agar and Poultry-Diseases

Avian pathogenic Escherichia coli (APEC) is responsible for colibacillosis in poultry. APEC remains a constant problem for the poultry industry, despite the use of antimicrobials and disinfectants in farms. The endemicity of APEC in poultry farms is associated with its biofilm-forming ability, which is further aggravated by various virulence factors and resistance to multiple drugs that help bacteria to thrive under different environmental conditions. To characterize APEC from affected broiler chickens and their environments, samples (n=114) from dead birds (heart, liver, lungs, and cloacal swab) and surrounding environments such as feeder, drinker, litter, PVC pipe, water tank wall, feed, and water were collected. The collected samples were subjected to microbial isolation using MacConkey Lactose agar (MLA) and Eosin Methylene Blue agar (EMB), which led to the isolation of 62 E. coli isolates. This was confirmed by uspA gene amplification and Vitek 2 Compact. These isolates were characterized using a set of five virulence genes (hlyF, ompT, iroN, iss, iutA), which yielded 47 (75.80%) isolates as APEC and the remaining as non-APEC. Furthermore, all the 62 isolates were subjected to microtiter plate assay for biofilm detection and the result showed that 36 (58.06%) isolates were able to form moderate to strong biofilms in Trypticase soy broth (TSB) at 72h of incubation. Of the 36 biofilm-producing isolates, 30 were APEC. Biofilm-related genes (crl, csgA, fimH, luxS, and papC) were also detected with higher prevalence among APEC isolates. Antimicrobial susceptibility test using Vitek 2 Compact revealed 43 (91.48%) of 47 APEC isolates as multiple drug resistant (MDR) and 8 (17.02%) as ESBL positive. This study reveals that APEC with biofilm formation ability is present in poultry farms. Further studies are needed to understand the role of biofilms in the pathogenesis and antimicrobial resistance of APEC.

Topics: Agar; Animals; Anti-Bacterial Agents; Biofilms; Chickens; Escherichia coli; Escherichia coli Infections; Poultry; Poultry Diseases; Water

Infectious coryza (IC) is an invasive upper respiratory disease caused by Avibacterium paragallinarum that affects birds, particularly chickens. The objective of this study is to isolate, characterize and molecularly identify the bacterium A. paragallinarum in poultry birds, as well as to determine its antibiotic sensitivity and resistance.. A total of 10 chickens from four different Iranian farms with typical IC symptoms were used in this study. The nasal swabs were streaked onto chocolate agar plates and blood agar plates and incubated at 37°C in 5% CO. By using biochemical assays and PCR analyses, seven strains of A. paragallinarum were isolated from samples of four chicken farms with typical IC clinical signs. Most isolates (4/7) showed the typical requirement for nicotinamide adenine dinucleotide (NAD) and an enriched CO. By discovering NAD-independent forms of A. paragallinarum, these species have a greater range than previously believed. A clear, cautious approach should be taken in diagnosing and possibly controlling IC.

Topics: Agar; Animals; Anti-Bacterial Agents; Carbon Dioxide; Chickens; Haemophilus paragallinarum; Iran; NAD; Poultry Diseases

The purpose of this study was twofold-first, to determine whether analysis of bacterial 16S ribosomal RNA (rRNA) in poultry litter corroborated standard

Topics: Agar; Animals; Bacterial Toxins; Chickens; Clostridium Infections; Clostridium perfringens; Enteritis; Enterotoxins; Farms; Polymerase Chain Reaction; Poultry Diseases; RNA, Ribosomal, 16S

Here, we describe the methods for isolation, purification, and propagation of Campylobacter jejuni bacteriophages from samples expected to contain high number of phages such as chicken feces. The overall steps are (1) liberation of phages from the sample material; (2) observation of plaque-forming units on C. jejuni lawns using a spot assay; (3) isolation of single plaques; (4) consecutive purification procedures; and (5) propagation of purified phages from a plate lysate to prepare master stocks.

