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PubMed2005

Liquid chromatographic tandem mass spectrometric determination of five coccidiostats in poultry eggs and feed.

A method is described which permits the quantitative detection of the chemical coccidiostats halofuginone, robenidine, diclazuril, nicarbazin and dimetridazole and its main metabolite 2-hydroxydimetridazole in poultry eggs and feed. Sample preparations were kept very simple and are based upon extraction with an organic solvent. Sample extracts were injected into the liquid chromatography tandem mass spectrometry (LC-MS/MS) system on a C18 column and a gradient elution was performed. Dimetridazole-D3 and diclazuril-bis, a structural analogue of diclazuril, were used as internal standards. Detection was performed on a triple quadrupole mass spectrometer in the selected reaction monitoring mode after ionisation in the positive or negative electrospray ionisation mode. Argon was applied as collision gas for collision induced dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC [Official Journal of the European Communities L221 (2002) 8].

Animal Feed↗
PubMed2005

Adsorption of trace elements from poultry litter by montmorillonite clay.

Poultry litter (PL) is used as fertilizer on agricultural lands because of its high nutrient content. However, the litter also contains trace elements such as As, Cd, Cu, Pb, and Zn. On land application of PL, these trace elements may be absorbed by crops, leach into groundwater, or enter the aquatic system as run-off. The objective of this research was to study the effect of the addition of montmorillonite clay-mineral (CM) in reducing the release of trace elements from PL. Cd, Cu, and Zn showed significant decreases of 29, 34, and 22%, respectively, in PL aqueous leachate (compared with the control-PL without CM) on mixing with 0.05 g CM but no change in As, Co, and Cr concentrations was observed. Lead showed a significant increase in PL aqueous leachate on mixing with 0.2 g CM but Pb concentration was two orders of magnitude less than in CM aqueous leachate alone. On washing, the settled precipitate (PL+CM) in the centrifuge tubes with water (desorption study) most of the adsorbed metals (Cd 85%, Cu 61%, and Zn 100%) were released. The results of this study show that the addition of CM resulted in significant adsorption of Cd and Cu from PL.

Adsorption↗
PubMed2004

Molecular characterization of multidrug-resistant Enterococcus spp. from poultry and dairy farms: detection of virulence and vancomycin resistance gene markers by PCR.

Thirty multidrug-resistant Enterococcus spp. strains, including two from the milk of cows with mastitis, nine from chicken litter and 19 from turkey litter, were isolated. Twenty-five were identified by biochemical methods as E. gallinarum and five as E. faecalis. Most of the isolates were resistant to vancomycin, gentamicin, streptomycin, tetracycline, erythromycin, bacitracin, kanamycin and nalidixic acid but sensitive to ciprofloxacin, sulfamethoxazole, chloramphenicol, ampicillin and ofloxacin. Attempts were made by partial amplification of the gene sequences to detect the vancomycin resistance markers vanA (734-bp), vanB (420-bp), vanC1 (531-bp), and vanC2-C3 (673-bp); virulence markers cylA (427-bp) and cylB (225-bp) for enterococcal cytolysin and a biofilm-forming surface protein (Esp). Individual and multiplex-PCR assays for vancomycin resistance markers revealed the vanC1 gene in 22 E. gallinarum strains. None of the remaining isolates including five E. faecalis strains (MIC=2 microg ml(-1)) and three E. gallinarum strains (MIC=8 microg ml(-1)) had any of the van genes tested. Analysis by pulsed-field gel electrophoresis (PFGE) and a comparison of smaI banding profiles showed 11 different patterns. Probing with a DIG-labeled vanC1 PCR product indicated a common 38.0 kb SmaI DNA fragment in all the E. gallinarum strains harboring the vanC1 gene. The genes cylA and cylB were detected only in one clinical E. gallinarum isolate and two quality control clinical strains of E. faecalis (ATCC 51299 and 29212). None of the virulence factors were found in milk or poultry isolates. Intermediate level resistance to vancomycin in enterococci from the US animal farms was predominantly due to the presence of vanC1 gene.

Animals↗
PubMed2005

Scanning electron microscopy of foreleg tarsal sense organs of the poultry red mite, Dermanyssus gallinae (DeGeer) (Acari:Dermanyssidae).

