Poultry Tissues Research Articles

Antibiotic Residues in Edible Poultry Tissues and Products in Nigeria: A Potential Public Health Hazard

Antibiotics are used by the veterinarians and poultry industry to enhance growth rates, health of the birds, feed efficiency, egg production and for therapeutic reasons to reduce the incidence of poultry diseases. Antibiotics had been used in poultry production for both therapeutic and prophylactic purposes. Most poultry farmers in Nigeria have employed the use of various antibiotics with or without the guidance of veterinarians. Although antibiotics benefit most of its uses, the illegal use of these drugs has led to the accumulation of toxic antibiotic residues in edible poultry products destined for human consumption. And this poses a major threat and hazard to the public that could be toxicological, microbiological or immunological. Several analytical techniques are available to screen edible poultry products with levels of toxic antibiotic residues. Drug withdrawal period and maximum residue level of antibiotics should be strictly observed in treated birds before any poultry product (meat or eggs) is passed “wholesome” for human consumption. There are still few researches addressing this major public health problem coupled with lack of awareness in Nigeria. Therefore, there is need for public enlightenment campaign on the dangers of antibiotic residues in edible poultry tissues and products to the teeming populace and restriction and regulation of indiscriminate use of antibiotics in poultry production and industry to curb the incidence of this public health hazard in Nigeria.

Molecular Epidemiology of Nontyphoidal Salmonella in Poultry and Poultry Products in India: Implications for Human Health.

Human infections with non-typhoidal Salmonella (NTS) serovars are increasingly becoming a threat to human health globally. While all motile Salmonellae have zoonotic potential, Salmonella Enteritidis and Salmonella Typhimurium are most commonly associated with human disease, for which poultry are a major source. Despite the increasing number of human NTS infections, the epidemiology of NTS in poultry in India has not been fully understood. Hence, as a first step, we carried out epidemiological analysis to establish the incidence of NTS in poultry to evaluate the risk to human health. A total of 1215 samples (including poultry meat, tissues, egg and environmental samples) were collected from 154 commercial layer farms from southern India and screened for NTS. Following identification by cultural and biochemical methods, Salmonella isolates were further characterized by multiplex PCR, allele-specific PCR, enterobacterial repetitive intergenic consensus (ERIC) PCR and pulse field gel electrophoresis (PFGE). In the present study, 21/1215 (1.73%) samples tested positive for NTS. We found 12/392 (3.06%) of tissue samples, 7/460 (1.52%) of poultry products, and 2/363 (0.55%) of environmental samples tested positive for NTS. All the Salmonella isolates were resistant to oxytetracycline, which is routinely used as poultry feed additive. The multiplex PCR results allowed 16/21 isolates to be classified as S. Typhimurium, and five isolates as S. Enteritidis. Of the five S. Enteritidis isolates, four were identified as group D Salmonella by allele-specific PCR. All of the isolates produced different banding patterns in ERIC PCR. Of the thirteen macro restriction profiles (MRPs) obtained by PFGE, MRP 6 was predominant which included 6 (21%) isolates. In conclusion, the findings of the study revealed higher incidence of contamination of NTS Salmonella in poultry tissue and animal protein sources used for poultry. The results of the study warrants further investigation on different type of animal feed sources, food market chains, processing plants, live bird markets etc., to evaluate the risk factors, transmission and effective control measures of human Salmonella infection from poultry products.

Screening for antibiotic residues in swine and poultry tissues using the STAR test

During a 12-month period, 138 tissue samples (muscle, liver, kidney) were screened for antibiotic residues (ARs), using the five-plate STAR test. Samples positive to one or more antibiotics were detected in 33.9% of the chicken samples and in 26% of the pig samples. All the chicken liver samples, 32.2% of the chicken muscle and 14% of the swine liver samples were positive to sulphonamides and β-lactams, 25% of the kidney, 11.1% of the liver and 4% of the muscle tissue of the swine samples and 1.7% of the chicken muscle samples were positive to tetracycline, 12.5% of the kidney and 4% of the muscle swine samples were positive to aminoglycosides and macrolides. No quinolone residues were detected in any samples. The use of a microbiological method such as the STAR-test, for ARs screening in food is an effective low cost scheme indicating potential contamination with ARs.

