agar and Swine-Diseases

Campylobacteriosis is a significant public health concern with Campylobacter jejuni and Campylobacter coli as main causative agents. Moreover, there is an increasing recognition of other pathogenic Campylobacter species and Campylobacter-like organisms as Arcobacter. However, current knowledge on presence of Arcobacter species in wild boars (Sus scrofa) is lacking, and knowledge on Campylobacter species is based on methods favoring growth of thermotolerant species. In this study, fecal samples originating from 76 wild boars hunted in Campania region (Italy) were examined for the presence of Campylobacter(-like) organisms by a culture dependent approach. Three isolation protocols were performed in parallel: Arcobacter-selective agar plates, mCCDA plates and isolation by passive filtration onto non-selective blood agar plates were used as quantitative isolation methods. Enrichment broths, i.e. Arcobacter selective enrichment broth, Preston broth and CAT broth were used for qualitative detection of low levels or stressed Campylobacter(-like) organisms. The Arcobacter and Campylobacter isolates were identified at species level using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and 16S ribosomal RNA (rRNA) sequence analysis. Overall, 41 (53.9%) of the animals excreted Arcobacter or Campylobacter while 38 (50.0%) shed Campylobacter and 8 (10.5%) Arcobacter. Campylobacter lanienae predominated and was isolated from 31 (40.8%) animals. No statistical difference between the age groups or gender with regard to the fecal excretion of Campylobacter(-like) organisms was observed. Thirty animals (39.5%) shed Campylobacter spp. exceeding levels of 10 ³ CFU g

Topics: Agar; Animals; Arcobacter; Campylobacter; Campylobacter Infections; Sus scrofa; Swine; Swine Diseases

Production-limiting diseases in swine caused by Brachyspira are characterized by mucohemorrhagic diarrhea (B. hyodysenteriae and "B. hampsonii") or mild colitis (B. pilosicoli), while B. murdochii is often isolated from healthy pigs. Emergence of novel pathogenic Brachyspira species and strains with reduced susceptibility to commonly used antimicrobials has reinforced the need for standardized susceptibility testing. Two methods are currently used for Brachyspira susceptibility testing: agar dilution (AD) and broth microdilution (BMD). However, these tests have primarily been used for B. hyodysenteriae and rarely for B. pilosicoli. Information on the use of commercial susceptibility testing products such as antibiotic gradient strips is lacking. Our main objective was to validate and compare the susceptibility results, measured as the minimum inhibitory concentration (MIC), of 6 antimicrobials for 4 Brachyspira species (B. hyodysenteriae, "B. hampsonii", B. pilosicoli, and B. murdochii) by BMD and AD (tiamulin, valnemulin, lincomycin, tylosin, and carbadox) or antibiotic gradient strip (doxycycline) methods. In general, the results of a high percentage of all 4 Brachyspira species differed by ±1 log2 dilution or less by BMD and AD for tiamulin, valnemulin, lincomycin, and tylosin, and by BMD and antibiotic gradient strip for doxycycline. The carbadox MICs obtained by BMD were 1-5 doubling dilutions different than those obtained by AD. BMD for Brachyspira was quicker to perform with less ambiguous interpretation of results when compared with AD and antibiotic gradient strip methods, and the results confirm the utility of BMD in routine diagnostics.

Topics: Agar; Animals; Anti-Bacterial Agents; Brachyspira; Diarrhea; Diterpenes; Drug Resistance, Bacterial; Gram-Negative Bacterial Infections; Microbial Sensitivity Tests; Swine; Swine Diseases

The primary aim of this study was to evaluate the level of agreement of the E-test for in vitro antimicrobial susceptibility testing of Campylobacter coli using the agar dilution technique, which is the approved method. A convenience sample of 80 Ontario swine farms was chosen for this study; each farm was visited from January to June 2004. A total of 233 isolates of C. coli were tested for susceptibility to 10 antimicrobials by agar dilution and the E-test. Performance of the tests was evaluated using 7 quality control strains: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, Campylobacter jejuni ATCC 33560, and Campylobacter coli ATCC 33559 for the E-test and E. coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and C. jejuni ATCC 33560 for the agar dilution test. Weighted Cohen's kappa and prevalence-adjusted bias-adjusted kappa (PABAK) tests were used for statistical analysis. The E-test and agar dilution test results had a strong agreement when resistance to streptomycin and tetracycline were evaluated (weighted kappa: 0.68 and 0.66, respectively). However, marked disagreement was detected when testing susceptibility to nalidixic acid and ampicillin (0.15 and 0.22, respectively). Almost perfect agreement was detected by PABAK when testing susceptibility to gentamicin (0.99). Agreement was found to be moderate for ciprofloxacin, azithromycin, clindamycin, erythromycin, and chloramphenicol. Although the level of agreement between the E-test and agar dilution depended on the antimicrobial being tested, the E-test always detected a lower proportion of resistant isolates compared to agar dilution.

