired properties. As some of them are attracting attention as potential antimicrobial agents that inhibit the growth of certain microorganisms, and the products of animal origin are controlled for the presence of residues of inhibitory (antimicrobial) substances, the aim of this work was to verify the presence of inhibitory substances in poultry meat (muscle, heart, liver, kidneys of broiler chickens) after the consumption of the feed mixture with addition of fermented feed (wheat bran fermented with the strain Umbelopsis isabellina CCF 2412) in a dose of 10 % of the total amount of the feed. The detection of residues was performed by two approved microbiological screening methods, the screening test for the detection of antibiotic residues (STAR) and the Premi®Test. Both methods detected the positive results and pre-identified the presence of residues of the inhibitory substances not only in the meat of broiler chickens but also in the investigated fermented feed. Due to the antimicrobial potential of the fermented feed and the possible presence of the false-positive results, each positive result must be confirmed by a confirmatory analysis

Comparison of four microbiological inhibition tests for the screening of antimicrobial residues in the tissues of food-producing animals

The study compares two existing microbiological inhibition tests, Screening Test for Antibiotic Residues (STAR) and Premi®Test with two recently introduced tests, Nouws Antibiotic Test (NAT) and Total Antibiotics for the screening of antimicrobial residues in the tissues of food-producing animals. In the negative or positive sample classification based on inhibition of the growth of test strain sensitive to many antibiotics and sulphonamides, out of 142 samples obtained from slaughterhouses and retail operations, 39 samples yielded a positive result in one or more tests: 4 samples in four tests, 14 samples in three tests, 13 samples in two tests, and 8 samples in one test. As for the numbers of observed positive samples, the descending sequence of tests was: STAR, Total Antibiotics, Premi®Test, NAT. The growth inhibition was observed in three out of seven test strains, namely Bacillus cereus ATCC 11778, Kocuria rhizophila ATCC 9341, and Bacillus stearothermophilus var. calidolactis. Considering the test strains sensitivity and no inhibition on the Bacillus pumilus NCIMB 10822 NAT test plates, our preliminary conclusion is that the animal samples are suspected for the presence of tetracycline, macrolide, and b-lactam antibiotics

DETECTION OF LASALOCID RESIDUES IN THE TISSUES OF BROILER CHICKENS BY A NEW SCREENING TEST TOTAL ANTIBIOTICS

<p class="MsoNormal" style="text-justify: inter-ideograph; margin: 0cm 0cm 0pt; line-height: 150%; text-align: justify; mso-layout-grid-align: none;"><span style="font-size: small;"><span style="font-family: Times New Roman;"><span style="mso-ansi-language: EN-GB;" lang="EN-GB">The aim of the present study was to evaluate the microbial growth inhibition test Total antibiotics for the screening of lasalocid residues in the tissues of broiler chickens after its oral administration in medicated feed. The residues were investigated throughout the 5-day withdrawal period /WP/) and also on day 6 representing the first day following the WP. </span><span style="mso-ansi-language: EN-GB; mso-fareast-font-family: Batang; mso-fareast-language: KO;" lang="EN-GB">All broiler chicken tissues were positive for lasalocid. The breast muscle was positive (the presence of residues at/above the detection limit /LOD/ of method) up to day 1 of the WP, the thigh muscle, gizzard, heart, skin and fat up to day 3 of the WP and the liver and kidneys</span><span style="mso-ansi-language: EN-GB;" lang="EN-GB"> up to day 4 of the WP. When evaluating the dubious results (</span><span style="mso-ansi-language: EN-GB; mso-fareast-font-family: Batang; mso-fareast-language: KO;" lang="EN-GB">the presence of residues just below the LOD of method), the breast muscle was suspect positive up to day 3 of the WP and the gizzard, skin and fat up to day 4 of the WP. </span><span style="mso-ansi-language: EN-GB;" lang="EN-GB">No positive or dubious results were detected on day 5 of the WP. </span><span style="mso-ansi-language: EN-GB; mso-fareast-font-family: Batang; mso-fareast-language: KO;" lang="EN-GB">The LOD of </span><em style="mso-bidi-font-style: normal;"><span style="mso-ansi-language: EN-GB;" lang="EN-GB">Bacillus stearothermophilus </span></em><span style="mso-ansi-language: EN-GB; mso-bidi-font-style: italic;" lang="EN-GB">var. <em>c</em></span><em style="mso-bidi-font-style: normal;"><span style="mso-ansi-language: EN-GB;" lang="EN-GB">alidolactis</span></em><span style="mso-ansi-language: EN-GB; mso-bidi-font-style: italic;" lang="EN-GB"> </span><span style="mso-ansi-language: EN-GB; mso-fareast-font-family: Batang; mso-fareast-language: KO;" lang="EN-GB">for maduramycin was </span><span style="mso-ansi-language: EN-GB;" lang="EN-GB">500 &micro;g.l^{<span style="text-transform: uppercase;">-1</span>}<span style="text-transform: uppercase;">.<br /><br /><span style="color: #111111; font-family: Verdana, Arial, Helvetica, sans-serif; font-size: 9px; line-height: normal; text-transform: none;"><strong>doi:10.5219/140</strong></span></span></span></span></span></p

