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Mikrobiyoloji Bulteni Oct 2021Increasing resistance to first-line antibiotics used in the treatment of infections caused by Salmonella and Shigella species is emerging. Azithromycin presents a good...
Increasing resistance to first-line antibiotics used in the treatment of infections caused by Salmonella and Shigella species is emerging. Azithromycin presents a good alternative treatment option for Salmonella and Shigella infections. However, there are limited data regarding the susceptibility of azithromycin in Turkey. In this study, we aimed to evaluate the susceptibility of Salmonella and Shigella species to azithromycin, to determine and compare the minimum inhibitory concentration (MIC) values and disk diffusion zone diameters. In addition, susceptibility to meropenem and first-line antibiotic options in isolates was also investigated. A total of 170 Salmonella, 76 Shigella clinical isolates collected between 2014 and 2018 in our hospital were tested for their susceptibility to azithromycin, meropenem, ampicillin, pefloxacin, trimetoprim-sulfamethoxazole, ceftazidime, and cefotaxime. Isolates were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry. The isolates were confirmed and serotyped by the reference laboratory using the conventional slide agglutination method. Susceptibility of the isolates to azithromycin and other antibiotics was evaluated by Kirby-Bauer disk diffusion method. MIC values of azithromycin were determined by the reference broth microdilution method. Combined disk diffusion test was used for the detection of extended spectrum beta-lactamase (ESBL) production. Polymerase chain reaction was performed for macrolide and carbapenem resistance genes and the detected resistance genes were confirmed by sequencing. Of the 76 Shigella isolates tested, 64 (84.2%) were identified as Shigella sonnei, 10 (13.2%) as Shigella flexneri, one (1.3%) as Shigella boydii, and one (1.3%) as Shigella dysenteriae. Among the 170 Salmonella isolates, 131 (77%) were identified as Salmonella enteritidis, 11(6.5%) as Salmonella Typhimurium, 8 (4.7%) as Salmonella Kentucky, 5 (2.9%) as Salmonella Paratyphi B, 4 (2.4%) as Salmonella Infantis, 3 (1.8%) as Salmonella Cholerasuis, and 8 (4.7%) as other serovars (Salmonella Agona, Salmonella Dabou, Salmonella Gallinarum, Salmonella Hadar, Salmonella Muenchen, Salmonella Newport, Salmonella Paratyphi C, Salmonella Senftenberg), respectively. ESBL production was determined as 7.9% (6/76) in Shigella isolates and 2.9% (5/170) in Salmonella isolates. A carbapenem resistant S.Senftenberg isolate positive for the blaOXA-48 resistance gene was detected in our study. Meropenem MIC value of the isolate was detected as > 32 µg/ml with gradient diffusion test. Among all isolates, only one S.boydii isolate was detected as resistant to azithromycin with a MIC value of 128 µg/ml. The isolate was positive for the existence of mphA gene by PCR. In the disk diffusion test, azithromycin inhibition zone diameters were ≥ 12 mm in all of the tested isolates, except for the azithromycin-resistant isolate, and the azithromycin MICs were determined as ≤ 16 µg/ ml by broth microdilution. Increasing resistance to commonly used antibiotics in Salmonella and Shigella species is emerging. The detection of a carbapenem-resistant Salmonella isolate in our study indicates that the spread of carbapenem resistance to other Enterobacterales species may cause global problems. Antimicrobial susceptibility testing of azithromycin for Salmonella and Shigella species has been difficult to establish due to the lack of approval in vitro breakpoints for all species. Consequently, our data shows that azithromycin exhibits as a good alternative therapeutic choice for the treatment of gastrointestinal diseases caused by Salmonella and Shigella species. Further studies are needed to provide appropriate in vitro breakpoints supported by clinical data.
Topics: Anti-Bacterial Agents; Azithromycin; Carbapenems; Drug Resistance, Bacterial; Humans; Microbial Sensitivity Tests; Salmonella; Shigella
PubMed: 34666650
DOI: 10.5578/mb.20219702 -
Frontiers in Bioengineering and... 2022Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of has been associated with various...