Topics: Agar; Animals; Anti-Bacterial Agents; Campylobacter coli; Colony Count, Microbial; Drug Resistance, Bacterial; Microbial Sensitivity Tests; Ontario; Reproducibility of Results; Sensitivity and Specificity; Swine; Swine Diseases

To compare three culture methods to detect Yersinia enterocolitica from oral or rectal swabs from experimentally infected pigs.. The three methods used were: direct plating on Cefsulodin-Irgasan-Novobiocin (CIN) agar, cold enrichment in phosphate buffered saline (PBS) followed by plating on CIN agar and selective enrichment with Luria-Bertani-Bile Salts Irgasan (LB-BSI) followed by plating on CIN agar. Selective enrichment with LB-BSI produced the highest recovery rate (63%), when compared with cold enrichment (52%) and plating on CIN agar alone (43%). Selective enrichment with LB-BSI was significantly (P < 0.02) more sensitive than direct plating on CIN agar and more sensitive than cold enrichment (P < 0.1).. Selective enrichment with LB-BSI was more sensitive than the widely accepted method of cold enrichment and it reduced the time required for detection of Y. enterocolitica by three weeks. Selective enrichment with LB-BSI was also compatible with a multiplex PCR technique.

Topics: Agar; Animals; Bacteriological Techniques; Colony Count, Microbial; Culture Media; DNA, Bacterial; Mouth; Polymerase Chain Reaction; Rectum; Sensitivity and Specificity; Swine; Swine Diseases; Time Factors; Yersinia enterocolitica; Yersinia Infections

The agar-gel migration technique has previously been described, however, aspects regarding the effect of timing on worm migration needed further scrutiny. In the first experiment, pigs inoculated with Oesophagostomum dentatum were slaughtered simultaneously and their intestines stored at 21-23 degrees C until processed pairwise 2, 4, 6, 8, 12 and 18 h after slaughter. More than 95% of the worms migrated out of the agar if processed within 6 h. In the second experiment, intestines were treated immediately after slaughter and the migratory speed of adult worms or 4th-stage larvae of O. dentatum or O. quadrispinulatum, or adult Hyostrongylus rubidus were studied. For both Oesophagostomum species, more than 90% of the worms were recovered within 1 h. H. rubidus was significantly slower; however, approximately 98% of the worms had migrated out of the agar-gel by 20 h. This information is essential in planning experiments where recovery of live worms is of value.

Topics: Agar; Animals; Female; Gels; Intestines; Larva; Male; Oesophagostomiasis; Oesophagostomum; Parasitology; Sex Characteristics; Swine; Swine Diseases; Time Factors; Trichostrongyloidea; Trichostrongyloidiasis

Four groups each of 3 pigs were inoculated with Ascaris suum eggs. Pigs in groups 1 and 3 were inoculated with 1000 eggs, and pigs in groups 2 and 4 with 10,000 eggs. On day 10 and 21 post-inoculation (p.i.), respectively, groups 1 + 2 and 3 + 4 were slaughtered, and the contents from the small intestines collected. The contents were mixed with agar to a final concentration of 1% agar and allowed to sediment. The larvae were allowed to migrate from the agar-gel into 38 degrees C 0.9% saline overnight, and were then collected on a sieve (20 microns mesh) and counted. The larvae retained in the agar-gel were counted after pouring the melted agar through a sieve (20 microns mesh). The results showed that more than 97% of the larvae migrated out of the agar-gel and were available for counting in an almost clean suspension. The inoculation dose level did not significantly affect the recovery percentage, neither did the larval stage (10 or 21 days old larvae). The variation in the time interval from slaughtering to start of incubation (interval 57-155 min) did not significantly affect the recovery percentage.

Topics: Agar; Animals; Ascariasis; Ascaris suum; Gels; Intestine, Small; Larva; Swine; Swine Diseases; Time Factors

Nine groups of 5 pigs were inoculated with Ascaris suum eggs on day 0. Groups 1, 2, and 3 were inoculated with 100 eggs, groups 4, 5, and 6 with 1,000 eggs, and groups 7, 8, and 9 with 10,000 eggs. On day 3, groups 1, 4, and 7 were slaughtered, on day 7 groups 2, 5, and 8, and on day 10 groups 3, 6, and 9. The liver (days 3 and 7) and lungs (days 3, 7, and 10) were removed and 2, 25% samples of both organs were collected. Larvae were recovered from 1 sample by the Baermann method and from the other by an agar-gel method. Overall there were no significant differences in the liver larval recovery between the 2 methods. The use of the agar-gel method resulted in a very clean suspension of larvae and thereby reduced the sample counting time by a factor of 5-10 compared to the Baermann method. With both methods larval recovery from the lungs resulted in a clean larval suspension that was easy to count, and there were overall no significant differences between the 2 methods, although there was a tendency toward the Baermann method recovering more larvae from the lungs than the agar-gel method. The tissue sample dry weight did not significantly influence larval recovery by the agar-gel method, and the time interval from slaughtering to start of incubation on day 3 (interval 51-92 min), day 7 (interval 37-114 min), and day 10 (interval 50-129 min) had no significant effect on recovery by either method.

