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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 -
Microbiology Spectrum Aug 2022Paratyphoid avian salmonellosis is considered one of the leading causes of poultry death, resulting in significant economic losses to poultry industries worldwide. In...
Genome-Based Assessment of Antimicrobial Resistance and Virulence Potential of Isolates of Non-Pullorum/Gallinarum Salmonella Serovars Recovered from Dead Poultry in China.
Paratyphoid avian salmonellosis is considered one of the leading causes of poultry death, resulting in significant economic losses to poultry industries worldwide. In China, especially in Shandong province, the leading producer of poultry products, several recurrent outbreaks of avian salmonellosis have been reported during the last decade where the precise causal agent remains unknown. Moreover, the establishment of earlier and more accurate recognition of pathogens is a key factor to prevent the further dissemination of resistant and/or hypervirulent clones. Here, we aim to use whole-genome sequencing combined with toolkits to provide the genomic features of the antimicrobial resistance and virulence potential of 105 regionally representative non-Pullorum/Gallinarum Salmonella isolates recovered from dead poultry between 2008 and 2019 in Shandong, China. Additionally, phenotypic susceptibility to a panel of 15 antibiotics representing 11 classes was assessed by the broth microdilution method. In this study, we identified eight serovars and nine multilocus sequence typing (MLST) types, with Salmonella enterica serovar Enteritidis sequence type 11 (ST11) being the most prevalent (84/105; 80%). Based on their phenotypic antimicrobial resistance, 77.14% of the isolates were defined as multidrug resistant (≥3 antimicrobial classes), with the detection of one S. Enteritidis isolate that was resistant to the 11 classes. The highest rates of resistance were observed against nalidixic acid (97.14%) and ciprofloxacin (91.43%), followed by ampicillin (71.43%), streptomycin (64.77%), and tetracycline (60%). Genomic characterization revealed the presence of 41 resistance genes, with an alarmingly high prevalence of (60%), in addition to genomic mutations affecting the DNA gyrase () and DNA topoisomerase IV () genes, conferring resistance to quinolones. The prediction of plasmid replicons detected 14 types, with a dominance of IncFIB(S)_1 and IncFII(S)_1 (87.62% for both), while the IncX1 plasmid type was considered the key carrier of antimicrobial resistance determinants. Moreover, we report the detection of critical virulence genes, including , , , , and , in addition to the typical determinants for Salmonella pathogenicity island 1 (SPI-1) and SPI-2. Furthermore, phylogenomic analysis revealed the detection of three intra-farm and five inter-farm transmission events. Overall, the detection of Salmonella isolates presenting high antimicrobial resistance and harboring different critical virulence genes is of major concern, which requires the urgent implementation of effective strategies to mitigate non-Pullorum/Gallinarum avian salmonellosis. Avian salmonellosis is one of the leading global causes of poultry death, resulting in substantial economic losses in China (constituting 9% of overall financial losses). In Shandong province, a top poultry producer (30% of the overall production in China, with 15% being exported to the world), extensive outbreaks of avian salmonellosis have been reported in the past decade where the causal agents or exact types remain rarely addressed. From approximately 2008 to 2019, over 2,000 Salmonella strains were isolated and identified from dead poultry during routine surveillance of 95 poultry farms covering all 17 cities in Shandong. Approximately 1,500 isolates were confirmed to be of non-Pullorum/Gallinarum Salmonella serovars. There is an urgent need to understand the mechanisms behind the implication of zoonotic Salmonella serovars in systemic infections of poultry. Here, we analyzed populations of clinically relevant isolates of non-Pullorum/Gallinarum Salmonella causing chicken death in China by a whole-genome sequencing approach and determined that antimicrobial-resistant Salmonella Enteritidis remained the major cause in the past decades.
