Molecular detection of Salmonella isolated from commercial chicken

Document Type : Full paper (Original article)


1 Ph.D. Student in Veterinary Pathology, Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh, 482001, India

2 Department of Veterinary Pathology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh, 482001, India

3 Ph.D. Student in Veterinary Pharmacology & Toxicology, Department of Veterinary Pharmacology & Toxicology, College of Veterinary Science and Animal Husbandry, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh, 482001, India

4 Department of Veterinary Microbiology, College of Veterinary Science and Animal Husbandry Rewa, Nanaji Deshmukh Veterinary Science University, Jabalpur, Madhya Pradesh, 482001, India


Background: Salmonella in chicken, specially, the motile salmonellae, causes the food chain unsafe from farm to table and is considered a significant global threat to public health. Aims: The present study was carried out for molecular detection of Salmonellae in commercial poultry using PCR. Methods: The study was conducted for eight months, from July 2019 to February 2020, and a total of 26 poultry farms, including 15 broiler and 11-layer farms, were visited individually. Pooled faecal samples were obtained from the sheds. A total of 189 necropsy cases were examined for gastrointestinal lesions. Isolation and identification of the organism were done using microbe culture method, and the molecular characterization was performed via PCR targeting invA and ent genes. Results: The prevalence of salmonellosis in the broiler and layer farms was recorded at 20.0% and 45.4%, respectively, through the traditional gold standard culture method. From 189 necropsy birds, salmonellosis was recorded at 1.58% dead cases. Molecular detection of Salmonella isolates by PCR targeting invA gene was confirmed in 13.33% of the broiler farms and 36.3% of the layer farms. Further detection of Salmonella enteritidis was performed by PCR targeting ent gene by which 11.11% positivity was determined. Conclusion: This study, focused on the Salmonella prevalence, highlighted the zoonotic importance of the bacterium in the commercial poultry farms, which can subsequently be dispersed into the human food chain causing harmful health effects.