Gallinarum causes fowl typhoid in poultry leading to a huge economic loss to the poultry industry. The large virulence plasmid of has been associated with various systemic infections in poultry. A five-gene spanning region (spv) of 7.8 kb on the large plasmid mainly confers virulence to the bacteria. However, the exact role of these genes in virulence has not been elucidated yet. SpvB exhibits delayed cell death by preventing actin polymerization followed by apoptosis during intracellular infection. The specific role of SpvB in causing the disease is not known yet. In the current study, the SpvB gene was deleted through CRISPR/Cas9 method from a large virulent plasmid of locally isolated strain (SG18). The homology-directed repair method was used for complete deletion of SpvB gene using the modified pCas9 plasmid. The SpvB-deleted strain (ΔSpvB_SG18), when tested for its virulence in broiler chicken showed no diseases signs and mortality. In addition, the avirulent strain does not affect the bird's weight and was rapidly cleared from the liver after infection. However, it cleared from the intestine only after 4-5 days, which suggests that the ΔSpvB_SG18 strain is unable to invade from the intestine to the liver. This is the first study to report a complete gene deletion from the virulent plasmid and its effect. This method will be useful for the deletion of virulent genes from , to study their role in pathogenesis, and to prepare an effective vaccine strain for controlling fowl typhoid in poultry.
PubMed: 35769104
DOI: 10.3389/fbioe.2022.885227 -
Data in Brief Apr 2023Gallinarum (SG) is a host-restricted enterobacteria and the causative agent of fowl typhoid in poultry. Here, we report the complete genomes of two strains belonging to...
Gallinarum (SG) is a host-restricted enterobacteria and the causative agent of fowl typhoid in poultry. Here, we report the complete genomes of two strains belonging to this serotype. SA68 is a field strain isolated from the livers of dead hen carcasses of a commercial layer farm presenting high mortality located in São Paulo city, Brazil, in 1990. Strain 9R corresponds to a live attenuated SG commercial vaccine. DNA was extracted from pure cultures and subjected to whole genome sequencing (WGS) using the Ion Torrent PGM System. The assemblies reached lengths of 4,657,435 (SA68) and 4,657,471 (9R) base pairs. Complete genomes were deposited in GenBank under the accession numbers CP110192 (SA68) and CP110508 (9R). Both genomes were analyzed and compared in terms of molecular typing, antibiotic resistance genes, virulence genes, Salmonella pathogenic islands (SPIs), insertion sequences and prophages. The data obtained show many similarities in the genetic content, with the exception of the SPI-12 and CS54 pathogenic islands, which are exclusive to the field strain. The information generated will help to understand the virulence differences of field and vaccinal SG strains and can be used to perform evolutionary and epidemiologic studies.
PubMed: 36865996
DOI: 10.1016/j.dib.2023.108959 -
Poultry Science Jul 2024An effective vaccine strategy is indispensable against infectious bronchitis virus (IBV) and fowl typhoid (FT), both of which threaten the poultry industry. This study...
A low-endotoxic Salmonella enterica Gallinarum serovar delivers infectious bronchitis virus immunogens via a dual-promoter vector system that drives protective immune responses through MHC class-I and -II activation in chickens.
An effective vaccine strategy is indispensable against infectious bronchitis virus (IBV) and fowl typhoid (FT), both of which threaten the poultry industry. This study demonstrates a vector system, pJHL270, designed to express antigens in prokaryotic and eukaryotic cells. The vector system stimulates immune responses via synchronized antigen presentation to MHC class-I and -II molecules to produce balanced Th1/Th2 responses. The vaccine antigens were crafted by selecting the consensus sequence of the N-terminal domain of the spike protein (S1-NTD) and a conserved immunogenic region of the nucleocapsid protein (N-) from IBV strains circulating in South Korea. The vaccine antigen was cloned and transformed into a live-attenuated Salmonella Gallinarum (SG) strain, JOL2854 (∆lon, ∆cpxR, ∆rfaL, ∆pagL, ∆asd). Western blot analysis confirmed concurrent antigen expression in Salmonella and eukaryotic cells. Oral immunization with the SG-based IBV vaccine construct JOL2918 induced IBV antigen and Salmonella-specific humoral and cell-mediated immune responses in chickens. PBMCs collected from immunized chickens revealed that MHC class-I and -II expression had increased 3.3-fold and 2.5-fold, respectively, confirming MHC activation via bilateral antigen expression and presentation. Immunization induced neutralizing antibodies (NAbs) and reduced the viral load by 2-fold and 2.5-fold in the trachea and lungs, respectively. The immunized chickens exhibited multifaceted humoral, mucosal, and cell-mediated responses via parallel MHC class-I and -II activation as proof of a balanced Th1/Th2 immune response. The level of NAbs, viral load, and gross and histological analyses provide clear evidence that the construct provides protection against IBV and FT.