Determination of Environmental Deposition of Lead (Pb) in Certain Selected Samples in the Industrial Areas of Gazipur District of Bangladesh

Hasty industrialization affects the environment in different ways by discharging the large quantity of effluents as dissipate water in the surrounding water bodies, causing the solemn problems to environment. Twenty (n=20) samples of each category were collected from different locations of Gazipur. 3-5 goat of each sample was considered for analytical procedure. The results of analysis of variance of mean Pb-concentration in different samples collected from different locations differed significantly (P<0.05). The detected mean value of lead (Pb) concentration in drinking water (ground water), surface water, industrial waste water and agricultural soil (crop land) was 0.048±0.005mg/L ml/L, 0.428±0.098 ml/L, and 38.859±5.766 ml/L and 79.059±16.261 mg/kg. The mean value of concentrations of lead (Pb) was also detected in poultry tissue and litter samples. The highest level of lead (Pb) concentration was found in bone followed by brain, kidney, liver and muscles. The detected mean value of lead (Pb) in bone, brain, kidney, liver and muscles were 0.082±0.005 mg/kg, 0.073±0.007 mg/kg, 0.071±0.007 mg/kg, 0.052±0.011 mg/kg and 0.038±0.005, respectively. As regards biological samples, the highest natural accumulation of lead (Pb) was found in chicken bone while the lowest concentration (Pb) was detected in muscles. The natural accumulation of Pb was highly and significantly (P<0.001). The poultry litter (0.580±0.059 mg/kg) was also detected significantly (P<0.05) as a major toxic agent in respect of lead (Pb) for environmental degradation due to the use of poultry litter in agricultural soil (crop land) to grow more crops and vegetables.

Detecção de resíduos de enrofloxacina por ensaio imunoenzimático e cromatografia líquida acoplada à espectrometria de massas em ovos comerciais de galinha após tratamento

A enrofloxacina é um dos antimicrobianos mais utilizados na avicultura industrial, e a deposição de resíduos em produtos avícolas, como os ovos, são de grande importância para a saúde pública. Na legislação brasileira não existe padronização do período de carência para o seu uso na produção avícola e não há Limite Máximo de Resíduo (LMR) fixado para enrofloxacina em ovos. Neste estudo, foi utilizado o kit de ELISA comercial (Bioo Scientific(r)) e a LC-MS/MS na pesquisa de enrofloxacina em ovos de 30 galinhas tratadas previamente via água de bebida, com 10mg/kg de enrofloxacina, durante cinco dias. Seis ovos foram coletados diariamente e analisados durante o tratamento e após a sua suspensão, durante 15 dias. A deposição de resíduos obteve níveis máximos no quinto dia de tratamento das aves, declinando gradativamente até não ser detectada a partir do nono dia de suspensão do tratamento. Considerando como base o LMR de 100µg/kg fixado pelo Brasil para tecidos comestíveis de aves e pela União Europeia para músculo, gordura e pele, após seis dias de suspensão do tratamento, os níveis de resíduos foram inferiores a esse limite, tendo como médias 37,43µg/kg na LC-MS/MS e 14,731µg/kg no ELISA. Dentro das condições deste estudo, um período de carência de seis dias seria mais adequado para utilização dos ovos para consumo humano. Foram detectados valores de resíduos nos ovos menores no ELISA em relação à LC-MS/MS para a mesma amostra, mas os dois métodos apresentaram concordância estatística entre si. A LC-MS/MS é o teste recomendado pela legislação brasileira para a análise de resíduos em alimentos; entretanto, pelos resultados obtidos, o kit de ELISA utilizado também pode ser aplicado na detecção de resíduos de enrofloxacina em ovos, com as vantagens de rapidez e simplicidade.

