Detection of Salmonella Heidelberg resistant to colistin in the intestinal content of pigs at slaughter

Salmonella Heidelberg has increasingly been reported as cause of human salmonellosis worldwide. In Brazil, S. Heidelberg has been reported in poultry but it is infrequently isolated from pigs. Here, we describe the isolation of S. Heidelberg resistant to colistin from slaughter pigs. Five pigs and their carcasses belonging to a same slaughter batch in ten consecutive days were sampled for fragment of intestine in the ileocecal region and sponges rubbed on the carcass surface (400 cm2) before chilling. Salmonella detection was performed according to the ISO 6579:2002. Intestinal content was also subjected to Salmonella enumeration by a miniaturized Most Probable Number (MPN) protocol. Salmonella isolates were characterized by antimicrobial resistance by the disk diffusion test, the minimum inhibitory concentration to colistin determination and to gene mcr-1 investigation by PCR. Salmonella was isolated from the intestinal content of 64% (32/50) of the pigs, in amounts that varied from 2.7 to \u3e1,400 MPN/g. Salmonella Heidelberg was the most frequent serovar identified in the intestinal content samples (20/50; 40%), and this serovar was present in eight of the ten pig batches sampled. At the prechill, Salmonella was isolated from 8% of carcasses, and S. Heidelberg was not detected. Salmonella Heidelberg strains were resistant against ampicillin (n=9), tetracycline (n=8), sulfonamide (n=8) and gentamicin (n=5). Nine multi-drug resistant strains were detected; among them four strains were positive for the gene mcr-1. In these strains the MIC value was 8 μg.mL-1, while in the strains without the mcr-1 gene it ranged from 2 μg.mL-1 to 4 μg.mL-1. Therefore, humans in contact with carrier pigs or their environment may be exposed to S. Heidelberg, including strains harboring the gene mcr-1

Detection of Genotipically Related Multi-resistant Escherichia coli Isolates in Pig Feces and Carcasses

Background: Antimicrobial resistant bacteria are considered a hazard not only for the treatment of animal diseases but also for public health. Commensal bacteria, such as Escherichia coli are considered a good indicator of antimicrobial resistance in the population, because it is a gut inhabitant and thus undergoes constant pressure of selection by the administration of antimicrobials. Regarding the public health, it is important to evaluate if resistant bacteria carried in the intestinal content of slaughter pigs can be found on the surface of pre chill carcasses. Therefore, the aims of this study were to evaluate the frequency of antimicrobial resistance in E. coli isolated from feces and pig carcasses; and to assess if multi-resistant isolates from both sources were phenotypically and genotypically related.Materials, Methods & Results: Two sampling cycles were conducted in three pig slaughterhouses (A, B and C). In each cycle, samples were collected form: i. feces deposited on the pen floor of the lairage; ii. surface of carcasses at the prechill step. Samples were submitted to a protocol of isolation and confirmation of Escherichia coli. Isolates were grouped according to the origin: feces (n = 355); carcasses (n = 319); and evaluated for antimicrobial resistance by agar diffusion test. Ninety two isolates presenting multidrug resistance profile were analyzed by pulsed-field gel eletrophoresis (PFGE). Among the 674 isolates of E. coli, 7.4% were susceptible to all tested antibiotics while 79.5% (536/674) were multi-resistant. The most frequent resistance patterns were displayed to tetracycline (Tet, 85.9%), ampicillin (Amp, 73.0%), sulfonamide (Sul, 70.0%), florfenicol (Flo, 65.0%) and nalidixic acid (Nal, 58.9%). The most frequent multi-resistance profile among isolates from both origins was [AmpFloNalSulTet]. Multiresistant isolates originated from feces and carcasses displaying genotypically related pulsotypes (≥70% similarity) were found in all three slaughterhouses.Discussion: In agreement with other studies, E. coli isolated from pig feces and carcasses demonstrated a high frequency of antimicrobial resistance and multi-resistance. The most frequent resistance profiles included antimicrobials frequently used on farm as well as drugs that have been banned as feed additives some years ago in Brazil. The selection of resistant strains may be related to the selection pressiondue to the use of antimicrobials in the pig production chain as well as the co-selection of resistance mediated by genes located in common genetic elements. Therefore, the ban of an individual drug is not always associated with the immediate disappearance of the resistance phenotype in the bacteria population. The fact that most multi-resistant E. coli isolates from carcasses belonged to pulsotypes related to those originated from feces samples indicates that resistant E. coli isolates selected on farm may be able to survive the slaughter process and be found on the carcass. In this case, the possibility of those strains being able to reach the population through the consumption of pork products may have to be considered. This hazard has motivated the ban of antimicrobial use in animals in some countries. However, the ban of antimicrobials use on farm is a controversial issue, due to the economical losses that may result from this measure. Therefore, the prudent use of antimicrobials on farm should be encouraged and its influence in the multi-resistance profile of the enteric microbiota should be further studied