Topics: Agar; Animals; Bacteriophages; Campylobacter Infections; Campylobacter jejuni; Chickens; Culture Media; Feces; Poultry Diseases; Viral Plaque Assay

Conventional culturing methods enable the detection of Campylobacter in broiler flocks. However, laboratory culture of Campylobacter is laborious because of its fastidious behavior and the presence of competing nontarget bacteria. This study evaluated different protocols to isolate Campylobacter from broiler litter, feces, and cloacal and drag swabs. Samples taken from commercial Brazilian broiler flocks were directly streaked onto Preston agar (PA), Campy-Line agar (CLA), and modified charcoal cefoperazone deoxycholate agar (mCCDA) and also enriched in blood-free Bolton broth (bfBB) for 24 and 48 h followed by plating onto the different selective media. Higher numbers of Campylobacter-positive cloacal and drag swab samples were observed using either direct plating or enrichment for 24 h before plating onto PA, compared with enrichment for 48 h (P < 0.05). Furthermore, direct plating was a more sensitive method to detect Campylobacter in broiler litter and feces samples. Analysis of directly plated samples revealed that higher Campylobacter levels were detected in feces streaked onto PA (88.8%), cloacal swabs plated onto mCCDA (72.2%), drag swabs streaked onto CLA or mCCDA (69.4%), and litter samples inoculated onto PA (63.8%). Preston agar was the best agar to isolate Campylobacter from directly plated litter samples (P < 0.05), but there was no difference in the efficacies of PA, mCCDA, and CLA in detecting Campylobacter in other samples. The isolated Campylobacter strains were phenotypically identified as Campylobacter jejuni or Campylobacter coli. The predominant contaminant observed in the Campylobacter cultures was Proteus mirabilis, which was resistant to the majority of antimicrobial agents in selective media. Together, these data showed that direct plating onto PA and onto either CLA or mCCDA as the second selective agar enabled the reliable isolation of thermophilic Campylobacter species from broiler samples. Finally, Campylobacter was detected in all broiler flocks sampled.

Topics: Agar; Animals; Bacteriological Techniques; Campylobacter; Campylobacter Infections; Chickens; Cloaca; Feces; Poultry Diseases

Ornithobacterium rhinotracheale (ORT) is a gram-negative staining rod. In chickens and turkeys ORT causes a respiratory disease. Between 2009 and 2011 some 714 dry swabs taken from diseased turkeys, broilers, broiler breeders, layers, or from unknown origin were investigated by PCR for the presence of ORT. Swabs that tested positive numbered 197 out of 481 from turkeys (41.0%), 10 out of 144 from broilers or broiler breeders (6.9%), 17 out of 28 from layers (60.7%), and 26 out of 61 from unknown origin (42.6%). The results of three swabs from turkeys were suspect. Furthermore, 310 isolates from turkeys and 62 isolates from unknown origin were typed using an agar gel precipitation (AGP) test. Of the isolates from turkeys, 56.1% belonged to serotype A and 20.6% to serotype E. The prevalence of other isolates was below 10%. Serotypes D, F, and K were not detected. Eleven isolates were not typable with reference sera against serotypes A-L. The three serotypes most often found in the isolates from unknown origin were A (35.5%), B (19.4%), and C (12.9%). The prevalence of other isolates was below 10%. Serotypes F and K were not detected. Seven isolates were not typable with reference sera A-L. Cross-reactions, especially of serotype A isolates with serotypes I, H, and J, were common. Additionally, the partial 16S ribosomal RNA (rRNA) and the complete Or01 genes of reference strains A-H and of nine field isolates were cloned and sequenced. Identity scores of 16S rRNA fragments were between 98% and 100%. Identities of the Or01 sequences were between 94% and 100%. Phylogenetic trees of both genes showed similarities. However, there was no apparent correlation between reference strains and isolates belonging to one serotype, so sequencing of 16S rRNA or of the Or01 gene does not seem to be a suitable method to replace the AGP for serotyping. Further investigations are necessary to clarify the cross-reactions between different serotypes and their real role in the pathogenicity and in consideration of vaccine production.