Antibiotic residues in broiler and layer meat in Chittagong district of Bangladesh

Aim: The present study was described thin layer chromatography (TLC) and ultra-high performance liquid chromatography (UHPLC) method for the detection of antibacterial substances in poultry muscle (breast and thigh), kidney, and liver. Materials and Methods: TLC method was used for screening detection of tetracycline, amoxicillin, ciprofloxacin, and enrofloxacin residues in poultry tissues. The samples were extracted with trichloroacetic acid (30%), diethyl ether, followed by detection in pre-coated TLC paper on ultraviolet detector. The UHPLC method was used for the quantification of antimicrobial residues in poultry tissues. Results: The residues of tetracycline were 48% in livers, 24% in kidneys, 20% in thigh muscles, and 24% in breast muscles. Ciprofloxacin residues were found 44% in liver, 42% in kidneys, 34% in thigh muscles and 30% in breast muscles. Enrofloxacin residues were found 40% in liver, 34% in kidneys, 22% in thigh muscles, and 18% in breast muscles. Amoxicillin residues were found 42% in liver, 30% in kidneys, 26% in thigh muscles and 22% in breast muscles. Most of the cases highest residues were found in liver such as tetracycline (48%), ciprofloxacin (44%), enrofloxacin (40%) and amoxicillin (42%) and almost lowest in breast muscles. In addition, nine positive samples from broiler were selected for amoxicillin residue quantification by UHPLC. It was observed that the concentration of amoxicillin residue in liver was ranging from 16.92 μg/kg to 152.62 μg/kg and in breast muscle was 45.38 μg/kg to 60.55 μg/kg, respectively. The maximum and minimum peak time was 4.7-5.2 min. Among the poultry tissues, liver had the highest level of antibiotic residues in comparison to other samples but the variation was not significant (p>0.05). Conclusions: Evidence suggests that more judicious use of antimicrobials in food animals will reduce the selection of resistant bacteria and help to preserve these valuable drugs for both human and veterinary medicine. The method described in this study is a simple, easy inexpensive which can be readily adopted by any laboratory for the detection of antibiotic residues in tissues of food-producing animals.

Anti-microbial resistance and molecular characterization of Salmonella serovars from man and animals

Varied samples (1,132) comprising poultry meat (212), poultry eggs (49), poultry droppings (60), autopsied poultry tissues (60), pork (156), pig faeces (189), cattle dung (105), buffalo dung (103), sheep faeces (11), goat faeces (31), deer faeces (2) and human stool (154) were screened for isolation of Salmonella spp. Total 50 Salmonella isolates were recovered from samples, viz. poultry meat (2), poultry eggs (1), poultry droppings (6), autopsied poultry tissues (16), pork (2), pig faeces (9), cattle dung (6), buffalo dung (3), sheep faeces (4) and human stool (1). The recovered Salmonella isolates belonged to 10 different serovars with most prevalent serovar S. Typhimurium (21) followed by S. Weltevreden (12), S. Ughelli (5), S. Essen (3), S. Elisabethville (2), S. Lagos (2), S. Drogana (2), S. Enteritidis (1), S. London (1) and un-typable Salmonella (1). Salmonella isolates showed extreme variation in resistance (0–100%) against anti-microbial agents. The maximum resistance was observed among Salmonella isolates against sulphamethizole and furazolidone (100% each) followed by kanamycin (50%), gentamicin (44%), nalidixic acid (14%), tobramycin (10%), amikacin, ampicillin, streptomycin and tetracycline (8% each), amoxicillin/ clavulanic acid (6%), norfloxacin (4%), ciprofloxacin and cefotaxime (2% each) and chloramphenicol (0%). A high level of sensitivity in Salmonella isolates was observed against chloramphenicol (100%) and tetracycline (92%). A high multiple antibiotic resistance (MAR) index in a range of 0.133–0.666 was observed in Salmonella serovars. Molecular characterization of Salmonella was performed by targeting amplification of invA gene primer fragment for confirmation of Salmonella spp. in which all isolates were found positive for this gene. Moreover, this gene encodes virulence factor in Salmonella spp. which was exhibited by all Salmonella isolates of the present study.

