Luciana Ruschel dos Santos

Antibiotic resistance in Salmonella Enteritidis isolated from broiler carcasses

Eighty Salmonella Enteritidis strains isolated from broiler carcasses between May 1995 and April 1996 in the State of Rio Grande do Sul, Brazil, were tested for antibiotic susceptibility using the disk diffusion method. Resistance to colistin, novobiocin, erythromycin and tetracycline was observed in 100% of the isolates. The strains showed intermediate resistance at different levels to kanamycin (1.25%), enrofloxacin (3.75%), neomycin (3.75%), fosfomycin (20%), sulphonamides (86.25%) and nitrofurantoin (90%). Resistance to ciprofloxacin, norfloxacin, gentamicin, polymyxin B, sulphametrim and sulphazotrim was not found. Since resistance to antibiotics especially those introduced in the last decades, was detected, it is recommended that their use must be based on the results of resistance tests or minimum inhibitory concentration tests.

Salmonella spp. in raw broiler parts: occurrence, antimicrobial resistance profile and phage typing of the Salmonella Enteritidis isolates

The present study was carried out to evaluate the occurrence of Salmonellae in raw broiler parts and to determine the antimicrobial resistance profile of the isolated strains. Twenty-four (39.3%) broiler parts samples were positive for Salmonella and twenty-five Salmonella strains were isolated, since two different serovars were detected in one single positive sample. Salmonella Enteritidis was the most prevalent serovar. Among Salmonella Enteritidis isolates, 95.2% belonged to Phage Type 4 (PT4) (20/21) and 4.8% to PT7 (1/21). Twenty-two (88%) strains of Salmonella were resistant to at least one antimicrobial agent, generating eight different resistance patterns. The S. Typhimurium (n: 1) and S. Hadar (n: 3) isolates presented multiple resistance. Three S. Enteritidis isolates were susceptible to all antimicrobials tested, two were resistant only to tetracycline. The high prevalence of Salmonella in the broiler parts strenghtens the importance of the use of good manufacturing practices (GMP), and HACCP. The results also emphasize the need for the responsible use of antimicrobials in animal production.

Inoculation of newly hatched broiler chicks with two Brazilian isolates of Salmonella Heidelberg strains with different virulence gene profiles, antimicrobial resistance, and pulsed field gel electrophoresis patterns to intestinal changes evaluation

Salmonella Heidelberg is one of the 3 most frequently isolated serovars from human Salmonella cases in Canada, and the fourth most commonly reported Salmonella serovar in human foodborne disease cases in the United States. Since 1962, Salmonella Heidelberg has been isolated and reported in poultry and poultry products in Brazil. The poultry industry has focused efforts on reducing salmonellae incidence in live production in an effort to reduce Salmonella in the processing plant. A better understanding of the initial infection in chicks could provide approaches to control Salmonella contamination. The objective of the present study was to evaluate 2 Salmonella Heidelberg strains that differed in the presence of virulence genes invA, agfA, and lpfA; antimicrobial resistance profiles; and epidemiologic profiles on aspects of pathogenicity and intestinal morphology. Newly hatched broiler chicks were inoculated with 2 strains (SH23 and SH35) of Salmonella Heidelberg and cecal morphometry, histopathology, electron microscopy, and bacterial counts in the liver and cecum were assessed. The SH23 and SH35 strains resulted in different changes in villi height and crypt depth and inflammatory cell infiltration in the cecum. The SH35 group had higher liver and cecum bacterial cell counts when compared with SH23 strains.

