• Title/Summary/Keyword: salmonella

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Rapid and Sensitive Detection of Salmonella in Chickens Using Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Dipstick

  • Liu, Zhi-Ke;Zhang, Qiu-Yu;Yang, Ning-Ning;Xu, Ming-Guo;Xu, Jin-Feng;Jing, Ming-Long;Wu, Wen-Xing;Lu, Ya-Dong;Shi, Feng;Chen, Chuang-Fu
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.454-464
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    • 2019
  • Salmonellosis is a highly contagious bacterial disease that threatens both human and poultry health. Tests that can detect Salmonella in the field are urgently required to facilitate disease control and for epidemiological investigations. Here, we combined loop-mediated isothermal amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) to rapidly and accurately detect Salmonella. LAMP primers were designed to target the Salmonella invA gene. LAMP conditions were optimized by adjusting the ratio of inner to outer primers, $MgSO_4$ concentration, dNTP mix concentration, amplification temperature, and amplification time. We evaluated the specificity of our novel LAMP-LFD method using six Salmonella species and six related non-Salmonella strains. All six of the Salmonella strains, but none of the non-Salmonella strains, were amplified. LAMP-LFD was sensitive enough to detect concentrations of Salmonella enterica subsp. enterica serovar Pullorum genomic DNA as low as $89fg/{\mu}l$, which is 1,000 times more sensitive than conventional PCR. When artificially contaminated feed samples were analyzed, LAMP-LFD was also more sensitive than PCR. Finally, LAMP-LFD gave no false positives across 350 chicken anal swabs. Therefore, our novel LAMP-LFD assay was highly sensitive, specific, convenient, and fast, making it a valuable tool for the early diagnosis and monitoring of Salmonella infection in chickens.

High-throughput sequencing-based metagenomic and transcriptomic analysis of intestine in piglets infected with salmonella

  • KyeongHye, Won;Dohyun, Kim;Donghyun, Shin;Jin, Hur;Hak-Kyo, Lee;Jaeyoung, Heo;Jae-Don, Oh
    • Journal of Animal Science and Technology
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    • v.64 no.6
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    • pp.1144-1172
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    • 2022
  • Salmonella enterica serovar Typhimurium isolate HJL777 is a virulent bacterial strain in pigs. The high rate of salmonella infection are at high risk of non-typhoidal salmonella gastroenteritis development. Salmonellosis is most common in young pigs. We investigated changes in gut microbiota and biological function in piglets infected with salmonella via analysis of rectal fecal metagenome and intestinal transcriptome using 16S rRNA and RNA sequencing. We identified a decrease in Bacteroides and increase in harmful bacteria such as Spirochaetes and Proteobacteria by microbial community analysis. We predicted that reduction of Bacteroides by salmonella infection causes proliferation of salmonella and harmful bacteria that can cause an intestinal inflammatory response. Functional profiling of microbial communities in piglets with salmonella infection showed increasing lipid metabolism associated with proliferation of harmful bacteria and inflammatory responses. Transcriptome analysis identified 31 differentially expressed genes. Using gene ontology and Innate Immune Database analysis, we identified that BGN, DCN, ZFPM2 and BPI genes were involved in extracellular and immune mechanisms, specifically salmonella adhesion to host cells and inflammatory responses during infection. We confirmed alterations in gut microbiota and biological function during salmonella infection in piglets. Our findings will help prevent disease and improve productivity in the swine industry.

Detection of invA and spvC in Salmonella spp. isolated from duck farms (오리 농장에서 분리한 Salmonella속 균에서 invA 및 spvC gene의 검출)

  • Cho, Jae-Keun
    • Korean Journal of Veterinary Service
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    • v.33 no.4
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    • pp.341-344
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    • 2010
  • Poultry and poultry products have been implicated as a major source of Salmonella infection in human, and infection due to Salmonella serotypes continue to be a major health problem. The presence of two virulence genes, invA and spvC, in 34 Salmonella isolates obtained from duck farms was investigated. All isolates contained the invA gene, and spvC gene was found in 20 (58.8%) of 34 Salmonella isolates : S. Typhimurium (n=8), S. Fyris (n=5), S. Enteritidis (n=3), S. Typhimurium var. copenhagen (n=1), S. Haardt (n=1) and S. Mbandaka (n=1). This study showed the presence of the spvC gene was widely distributed in between different Salmonella enterica isolates.