Scanning electron microscopy (SEM) studies on the poultry red mite, Dermanyssus gallinae (DeGeer), showed that the tip of the foreleg tarsus of both sexes and early instar stages (larvae, protonymph and deutonymph) bears a cluster of 10 blunt peg-type sensillae. Diversity in size, shape and surface features of the wall of these sensillae characterize the sensilla complex (sensory field). Two short setae (1 and 2) with smooth walls lack detectable pores, whereas three medium-sized setae (4, 5 and 8) bear pores in the wall. Four medium or long setae (3, 6, 7 and 9) have longitudinal grooves or ribs in the wall, whilst one seta (10) possesses both pores and grooves in the wall. Based on the morphology, it is suggested that the porous setae could be olfactory receptors, and the grooved setae could be mechanoreceptors.

Animals↗
PubMed2004

Evaluation of an automated immunomagnetic separation method for the rapid detection of Salmonella species in poultry environmental samples.

Automated immunomagnetic separation (AIMS), using Dynabeads anti-Salmonella (Dynal, Oslo), was evaluated for its ability to detect Salmonella spp. in poultry environmental samples in comparison with standard, culture-based method (Health Canada, Health Protection Branch, MFHPB-20). AIMS was found to be more reliable in detecting Salmonella from artificially inoculated enrichment broths at low levels and exhibited a 15.5% higher sensitivity value than the culture method.

Animals↗
PubMed2005

Evaluation of an antimicrobial resistance monitoring program for Campylobacter in poultry by simulation.

An ideal national resistance monitoring program should deliver a precise estimate of the resistance situation for a given combination of bacteria and antimicrobial at a low cost. To achieve this, decisions need to be made on the number of samples to be collected at each of different possible sampling points. Existing methods of sample size calculation can not be used to solve this problem, because sampling decisions do not only depend on the prevalence of resistance and sensitivity and specificity of resistance testing, but also on the prevalence of the bacteria, and test characteristics of isolation of these bacteria. Our aim was to develop a stochastic simulation model that optimized a national resistance monitoring program, taking multi-stage sampling, imperfect sensitivity and specificity of diagnostic tests, and cost-effectiveness considerations into account. The process of resistance testing of Campylobacter spp. isolated from cloacal swab samples from poultry was modeled using a Markov Chain Monte Carlo model. Different sampling scenarios on the number of flocks to be tested, the number of birds from each flock, and the number of campylobacter colonies submitted to susceptibility testing were evaluated regarding the precision of the resulting prevalence estimate. Precision of the prevalence estimate was defined as the absolute difference between apparent and true prevalence of resistance. A partial budget approach was utilized to find the most cost-effective combination of samples to obtain a defined precision of the prevalence estimate. For a sampling scenario testing 100 flocks, five birds per flock, and one campylobacter colony per sample, the median error of the prevalence estimate was 2.5%, and 95% of the simulations resulted in an error of 7% or less. When the total number of samples was kept constant, maximizing the number of flocks tested, and only testing one bird per flock resulted in the most precise prevalence estimate. Submitting more than one campylobacter colony to resistance testing did not improve the prevalence estimate. Partial budget analysis indicated that the most cost-effective strategy was testing of two birds per flock, and submitting one colony per sample to resistance testing.

Animals↗
PubMed2026

Population structure and antibiotic resistance of Salmonella isolates from diseased poultry in Jiangxi Province, China.

Salmonella poses a significant threat to human and animal health. However, the relationship among population diversity, antibiotic resistance, and infection risk remains largely unexplored. In this study, 69 Salmonella strains were isolated from diseased poultry in Jiangxi Province from 2021 to 2024. Using whole-genome sequencing, serotype prediction, MLST, virulence and resistance gene analysis, antibiotic susceptibility testing, and mobile genetic element annotation, we characterized the diversity, resistance profiles, and transmission mechanisms of these strains. The results showed high diversity, with Salmonella enterica subsp. enterica serovar Typhimurium (>60%) and ST19 (62.31%) as the dominant serovar and sequence type, respectively. Several avian isolates were genomically similar to human isolates, indicating potential zoonotic risk. All strains harbored conserved core virulence modules, whereas accessory modules (e.g., cdtB, astA, pefA) varied and may affect pathogenicity. The multidrug resistance rate was 97.1%, with 100% resistance to erythromycin, tilmicosin and tiamulin, and resistance rates of 91.3%, 84.1%, and 71.0% to sulfonamides, enrofloxacin, and ceftiofur, respectively. Sixty-eight resistance genes were identified. Highly conserved antimicrobial resistance gene (ARG) modules (e.g., sul2-aph(3″)-Ib-aph(6')-Id-tet(A)) were shared between chromosomes and plasmids and were flanked by mobile elements such as Tn3 and IS3. Genomic islands (GIs) and plasmids in some strains carried resistance gene clusters highly homologous to those in pathogens from humans, pigs, and chickens, suggesting active horizontal transfer of resistance genes across hosts. This study revealed high diversity, prevalent multidrug resistance, and active horizontal transfer of resistance genes in avian-derived Salmonella from Jiangxi Province, emphasizing the need for cross-host resistance monitoring and antibiotic management within the 'One Health' framework.