Topics: Agar; Animals; Ascariasis; Ascaris suum; Female; Gels; Larva; Liver; Lung; Male; Swine; Swine Diseases

A method has been developed for separating Serpulina hyodysenteriae, a large spirochete and the causative agent of swine dysentery (SD), from other fecal anaerobic bacteria in rectal and colonic swabs. This was done by cutting the blood agar in parallel cuts and streaking perpendicular to the cuts in the center of the petri dish. Migration of S. hyodysenteriae from the central streak was apparent by the presence of strong beta-hemolysis along the edges of the cuts. If only S. hyodysenteriae migrated in the cut, they migrated to the end of the cut. However, if both motile bacteria and S. hyodysenteriae migrated in the cut, the motile bacteria migrated to the end of the cut where they formed colonies and the S. hyodysenteriae located along the edges of the cut between the colonies of motile bacteria and the central streak. Although motile bacteria were present where S. hyodysenteriae located, the growth of the motile bacteria was partially inhibited since they rarely formed visible colonies and were low in number. The cut in the agar was thought to improve traction for the serpentine movement of the S. hyodysenteriae and for the flagellar movement of the motile bacteria. Use of sliced blood agar was superior to conventionally streaked blood agar in that (i) it was easier to see strong beta-hemolysis on sliced agar; (ii) frequently, a confirmatory diagnosis could be made using only one petri dish with sliced agar, thereby saving time and media; (iii) S. hyodysenteriae could sometimes be isolated free of other bacteria; and (iv) sliced agar was more effective in isolating S. hyodysenteriae from swine with chronic diarrhea and nondiarrhetic carriers of SD in which the shedding of S. hyodysenteriae was low.

Topics: Agar; Animals; Bacteriological Techniques; Brachyspira hyodysenteriae; Colon; Dysentery; Evaluation Studies as Topic; Hemolysis; Rectum; Spirochaetales Infections; Swine; Swine Diseases

Four groups with three pigs in each group were inoculated with Oesophagostomum dentatum larvae (L3 larvae). Groups 1 and 3 were inoculated with 20,000 larvae, and Groups 2 and 4 with 200,000 larvae. On Days 11 and 34, respectively, Groups 1 and 2 and Groups 3 and 4 were slaughtered, and the contents from the large intestines collected. Subsamples of intestinal contents were mixed with agar to a final concentration of 1% agar and allowed to set. The worms were allowed to migrate from the agar-gel into 38 degrees C 0.9% saline overnight. Then the worms were collected on a sieve (38 microns mesh) and counted. The worms retained in the agar-gel were counted after pouring the melted agar through a sieve (38 microns mesh). The results showed that more than 95% of the worms migrated out of the agar-gel, and subsequently were available for counting in an almost clean suspension. Additionally the method yielded a high worm recovery; all stages were recovered. The recovery percentage was not significantly affected by either the dose of parasites or the time interval from slaughtering to start of incubation (37-128 min).

Topics: Agar; Animals; Female; Gels; Intestine, Large; Larva; Male; Oesophagostomiasis; Oesophagostomum; Parasitology; Swine; Swine Diseases; Time Factors

Crystal violet, lincomycin, spectinomycin and bacitracin were evaluated as selective agents in media for isolation of Haemophilus pleuropneumoniae. No single antimicrobial agent or combination of two or more inhibited all non-Haemophilus strains (Escherichia coli, Pasteurella haemolytica, Pasteurella multocida, Streptococcus faecalis, Streptococcus equisimilis and Staphylococcus aureus) without marked suppression of 16 H. pleuropneumoniae strains. A medium containing 1 micrograms/mL of crystal violet, 1 microgram/mL of lincomycin, 8 micrograms/mL of spectinomycin and 128 micrograms/mL of bacitracin inhibited one E. coli strain and the Gram-positive strains while H. pleuropneumoniae strains were suppressed to a minor degree only. Haemophilus pleuropneumoniae was isolated on the selective medium on three occasions from the nose or pharynx of two out of eight experimentally inoculated pigs. Haemophilus pleuropneumoniae was recovered from the nose of only two pigs at necropsy and from tonsil of one, whereas the lower airways in most pigs and the lung lesions in all pigs were positive. There was no advantage to using the selective medium for the recovery of H. pleuropneumoniae at necropsy from these eight experimentally infected pigs, probably because other bacteria were absent or present in very low numbers in the tissues with H. pleuropneumoniae. The isolation rate on selective medium was higher than the rate on non-selective medium (p less than or equal to 0.1; chi 2 test) when the airways of slaughtered pigs were cultured. This was likely due to a high degree of contamination. Dry swabs placed in tryptone yeast extract with nicotinamide-adenine-dinucleotide gave a significantly higher recovery rate than commercial Culturette swabs in modified Stuart's transport medium.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Agar; Animals; Bacitracin; Bacteriological Techniques; Carrier State; Caseins; Culture Media; Gentian Violet; Haemophilus; Haemophilus Infections; Lincomycin; Pleuropneumonia; Protein Hydrolysates; Serotyping; Spectinomycin; Swine; Swine Diseases