Prevalence of Salmonella Enterotoxin Gene(stn) among Clinical Strains Isolated in Korea and Regulation of stn Expression (한국에서 분리된 병원성 Salmonella 균주의 장독소 유전자(stn) 분포와 발현조절 기작)

  • 임상용;유상렬
    • Microbiology and Biotechnology Letters
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    • v.28 no.6
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    • pp.316-321
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    • 2000
  • Role of enterotoxin from Salmonella in pathogenesis is not know. Enterotoxin gene from Salmonella typhimurium(stn) encodes a 29kDa toxin that has no homology to any other known enterotoxins. Expression of stn is enthanced upon contact with epithelial cell but not all strains having the stn gene express Stn, Based on PCR analysis, we found that all 36 clinical strains of Salmonella isolated in Korea tested carried the stn gene. To understand the trgulation of the stn transcription, the expression of stn was studies in vitro. RNA polymerase was purified by polymin P fraction-ation, DNA-agarose affinity chromatography, and Mono-Q ion exchange chromatography from Salmonella. The expression of stn was inhibited by cAMP·CRP complex by about 50%.

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Characteristics of Salmonella spp isolated from poultry carcasses (닭 도체에서 분리한 Salmonella spp의 특성 분석)

  • Lee, Ho-Won;Hong, Chong-Hae;Jung, Byeong-Yeal
    • Korean Journal of Veterinary Service
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    • v.30 no.3
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    • pp.339-351
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    • 2007
  • Salmonella infections cause the diseases in poultry and some zoonotic Salmonella can be transmitted to human through poultry products, resulting in food-borne disease. This study was conducted to obtain some useful information for the control of salmonellosis in human. Twenty four Salmonella spp were isolated from poultry carcasses, and they were examined with several methods such as serotyping, antimicrobial resistance test and random amplified polymorphic DNA(RAPD) to identify their characteristics. In serotyping test of 24 strains S enteritidis was 17 (70.8%), followed by S schwarzengrund 3 (12.5%), untyped strain 4 (16.7%). In the results of antimicrobial resistance test, 23 (95.8%) isolates were resistant to at least one antimicrobial agent, generating eight different resistance patterns. In RAPD analysis using URP-6 primer to differentiate Salmonella isolates within a serotype, 4 serogroups were divided into 10 RAPD types: 5 types in S enteritidis, 2 types in S schwarzengrund and 3 types in the remainder.

Detection of virulence, specific genes and antibiotic resistance of isolated Salmonella spp. strains from rabbits infected with salmonellosis

  • Huynh Van Chuong;Nguyen Minh Tuan;Nguyen Thi Nhu Anh;Le Thi Lan Phuong;Nguyen Xuan Hoa
    • Korean Journal of Veterinary Research
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    • v.63 no.2
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    • pp.16.1-16.6
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    • 2023
  • Salmonella spp. are pathogens involved in most salmonellosis in rabbits. This study examined Salmonella disease in rabbits raised in Thua Thien Hue, Vietnam. Two hundred and 56 rectal swabs of rabbits were taken, and a carrier rate of 33.98% was found. In addition, all the isolated Salmonella spp. strains were 100% motile; positive for H2S, catalase, Voges Proskauer, coagulase, citrate, maltose, and dextrose; and negative for indole, methyl red, urease, oxidase, sucrose, and lactose. The Kirby-Bauer method showed that these Salmonella strains were susceptible to doxycycline (93.2%), tetracycline (84.1%), and levofloxacin (65.9%). On the other hand, they were highly resistant to streptomycin (95.5%), ampicillin (93.2%), colistin (40.9%), and gentamicin (34.1%). Furthermore, polymerase chain reaction used to screen for virulence and specific genes of Salmonella strains showed that all Salmonella strains isolated carried InvA, fimA, and Stn.

Phenotypic and Genotypic Characterization of Salmonella spp. Isolated from Pigs and their Farm Environment in Korea

  • Lim, Suk-Kyung;Byun, Jung-Ryul;Nam, Hyang-Mi;Lee, Hee-Soo;Jung, Suk-Chan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.50-54
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    • 2011
  • This study's objective was to determine the prevalence of Salmonella spp. in pigs and their farm environments in Korea, and to investigate the relationship between the strains based on their phenotypic and genotypic characteristics. A total of 36 Salmonella spp. were isolated in this study: 18 isolates from 492 pigs (3.7%) and 18 isolates from 418 (4.3%) farmhouse environmental samples from 16 different pig farms. Of the Salmonella strains isolated from the numerous environmental samples, the highest prevalence was observed in slurry or manure, followed by partitions, farmer's hands, floors, water/nipples, ventilation sources, and feed, respectively. All the Salmonella isolates originating from different farms were genetically distinct. In three farms, however, identical phage types and pulse-field gel electrophoresis patterns were observed among Salmonella isolates from pig feces and environmental samples. This study suggests that environments contaminated with Salmonella could pose an infection risk to pigs on pig farms.