Main Subjects

Alvarez, J; Sota, M; Vivanco, AB; Perales, I; Cisterna, R; Rementeria, A and Garaizar, J (2004). Development of a multiplex PCR technique for detection and epidemiological typing of Salmonella in human clinical samples. J. Clin. Microbiol., 42: 1734-1738.
Anjay, AK; Agarwal, RK; Ramees, TP; Dubal, ZB; Kaushik, P; Kumar, MS; Dudhe, NC; Milton, AAP; Abhishek, BK and Shagufta, B (2015). Molecular typing of Salmonella typhimurium and S. enteritidis serovars from diverse origin by ERIC-PCR. J. Pure Appl. Microbiol., 9: 2627-2634.
Aseel, AS; Mayada, FH and Majed, HM (2011). Isolation and molecular identification of Salmonella typhimurium from chicken meat in Iraq. J. World’s Poult. Res., 3: 63-67.
Balachandran, P; Balasubramaniam, GA and Sivaseelan, S (2013). Status on prevalence of gastrointestinal diseases in laying chicken. Shanlax Int. J. Vet. Sci., 1: 4-7.
Barrow, PA (2000). The paratyphoid salmonellae. Rev. Sci. Tech. Off. Int. Epizoot., 19: 351-366.
Bordoloi, S; Nayak, A; Sharma, V and Singh, RV (2017). Identification of biofilm forming Salmonella species of layers from tribal area of Hoshangabad district of Madhya Pradesh. J. Anim. Res., 7: 1001-1007.
Carrique-Mas, JJ and Davies, RH (2008). Salmonella enteritidis in commercial layer flocks in Europe: Legislative background, on-farm sampling and main challenges. Braz. J. Poult. Sci., 10: 1-9.
Chowdhury, A; Iqbal, A; Uddin, MG and Uddin, M (2011). Study on isolation and identification of Salmonella and Escherichia coli from different poultry feeds of Savar region of Dhaka, Bangladesh. J. Sci. Res., 3: 403-411.
Dewaele, I; Van Meirhaeghe, H; Rasschaert, G; Vanrobaeys, M; De Graef, E; Herman, L; Ducatelle, R; Heyndrickx, M and De Reu, K (2012). Persistent Salmonella enteritidis environmental contamination on layer farms in the context of an implemented national control program with obligatory vaccination. Poult. Sci., 91: 282-291.
Dogru, AK; Ayaz, ND and Gencay, YE (2010). Serotype identification and antimicrobial resistance profiles of Salmonella spp. isolated from chicken carcasses. Trop. Anim. Health Prod., 42: 893-897.
Freitas, CG; Santana, AP; da Silva, PHC; Goncalves, VSP; Barros, MDAF; Torres, FAG; Murata, LS and Perecmanis, S (2010). PCR multiplex for detection of Salmonella enteritidis, typhi and typhimurium and occurrence in poultry meat. Int. J. Food Microbiol., 139: 15-22.
Galan, JE; Ginocchio, C and Costeas, P (1992). Molecular and functional characterization of the Salmonella invasion gene invA: homology of invA to members of a new protein family. J. Bacteriol. Res., 174: 4338-4349.
Gelinski, JMLN; Martin, G; Destro, MT; Landgraf, M and Franco, BDGDM (2002). Rapid detection of Salmonella in foods using a combination of SPRINTTM, MSRVTM and Salmonella Latex TestTM. Rev. Bras. Cienc. Farm., 38: 315-322.
Gorski, L (2012). Selective enrichment media bias the types of Salmonella enterica strains isolated from mixed strain cultures and complex enrichment broths. PLoS One., 7: e34722.
Hyeon, JY; Li, S; Mann, DA; Zhang, S; Kim, KJ; Lee, DH; Deng, X and Song, CS (2021). Whole-genome sequencing analysis of Salmonella enterica serotype Enteritidis isolated from poultry sources in South Korea, 2010-2017. Pathogens. 10: 1-12.
Jinu, M; Agarwal, RK; Sailo, B; Wani, MA; Kumar, A; Dhama, K and Singh, MK (2014). Comparison of PCR and conventional cultural method for detection of Salmonella from poultry blood and faeces. Asian J. Anim. Vet. Adv., 9: 690-701.
Jofre, A; Martin, B; Garriga, M; Hugas, M; Pla, M; Rodriguez-Lazaro, D and Aymerich, T (2005). Simultaneous detection of Listeria monocytogenes and Salmonella by multiplex PCR in cooked ham. Food Microbiol., 22: 109-115.
Kaushik, P; Kumari, S; Bharti, SK and Dayal, S (2014). Isolation and prevalence of Salmonella from chicken meat and cattle milk collected from local markets of Patna, India. Vet. World. 7: 62-65.
Kumari, D; Mishra, SK and Lather, D (2013). Pathomicrobial studies on Salmonella gallinarum infection in broiler chickens. Vet. World. 6: 725-729.
Malorny, B; Tassios, PT; Radstrom, P; Cook, N; Wagner, M and Hoorfar, J (2003). Standardization of diagnostic PCR for the detection of foodborne pathogens. Int. J. Food Microbiol., 83: 39-48.
Meteab, BK and Abed, AAA (2018). Isolation and identification of Salmonella serotypes in poultry. Al-Qadisiyah J. Vet. Med. Sci., 17: 75-80.
Mir, IA; Kashyap, SK and Maherchandani, S (2015). Isolation, serotype diversity and antibiogram of Salmonella enterica isolated from different species of poultry in India. Asian Pac. J. Trop. Biomed., 5: 561-567.
Mshelbwala, FM; Saidu, SNA; Kwanashie, CN; Kadiri, AKF; Anise, EO; Nyillah, EL and Ibrahim, NDG (2018). Comparative study on the sensitivity of bacteriology and immunohistochemical technique in the diagnosis of natural salmonellosis in chickens. Sokoto J. Vet. Sci., 16: 60-70.
Nahar, A; Siddiquee, M; Nahar, S; Anwar, KS and Islam, S (2014). Multidrug resistant-Proteus mirabilis isolated from chicken droppings in commercial poultry farms: Bio-security concern and emerging public health threat in Bangladesh. J. Biosaf. Health Edu., 2: 1-5.
Naik, VK; Shakya, S; Patyal, A and Gade, NE (2015). Isolation and molecular characterization of Salmonella spp. from chevon and chicken meat collected from different districts of Chhattisgarh, India. Vet. World. 8: 702-706.
Nazir, S; Kamil, SA; Darzi, MM; Mir, MS and Amare, A (2012). Pathology of spontaneously occurring salmonellosis in commercial broiler chickens of Kashmir valley. J. World’s Poult. Res., 2: 63-69.
