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Comparison of pulsed-field gel electrophoresis and phage typing for discriminating poultry strains of Staphylococcus aureus.

OBJECTIVE: To compare pulsed-field gel electrophoresis (PFGE) patterns of Staphylococcus aureus from chickens in England, Belgium, Bulgaria, Argentina, and Japan, to assess the value of PFGE for discriminating strains, and to compare results obtained by PFGE with those obtained by biotyping and phage typing. SAMPLE POPULATION: 78 S aureus isolates from diseased and healthy chickens. PROCEDURE: Chromosomal DNA of S aureus was digested with restriction endonuclease Sma I, and fragments were separated by PFGE in 1% agarose gel. RESULTS: All 78 strains from 5 countries were classified as poultry ecovar according to a previously established biotyping system. Chromosomal DNA was cut by Sma I into 18 to 23 fragments ranging from about 3 to 685 kb. Seventy-eight strains produced 15 types, arbitrarily designated A to O, and 45 subtypes. Some differences were observed in PFGE patterns among countries. However, 10 fragments (333, 190, 110, 63, 55, 42, 34, 19, 10, and 3 kb) were highly conserved and were shared by almost all (> 78%) of the strains examined. The PFGE patterns were compared with those obtained by phage typing. All 29 strains belonging to avian phage-group II produced type A and 19 subtypes. Of the 15 strains belonging to phage-group I, 11 produced 8 types (B to H, O) and 5 subtypes that were different from those of type A. CONCLUSIONS: Genomic DNA fingerprinting by PFGE is an effective technique for discriminating poultry S aureus strains and appears to be a useful method for subtyping strains of avian phage groups or the poultry-specific ecovar.

Animals↗

Analysis of aflatoxins in poultry and pig feeds and feedstuffs used in Colombia.

Feedstuffs and mixed feeds used for poultry and pig nutrition in Colombia were analyzed for aflatoxins by using a liquid chromatographic technique with a limit of detection of 1 microgram/kg for each aflatoxin (B1, B2, G1, and G2). Samples of grain sorghum, maize, processed soybean, rice meal, cottonseed meal, and poultry and pig feeds, representative of Colombian production for the 1995-1996 harvest, were taken from feed-manufacturing plants in various cities. Aflatoxins were detected in 11 of 45 samples of sorghum, 4 of 33 samples of maize, 8 of 22 samples of rice meal, 15 of 17 samples of cottonseed meal, 1 of 12 samples of other feedstuffs, 12 of 30 samples of poultry feed, and 7 of 16 samples of pig feed. Aflatoxins were not detected in soybean. Only 9 of 58 positive samples contained total aflatoxin levels exceeding maximum tolerable limits in Colombia.

Aflatoxins↗

Genomic characterization of avian metapneumovirus subtypes A and B in United States poultry by targeted amplicon sequencing.

Avian metapneumovirus (aMPV) subtypes A and B emerged in United States poultry in late 2023 and early 2024, prompting genome-scale surveillance from clinical samples. Here, we developed and optimized targeted amplicon sequencing (TAS) assays for both subtypes and applied them to 104 subtype-positive clinical samples collected from chicken and turkey farms across nine US states between early 2024 and early 2026. TAS recovered 91 genomes suitable for comparative analysis, including 44 aMPV-A and 47 aMPV-B sequences, with successful recovery extending to Ct values of 34.6 for aMPV-A and 31.2 for aMPV-B. Recovered genomes showed near-complete breadth and high mapping efficiency. Phylogenetic analyses of both G-gene and whole-genome datasets showed that US aMPV-A field strains formed a distinct monophyletic lineage within group IV and resolved into three closely related clusters. Cluster 2, first recognized in North Carolina and later detected in Ohio, spread across 30 turkey and chicken farms. Cluster 2 genomes were defined by a concentrated G-protein hotspot within residues 209-275, and most North Carolina Cluster 2 genomes carried 10-11 nonsynonymous substitutions in this region, including multiple proline substitutions suggestive of local structural change. Missouri Cluster 3 remained cohesive but distinct from both Cluster 1 and Cluster 2 in the G-gene and whole-genome trees. In contrast, US aMPV-B field strains remained highly homogeneous across hosts and states, with more than 99% nucleotide identity by both G-gene and WGS analyses. We also identified 12 vaccine-derived genomes on both vaccinated and nonvaccinated farms. These included six genomes related to aMPV-A vaccine and six related to aMPV-B vaccines (VCO3/50 and 1062), all of which retained vaccine-defining markers together with additional substitutions consistent with continued circulation after vaccine use in the field. Selection analyses showed that the G gene had the highest gene-wise dN/dS ratio in both subtypes. Additional elevated signal was observed in SH and M2, and candidate positively or episodically selected codons were concentrated in the subtype A Cluster 2 G-gene hotspot. These findings show that TAS supports direct-from-sample aMPV genomic surveillance and provides genomic context for field clusters, vaccine-derived lineages, and continued adaptive change in aMPV in US poultry.