Simultaneous determination of six aflatoxins and sterigmatocystin in livestock and poultry tissues by QuEChERS-ultra high performance liquid chromatography-tandem mass spectrometry

A method for the simultaneous determination of six aflatoxins (AFTs) and sterigmatocystin (SMC) in livestock and poultry tissues by QuEChERS-ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was developed and validated. The samples were extracted with 10 mL acetonitrile-water (84:16, v/v) after enzymatic hydrolysis with 20 μL β-glucosidase. The extracts were cleaned with 1.0 g each of sodium chloride and anhydrous magnesium sulfate, and then degreased with 5 mL hexane. The residues were re-dissolved in 1 mL acetonitrile-water (80:20, v/v) containing 5 mmol/L ammonium acetate. Methanol and 5 mmol/L ammonium acetate were used as the mobile phases. Six AFTs and SMC were analyzed by multiple reaction monitoring in the positive ion mode. Matrix-matched calibration curves and external standards were used for the accurate quantification of the six AFTs and SMC. Good linear relationships were obtained, the correlation coefficients (R2) were greater than 0.99. The limits of detection (LODs, S/N=3) and the limits of quantification (LOQs, S/N=10) ranged from 0.007 to 0.30 μg/kg, and 0.02 to 0.91 μg/kg, respectively. The recoveries at different spiked levels were satisfactory, and ranged from 77.3% to 118.5%. This method is simple, easy and sensitive, and is suitable for the rapid determination of the six AFTs and SMC in different pork, pork liver and chicken samples.

Performance and Fatty Acid Composition of Adipose Tissue, Breast and Thigh in Broilers Fed Flaxseed: A Review

Interest on the enrichment of poultry meat with n-3 fatty acids has increased given its important role in human health. Flaxseed is the main sources of n-3 fatty acids, and contains between 45 to 71% of total fatty acids of the oil as α-linolenic acid. This paper reviews the plausibility of n-3 enrichment. Its focus is on the processing of flaxseed and begins by summarizing the benefits of supplementation on broiler performance. The literature on altering the FA deposition in different tissues is then reviewed, and the factors that affect the incorporation of n-3 PUFA into edible tissues of poultry are investigated. Flaxseed supplementation caused a reduction in the abdominal fat pad, and the main fatty acid deposited in the tissue is LNA. The use of fold-change analysis allowed interpreting and determining the variation of results within experiments that do not report data in similar units of measure. The fold change analysis identified three categories of desaturation response to feeding flaxseed to broilers, resulting in different values for EPA and DHA in both breast and thigh tissues: high, medium and low fold-changes. The use of flaxseed oil, whole or ground flaxseed 14 to 21 day before slaughter is recommended to poultry producers as feeding strategies to optimize n-3 enrichment, without compromising animal performance. Enriched DHA deposition could be accomplished feeding whole flaxseed.

Efficient molecular and genetic technique for identification of Salmonella and Proteus strains

Fast microorganism genotyping technique has been developed with the use of the double distribution and selective labeling method (DDSL) for identification of disease-associated serotypes of Salmonella and Proteus strains. Isolates have been obtained from poultry tissues and dung from different places in various years. The results of strain genotyping conform to epizootological data. The method can be recommended for practical application in ascertainment of transmission of infectious disease and localization of bacterial pathogen source.

Can the unauthorised use of ceftiofur be detected in poultry?