Salmonella clinical isolates from Brazilian pig herds: genetic relationship and antibiotic resistance profiling

In Brazil, since 2011 clinical cases of salmonellosis has been increasing substantially. Nevertheless, few information is available about the antimicrobial profile, distribution, serotypes and genetic relationship among the strains. The objectives of this study were: to identify the Salmonella serotypes, to characterize the in vitro antimicrobial resistance profiles and to determine the genetic relationship of clinical isolates in Brazil. During 2016, clinical isolates of Salmonella (111) from nine States were sent to Embrapa Swine and Poultry for complementary analysis. First, isolates were serotyped by Kauffmann White Scheme. In parallel, the strains were tested against fifteen antimicrobials by disk diffusion method and genotyping was performed by Pulsed Field Gel Electrophoresis (PFGE) using the XbaI restriction enzyme. As expected, the main serovars found were Typhimurium and Choleraesuis. Four strains showed resistance to only one antimicrobial and 76.5% (85/111) were considered multi-resistant. The highest level of resistance was found against to tetracycline. More than 80% of the strains were susceptible to fosfomycin, lincomycin/spectinomycin and norfloxacin. It was possible to identify one major Choleraesuis clonal group present in different Brazilian States. Further, several small clonal groups were obtained for Typhimurium. In conclusion, clinical salmonellosis caused by Typhimurium and Choleraesuis is endemic in pig production areas and the majority of the strains are multi-resistant

Escherichia coli in chicken carcasses in southern Brazil : absence of shigatoxigenic (STEC) and isolation of atypical enteropathogenic (aEPEC)

The aim of this study was to investigate the presence of shiga toxin-producing Escherichia coli (STEC) and atypical enteropathogenic Escherichia coli (aEPEC) in frozen chicken carcasses sold at stores in southern Brazil. Typical E. coli colonies were enumerated in 246 chicken carcasses, and the presence of stx1, stx2, eae genes was investigated in their rinse liquid and in E. coli strains isolated from those carcasses. Strains of E. coli were also investigated for the presence of bfp gene. A median of 0.6 cfu.g-1(ranging from <0.1 to 242.7 cfu.g-1) of typical E. coli colonies was found in the carcasses. Shiga toxin-encoding genes (stx1 and stx2) were not detected, indicating that the chicken carcasses were negative for STEC. The intimin protein gene (eae) was detected in E.coli isolated from 4.88% of the carcasses; all tested strains were negative for the bfp gene and were classified as aEPEC. Twenty-two aEPEC strains were tested for resistance to ten antimicrobials and subjected to macrorestriction (PFGE). All the tested aEPEC strains were fully susceptible to cephalosporins, ciprofloxacin and colistin. Resistance to sulfonamide (65%), ampicillin (55%), tetracycline (50%) and gentamicin (45%) were the most frequent. The PFGE profile demonstrated a low level of similarity among the resistant strains, indicating that they were epidemiologically unrelated. The results indicate that aEPEC strains can contaminate chicken meat, and their association with strains implicated in human diarrhea needs to be further investigated

Antimicrobial resistance and genetic relationships of enterococci from siblings and non-siblings Heliconius erato phyllis caterpillars