Topics: Animals; Chickens; Infectious bronchitis virus; Poultry Diseases; Coronavirus Infections; Salmonella enterica; Viral Vaccines; Serogroup; Genetic Vectors; Promoter Regions, Genetic; Histocompatibility Antigens Class I
PubMed: 38795516
DOI: 10.1016/j.psj.2024.103844 -
International Journal of Molecular... Sep 2022serovar Gallinarum, biovar Pullorum, is an avian-specific pathogen which has caused considerable economic losses to the poultry industry worldwide. Two-component...
serovar Gallinarum, biovar Pullorum, is an avian-specific pathogen which has caused considerable economic losses to the poultry industry worldwide. Two-component systems (TCSs) play an essential role in obtaining nutrients, detecting the presence of neighboring bacteria and regulating the expression of virulence factors. The genome analysis of . Pullorum strain S06004 suggesting the carriage of 22 pairs of TCSs, which belong to five families named CitB, OmpR, NarL, Chemotaxis and LuxR. In the CitB family, three pairs of TCSs, namely CitA-CitB, DcuS-DcuR and DpiB-DpiA, remain unaddressed in . Pullorum. To systematically investigate the function of the CitB family in . Pullorum, four mutants, Δ (abbreviated as Δ), Δ (Δ), Δ (Δ) and ΔΔΔ (Δ3), were made using the CRISPR/Cas9 system. The results demonstrated that the CitB family did not affect the growth of bacteria, the results of biochemical tests, invasion and proliferation in chicken macrophage HD-11 cells and the expression of fimbrial protein. But the mutants showed thicker biofilm formation, higher resistance to antimicrobial agents, enhanced tolerance to inhibition by egg albumen and increased virulence in chicken embryos. Moreover, the deletion of Dpi TCS was detrimental to survival after exposure to hyperosmotic and oxidative environments, as well as the long-term colonization of the small intestine of chickens. Collectively, we provided new knowledge regarding the possible role of the CitB family involved in the pathogenic processes of . Pullorum.
Topics: Animals; Chick Embryo; Chickens; Poultry Diseases; Salmonella; Salmonella Infections, Animal; Salmonella enterica
PubMed: 36077599
DOI: 10.3390/ijms231710201 -
Poultry Science Dec 2023This study aimed to assess the effects of a Lactobacillus helveticus ATCC 15009-derived postbiotic in mitigating experimental Salmonella Gallinarum infection. For this...
This study aimed to assess the effects of a Lactobacillus helveticus ATCC 15009-derived postbiotic in mitigating experimental Salmonella Gallinarum infection. For this purpose, a sample of Lactobacillus sp. was inoculated in 2 different media, each containing different postbiotics (sensitized and nonsensitized). Both inocula had their antagonistic effect over S. Gallinarum tested through the spot-on-the-lawn method. It revealed that the sensitized postbiotic had a higher action potential over Lactobacillus sp. than the nonsensitized one (P < 0.05). Then, 48 day of hatch chicks were divided into 4 groups: A = Lactobacillus sp. (10 CFU/mL) inoculum on the 18th day; B = Lactobacillus sp. (10 CFU/mL) inoculum on the 18th day and postbiotic inoculum on the 19th day; C = postbiotic inoculum on the 19th day; and D = sterile saline inoculum on 18th and 19th days. On the 21st day, all chicks were infected with S. Gallinarum (10 CFU/mL). On the 23rd day, the animals were euthanized by cervical dislocation, and the ceca and liver were aseptically removed. Bacterial count of S. Gallinarum with serial decimal dilution was performed with these organs. It revealed that the prophylactic treatment with the postbiotic that modulates the intestinal microbiota was as efficient as the probiotic administration in reducing S. Gallinarum in the cecum and liver of chicks (P < 0.05). These data point to a new range of alternatives for preventing S. Gallinarum, which might help the poultry industry produce safer food for human consumption.