Topics: Agar; Animals; Anti-Bacterial Agents; Bacillus subtilis; Biological Assay; Cattle; Cattle Diseases; Diffusion; Kidney; Microbial Sensitivity Tests; Micrococcus; Sulfonamides; Swine; Swine Diseases

Optimal culture conditions in artificial nutritive media were determined for a defined avirulent strain of Treponema hyodysenteriae and for four field strains of treponemas in pigs with clinical dysentery. The treponemas were isolated with the use of milliporous filters with pores of 0.3 micrometer in diameter, which were located on the surface of blood agar. No significant difference in the influence of equine, bovine or sheep blood on the growth of treponemas was determined. The commercial amount of glucose in the used media, 2.0 to 2.5 g per 1,000 ml, was quite sufficient for the growth of the treponemas and it was not necessary to increase the amount. After reaching the optimal rate of growth the oxidoreduction potential was diminished by adding cystein or cystein hydrochloride and placing the Petri dishes with the media, prior to inoculation, into an anaerobic medium filled with hydrogen. The suitable composition of the culture atmosphere created in a special anaerostat comprised 0.4 to 1.0% carbon dioxide and the rest being hydrogen. Treponemas grew on the blood agar in zones with very slight hemolysis without forming separated colonies.

Topics: Agar; Animals; Blood; Culture Media; Dysentery; Horses; Intestine, Large; Sheep; Swine; Swine Diseases; Treponema

Topics: Agar; Animals; Bacteriological Techniques; Blood; Culture Media; Intestines; Pasteurella; Pasteurella Infections; Swine; Swine Diseases

Topics: Agar; Animals; Blood Proteins; Electrophoresis; gamma-Globulins; Gels; Globulins; Immunoelectrophoresis; Serum Globulins; Sheep; Sheep Diseases; Species Specificity; Swine; Swine Diseases; Trichinella; Trichinellosis

Topics: Agar; Agglutination Tests; Animals; Antigens; Culture Media; Fluorescent Antibody Technique; Glucose; Haemophilus; Haemophilus Infections; Horses; Immune Sera; Methods; Pleuropneumonia; Precipitin Tests; Rabbits; Saccharomyces; Serotyping; Sheep; Swine; Swine Diseases

Topics: Agar; Animals; Bacteriological Techniques; Blood; Horses; Immune Sera; Mycoplasma; Mycoplasma Infections; Serotyping; Swine; Swine Diseases

Topics: Agar; Animals; Antibodies; Antibody Formation; Antigen-Antibody Reactions; Cattle; Cattle Diseases; Foot-and-Mouth Disease; Goats; Immunodiffusion; Mice; Retrospective Studies; Sheep; Sheep Diseases; Swine; Swine Diseases; Viral Vaccines

Topics: Agar; Animals; Anti-Bacterial Agents; Cattle; Cattle Diseases; Culture Techniques; Drug Resistance, Microbial; Goats; Mycoplasma; Mycoplasma Infections; Sheep; Sheep Diseases; Swine; Swine Diseases; Virulence

Topics: Agar; Animals; Anti-Bacterial Agents; Chloramphenicol; Colistin; Diffusion; Dihydrostreptomycin Sulfate; Drug Resistance, Microbial; Escherichia coli; Escherichia coli Infections; Furazolidone; Hemolysis; Kanamycin; Microbial Sensitivity Tests; Neomycin; Paper; Serotyping; Swine; Swine Diseases; Tetracycline

Topics: Agar; Animals; Brucella; Brucellosis; Disease Reservoirs; Female; Genitalia; Germany, East; Glycerol; Male; Serologic Tests; Swine; Swine Diseases

Topics: Agar; Animals; Antigen-Antibody Reactions; Chemical Precipitation; Classical Swine Fever; Gels; Swine; Swine Diseases

Topics: Agar; Animals; Birds; Disease; Erysipelas; Erysipeloid; Iron; Meat; Poultry; Swine; Swine Diseases