Effective screening of antibiotic and coccidiostat residues in food of animal origin by reliable broad-spectrum residue screening tests

Foods of animal origin are controlled for antibiotic and coccidiostat residues. The rapid residue detection is possible using reliable broad-spectrum screening tests. This study’s objective using four microbial inhibition tests for the detection and identification of antibiotic and coccidiostat residues in different foods of animal origin: Premi®Test, EXP Ampulle test, Milchtest and Screening Test for Antibiotic Residues (STAR). Four hundred and thirty (430) food samples (165 animal tissues, 152 raw cow’s milk and 113 eggs) were randomly collected and screened. Using the Premi®Test, 18 samples were positive and 6 samples dubious. Using the EXP Ampulle test, 31 samples were positive and 2 samples dubious. Using the Milchtest, 15 samples were positive and 12 samples dubious. Using the STAR, 65 samples were positive with 62 samples positive on plates specific for beta-lactams and sulphonamides, 4 samples on plates specific for aminoglycosides, 8 samples on plates specific for macrolides and beta-lactams; and 7 samples on the plates specific for tetracyclines. Retesting using penicillinase and para-aminobenzoic acid (PABA) to confirm the presence of beta-lactams or sulphonamides all potentially positive tube test samples revealed 21 samples positive for beta-lactams and 27 samples positive for sulphonamides. Further testing of sulphonamide positive chicken samples revealed the positivity for coccidiostat salinomycin which was confirmed by testing with PABA, which counteracting salinomycin inhibition. Three hundred and sixty six (366) animal food samples were negative for antibiotic and coccidiostat residues. Microbial inhibition tests are preferred for initial antibiotic screening and have also proven useful for coccidiostat screening and post-screening.HIGHLIGHTS Foodstuffs of animal origin are subject to controls on antibiotic and coccidiostat residues. Microbial inhibition tests are still the preferred choice for the initial screening of antibiotic residues in food matrices and could become a useful tool for the screening of coccidiostat residues also. More specific post-screening analysis with PABA proved unexpectedly to be a reliable tool in the preliminary detection of coccidiostat residues in poultry meat and eggs

Humic Substances as a Versatile Intermediary

Humic substances are organic ubiquitous components arising in the process of chemical and microbiological oxidation, generally called humification, the second largest process of the carbon cycle. The beneficial properties of these various substances can be observed in many fields of life and health, whether it is the impact on the human organism, as prophylactic as well as the therapeutic effects; animal physiology and welfare, which is widely used in livestock farming; or the impact of humic substances on the environment and ecosystem in the context of renewal, fertilization and detoxification. Since animal health, human health and environmental health are interconnected and mutually influencing, this work brings insight into the excellence of the use of humic substances as a versatile mediator contributing to the promotion of One Health

Detection of Gluten in Gluten-Free Foods of Plant Origin

The work deals with the issue of standardization and more accurate methodology for the isolation of gluten DNA in gluten-free products of plant origin, which is more demanding due to the more complex structure of plant cells. Three isolation methods were compared, of which the combination of glass and zirconium beads, Proteinase K and a commercially produced isolation kit was confirmed to be the most effective procedure. The given isolation procedure was more effective in one-component gluten-free foods, where the concentration of the obtained DNA ranged from 80.4 &plusmn; 0.7 to 99.0 &plusmn; 0.0 ng/&micro;L. The subsequent PCR reaction revealed the presence of gluten not only in guaranteed gluten-free products (40%), but also in naturally gluten-free foods (50%). These were mainly gluten-free sponge cakes, gluten-free biscuits &ldquo;Cranberries&rdquo;, cocoa powder, coffee &ldquo;3in1&rdquo;, and instant coffee

Effect of Cladosporium cladosporioides on the Composition of Mycoflora and the Quality Parameters of Table Eggs during Storage

The eggshells of 120 experimental one-day-old table eggs were contaminated with the spore suspension of Cladosporium cladosporioides, divided into three groups (A–C) and stored at three different temperatures (3 °C, 11 °C and 20 °C) for 28 days. Visible growth of molds on/in experimental eggs was not observed within the entire storage period. No significant differences in the numbers of molds were found between particular groups of eggs. However, the composition of egg mycoflora was greatly influenced by storage conditions. Three mold genera were identified using the PCR method. The highest mold numbers were determined on Day 14 (Groups A and C) and Day 21 (Group B) when the maximum relative humidity and dew point temperature were recorded. On the same days, the dominance of Penicillium spp. and the minimum eggshell firmness were observed. Noticeable changes in egg quality were observed in eggs stored at 20 °C, and most of these eggs were downgraded at the end of storage period. The growth ability differed significantly among three mold genera. Penicillium spp. and Fusarium spp. showed better growth intensity at increased values (0.91–0.94) of water activity (aw) indicating a possible risk associated with the occurrence of mycotoxins in the egg contents