Topics: Animals; Anti-Bacterial Agents; Chickens; Drug Resistance, Bacterial; Multilocus Sequence Typing; Poultry; Poultry Diseases; Salmonella; Salmonella Food Poisoning; Salmonella Infections, Animal; Salmonella enterica; Salmonella enteritidis; Serogroup; Virulence
PubMed: 35727054
DOI: 10.1128/spectrum.00965-22 -
Frontiers in Microbiology 2022serovar Gallinarum (. Gallinarum) is a host-specific pathogen causing fowl typhoid, a severe systemic infection in poultry, which leads to substantial economic losses...
serovar Gallinarum (. Gallinarum) is a host-specific pathogen causing fowl typhoid, a severe systemic infection in poultry, which leads to substantial economic losses due to high morbidity and mortality in many developing countries. However, less is known about the pathogenic characteristics and mechanism of . Gallinarum-induced systemic infection in chickens. In this study, we deleted the . Gallinarum UDP--acetylglucosamine-1-phosphate transferase gene, which contributes to the biosynthesis of enterobacterial common antigen (ECA), and studied the pathogenicity of this ::Cm strain in a chicken model of systemic infection. The ::Cm mutant strain showed comparable growth but lower resistance to bile acid and nalidixic acid than the wild-type strain . In the oral infection model of chickens, the virulence of the ::Cm strain was significantly attenuated . Chickens infected with wild-type strain showed typical clinical signs and pathological changes of fowl typhoid and died between 6 and 9 days post-infection, and the bacteria rapidly disseminated to systemic organs and increased in the livers and spleens. In contrast, the ::Cm mutant strain did not cause chicken death, there were no significant clinical changes, and the bacterial numbers in the liver and spleen of the chickens were significantly lower than those of the chickens infected with the wild-type strain. In addition, the expression of interleukin (IL)-1β, tumor necrosis factor (TNF)-α, and CXCLi1 in the livers of ::Cm-infected chickens was significantly lower than that of the chickens infected with the wild-type strain. Furthermore, the attenuated ::Cm strain could persistently colonize the liver and spleen at low levels for up to 25 days post-infection and could induce a protective immune response in the chickens. These results indicate that the gene is an important virulence factor of . Gallinarum in the chicken model of systemic infection, and the avirulent ::Cm mutant could possibly be used as a live-attenuated vaccine strain for controlling fowl typhoid.
PubMed: 35694286
DOI: 10.3389/fmicb.2022.880932 -
Veterinary Medicine International 2022Medicinal plants have been the good source of treatment for different ailments of humans as well as animals for centuries. However, in Tanzania, few plants were...
Medicinal plants have been the good source of treatment for different ailments of humans as well as animals for centuries. However, in Tanzania, few plants were investigated for their efficacy against various diseases of chickens. In the present study, four medicinal plants were investigated against isolated from chickens. The minimum inhibitory concentration (MIC) using the broth microdilution methods and minimum bactericidal concentration (MBCs) were used to evaluate the activities of plants against chicken salmonellosis. For the safety of chickens against the toxicity of plants, the cytotoxicity assay was determined using a brine shrimp lethality test. leaf ethyl acetate (ALEA), leaf methanolic (ArM), leaf ethyl acetate (ArLEA), and leaf ethyl acetate (PGLEA) extracts exhibited the highest MIC (0.3906 mg/mL) and MBC (3.125 mg/mL), respectively. The tuber ethyl acetate (DTEA) and tuber pet ether (DTPE) extracts displayed MIC of 1.563 mg/mL and 12.50 mg/mL and MBC of 12.50 mg/mL and 25.50 mg/mL, respectively. The highest LC values exhibited in ranged from 7.937 × 10 mg/mL to 7.242 × 10 mg/mL for pet ether and methanolic extracts, respectively, while ALEA extract exhibited LC of 7.645 × 10 mg/mL. Generally, the extracts with MIC 0.3906 mg/mL and MBC 3.125 mg/mL demonstrated the highest antibacterial activity with low toxicity efficient to manage chicken salmonellosis. , which exhibited higher toxicity, warrants further investigation on insecticidal and anticancer agents.