Horizontal gene transfer↗
PubMed2004

High prevalence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain.

Salmonellosis is a major foodborne infection in Spain, and strains that are resistant to a great variety of antibiotics have become a major public health concern. The aim of this study was to determine the level of antibiotic resistance in 133 Salmonella isolates obtained from a poultry slaughterhouse in Zaragoza (NE Spain). Antimicrobial resistance testing was performed by disk diffusion method using 19 antibiotics. Results were interpreted following the NCCLS criteria. Overall, the highest percentage of resistance was found to the following antimicrobial agents: sulfadiazine (96.2%), neomycin (53.4%), tetracycline (21.8%), and streptomycin (11.3%). All isolates were found to be resistant to one or more of the antibiotics tested. Multiple resistance was observed in 87 strains (65.4%). We found 23 different patterns of resistance in Salmonella Enteritidis. Resistance to sulfadiazine was the most common single resistance. The most frequent patterns of multiresistant strains were neomycin+sulfadiazine and neomycin+tetracycline+sulfadiazine. S. 4,5,12:b: showed the highest percentages of resistance to the tested drugs, with five different resistance patterns found. Ampicillin+chloramphenicol+streptomycin+sulphonamides+tetracycline (ACSSuT) resistance pattern, commonly associated with S. Typhimurium DT 104, was not detected in strains of the same phage type from broilers. The appearance of substantial multiresistance in foodborne Salmonella isolates suggests the need for more prudent use of antibiotics by farmers, veterinarians, and physicians.

Abattoirs↗
PubMed2004

Molecular epidemiology of avian pathogenic Escherichia coli (APEC) isolated from colisepticemia in poultry.

The molecular biology and epidemiology of 150 avian pathogenic Escherichia coli strains (APEC) isolated from septicemic poultry in Germany was investigated by serotyping, pulsed field gel electrophoresis (PFGE), and polymerase chain reaction (PCR). Only 49.6% of the isolates could be grouped to serogroups O1, O2, and O78. Macrorestriction analyses data revealed two large clonal groups (clusters I and II) among the APEC strains with a similarity of 60.9% to each other. An association between restriction pattern and serogroup or origin of the strains was only present in a few subgroups of each clusters I and II, but was not evident. In contrast, our data revealed distinct combinations of virulence-associated genes in that 51.2% of the O2-strains harboured a combination of the genes fyuA, irp2, iucD, tsh, vat, fimC, and colV and 36.4% of the O78-strains possessed the same gene combination with exception of vat. With 34 different gene combinations the non-O1, -O2, -O78 isolates revealed a higher variability in their virulence gene pattern than O1-, O2-, and O78-strains with 6, 13, and 9 patterns, respectively. Our data indicate only a limited association between the virulence gene pattern and the serogroup of APEC strains and question the sensitivity of O-typing for APEC identification without the application of further diagnostic tools. Although a limited number of APEC clones exist, horizontal gene transfer seems to be common in these pathogens. These findings strengthen further research on the population structure of APEC and may be the reason for the lack of clear definition of this common E. coli pathotype.

Animals↗
PubMed2005

Sandwich ELISA detection of Clostridium perfringens cells and alpha-toxin from field cases of necrotic enteritis of poultry.

Sandwich ELISAs (sELISAs) for the detection of Clostridium perfringens cells and alpha-toxin were developed and used to screen intestinal samples from normal broiler chickens and from clinical cases of necrotic enteritis. The assays clearly distinguished between the two sets of samples. The sELISA absorbance values from samples obtained from the majority of healthy birds were low and those from the majority of necrotic enteritis cases were high. Together, the assays provide a suitable test for the rapid screening for the diagnosis of necrotic enteritis in poultry.

Animals↗
PubMed2000

Determination of four nitroimidazole residues in poultry meat by liquid chromatography-mass spectrometry.

A multi-residue liquid chromatography-mass spectrometry (LC-MS) assay method is described for the determination of four nitroimidazoles in poultry muscle. The extraction procedure is based on liquid-liquid extraction with ethyl acetate followed by an evaporation step. A deuterated internal standard is used. The LC separation was made on a C18 bonded silica column with an aqueous formic acid (0.2%) solution-methanol-acetonitrile (81:13:6) mobile phase. Following electrospray ionization, the protonated molecular ion [M+H]+ is obtained for each compound. Monitoring several ions for each nitroimidazole provides the specificity required for confirmatory assay. Validation of the method was performed to estimate linearity, intra-day and inter-day repeatability, accuracy and detection limit. The present method is capable of identifying nitroimidazole residues in muscle at levels below 5 microg/kg.