Background: Studies evaluating bacteria in insects can provide information about host–microorganism–environment interactions. The gut microbial community has a profound effect on different physiological functions of insects. Enterococcus spp. are part of the gut community in humans and other animals, as well as in insects. The presence and antimicrobial resistance profile of enterococci are well studied in different animals; however, data for Heliconius erato phyllis (Lepidoptera: Nymphalidae) do not yet exist. Therefore, the aims of this study were to evaluate the distribution of enterococcal species, their antimicrobial resistance profile and virulence genes, and the genetic relationships between enterococci isolated from fecal samples from sibling and non-sibling H. erato phyllis caterpillars collected from different sites in South Brazil. Methods: Three H. erato phyllis females were captured (two from a forest fragment and one from an urban area), and kept individually in open-air insectaries. Eggs were collected and caterpillars (siblings and non-siblings) were fed daily with Passiflora suberosa leaves. Fecal samples (n = 12) were collected from fifth-instar caterpillars, inoculated in selective medium, and 15 bacterial colonies were randomly selected from each sample. Enterococci were identified by PCR and MALDI-TOF, analyzed by disk diffusion antimicrobial susceptibility tests, and screened for resistance and virulence genes by PCR. The genetic relationships between the strains were determined using pulsed-field gel electrophoresis (PFGE). Results: A total of 178 enterococci strains were identified: E. casseliflavus (74.15%; n = 132), E. mundtii (21.34%; n = 38), E. faecalis (1.12%; n = 2) and Enterococcus sp. (3.37%; n = 6). High rates of resistance to rifampicin (56%) and erythromycin (31%) were observed; 120 (67.41%) of the isolates showed resistance to at least one antibiotic and six (3.37%) were multidrug-resistant. None of the erythromycin-resistant strains was positive for the erm(B) and msrC genes. The virulence genes esp, ace, and gelE were observed in 35%, 7%, and 1% of the strains, respectively. PFGE separated the enterococci into 22 patterns, four being composed of strains from sibling caterpillars. Conclusion: Enterococcus casseliflavus was the dominant species in fecal samples of fifth-instar caterpillars. Resistant enterococci strains may be related to environmental pollution or the resistome. The PFGE analysis showed genetic relationships between some strains, suggesting that the enterococci isolated from fecal samples of the sibling caterpillars might have come from common sources, e.g., via diet (herbivory) and/or vertical transmission (through the egg surface). Further studies will be conducted to better understand the role of Enterococcus in the microbial community of the gastrointestinal tract of these insects, and the mechanisms involved in acquisition and maintenance of enterococci

Quality Of Life Of Patients With Breast And Gynecological Cancer Faced With Anticancer Chemotherapy

Quality of life of patients with breast and gynecological cancer faced with anticancer chemotherapy &nbsp;Backgroud: the evaluation of quality of life in cancer patients is of interest to researchers due to its impact on health policies. The aim of this study was to assess the quality of life of patients with breast and gynecological cancer, undergoing chemotherapy. &nbsp;Methods and Findings: this was a quantitative, descriptive and longitudinal study, conducted in the Clinical Hospital of the Triangulo Mineiro Federal University. To assess the quality of life, the Portuguese brief version of the World Health Organization Quality of Life (WHOQOL-BREF) instrument was applied. The paired Student’s t-test was used for the analysis of the means of/ the scores of the domains. The study included 14 women. The lowest means were observed in the physical domain, 51.78±15.89 and 50.25±15.37, respectively before and after treatment. There was also a compromise of the psychological and environment domains, however, the data analyzed did not show statistical significance. The main limitation of this study is the sample size. &nbsp;Conclusion: quality of life presents itself as an important factor for the measurement of results in clinical studies. Keywords: Quality of Life; Breast Neoplasms; Genital Neoplasms, Females; Chemotherapy

Porcine circovirus type 3 : immunohistochemical detection in lesions of naturally affected piglets

This study aimed to evaluate the relationship between porcine circovirus type 3 (PCV3) viral load and histopathological findings in perinatal piglet tissues and to develop an immunohistochemical method for detecting the virus in lesions. The quantitative polymerase chain reaction (qPCR) cycle threshold (Ct) when amplifying PCV3 DNA and the area of perivascular inflammatory infiltrates in different organs [central nervous system (CNS), lung, heart, liver, spleen, and lymph nodes] were compared. To develop an immunohistochemistry technique, rabbit sera were produced against PCV3-capsid protein peptides selected using bioinformatic analyses. The assay was initially implemented using a tissue sample previously tested using qPCR and in situ hybridization to optimize the procedure and reagent dilutions. To evaluate immunohistochemistry performance, tissue samples from another 17 cases were analyzed using standardized parameters. The most common microscopic lesion was multisystemic periarteritis, with associated vasculitis, as the mesenteric vascular plexus is one of the most affected organs. Other tissues, such as the heart, lung, CNS, and skeletal muscle, were also affected. Comparison of the Ct values for different tissues showed no significant difference, except in lymphoid organs (spleen and lymph nodes), which had significantly higher viral loads than the CNS tissues. There was no correlation between Ct values and perivascular inflammatory infiltrates. PCV3 immunohistochemistry revealed granular immunolabeling, mainly in the cytoplasm of cells in the vascular mesenteric plexus, heart, lung, kidney, and spleen