PubMed: 35265313
DOI: 10.1155/2022/2294120 -
Journal of Food Science and Technology Mar 2022spp causes foodborne diseases related to the consumption of contaminated foods, especially poultry products. This study aimed to investigate the occurrence of spp....
spp causes foodborne diseases related to the consumption of contaminated foods, especially poultry products. This study aimed to investigate the occurrence of spp. serovars in raw eggs from supermarkets and street food markets in southern Brazil, to analyze virulence genes, resistance profiling to antimicrobials and sanitizers, and to determine the susceptibility of the isolates to extract. Among 160 samples analyzed, just two (1.25%) were positive for spp.. One positive sample was from egg yolk (. serovar Gallinarum, isolate S28), and another one was from eggshell (. serovar Panama, isolate S37). Regarding the virulence genes, the isolate S37 harbored all the genes evaluated ( and ), while the isolate S28 did not harbor the gene. The isolate S28 was resistant to tobramycin, azithromycin, and trimethoprim, while the isolate S37 showed resistance profile just to nalidixic acid. However, none of the resistance genes evaluated were identified. Both isolates showed resistance to benzalkonium chloride, chlorhexidine digluconate, sodium hypochlorite, and peracetic acid, presenting high MIC values for these sanitizers. In contrast, extract showed antimicrobial activity against the isolates S28 and S37, however, more studies are needed to prove its potential as a natural antimicrobial compound.
PubMed: 35153327
DOI: 10.1007/s13197-021-05113-5 -
Immunobiology Mar 2022Poultry animals act as natural reservoirs of invasive non-typhoidal Salmonella [iNTS] serovars and consumption of iNTS contaminated poultry meat and eggs is one of the...
Poultry animals act as natural reservoirs of invasive non-typhoidal Salmonella [iNTS] serovars and consumption of iNTS contaminated poultry meat and eggs is one of the major sources of iNTS infection in developed and developing countries. Irrational use of antibiotics in the poultry industry gives rise to the global emergence of multi drug resistant iNTS strains. Among different strategies to control iNTS infection in poultry farms, vaccination is now being widely used. There are several licensed vaccines available in the market for poultry animals to ameliorate iNTS infection but none of them have broad spectrum protective efficacy. In this study we have formulated a single novel trivalent iNTS outer membrane vesicles [OMVs] based immunogen which can confer long term broad spectrum protection against most prevalent iNTS serovars. We have isolated OMVs from Salmonella Typhimurium [ST], Salmonella Enteritidis [SE], and Salmonella Gallinarum [SG] and formulated the trivalent immunogen by mixing OMVs in a 1:1:1 ratio. One day old chicks were immunized thrice via oral route at two week intervals. Vaccination significantly induced serovar specific antibodies detected up to 180 days post immunization. Post challenge with both homologous and heterologous [S. Infantis] serovars, immunized birds showed reduced level of fecal shedding and organ invasion. A long term efficacy study also showed reduced levels of tissue invasion up to one year post immunization. These results demonstrate that our novel formulation of immunogen could be a broad spectrum potential vaccine for both layer and broiler breeds against iNTS mediated salmonellosis and fowl typhoid.
Topics: Animals; Chickens; Poultry Diseases; Salmonella Infections, Animal; Salmonella Vaccines; Salmonella enteritidis; Typhoid Fever
PubMed: 35121180
DOI: 10.1016/j.imbio.2022.152183 -
Frontiers in Microbiology 2021is a poultry restricted-pathogen causing fowl-typhoid disease in adult birds with mortality rates up-to 80% and exhibit resistance against commonly used antibiotics. In...