Ceftiofur is a third-generation cephalosporin antibiotic used to treat cattle and swine for bacterial infection of the respiratory tract. It is not authorised for use in poultry within the European Union. Due to the complexity of the chemistry and metabolism of ceftiofur, maximum residue limits (MRLs) are based on desfuroylceftiofur (DFC) equivalents after chemical conversion of all compounds that have an intact ß-lactam ring. In practice the DFC is usually stabilised as the acetamide (desfuroylceftiofur acetamide – DFCA) for analysis. Because of recent evidence of off-label use in the European Union, a policy need emerged to develop a cost-effective method for the detection of ceftiofur residues in poultry tissues. One-day-old chicks were each dosed subcutaneously with ceftiofur and samples taken from day 1 to day 44 post-dosing. Residues of ceftiofur parent compound were detected in whole chicks, wing feathers and faeces. On the basis of this finding it was decided to evaluate ceftiofur parent, as the marker, instead of proceeding with the time-consuming conversion to DFCA. Expected metabolites, DFC and desfuroylceftiofur cysteine disulfide (DCCD), were not detected in whole chicks, muscle or liver, but DFC was found in wing feathers. These results indicate that determination of ceftiofur parent compound in either whole chicks or possibly wing feathers and faeces may allow the detection of the misuse of ceftiofur.

In-house quality control material of nicarbazin and narasin in eggs: preparation and inter-laboratory evaluation

Coccidiostats are a group of pharmacologically active substances widely used in veterinary practice. Their residues are detected relatively often in poultry tissues and egg samples analyzed as part of official residue control programs in the European Union. Therefore, accuracy of quantitative results needs to be monitored through internal and external quality control studies. In addition, the use of materials containing incurred residues would be welcome to for ongoing monitoring of the method accuracy. Unfortunately, in the field of veterinary drug residues, certified reference materials are often unavailable. Therefore, in-house quality control material of incurred lyophilized eggs containing narasin and nicarbazin has been produced and characterized. The eggs originated from hens receiving feed with coccidiostat premix Maxiban were mixed to obtain presumed concentrations of residues and freeze-dried. Homogeneity of the material was verified by the duplicate analysis of ten random samples, and the results proved that the between samples variation was negligible in comparison with the method repeatability. No measurable loss of analytes was observed within 1 year; the slope of the regression line of the results of stability measurements was not significantly different from zero. The assigned values were expressed as medians of the results of inter-laboratory comparison performed in four different European laboratories; the uncertainty of the material was estimated, taken into consideration all above tests, resulting in (14.4 ± 2.53) µg/kg for nicarbazin and (7.91 ± 1.52) µg/kg for narasin.

Simultaneous immunochemical detection of four banned antibiotic growth promoters in raw and cooked poultry tissue

Spiramycin, tylosin, bacitracin and virginiamycin are among a group of antibiotic growth promoters that have been banned in the European Union since the 1999 Council. This was due to concerns over the development of resistant bacteria emerging between humans and animals with the threat of antibiotics no longer being able to be used effectively to treat human infections. A sensitive and fast immunochemical method is presented for the determination of these four antibiotic growth promoters simultaneously in poultry tissue. The method employs methanol extraction followed by sample clean-up by solid-phase extraction (SPE) with determination by enzyme-linked immunoabsorbant assay (ELISA). The limit of detection (LOD) was less than 1 ng g−1 and the detection capability (CCβ) was 3 ng g−1 or less for all four antibiotic growth promoters. Validation was completed with both raw and cooked chicken, therefore either matrix could be used for the monitoring of these banned drugs. In a feeding trial no residues of either bacitracin or virginiamycin were found in medicated birds even without a withdrawal period. In the case of tylosin and spiramycin much higher residues level were detected immunochemically than was the case by mass spectrometry.