Nierop, W; Duse, AG; Marais, E; Aithma, N; Thothobolo, N; Kassel, M; Stewart, R; Potgieter, A; Fernandes, B; Galpin, JS and Bloomfield, SF (2005). Contamination of chicken carcasses in Gauteng, South Africa, by Salmonella, Listeria monocytogenes and Campylobacter. Int. J. Food Microbiol., 99: 1-6.
Oliveira, SD; Santos, LR; Schuch, DMT; Silva, AB; Salle, CTP and Canal, CW (2002). Detection and identification of salmonellas from poultry-related samples by PCR. Vet. Microbiol., 87: 25-35.
O’Regan, E; McCabe, E; Burgess, C; McGuinness, S; Barry, T; Duffy, G; Whyte, P and Fanning, S (2008). Development of a real-time multiplex PCR assay for the detection of multiple Salmonella serotypes in chicken samples. BMC Microbiol., 8: 1-11.
Pantin-Jackwood, MJ (2013). Multicausal enteric diseases. In: Swayne, DE (Ed.), Diseases of poultry. (13th Edn.), Ames, Iowa, Iowa State University Press. PP: 1322-1325.
Park, SH; Ryu, S and Kang, DH (2012). Development of an improved selective and differential medium for isolation of Salmonella spp. J. Clin. Microbiol., 50: 3222-3226.
Poppe, C; Irwin, RJ; Messier, S; Finley, GG and Oggel, J (1991). The prevalence of Salmonella enteritidis and other Salmonella spp. among Canadian registered commercial chicken broiler flocks. Epidemiol. Infect., 107: 201-211.
Rahn, K; De Grandis, SA; Clarke, RC; McEwen, SA; Galan, JE; Ginocchio, C; Curtiss Iii, R and Gyles, CL (1992). Amplification of an invA gene sequence of Salmonella typhimurium by Polymerase Chain Reaction as a specific method of detection of Salmonella. Mol. Cell. Probes. 6: 271-279.
Rall, VLM; Rall, R; Aragon, LC and Silva, MGD (2005). Evaluation of three enrichment broths and five plating media for Salmonella detection in poultry. Braz. J. Microbiol., 36: 147-150.
Ramya, P; Madhavarao, T and Rao, LV (2012). Study on the incidence of Salmonella enteritidis in poultry and meat samples by cultural and PCR methods. Vet. World. 5: 541-545.
Ranjbar, VR; Basiri, S and Abbasi-Kali, R (2020). Paratyphoid infection caused by Salmonella typhimurium in a pigeon flock (Columbia livia) in Iran. J. Zoonotic Dis., 4: 43-48.
Salem, WM; El-Hamed, DS; Sayed, W and Elamary, R (2017). Alterations in virulence and antibiotic resistant genes of multidrug-resistant Salmonella serovars isolated from poultry: The bactericidal efficacy of Allium sativum. Microb. Pathog., 108: 91-100.
Samanta, I; Joardar, SN; Das, PK; Sar, TK; Bandyopadhyay, S; Dutta, TK and Sarkar, U (2014). Prevalence and antibiotic resistance profiles of Salmonella serotypes isolated from backyard poultry flocks in West Bengal, India. J. App. Poult. Res., 23: 536-545.
Saravanan, S; Purushothaman, V; Murthy, TRGK; Sukumar, K; Srinivasan, P; Gowthaman, V; Balusamy, M; Atterbury, R and Kuchipudi, SV (2015). Molecular epidemiology of Non-typhoidal Salmonella in poultry and poultry products in India: implications for human health. Indian J. Microbiol., 55: 319-326.
Scholz, HC; Arnold, T; Marg, H; Rosler, U and Hensel, A (2001). Improvement of an invA-based PCR for the specific detection of Salmonella typhimurium in organs of pigs. In: Fourth International Symposium on the Epidemiology and Control of Salmonella and Other Food Borne Pathogens in Pork. January 2001. PP: 401-403.
Shanmugasamy, M; Velayutham, T and Rajeswar, J (2011). InvA gene specific PCR for detection of Salmonella from broilers. Vet. World. 4: 562-564.
Sharma, I and Das, K (2016). Detection of invA gene in isolated Salmonella from marketed poultry meat by PCR assay. J. Food Process. Technol., 7: 1-4.
Shekhar, C; Upadhyay, AK and Singh, SP (2013). Prevalence of Salmonella in foods of animal origin and its public health significance. J. Vet. Public Health. 11: 57-60.
Sojka, MG; Carter, MA and Thorns, CJ (1998). Characterisation of epitopes of type 1 fimbriae of Salmonella using monoclonal antibodies specific for SEF21 fimbriae of Salmonella enteritidis. Vet. Micro., 59: 157-174.
Soria, MC; Soria, MA; Bueno, DJ; Godano, EI; Gomez, SC; ViaButron, IA; Padin, VM and Roge, AD (2017). Salmonella spp. contamination in commercial layer hen farms using different types of samples and detection methods. Poult. Sci., 96: 2820-2830.
Suresh, Y; Bindukiranmayi, C; Rao, TS; Srivani, M; Uday, A and Sankar, SN (2019). Prevalence and virulence gene profiles of Salmonella serovars isolated from animal origin. J. Pharm. Innov., 8: 139-145.
Upadhyay, A; Swamy, M; Dubey, A and Shrivastava, DP (2015). Prevalence of bovine salmonellosis in organised dairy farm. The Indian J. Vet. Sci. Biotech., 10: 60-62.
Vaez, H; Ghanbari, F; Sahebkar, A and Khademi, F (2020). Antibiotic resistance profiles of Salmonella serotypes isolated from animals in Iran: a meta-analysis. Iran. J. Vet. Res., 21: 188-197.
Vassiliadis, P (1983). The Rappaport-Vassiliadis (RV) enrichment medium for the isolation of salmonellas: an overview. J. Appl. Bacteriol., 54: 69-76.
Veen, C; De Bruijn, ND; Dijkman, R and De Wit, JJ (2017). Prevalence of histopathological intestinal lesions and enteric pathogens in Dutch commercial broilers with time. Avian Pathol., 46: 95-105.
Yang, Y; Tellez, G; Latorre, JD; Ray, PM; Hernandez, X; Hargis, BM; Ricke, SC and Kwon, YM (2018). Salmonella excludes Salmonella in poultry: confirming an old paradigm using conventional and barcode-tagging approaches. Front. Vet. Sci., 5: 1-7.
Yhiler, NY; Bassey, EB and Useh, MF (2015). Evaluation of the performance of two selective enrichment media and two selective plating media for the detection of Salmonella from primary poultry production, according to ISO 6579: 2002. Open J. Med. Microbiol., 5: 128-135.