Phenotypic and genotypic characterization of Salmonella enteritidis isolates

In order to study the epidemiology of Salmonella Enteritidis outbreaks and determine the source of contamination so that a recurrence can be avoided, detailed characterization is necessary. Thus, the purpose of this study was to verify whether rep-PCR was able to discriminate among Salmonella Enteritidis isolates. Phage typing, detection of virulence genes and antimicrobial resistance testing were also associated to repPCR results. One hundred and two S. Enteritidis isolates from broiler carcasses, food, human, pigs, poultryrelated samples, and nine isolates from other countries were genotypically typed by REP-PCR, ERIC-PCR and BOX-PCR, collectively called rep-PCR. Phage typing, detection of virulence genes and antimicrobial resistance testing were also performed. Only three fingerprinting profiles were obtained with each rep-PCR method, with the majority of isolates belonging to the same profile. No relationship was observed between genotypic profile and year, place of isolation or source of infection. However, the less frequent rep-PCR profiles showed single antimicrobial resistance patterns. Although few strains isolated from swine were analyzed, different antimicrobial resistance patterns were observed. Furthermore, phage type 4 was not found in swine isolates. rep-PCR showed a lower discriminatory power as compared with antimicrobial resistance and phage typing, but the combination of genotypic and phenotypic methods was more discriminatory than any method alone, resulting in 48 different types.

Quantification of biofilm production on polystyrene by Listeria, Escherichia coli and Staphylococcus aureus isolated from a poultry slaughterhouse

This study assessed biofilm formation on polystyrene by Staphylococcus aureus, Listeria monocytogenes, L. welshimeri and Escherichia coli, isolated from a slaughtering plant, grown on tryptic soy broth (TSB) using different glucose concentrations. The tested bacteria produced biofilm in at least one of the concentrations used, and some of them were strong biofilm producers.

Behavior of Salmonella heidelberg and Salmonella enteritidis strains following broiler chick inoculation: evaluation of cecal morphometry, liver and cecum bacterial counts and fecal excretion patterns

Over the years, Salmonella Heidelberg (SH) has gained prominence in North America poultry production and in the poultry production of other countries. Salmonella Heidelberg has been isolated and reported from poultry and poultry products in Brazil since 1962, whereas Salmonella Enteritidis (SE) has only emerged as a serious problem in poultry and public health since 1993. These strains of Salmonella can cause intestinal problems in newly hatched chicks, and infection may persist until adulthood. Upon slaughter of chickens, Salmonella can contaminate carcasses, a condition that poses a threat to human health. The aim of this study was to compare the fecal excretion of Salmonella Enteritidis and Salmonella Heidelberg in newly hatched chicks (orally inoculated with 105ufc/mL each) until 20 days of age. In addition, the ratio of cecal villus height:crypt depth (morphometry) and liver and cecum cell counts was analyzed in chicks ranging from 0 to 3 days of age and infected with these two Salmonella strains. One hundred seventeen chicks were separated into one of three experimental groups: a control group, an SE-infected group and an SH-infected group. Eight chicks per group were euthanized at 6, 12 and 72 hours post-inoculation (pi) to allow for Salmonella isolation from the liver and cecum and for the collection of the cecum for villi and crypt analysis. Other birds were allowed to mature to 20 days of age and cloacal swabs were taken at 2, 6, 13 and 20 days pi to compare the fecal excretion of inoculated strains. The Salmonella Enteritidis group had a higher number of cells excreted during the trial. Both strains were isolated from the liver and cecum by 6h pi. At 12h pi the Salmonella Heidelberg group had high cell counts in the cecum. No difference was found in liver cell counts. Both strains showed lower villus height:crypt depth ratio than the control group post-infection.

Antimicrobial resistance in Campylobacter spp isolated from broiler flocks

The aim of this study was to assess the antimicrobial susceptibility of 62 Campylobacter spp. strains obtained from broiler flocks using the agar diffusion method. The Campylobacter spp strains were isolated from 22 flocks aged between 3 and 5 weeks of life, isolated from cloacae swabs, stools and cecal droppings in the farm and from the carcass rinsing in the slaughterhouse. Campylobacter spp strains were tested on Mueller-Hilton (MH) agar (27 samples) and MH plus TTC agar (35 samples). The antimicrobial susceptibility test revealed a 62.5% resistance to at least one drug, especially to enrofloxacin (71%), neomycin (50%), lincomycin (50%), tetracycline (43%), penicillin (42%), ceftiofur (33%) amoxicillin (27%), spiramycin (20%), ampicillin (18%) and norfloxacin (14%), whereas a lower percentage of strains was resistant to erythromycin (10%) and doxycycline (10%). All strains were sensitive to gentamicin and lincomycin-spectinomycin and 80% of them to colistin. These results indicate that it is necessary to reduce the use of antimicrobials in veterinary and human medicine.