Chicken Cytochrome P450 1A5 Is the Key Enzyme for Metabolizing T-2 Toxin to 3'OH-T-2

The transmission of T-2 toxin and its metabolites into the edible tissues of poultry has potential effects on human health. We report that T-2 toxin significantly induces CYP1A4 and CYP1A5 expression in chicken embryonic hepatocyte cells. The enzyme activity assays of CYP1A4 and CYP1A5 heterologously expressed in HeLa cells indicate that only CYP1A5 metabolizes T-2 to 3′OH-T-2 by the 3′-hydroxylation of isovaleryl groups. In vitro enzyme assays of recombinant CYP1A5 expressed in DH5α further confirm that CYP1A5 can convert T-2 into TC-1 (3′OH-T-2). Therefore, CYP1A5 is critical for the metabolism of trichothecene mycotoxin in chickens.

Development and Validation of Electromigration Technique for the Determination of Lincomycin and Clindamycin Residues in Poultry Tissues

An analytical method to quantify lincosamide residues (lincomycin and clindamycin) in poultry tissue samples by micellar electrokinetic capillary chromatography combined with UV detection has been developed and validated. The clean-up procedure with a simple deproteinization and solid-phase extraction was successfully applied for the determination of antibiotics in different edible tissue samples of poultry. The electrophoretic separation was achieved using an uncoated fused silica capillary (57 cm × 75 μm i.d.), voltage 16 kV, temperature 22 °C, and the optimal wavelength at 200 nm. The running buffer consisting of 10 mM sodium tetraborate decahydrate (pH 9.3) and 25 mM sodium dodecylsulfate was employed. The validation of the method was performed by evaluation of the analytical parameters: linearity, specificity, precision, and accuracy with limits of detection of 13.2 and 18.5 ng/g, for clindamycin and lincomycin, respectively. Good intermediate precision was obtained with relative standard deviation values less than 12 %. The mean recoveries were 91.2 and 87.7 % for lincomycin and clindamycin, respectively. The obtained results confirm that the proposed method is capable to identify and quantify of lincosamide residues in animal tissues far below maximum residue limit values and could be suitable for routine application in commercial of poultry samples.

Antioxidant activity of carnosine extracted from various poultry tissues

The aim of the present research was 1) to extract carnosine from different low economic value poultry tissues and 2) to measure their antioxidant activities using different analytical methods. Low economic value poultry tissues such as the head, liver, lungs, tail, gizzard, brain, and heart were used in this study. Results have indicated that carnosine was present in all the tissue samples investigated. The liver had the highest (102.29 mg/g) and brain the lowest carnosine content (0.95 mg/g; P ≤ 0.05). Except for the brain, all tissue ultrafiltrates and reconstituted dry powders showed TBA reactive species inhibition ranging from 20.87 to 39.57% and 5.66 -14.47%, respectively. Free radical scavenging activity of ultrafiltrate from all tissues samples ranged from 25.11 to 79.38%, whereas this activity was higher (29.76 to 84.05%) in the reconstituted dry powder of all tissue samples. Conclusions include that extraction of bioactive dipeptide carnosine can be exploited from low economic value poultry tissues to increase the economy of the poultry industry.

Investigation of avian influenza viral ribonucleic acid destruction in poultry co-products under rendering conditions

SUMMARY This study was conducted to determine the time and temperature requirements needed to destroy avian influenza viral RNA in high-fat poultry tissues and to determine whether those conditions are met by rendering cooking processes. Because rendered poultry products are used worldwide for feed ingredients, it is imperative to validate the destruction of avian influenza viral RNA to ensure the safety of rendering co-products and avoid cyclic reinfection and disease in poultry, livestock, and potentially humans. Typical high-fat poultry offal was spiked with a known quantity of chemically inactivated, intact H5N9 low-pathogenicity avian influenza viral RNA. After subjecting the material to a variety of thermal doses, RNA was extracted from the material and assayed for the presence of the virus by using real-time reverse-transcription PCR. With thermal treatment at 100°C for 30 s or longer or at 110°C and above for 15 s or longer, the RNA of low-pathogenicity avian influenza virus A/Turkey/Wisconsin/68 H5N9, equivalent to 6 log10 of viable virus, was destroyed within the poultry rendering materials. These thermal treatment conditions are well below the range of temperatures and times used for rendering poultry carcasses and offal in the United States and Canada, where rendered materials are subjected to a heat treatment for a minimum of 30 min with a cooker exit temperature of not less than 118°C.