Animals↗
PubMed1997

Procedure for the determination of eight cholesterol oxides in poultry meat using on-column and solvent venting capillary gas chromatography.

A procedure for the determination of eight relevant cholesterol oxides in poultry meat has been developed. The method consists of the enrichment of cholesterol oxides by means of the combined use of solid-phase fractionation and thin-layer chromatography. Florisil and silica columns of 10 g permitted the handling of the total cholesterol oxides content included in the lipid bulk obtained after the Folch's extraction of 20 g of muscle meat. The determination of cholesterol oxides under their trimethylsilyl derivatives was performed by using capillary gas chromatography. The use of a fused-silica open tubular capillary column 30 m x 0.25 mm I.D. coated with 5% phenylmethylsilicone and with a film width of 0.25 micron permitted the separation of all the species. Two modes of injection (on-column and solvent venting) were evaluated and compared for the analysis of cholesterol oxides. On-column capillary gas chromatography (cGC) gave better absolute areas relative standard deviation (R.S.D.) values: 3% to 6% vs. 5% to 7% for solvent venting cGC. Regression analysis for each cholesterol oxide was performed for the two modes of injection. The possibility of large volume injection (10 microliters) by using the solvent venting mode was also evaluated in order to increase the sensitivity of the detection of cholesterol oxides. R.S.D. values for absolute areas ranging from 6% to 14% were obtained. The validation of the method was carried out within the range of 0.1-1 ppm. Absolute and relative recovery values ranging from 80% to 100% were obtained. Statistical analysis revealed that the method was reproducible. cGC-mass spectrometry was also used to confirm the peaks detected by cGC: the total ion chromatogram mode was used for the analysis of samples containing concentrations down to 0.1 ppm of cholesterol oxides. The analysis of fresh and cooked chicken meat revealed the presence of cholesterol oxides proceeding from the autoxidation of the cholesterol B-ring. Finally, saponification was found to be not as accurate as the described procedure for cholesterol oxides analysis.

Animals↗
PubMed1999

Liquid chromatography with ultraviolet detection of lasalocid, monensin, salinomycin and narasin in poultry feeds using pre-column derivatization.

A rapid and very effective analytical procedure for the simultaneous determination of four polyether antibiotics (PEs) lasalocid, monensin, salinomycin and narasin in poultry feeds was tested. PEs were extracted from samples using methanol and without and clean-up derivatized with 2,4-dinitrophenylhydrazine (DNP) in an acidic medium at 55 degrees C. The derivatization mixture was analyzed directly on an ODS column (150 x 4.6 mm, 5 microns) with methanol--1.5% aqueous acetic acid (90:10, v/v) as eluent and UV detection was carried out at 305/392 nm. The recoveries of the PEs from spiked samples were 85-100% with RSDs of 4-10% in a concentration range of 50-150 mg/kg.

Animal Feed↗
PubMed2000

Determination of nicarbazin and clopidol in poultry feeds by liquid chromatography.

A rapid and very effective analytical procedure for the simultaneous liquid chromatographic determination of two coccidiostats, clopidol (CLOP) and nicarbazin (NICA), in poultry feeds was developed and tested. The ground feed samples were extracted using aqueous dimethylformamide after moistening with water. Co-extracted feed constituents were removed with a solid-phase extraction alumina-basic column and the eluates were directly analyzed on an ODS column (250x4.6 mm, 5 microm) with acetonitrile-0.01 M acetate buffer (pH 4.6) as eluent. UV detection of CLOP and the 4,4'-dinitrocarbanilide portion of NICA was carried out at 265 and 345 nm, respectively. The mean recovery from NICA spiked samples was 95% with a RSD of 4% in a concentration range of 2-150 mg/kg while for CLOP it was 98% with a RSD of 5% in a concentration range of 5-150 mg/kg. The limits of detection of NICA and CLOP in feed, based on a detector signal-to-noise ratio of 3, were estimated to be 1 mg/kg and 2.5 mg/kg, respectively, and the lowest levels tested in feeds by this procedure were 2 mg/kg and 5 mg/kg, respectively.

Animal Feed↗
PubMed2002

Semi-continuous anaerobic digestion of solid poultry slaughterhouse waste: effect of hydraulic retention time and loading.