Rapid Detection Kit for Salmonella typhimurium (살모넬라 식중독균 신속 검출용 간이 진단키트)

  • Kim, Gi-Young;Yang, Gil-Mo;Park, Saet-Byeol;Kim, Yung-Hwun;Lee, Kang-Jin;Son, Jae-Yong;Kim, Hyuck-Joo;Lee, Sae-Rom
    • Journal of Biosystems Engineering
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    • v.36 no.2
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    • pp.140-146
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    • 2011
  • This study was performed to develop a rapid test kit for pathogenic Salmonella in various samples. The rapid detection kit has been fabricated based on nitrocellulose lateral-flow strip. Colloidal gold and biotin conjugated Salmonella antibodies were used as a tag and a receptor, respectively. Manually spotted Salmonella antibody and Neutravidin on nitrocellulose membrane were used as test and control lines, respectively. Feasibility of the rapid kit to detect Salmonella typhimurium in samples were evaluated. The intensity of the color of the test line started to increase with the samples in which higher concentration of the cells were contained. The sensitivity of the sensor was $10^6$ cfu/mL Salmonella spiked in PBS. Also, the rapid test kit could detect $10^6$ cfu/mL of Salmonella in chicken meat extract.

Studies on Salmonella isolated from chicks (초생추 유래 Salmonella속균의 생물학적 특성)

  • Oh, Gang-hee;Choi, Won-pil
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.501-510
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    • 1994
  • This paper deals with the distribution, reservoir and mode of spread of infection on 6 hatcheries in Taegu, Kyungpook and 5 broiler farms in Kyungpook during the period from June 1991 to June 1992. Isolated Salmonella were examined for serotypes, biotyping of Salmonella(S) typhimurtum, antibiotic susceptibility and some biochemical characteristics. Forty two Salmonella strains were isolated from 42(2.7% of 1,577 caecal samples of chicks, and their serotypes were S typhimurium 10, S typhimurium var Copenhagen 5, S infantis 4, S thompson 3, and untypable 20. The isolation rate of Salmonella varied from 0 to 5.1% in 6 hatcheries and that of Salmonella from 5 broiler farms was 10. 5%. Biotypes of 10 S typhimurium and 5 S typhimurium var Copenhagen strains isolated from chicks of hatcheries and broiler farms were biotype 2(86.6%), 8(6.7%), and 10a(6.7%), and 26i(6.7%) according to Duguid's scheme. Antibiotic susceptibility test of Salmonella isolated were performed by agar dilution method, using 9 antibiotics as follows: ampicillin(Am), chloramphenicol(Cm), gentamicin(Gm), kanamycin(Km), nalidixic acid(Na), rifampicin(Rf), streptomycin(Sm), sulfadimethoxine(Su), and tetracycline(Tc), All the strains were sensitive to RF. But 8 strans(23.8%) were resistant to one or more drugs and the most common resistance patterns of transferred R plasmids were SmSuGm and SmSu. Among 42 isolates, one had transferable citrate utilizing plasmid. S typhimurium and S typhimurium var copenhagen strains were resistant to killing by 90% normal guinea pig serum.

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Evaluation of the extract from lymph nodes for Salmonella monitoring in pig herds (돈군의 Salmonella 모니터링을 위한 림프절 추출액 사용에 대한 평가)

  • Jung, Byeong-Yeal;Choo, Ji-Hoon;Kim, Ji-Hun;Jung, Jae-Yun
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.119-125
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    • 2006
  • The objective of this study was to investigate the use of extract from mesenteric lymph nodes as an alternative to serum for ELISA to detect Salmonella antibodies in slaughter pigs. Among 324 slaughter pigs, 65 (20.1 %) were positive in the serum ELISA and 76 (23.5%) were positive in the ELISA with extract from lymph nodes. A total of 24 (7.4%) Salmonella representing 6 serotypes were isolated from mesenteric lymph nodes and 35 (10.8%) Salmonella belonging to 2 serotypes were also recovered from cecal contents of slaughter pig samples, respectively. The most prevalent serogroup was B (55.9% of isolates) and serotype was Typhimurium (52.5% of isolates). In the comparison of the results of between the serum ELISA and Salmonella isolation, kappa value was 0.28 with mesenteric lymph nodes and 0.37 with cecal contents, respectively. However, the extract ELISA had sensitivity of 98.5%, specificity of 95.4% and kappa value of 0.88 as compared with the serum ELISA. Because high degree of concordance between the serum ELISA and the extract ELISA was observed (P=0.24), extract from lymph nodes could be used as an alternative to serum for the detection of Salmonella antibodies in the ELISA.