Avaliação in vitro de clorexidina, amônia quaternária e ácido peracético frente a amostras de Salmonella Heidelberg isoladas de abatedouro avícola em 2005 e 2009

The objective of this investigation was to evaluate the in vitro sensibility of Salmonella Heidelberg to three commercially available disinfectants used for sanitization in poultry slaughterhouses. A total of 20 S. Heidelberg were tested (14 isolated in 2005 and six in 2009), and as active ingredients were used chlorhexidine (0.5%), quaternary ammonium (0.5 %) and peracetic acid (1%) at contact intervals of 5, 10, 15 e 20 minutes. All isolates were found to be sensitive to peracetic acid at four specific contact intervals. One hundred percent of S. Heidelberg isolated in 2005 was found to be sensitive to quaternary ammonium, while 33% of 2009 isolates were resistant at a 5-minute contact interval and 16.6% at 10-minutes. With respect to chlorhexidine, 25% of the 2005 isolates were resistant at a 5-minute contact interval, 33% of the 2009 isolates were resistant with the same time, and 17% at a 10-minute contact interval. It can be concluded that the highest disinfectant activity in vitro was found to be with peracetic acid for S. Heidelberg isolates in 2005 and 2009, whereas chlorhexidine and quaternary ammonium had a reduced action against 2009 isolates, indicating the progression of bacterial resistance against these sanitizers and the need for periodic evaluation and rotation of active principles for sanitization.

Salmonella fecal excretion control in broiler chickens by organic acids and essential oils blend feed added

Salmonellosis is an important disease with economic impact as it may affect animal performance and may result in foodborne disease in humans through the eggs and carcass contamination. Regarding the Salmonella control, it is possible to decrease its fecal excretion and the contamination of chicken carcasses by adding organic acids to the feed or drinking water at appropriate times. The aim of this study was to test a blend of organic acids and essential oils in broilers challenged with Salmonella Enteritidis (SE), and to verify the fecal excretion of Salmonella. Sixty broilers were placed in four groups. One group was the negative control. Another group was orally inoculated at 1 day-old with 10 5 CFU/mL of SE as a positive SE control. Two groups (T3 and T4) were orally inoculated at 1 day-old with 10 5 CFU/mL of SE and their feed was separately treated with 0.5 and 1% of organic acids and essential oils, respectively. To assess the fecal excretion of SE, cloacal swabs were collected from all birds at 2, 6, 13 and 20 days after inoculation. The T3 and T4 groups showed a reduction in fecal excretion of SE at 6 and 20 days after inoculation.

Antiseptics and microcosm biofilm formation on titanium surfaces

Oral rehabilitation with osseointegrated implants is a way to restore esthetics and masticatory function in edentulous patients, but bacterial colonization around the implants may lead to mucositis or peri-implantitis and consequent implant loss. Peri-implantitis is the main complication of oral rehabilitation with dental implants and, therefore, it is necessary to take into account the potential effects of antiseptics such as chlorhexidine (CHX), chloramine T (CHT), triclosan (TRI), and essential oils (EO) on bacterial adhesion and on biofilm formation. To assess the action of these substances, we used the microcosm technique, in which the oral environment and periodontal conditions are simulated in vitro on titanium discs with different surface treatments (smooth surface - SS, acid-etched smooth surface - AESS, sand-blasted surface - SBS, and sand-blasted and acid-etched surface - SBAES). Roughness measurements yielded the following results: SS: 0.47 µm, AESS: 0.43 µm, SB: 0.79 µm, and SBAES: 0.72 µm. There was statistical difference only between SBS and AESS. There was no statistical difference among antiseptic treatments. However, EO and CHT showed lower bacterial counts compared with the saline solution treatment (control group). Thus, the current gold standard (CHX) did not outperform CHT and EO, which were efficient in reducing the biofilm biomass compared with saline solution.