Identification and characterization of microRNA from chicken adipose tissue and skeletal muscle

MicroRNA (miRNA) is small noncoding RNA that is extensively expressed in organisms. Different types play important roles in various biological processes, such as growth and development. In this study, we identified 47 miRNA in chicken adipose tissue and skeletal muscle, of which 38 were known chicken miRNA, 4 were known miRNA homologous to other species, and 5 were potentially novel miRNA. The target genes from adipose tissue and skeletal muscle were predicted. The expression assay indicated that the 10 selected miRNA were differentially expressed in different developmental stages. Both miRNA-133a and miR-1a were muscle-related, whereas miR-122 was adipose-related miRNA. Certain identified miRNA may be essential to growth and development of chicken adipose tissue and skeletal muscle. Further studies of these miRNA will help to understand their functions in growth and development of adipose tissue and skeletal muscle of poultry.

Assessment of liquid chromatography–tandem mass spectrometry approaches for the analysis of ceftiofur metabolites in poultry muscle

The use of cephalosporin antibiotics in veterinary practice is likely to play an important role in the development of β-lactam-resistant bacteria. To detect off-label cephalosporin antibiotic usage, an analytical method is needed that, besides the native compound, also detects their active metabolites. In this paper, the applicability of three approaches for the quantitative analysis of ceftiofur using LC–MS/MS is assessed, viz. (A) analysis of ceftiofur, desfuroylceftiofur and/or desfuroylceftiofur cystein disulfide, (B) derivatisation of ceftiofur metabolites to desfuroylceftiofur acetamide and (C) chemical hydrolysis using ammonia, to produce a marker compound for ceftiofur. We found that approach A was not suited for quantitative analysis of total ceftiofur concentration or for effectively detecting off-label use of ceftiofur. Approach B resulted in adequate quantitative results, but was considered a single compound method because it depends on cleavage of a thioester group, which is present in only a limited number of cephalosporin antibiotics. Approach C showed adequate quantitative results but, in contrast to approach B, it is applicable to a range of cephalosporin antibiotics. Therefore, it is applicable as a broad quantitative screening of cephalosporin compounds in poultry tissue samples to indicate off-label use of cephalosporins in poultry breeding. Based on this study, it was concluded that approach C is the most suitable to detect off-label use of a range of cephalosporin antibiotics.

Determination of some Virulence Factors of Pasteurella multocia isolated from Human and Farm Animals infections

Twenty one Isolates of Pasteurella multocida were obtained from different clinical specimens of human and farm animals' infections. Human specimens included wounds swabs taken after cats and dog's bits beside the urine and sputum samples. The animal samples were nasal swabs and blood samples taken from chattels, also the poultry tissues of infected chicken with fowl cholera were collected. Bacterial isolates were isolated using Pasteurella multocida selective agar (PMSA) then identified doing different morphological, biochemical tests followed by api 20E diagnosis. The ability of the bacterial isolates to produce different virulence factors were studied, 18 isolates 85.7% were able to produce Lipase enzyme. Results of pathogenicity study on Lab. animals (mice) showed that there were 9 highly virulent isolates among the 21 (42.8%). The killing time was in between (10-24) hrs after injected the mice’s intraperitoneally. Some of the isolates showed their ability to produce the dermonecrotic toxin and the positive result appeared as highly purulent, dermonecrotic lesions after injection Genia pig intradermally. Results of antibiotic sensitivity test revealed that there were considerable variations in isolates susceptibility. Some of the isolates were highly resisted most of the used antibiotics while others were not. The crude bacterial extract from PMA20 isolate was tested to determine its ability to stimulate human lymphocytes division in vitro, results showed that the extract was able to stimulate cells division when the mitotic index was 1.2% as compared with the positive control.