Animals↗

Ammonia emissions from cattle, pig and poultry wastes applied to pasture.

In four field experiments, carried out in The Netherlands, small wind-tunnels were used to make direct measurements of ammonia (NH(3)) volatilization from different types of slurry and manure applied to the surface of grassland. During periods of up to six days following application, losses of NH(3)-N often amounted to more than 40% of the NH(4)-N applied. Percentage loss was highest (83%) from a poultry slurry and least (21%) from an air-dried poultry manure. Losses of NH(3)-N were generally greater from pig slurry (36-78%) than from cattle slurry (41%). In most cases 80% or more of the total NH(3)-N loss occurred within 48 h of application. Estimates were made of total annual NH(3) emissions from four systems of poultry housing. The highest total loss (50% of the N voided in droppings) occurred with a battery house producing a slurry with a low content of dry-matter; most of the loss took place after spreading. With a second battery house, in which the droppings were air-dried, the total loss was only 12%, with much lower emissions from the housing and during spreading.

Journal Article↗

Chlorinating poultry chiller water: the generation of mutagens and water re-use.

The purpose of this study was to assess the potential generation of mutagens by chlorination of water used to chill poultry carcasses. Unchlorinated chiller water from a nearby poultry processing plant was chlorinated to different levels in the laboratory. Residues of dichloromethane extracts of the treated water were tested for mutagenicity with the Ames Salmonella reverse mutation system. The rate of disappearance of added chlorine from the chiller water was also monitored using ferrous ammonium sulphate titration. The results showed a fast rate of disappearance of added free chlorine, only some of which reappeared as combined available chlorine. No mutagenic activity could be demonstrated at chlorination levels up to 100 ppm, but activity rose above 250 ppm, and a dose-response relationship was demonstrated. The significance of these findings for poultry processing is discussed.

Animals↗

Copper and Zn uptake by radish and pakchoi as affected by application of livestock and poultry manures.

Environmental safety of agricultural utilization of livestock and poultry manures from intensive farming is attracting great attention because the manures often contain high concentrations of heavy metals and organic pollutants. Pot experiments, in which a pig manure (PM), a chicken manure (CM) and a commercial organic manure (OM) with different concentrations of Cu and Zn to simulate soil metal accumulation by manure application for different times were utilized in a garden soil at a rate of 2% (W/W), were conducted to study the effect of application of these livestock and poultry manures on growth of radish (Raphanus sativus L.) and pakchoi (Brassica chinensis L.) as well as their Cu and Zn uptake. The results exhibit that the manures except the PM improved the growth of radish and pakchoi. The difference of biomass among the same manure treatments containing different concentrations of Cu and Zn, however, was insignificant. In addition, application of the livestock and poultry manures significantly increased soil pHs and electric conductivities (EC) compared with the control, which is ascribed that these manures had high pH and contained large amounts of inorganic ions. The available soil Zn concentrations in the PM were higher than that in the CM and OM, and the extractable soil Cu concentrations in the three manures were almost the same after radish growth in the garden soil but were different after pakchoi growth. Zinc and Cu concentrations in the radish and pakchoi tissues increased when the soil Zn and Cu concentrations increased by manures application, but were still within a safe value. An except is the treatment PM4 in which the Zn concentration of the above-ground part of radish was 28.7 mg kg-1, exceeding the Chinese Food Hygiene Standard of 20 mg kg-1 based on fresh weight. Good correlation was obtained between the extractable soil Zn (or Cu) concentrations extracted by 1.0 mol l-1 NH4NO3 and the Zn (or Cu) concentrations in radish and pakchoi tissues, which was expected to be effective in forecasting Cu and Zn availability to radish and pakchoi in manure agronomic utilization.