Topics: Humans; Animals; Chickens; Lactobacillus helveticus; Salmonella; Cecum; Salmonella Infections, Animal; Poultry Diseases
PubMed: 37832187
DOI: 10.1016/j.psj.2023.103095 -
Poultry Science Nov 2019Fowl typhoid (FT), which is caused by Salmonella enterica serovar Gallinarum (S. Gallinarum), leads to high morbidity and acute or subacute mortality in chickens of all...
Fowl typhoid (FT), which is caused by Salmonella enterica serovar Gallinarum (S. Gallinarum), leads to high morbidity and acute or subacute mortality in chickens of all ages. Although a live S. Gallinarum 9R vaccine was introduced in 2001 for commercial layer chickens in Korea, until recently, a variety of antimicrobials were widely used to prevent or treat FT. In this study, we investigated antimicrobial resistance in S. Gallinarum strains isolated from 2014 to 2018 and characterized the multidrug-resistant (MDR) strains to better understand the resistance trends in recent isolates. A total of 130 S. Gallinarum isolates were collected from chickens with FT, and the isolates showed highest rates of resistance to nalidixic acid (78.5%), followed by gentamicin (52.3%), ciprofloxacin (26.9%), and ampicillin (14.6%). Particularly, significant increases (P < 0.05) in the frequencies of resistance to the following antimicrobials were observed: ampicillin (from 7.7 to 28.6%), amoxicillin-clavulanate (from 0.0 to 10.7%), nalidixic acid (from 69.2 to 100.0%), ciprofloxacin (from 15.4 to 50.0%), chloramphenicol (from 0.0 to 17.9%), and colistin (from 0.0 to 14.3%). The prevalence of MDR isolates also rapidly increased from 23.1% in the 2014 to 60.7% in the 2018 (P < 0.05). The distribution of antimicrobial resistance genes in the 39 MDR S. Gallinarum isolates was as follows: ant(2")-I gene (22 isolates), blaTEM-1 gene (13 isolates), sul1 (9 isolates), sul2 (3 isolates), cmlA (3 isolates), and qnrB (3 isolates). Of 39, 25 (64.1%) MDR S. Gallinarum isolates also carried class 1 integrons, and these showed 5 types of resistance gene cassettes: dfrA12+aadA2 (36.0%), aadA2 (36.0%), aadA1-aadA2 (20.0%), dfrA12+catB3+aadA2 (4.0%), and dfrA12 (4.0%). Among the plasmid replicons, B/O (33.3%) was more prevalent than the other replicon types, followed by Frep (25.0%), FIIA (19.4%), FIB (13.9%), and I1 (8.3%). Antimicrobial resistance may become a serious problem because many drugs are likely ineffective for the treatment of FT. Therefore, these data support the critical need for comprehensive surveillance of antimicrobial resistance in poultry.
Topics: Animals; Anti-Bacterial Agents; Chickens; Drug Resistance, Multiple, Bacterial; Poultry Diseases; Prevalence; Republic of Korea; Salmonella Infections, Animal; Salmonella enterica
PubMed: 31350992
DOI: 10.3382/ps/pez376 -
Veterinary World Dec 2020Food poisoning caused by is among the most common gastrointestinal discomfort resulted from egg consumption which can produce various syndromes. The present study is a...
BACKGROUND AND AIM
Food poisoning caused by is among the most common gastrointestinal discomfort resulted from egg consumption which can produce various syndromes. The present study is a systematic review and meta-analysis investigation on the published studies about the prevalence of contamination in the consumed eggs in Iran.
MATERIALS AND METHODS
The data were collected and analyzed from four international search databases, including PubMed, Scopus, Science Direct, and Google Scholar and four Iranian databases comprising SID, MagIran, Civilica, and IranDoc. After searching all the databases, 303 articles were found, from which 31 articles were included in the final analysis.
RESULTS
According to the data analysis, the highest rate of contamination was belonged to the industrial eggs (7.49%), however, the prevalence rate was reported 13.61% in the eggshell part. The overall prevalence of contamination in consumed eggs of Iran using culture of microbial, molecular, molecular-serological, culture-molecular, culture-serological, and culture -molecular-serological methods was obtained 11.33%, 5.52%, 0.37%, 1.91%, 5.52%, and 0.73%, respectively. Prevalence in the 21 geographical areas, where studies have been conducted, ranged from 0% (Zahedan) to 29.06% (Tabriz). The studies have also showed that eight different serotypes were among the major cause of contamination in eggs. The most common serotype was Enteritidis and the highest diversity in contaminant serotypes was recorded in Talesh (including . Enteritidis, Gallinarum, Vircho, and Newport).