Topics: Agar; Animals; Bacterial Proteins; Chickens; Female; Flavobacteriaceae Infections; Germany; Ornithobacterium; Phylogeny; Polymerase Chain Reaction; Poultry Diseases; Prevalence; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Serotyping; Turkeys

Topics: Agar; Animals; Avian Leukosis; Avian Leukosis Virus; Bone Marrow; Bone Marrow Cells; Cell Division; Chick Embryo; Chickens; Clone Cells; Female; Neoplasm Regression, Spontaneous; Poultry Diseases; Vitelline Membrane

Topics: Agar; Animals; Chemical Precipitation; Chickens; Escherichia coli; Escherichia coli Infections; Gels; Poultry Diseases; Serotyping; Turkeys

Topics: Agar; Animals; Antibody Formation; Antigens; Bronchitis; Chemical Precipitation; Chickens; Newcastle Disease; Poultry Diseases; RNA Viruses; Virus Diseases

Topics: Agar; Animals; Avian Leukosis; Chick Embryo; Chickens; Culture Media; Culture Techniques; Cytopathogenic Effect, Viral; Ducks; Embryo, Nonmammalian; Fibroblasts; Herpesviridae; Kidney; Peripheral Nervous System Diseases; Poultry Diseases

Topics: Adenoviridae; Adenoviridae Infections; Agar; Animals; Bronchitis; Chickens; Culture Techniques; Electrophoresis; Hemagglutination Tests; Hemolytic Plaque Technique; Microscopy, Electron; Mycoplasma; Mycoplasma Infections; Poultry Diseases; Precipitin Tests; RNA Viruses; Trachea; Virus Diseases

Cell fractions of Pastuerella multocida (P-1059) were tested as vaccines against fowl cholera in turkeys. These fractions were culture filtrate, cell wall, and cytoplasm. A second culture filtrate preparation made from cells grown on blood-agar rather than the standard medium was also tested along with a "combination" preparation made by recombination of the cell fractions. Each preparation was tested in three vehicles: saline, alum (0.5%), and Freund Incomplete Adjuvant (50%). The turkeys vaccinated with these preparations were challenged by exposure to an experimental epornitic of fowl cholera. The combination fraction appeared to be the most promising vaccine when compared to the protective action of the commercial bacterin included in the test as a positive control.

Topics: Agar; Aluminum; Animals; Bacterial Vaccines; Blood; Cell Wall; Centrifugation, Density Gradient; Cytoplasm; Filtration; Freund's Adjuvant; Immunization; Immunochemistry; Injections, Subcutaneous; Pasteurella; Pasteurella Infections; Poultry Diseases; Sodium Chloride; Sulfates; Turkeys

Turkeys vaccinated with various experimental vaccines and a commerical bacterin for fowl cholera and surviving an artificially induced epornitic were killed, and their carcasses were examined for wholesomeness. It was evident that, if the "fitness for human consumption" judgement was considered in addition to mortality, efficacy ratings of the various vaccines changed. This suggests that the "fitness for human consumption" factor be considered in future evaluation of biologicals for use in meat-producing birds.

Topics: Agar; Aluminum; Animals; Bacterial Vaccines; Blood; Cell Wall; Cytoplasm; Filtration; Food Inspection; Freund's Adjuvant; Immunization; Pasteurella; Pasteurella Infections; Poultry Diseases; Sodium Chloride; Sulfates; Turkeys

Topics: Agar; Animals; Antibodies; Antigens; Avian Leukosis; Buffers; Chemical Precipitation; Chickens; Herpesviridae; Hydrogen-Ion Concentration; Immune Sera; Phosphates; Poultry Diseases; Precipitin Tests; Sodium Chloride; Temperature; Time Factors

Topics: Adsorption; Agar; Animals; Chickens; Erythrocytes; Hemagglutination; Hemagglutination Tests; Mycoplasma; Mycoplasma gallisepticum; Mycoplasma Infections; Poultry Diseases; Research