Animals↗

Use of poultry litter for biodegradation of soil contaminated with 2,4- and 2,6-dinitrotoluene.

Pseudomonas sp. and Pseudomonas putida can utilize dinitrotoluene (DNT) as N-source after the enzymatic removal of nitro groups from the aromatic ring. Addition of nutrients is known to stimulate the biodegradation process. Poultry litter has consortia of microorganisms (including Pseudomonas) along with many nutrients. The objective of this research was to study the biodegradation of 2,4- and 2,6-DNT contaminated soil (from Badger Army Ammunition Plant) using poultry litter. Complete biodegradation of both 2,4- and 2,6-DNT in the soil was observed after 1-day interaction with poultry litter. No degradation was observed using autoclaved litter.

Air Pollutants, Occupational↗

Evaluation of aluminosilicate compounds to reduce aflatoxin residues and toxicity to poultry and livestock: a review report.

The aflatoxins (AFs) are reported to be hepatotoxic, mutagenic, immunosuppressive, and carcinogenic. Methods to prevent, reduce, or remediate AF toxicity and residues in the environment are in great demand. Various AF-detoxification procedures are reviewed with particular emphasis on ammoniation and the use of adsorbent compounds to bind AF. A series of in vivo experiments by the authors are reviewed that evaluated the ability of a specific hydrated sodium calcium aluminosilicate (HSCAS) adsorbent to reduce the toxicity of AF to poultry and livestock and to reduce AF residues in milk. These studies showed that HSCAS forms stable bonds with AF in vitro, and when added to AF-contaminated poultry and livestock feeds, HSCAS is able to protect chickens, swine, and lambs from the deleterious toxic effects of AF and to reduce AF residues in milk of dairy cows and goats. These results indicate that HSCAS, when used in conjunction with other mycotoxin management practices, may prove effective for the preventive management of AF-contaminated feedstuffs in livestock and poultry and may reduce AF residues in the food-chain.

Adsorption↗

Ornithosis in poultry workers.

An outbreak of ornithosis in duck workers in the winter of 1979 and spring of 1980 was discovered by the investigation of a cluster of cases in Norfolk. A serological survey showed that 61% of duck workers but only 23% of control poultry workers had chlamydia group antibody titres of greater than or equal to 1:8. Altogether 9% of duck workers in the survey had antibody titres greater than or equal to 1:32 and a clinical illness suggestive of ornithosis. The proportions of seropositive tests and clinical attack rates were highest in workers eviscerating ducks and lowest in farm workers. It is suggested that a clinical history of contact with poultry should be considered relevant in the diagnosis of ornithosis and that clinicians caring for poultry workers should consider the possibility of ornithosis as an occupational disease.

Adult↗

VanA-type vancomycin-resistant enterococci (VRE) remain prevalent in poultry carcasses 3 years after avoparcin was banned.

Avoparcin was used as a growth promoting feed additive in Norwegian broiler and turkey production from 1986 until it was banned in 1995, when an association between vancomycin-resistant enterococci (VRE) and avoparcin use became apparent. A recent study regarding faecal samples documented a continuing high prevalence of VRE among Norwegian poultry 3 years after avoparcin was banned. In the present study, carcasses of broilers and turkeys from farms where avoparcin had previously been in use and carcasses of layer chickens from farms where avoparcin had never been used were examined for the presence of VRE. One carcass from each of 150 different farms was included. By a direct plating method, VRE were isolated from 30 of 100 samples of broilers and turkeys, but not from any samples of layer chickens. When an enrichment step was included, VRE were isolated from a total of 81 of the 100 samples of broilers and turkeys and from nine of the 50 samples of layer chickens. All VRE isolated were highly resistant to vancomycin (MIC > or = 256 microg/ml) and possessed the vanA gene. These results correspond to the prevalence of VRE recently documented in faecal samples from Norwegian poultry. The present study reveals a high prevalence of VRE in broiler and turkey carcasses. Consequently, consumers are exposed to VRE when handling raw poultry meat, although the public health significance of such exposure is unclear.