CONCLUSION
Results of this study revealed the high prevalence of contamination in eggs, in Iran. Therefore, disinfection and cleaning bed, cleaning of equipment and supplies, and proper maintenance temperature and humidity of the eggs are recommended. In addition, proper personal hygiene and prohibition of consuming raw egg products are essential.
PubMed: 33487993
DOI: 10.14202/vetworld.2020.2743-2751 -
Translational Animal Science Jan 2022subs. serovar Enteritidis is a potential biological pathogen of concern in the poultry industry. Contamination of the bacterium on eggshells has led to human illnesses....
subs. serovar Enteritidis is a potential biological pathogen of concern in the poultry industry. Contamination of the bacterium on eggshells has led to human illnesses. With the implementation of new regulations, animal feed manufacturing continues to be under more stringent requirements. Specifically, there is zero tolerance for Pullorum, Gallinarum, or Enteritidis in poultry feed. For this reason, it is important to determine an effective method of reducing or preventing contamination in feed for poultry. Therefore, the objective of this study was to evaluate the impact of sodium bisulfate (SBS; Jones-Hamilton, Co., Walbridge, OH) added to poultry mash to reduce or prevent growth over time. A single, commercially produced all-flock poultry mash was mixed with four different levels of SBS: 0.0%, 0.25%, 0.50%, and 0.70%. After SBS addition, the treated mash was inoculated with subsp, serovar Enteritidis (ATCC 13076) and enumerated for on days 0, 1, 2, 7, and 14 post-inoculation by plating on xylose lysine deoxycholate agar. There was no significant effect of SBS inclusion level on the reduction of (= 0.23); however, there was a significant effect of time across treatments (< 0.0001). Additionally, there was no inclusion level × time interaction (= 0.68). These results suggest that while SBS inclusion has no effect on concentrations, storage time is effective at reducing or eliminating contamination in poultry feed.
PubMed: 35088042
DOI: 10.1093/tas/txab232 -
Brazilian Journal of Microbiology :... Mar 2023The purpose of this research was the genotypic identification of lactic acid bacteria (LAB), isolated from the gastrointestinal tract (GIT) of healthy adult birds, and...
The purpose of this research was the genotypic identification of lactic acid bacteria (LAB), isolated from the gastrointestinal tract (GIT) of healthy adult birds, and the study of their safety regarding antibiotic resistance, physiological and functional properties involved in the colonization of the GIT of poultry, and Salmonella exclusion, as members of a potential mixed probiotic supplement for poultry. The nucleotidic sequence from Lactobacillus crispatus P1, L. animalis L3, and Enterococcus faecium CRL 1385 (ex-J96) showed 100, 99.8, and 99.3% identity with L. crispatus DSM 20584, Ligilactobacillus salivarius ATCC 11741, and E. faecium ATCC 19434, respectively. These strains showed no resistance to relevant antibiotics usually administered to animals proposed by the European Food Safety Authority. They could endure the detrimental conditions of the gastrointestinal tract (pH 2.6 and oxgall 0.1 and 0.4% w/v). In an ex vivo assay, the LAB showed high adherence to the three sections of the GIT, reaching values higher than 70%. The adhesion to mucus was strain-dependent: L. crispatus CRL 1453 evidenced the highest adhesion (> 19%) while Lig. salivarius subsp. salivarius CRL 1417 and E. faecium CRL 1385 adhered to a lower extent (> 9 and 2%, respectively). Moreover, the LAB elicited remarkable anti-Salmonella activity, taking into account that they could inhibit elevated counts of different Salmonella serovars, especially the host-specific serovars S. Gallinarum and S. Pullorum (up to 8 log CFU/mL decrease in Salmonella counts).
Topics: Animals; Lactobacillales; Poultry; Chickens; Salmonella; Gastrointestinal Tract; Probiotics
PubMed: 36333643
DOI: 10.1007/s42770-022-00860-9