is a poultry restricted-pathogen causing fowl-typhoid disease in adult birds with mortality rates up-to 80% and exhibit resistance against commonly used antibiotics. In this current study, a temperate broad host range bacteriophage SGP-C was isolated against from poultry digesta. It showed infection ability in all the 15 tested field strains of . The SGP-C phage produced circular, turbid plaques with alternate rings. Its optimum activity was observed at pH 7.0 and 37-42°C, with a latent period of 45 min and burst size of 187 virions/bacterial cell. The SGP-C lysogens, and exhibited super-infection immunity against the same phage, an already reported feature of lysogens. A virulence index of 0.5 and 0.001 as MV50 of SGP-C suggests its moderate virulence. The genome of SGP-C found circular double stranded DNA of 42 Kbp with 50.04% GC content, which encodes 63 ORFs. The presence of repressor gene at ORF49, and absence of tRNA sequence in SGP-C genome indicates its lysogenic nature. Furthermore, from NGS analysis of lysogens we propose that SGP-C genome might exist either as an episome, or both as integrated and temporary episome in the host cell and warrants further studies. Phylogenetic analysis revealed its similarity with temperate phages belonging to family . The encoded proteins by SGP-C genome have not showed homology with any known toxin and virulence factor. Although plenty of lytic bacteriophages against this pathogen are already reported, to our knowledge SGP-C is the first lysogenic phage against reported so far.
PubMed: 35095790
DOI: 10.3389/fmicb.2021.768931 -
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 -
Parasites & Vectors Jan 2022The possibility that Dermanyssus gallinae, the poultry red mite, could act as a vector of infectious disease-causing pathogens has always intrigued researchers and... (Review)
Review
The possibility that Dermanyssus gallinae, the poultry red mite, could act as a vector of infectious disease-causing pathogens has always intrigued researchers and worried commercial chicken farmers, as has its ubiquitous distribution. For decades, studies have been carried out which suggest that there is an association between a wide range of pathogens and D. gallinae, with the transmission of some of these pathogens mediated by D. gallinae as vector. The latter include the avian pathogenic Escherichia coli (APEC), Salmonella enterica serovars Enteritidis and Gallinarum and influenza virus. Several approaches have been adopted to investigate the relationship between D. gallinae and pathogens. In this comprehensive review, we critically describe available strategies and methods currently available for conducting trials, as well as outcomes, analyzing their possible strengths and weaknesses, with the aim to provide researchers with useful tools for correctly approach the study of the vectorial role of D. gallinae.
Topics: Animals; Chickens; Disease Vectors; Mite Infestations; Mites; Poultry; Poultry Diseases
PubMed: 35057849
DOI: 10.1186/s13071-021-05142-1 -
Animals : An Open Access Journal From... Dec 2021Antimicrobial resistance and pulsed-field gel electrophoresis (PFGE) genotypes of collected ser. Gallinarum isolates were investigated to examine the epidemiological...
Antimicrobial resistance and pulsed-field gel electrophoresis (PFGE) genotypes of collected ser. Gallinarum isolates were investigated to examine the epidemiological relationship between field outbreak isolates of ser. Gallinarum. Thirty ser. Gallinarum isolates collected from poultry farms with FT outbreaks from 2013 to 2018 in South Korea were analyzed. All isolates were resistant to at least 3 of the 18 antimicrobials tested and exhibited an MDR phenotype. All isolates showed resistance to streptomycin, sulfisoxazole, and colistin. One isolate was resistant to 9 antimicrobials. The antimicrobial resistance profile, streptomycin-sulfisoxazole-colistin-nalidixic acid-ciprofloxacin-gentamicin (18/30, 60.0%), was the most prevalent. PFGE types were classified into 10 groups with a 100% correlation cutoff in dendrograms for 30 field isolates. The dominant PFGE types were 1 (8/30, 26.7%), 4 (7/30, 23.3%), and 9 (5/30, 16.7%). Interestingly some isolates collected from the same and different companies had the same PFGE type. We reported a high MDR rate in ser. Gallinarum isolates. The present study highlights the occurrence of horizontal spread and cyclic contamination of MDR ser. Gallinarum within the same company. Furthermore, we showed cross-contamination between different companies. The characterization of these isolates would be helpful in the development of prevention and control strategies for MDR ser. Gallinarum infection in South Korea.
PubMed: 35011189
DOI: 10.3390/ani12010083