Animals↗

Modeling non-linear survival curves to calculate thermal inactivation of salmonella in poultry of different fat levels.

Survival curves of a cocktail of eight serotypes of Salmonella in ground poultry of different fat levels (1-12%), when heated rapidly to specified temperatures (58-65 degrees C), were examined. Because many of the survival curves were concave, values for two parameters: the asymptotic D-value and the "lag" times were estimated and used to develop secondary models for estimating the time needed to obtain a 7 log10 relative reduction as a function of fat level and temperature. To compute the necessary time, at a given temperature and fat level, the estimated lag time should be added to the product of 7 and the estimated asymptotic D-value. A model was also developed for estimating the standard error of the estimated times, so that upper confidence bounds for the necessary times can be computed. It was found that lag times increase with higher fat levels. The effect of fat on D-values depended on the species; it is estimated that, for a given increase of fat level, the increase of the D-value would be greater for ground chicken than that for ground turkey. In addition, there was a statistically significant species effect on D-values, with higher D-values for ground turkey than for ground chicken at the higher temperatures studied. The thermal death curves displayed a non-linear tendency, however, for estimation purposes, a linear curve was assumed. There was not a statistically significant interaction effect of fat levels and temperatures on D-values, thus, for modeling, it was assumed that z-values were not dependent on the fat levels. The z-values for ground chicken and turkey were estimated to be 5.5 degrees C and 6.1 degrees C, respectively, and are statistically significantly different. These findings should have substantial practical importance to food processors of cooked poultry, allowing them to vary their thermal treatment of ready-to-eat poultry products in a safe manner.

Animals↗

Mobility of Zn, Cd and Pb in soils as affected by poultry litter extract--II. Redistribution among soil fractions.

The application of poultry litter to metal-contaminated soils may influence metal leaching and distribution of metals among soil fractions. Soil columns (one uncontaminated control, one metal-amended, and two metal-contaminated soils) were leached with H2O, CaCl2, EDTA, and poultry litter extract (PLE) solutions. After leaching, the soil samples in the columns were sequentially extracted for water soluble (WS), exchangeable (EXC), organic matter (OM), Mn oxide (MNO), amorphous Fe oxide (AFEO), crystalline Fe oxide (CFEO) and residual (RES) fractions. The OM fraction showed high retention for Zn from the PLE. The EDTA redistributed Zn, Cd and Pb from the EXC, OM and MNO fractions to the WS fraction. The PLE usually solubilized more Zn and Cd from the EXC fraction than CaCl2. Neither PLE nor CaCl2 mobilized Pb. The application of poultry litter on metal-contaminated soils might cause Zn and Cd redistribution from the EXC to the WS fraction and enhance metal mobility.

Journal Article↗

MALDI-TOF MS quantification of coccidiostats in poultry feeds.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a relatively new technique that is having a great impact on analyses. This study is the first to demonstrate the use of linear MALDI-TOF MS to identify and quantify coccidiostats in poultry feeds. 2,5-Dihydroxybenzoic acid (DHB) was found to be the best matrix. In MALDI-TOF MS, coccidiostats form predominantly [M + Na](+) ions, with additional small amounts of [M + K](+) and [M - H + 2Na](+) ions, and no obvious fragment ions. Salinomycin and narasin were unstable in the concentrated DHB matrix solution but were stable when dried on the MALDI-TOF MS probe. A simple fast Sep-pak C18 cartridge purification procedure was developed for the MALDI-TOF MS quantification of coccidiostats in poultry feeds. The MALDI-TOF MS limit of detection for lasalocid, monensin, salinomycin, and narasin standards was 251, 22, 24, and 24 fmol, respectively. The method detection limit for salinomycin and narasin in poultry feeds was 2.4 microgram/g.