We studied the effect of hydraulic retention time (HRT) and loading on anaerobic digestion of poultry slaughterhouse wastes, using semi-continuously fed, laboratory-scale digesters at 31 degrees C. The effect on process performance was highly significant: Anaerobic digestion appeared feasible with a loading of up to 0.8 kg volatile solids (VS)/m3 d and an HRT of 50-100 days. The specific methane yield was high, from 0.52 to 0.55 m3/kg VS(added). On the other hand, at a higher loading, in the range from 1.0 to 2.1 kg VS/m3 d, and a shorter HRT, in the range from 25 to 13 days, the process appeared inhibited and/or overloaded, as indicated by the accumulation of volatile fatty acids and long-chain fatty acids and the decline in the methane yield. However, the inhibition was reversible. The nitrogen in the feed, ca. 7.8% of total solids (TS), was organic nitrogen with little ammonia present, whereas in the digested material ammonia accounted for 52-67% (up to 3.8 g/l) of total nitrogen. The TS and VS removals amounted to 76% and 64%, respectively. Our results show that on a continuous basis under the studied conditions and with a loading of up to 0.8 kg VS/m3 d metric ton (wet weight) of the studied waste mixture could yield up to 140 m3 of methane.

Abattoirs↗
PubMed2002

Restoration of drastically eroded land using coal fly ash and poultry biosolid.

A 3-year field study was conducted at a 12 ha soil-borrow area adjacent to the Columbia Metropolitan Airport, South Carolina to investigate the restorative effects of co-application of coal fly ash (FA) and a poultry biosolid (PB). FA was applied at 0, 22, 280, 560 and 1120 Mg (tonne) ha(-1), and PB at 5 and 10 Mg ha(-1). The area was seeded with erosion-control species Atlantic Coastal panic grass (Panicum amarum var amarum L.), sericea (Lespedeza cuneata var. appalow [Dumont] G. Don.) and weeping love grass (Eragrostis curvula Wolf.). Plant biomass and elemental composition were analyzed in sequential harvests. Soil and groundwater quality characteristics including pH, EC and elemental composition were also monitored throughout the study. In addition, the effect of amendments on the water holding capacity and bulk density of the soil was investigated. Amendment addition significantly increased plant biomass production by a maximum of 26% using 1120 Mg ha(-1) FA and 10 Mg ha(-1) PB. Application of the highest rate of FA significantly increased the plant tissue concentrations of Mn, As, Se and B. Soil pH was initially increased from 4.6 to 6.1 by amendments. Soil salinity was increased in the initial year only. Amended soils had higher concentrations of Ca, Mg, P and K, higher organic matter content and water holding capacity than unamended soil. Concentrations of plant-essential trace elements (B, Cu and Zn) that were marginally deficient in the unamended eroded soil increased to within typical soil concentrations following amendment with FA and PB. Groundwater quality was unaffected throughout the study. The co-application of FA and PB successfully promoted the revegetation of the eroded borrow area with no apparent adverse environmental side effects.

Animals↗
PubMed2003

Comparison of phenotypic and genotypic characteristics of Salmonella bredeney associated with a poultry-related outbreak of gastroenteritis in Northern Ireland.

OBJECTIVES: To employ a combination of phenotypic and genotypic subspecies typing methods to aid in an epidemiological investigation of an outbreak of Salmonella bredeney involving ten persons. METHODS: Isolates were characterised by employing antibiogram typing, in addition to two genotyping techniques, including pulsed field gel electrophoresis (PFGE), random amplification of polymorphic DNA (RAPD) with two oligonucleotide primers. RESULTS: An outbreak of gastroenteritis associated with S. bredeney (serovar O:4 H:Lv 1,7) occurred in Belfast, Northern Ireland in November 1997. In total, ten cases were confirmed, of which eight had consumed chicken cooked at local butchers and retailed through one of two local bakeries. One of the remaining cases was secondarily infected within her home and the final case had eaten a product other than cooked chicken from one of the bakeries. Food preparation practices were inadequate in one of the bakeries in question and record keeping and possibly cooking procedures were inadequate in the butchers. S. bredeney was isolated from an uncooked chicken supplied to the butchers confirming that improperly cooked chicken was most likely the source of the outbreak. All outbreak clinical isolates were indistinguishable from each other and were similar to the isolate obtained from the uncooked poultry demonstrating that these DNA-based methods were valuable in the molecular characterization of S. bredeney. CONCLUSIONS: This report emphasises the importance and maintenance of an effective hazard analysis critical control point (HACCP) approach to the processing and retailing of foodstuffs containing chicken in order to help eliminate hazards to public health.

Adult↗