Animal Feed↗

Rapid time-resolved fluoroimmunoassay for the screening of narasin and salinomycin residues in poultry and eggs.

Anticoccidial drugs are extensively used in the poultry industry to control the infection of the single-cell protozoa of the genus Eimeria. The most commonly used coccidiostats in poultry are the polyether ionophores such as narasin and salinomycin. This paper presents a rapid and simple method for the screening of residues of these two coccidiostatic compounds in poultry and eggs. The method is based on time-resolved fluoroimmunoassay. Sample preparation of eggs consists only of one extraction and evaporation step, and a solid phase extraction step is needed only for the muscle sample preparation. Mean recoveries were 91.0% from muscle tissue and 81.1% from eggs for both narasin and salinomycin. The performance of the assay was evaluated only for narasin because salinomycin had a cross-reactivity of 100% in the assay, and the recoveries of the compounds were not significantly different (P >0.05). The limits of detection [mean + 3 x standard deviation (SD)] of narasin were 0.56 and 0.28 microg/kg, and the limits of quantification (mean + 9 x SD) were 1.80 and 0.57 microg/kg for muscle and eggs, respectively. The coefficients of variation (CV) of the interassay precision of the method, evaluated by five replicate analyses of muscle samples spiked with 2 microg/kg of narasin and egg samples spiked with 1 microg/kg of narasin, were 4.1 and 6.4%, respectively. The CVs of intra-assay precision tests, determined by 10 replicate analyses at the above-mentioned concentration levels, were 3.8 and 4.5%, respectively.

Animals↗

Effect of replacing cotton seed cake with poultry droppings on weight gain of growing cattle at Bambui, Cameroon.

To investigate the effect of replacing cotton seed cake by poultry droppings in the diet of cattle, 30 animals were randomly allocated to three dietary treatments in a 2 x 3 factorial design, sex x treatments. The three treatments consisted of replacing 0%, 25% and 50% (T0, T25, T50 respectively) of the crude protein in 500 g of cotton seed cake with an equivalent amount of protein in poultry droppings. The concentrate offered was 615 g in T0, 652.5 g in T25 and 690 g in T50 per animal per day. After an adaptation period of 14 days, followed by 65 days of the experiment, the intake of concentrate and the average daily weight gains were recorded. There was no significant difference (p > 0.05) in the concentrate intake or average daily weight gain of bulls subjected to the three treatments. However, heifers tended to grow more slowly on T50 compared to T0 (p < 0.05), though the difference in their intake of concentrate was not significant (p > 0.05). Although T50 (47% inclusion in total concentrate), was less palatable, it seemed to have increased forage intake from pasture. These results suggest that poultry droppings can effectively substitute cotton seed cake in the diets of suckler cattle.

Animal Feed↗

Serum profiles of luteinizing hormone, oestradiol and cholesterol and ovarian functions in layer poultry birds (Gallus domesticus) fed diets containing furazolidone.

This study was carried out to assess the serum profiles of luteinizing hormone (LH), oestradiol, cholesterol and ovarian functions in layer poultry birds (Rhode Island Red: Gallus domesticus) fed a diet containing various concentrations of furazolidone (FZ). A total of 40 birds were randomly assigned to receive FZ 0, 200, 400 or 800 mg/kg feed (ppm) daily during the pre-laying age, i.e. 13-18 weeks (for 5 weeks). Blood samples were collected at weekly intervals. Concentrations of LH and oestradiol in serum were estimated at alternate weeks using radioimmunoassays. Serum cholesterol levels were analysed by an enzymatic calorimetric method. Furazolidone administration was terminated at the 18th week of age. The birds were sacrificed at 22nd week of age and ovarian tissues were processed for morphometric studies. Serum LH, oestradiol and cholesterol levels were affected by age (p < 0.001) and FZ dose (p < 0.001). Serum LH and oestradiol levels were lower (p < 0.05) in birds receiving FZ 800 mg/kg feed daily compared with the controls, whereas serum cholesterol profiles were lower (p < 0.05) in all FZ-administered groups than in the control group. The mean weight of ovaries having no yolky follicles observed in the group receiving FZ 400 or 800 mg/kg feed per day was reduced (p < 0.05) compared with the control group. Dosing FZ at 800 mg/kg feed per day reduced (p < 0.05) the mean volume of ovaries having no yolky follicles compared with the control group. In birds receiving FZ 800 mg/kg feed per day, the mean length of the oviduct was reduced (p < 0.05) as compared with the control group. Morphometric studies revealed that the mean number of oocytes with diameter in the range 401-800 microm decreased (p < 0.05) in birds fed FZ 400 or 800 mg/kg feed per day. Initial egg production was affected by age (p < 0.001) and dose (p < 0.001) of FZ. The mean number of eggs laid by different groups revealed that egg production was reduced (p < 0.05) in birds receiving FZ 800 mg/kg feed per day as compared with the controls. The present data suggest that FZ causes suppression in serum profiles of LH, oestradiol, cholesterol and ovarian functions in Rhode Island Red layer poultry birds. Therefore, great care must be taken with use of FZ in layer poultry birds (Gallius domesticus) with regard to dosage and duration of administration.

Animals↗

Rapid screening for monensin residues in poultry plasma by a dry reagent dissociation enhanced lanthanide fluoroimmunoassay.

Monensin, a carboxylic acid ionophore, is commonly fed to poultry to control coccidiosis. A method for rapid analysis of unextracted poultry plasma samples has been developed based on a novel immunoassay format: one-step all-in-one dry reagent time resolved fluorimetry. All assay specific components were pre-dried onto microtitration plate wells. Only addition of the serum sample diluted in assay buffer was required to perform analysis. Results were available one hour after sample addition. The limit of detection (mean +/- 3s) of the assay calculated from the analysis of 23 known negative samples was 14.2 ng ml-1. Intra- and inter-assay RSD were determined as 15.2 and 7.4%, respectively, using a plasma sample fortified with 50 mg ml-1 monensin. Eight broiler chickens were fed monensin at a dose rate of 120 mg kg-1 feed for one week, blood sampled then slaughtered without drug withdrawal. Plasma monensin concentrations, as determined by the fluoroimmunoassay ranged from 101-297 ng ml-1. This compared with monensin liver concentrations, determined by LC-MS, which ranged from 13-41 ng g-1. The fluoroimmunoassay described is extremely user friendly, gives particularly rapid results and is suitable for the detection and quantification of plasma monensin residues. Data from medicated poultry suggest that analysis of plasma may be useful in predicting the extent of monensin liver residues.

Animals↗

Characterization of lactobacilli towards their use as probiotic adjuncts in poultry.

AIMS: A total of 112 strains of lactic acid bacteria of duck origin were studied for their use as a probiotic feed supplement. METHODS AND RESULTS: In vitro studies included aggregation, co-aggregation, cell surface hydrophobicity and adhesion activities on poultry crop cells and human Hep2-cells. Additionally, growth with bile acids (chicken bile, ox gall and taurocholic acid) and tolerance to acidic pH were tested. Among all the isolates, two strains (Lactobacillus animalis TMW 1.972 and Lactobacillus salivarius TMW 1.992) were selected for a survival test in poultry. Monitoring and differentiation of these strains was achieved by selective detection as rifampicin and erythromycin double-resistant mutants. After a single feed administration, both micro-organisms were shown to persist in the crop and caecum of ducks for a period of 18 and 22 days, respectively. For identification of Lact. animalis and Lact. salivarius, two specific PCRs targeted against 16S rDNA were developed. CONCLUSIONS: Within the autochtoneous microflora of ducks, two strains of lactobacilli exhibited strong potential as probiotic adjuncts. The results indicate that the natural gut microflora of poultry serves as an excellent source for optimal strains. SIGNIFICANCE AND IMPACT OF THE STUDY: A general strategy for the selection of probiotic strains is presented. The suggested sequence of tests allows identification of the most promising candidates within complex ecosystems or large strain collections with minimal expenditure.

Animal Feed↗