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Govind Ballabh Pant University of Agriculture and Technology, Pantnagar

After independence, development of the rural sector was considered the primary concern of the Government of India. In 1949, with the appointment of the Radhakrishnan University Education Commission, imparting of agricultural education through the setting up of rural universities became the focal point. Later, in 1954 an Indo-American team led by Dr. K.R. Damle, the Vice-President of ICAR, was constituted that arrived at the idea of establishing a Rural University on the land-grant pattern of USA. As a consequence a contract between the Government of India, the Technical Cooperation Mission and some land-grant universities of USA, was signed to promote agricultural education in the country. The US universities included the universities of Tennessee, the Ohio State University, the Kansas State University, The University of Illinois, the Pennsylvania State University and the University of Missouri. The task of assisting Uttar Pradesh in establishing an agricultural university was assigned to the University of Illinois which signed a contract in 1959 to establish an agricultural University in the State. Dean, H.W. Hannah, of the University of Illinois prepared a blueprint for a Rural University to be set up at the Tarai State Farm in the district Nainital, UP. In the initial stage the University of Illinois also offered the services of its scientists and teachers. Thus, in 1960, the first agricultural university of India, UP Agricultural University, came into being by an Act of legislation, UP Act XI-V of 1958. The Act was later amended under UP Universities Re-enactment and Amendment Act 1972 and the University was rechristened as Govind Ballabh Pant University of Agriculture and Technology keeping in view the contributions of Pt. Govind Ballabh Pant, the then Chief Minister of UP. The University was dedicated to the Nation by the first Prime Minister of India Pt Jawaharlal Nehru on 17 November 1960. The G.B. Pant University is a symbol of successful partnership between India and the United States. The establishment of this university brought about a revolution in agricultural education, research and extension. It paved the way for setting up of 31 other agricultural universities in the country.

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  • ThesisItemOpen Access
    Isolation, identification and molecular characterization of nontyphoidal Salmonella and listeria SPP. from foods of animal origin
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Gunjiyal, Harshita; Maansi
    Non-Typhoidal Salmonella and Listeria are the two bacterial food-borne organisms that pose a major impact to the food sector worldwide. In addition, the emergence of multi-drug resistant organisms makes food-borne illnesses more severe. In view of this, the present study aims to ascertain the occurrence of non-typhoidal Salmonella and Listeria organisms in foods and their antimicrobial resistance profiles isolated from animal origin foods of four districts belonging to Kumaon region of Uttarakhand. A total of 250 samples comprising raw milk (n=141), milk products (n=59) and poultry meat (n=50) were collected randomly from multiple vendors, dairy farms, locality, butcher shops and screened for the presence of non-typhoidal Salmonella and Listeria organisms. The bacteria were isolated using culture method and biochemical identification was performed as per conventional method. Further, molecular characterization was done for confirmation. Antibiotic susceptibility testing was performed for the obtained isolates against a set of 12 antibiotics belonging to 9 different classes for Salmonella spp. and 8 different classes for Listeria spp. using the standard Kirby Bauer disc diffusion method. Salmonella spp. was detected in 7.2%; 18/250 and Listeria spp. in 2.4%; 6/250 of the 250 food samples studied. None of the Listeria isolates was found to be belonging to L. monocytogenes. Serotyping of Salmonella isolates revealed that S. Typhimurium and S. Weltevreden correspond to the dominant serotypes recording (4/18; 22.22%) higher serovar occurrence than S. Kentucky (2/18; 11.11%), S. Infantis (2/18; 11.11%). Rest were untypable (6/18; 33.33%). U.S Nagar harbored more Salmonella spp. (12.5%) followed by Nainital district (3.90%). On the other hand, Nainital district (3.12%) was found to harbor more Listeria spp. than U.S Nagar (1.9%). On subjection to antimicrobial susceptibility testing, Salmonella isolates showed varying degree of sensitivity to Co-trimoxazole (55.55%), co-resistance to Gentamicin and Chloramphenicol (44.44%). Complete phenotypic resistance (100%) was found for Cefotaxime and Erythromycin followed by Nalidixic acid (72.22%). Out of 18 obtained Salmonella isolates, 14 isolates (77.77%) were multi-drug resistant. A total of 12 different antimicrobial resistance patterns were observed. On the other hand, Listeria spp. were completely susceptible (100%) to Vancomycin, Chloramphenicol and Ampicillin. Complete resistance (100%) was found for Kanamycin and Tetracycline followed by Amikacin (83.33%). Out of six obtained isolates for Listeria spp. five isolates (5/6; 83.3%) were multi-drug resistant. A total of 5 different antimicrobial resistance patterns were observed which can be related to the non-judicious administration of antibiotics during both prophylaxis and treatment. Therefore, this study warrants careful consideration towards adopting hygienic measures and consumption of properly cooked food along with judicious use of antibiotics.
  • ThesisItemOpen Access
    Epidemiology and molecular characterization of Cystic echinococcosis in cattle and Cysticercosis in pigs
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-06) Pathak, Ayushi; Upadhyay, A.K.
    The present study was undertaken to study the epidemiology of Echinococcosis in cattle and Cysticercosis in pigs through slaughter house study of Uttarakhand. During this study, 264 buffaloes and 100 pigs were studied. The overall prevalence rate of Hydatidosis was 13.26% {Nainital (n=172)- 14.53%, Bageshwar (n=25), 12%, Almora (n=18)-11.11%, US Nagar (n=38), 7.89% and Rampur (n=11)- 18.18%}. For Cysticercosis, the overall prevalence rate was 3% {US Nagar (n=60)-1.66%, Nainital (n=30)-3.33% and Bageshwar (10)-10%}. Both the diseases were found to be independent of the geographical location of the animals. The average prevalence recorded in lungs, liver and both lungs & liver was 14.28%, 71.42% and 14.28%, respectively without significant statistical difference. Genderwise prevalence revealed no significant difference statistically and was higher in females in both the diseases (hydatidosis- 15.43% and cysticercosis-3.03%) and lower in males (hydatidosis-9.8% and cysticercosis-2.94%). Statistical significant difference could not be observed on age-wise prevalence considerations {Hydatidosis- 6-7y (16.66%), 4-5y (14.63%) and 2-3y (8%); cysticercosis-1-2y (4.16%) and 3-4 years (2.77%)}. Various types of cysts were recovered from different organs without statistical significant difference in rate of infection {single (lungs-80%, liver-80%) as well as multiple (lungs-20%, liver-20%)}. The overall fertility rates found in the cysts were 71.42% (lungs-40%, liver-80%, lungs & liver-60%). The overall viability rate of the cysts recovered from different organs was 80% (lungs-50%, liver- 80%, lungs & liver-100%). Significant difference was not observed in sizes of cyst and their viability {3-6cm (85.71%); >6cm (77.78%)}. DNA was extracted from 25 Echinococcus samples and 3 T. solium samples. Cox1 gene amplification fragment length observed was 440bp in Echinococcus and 420bp in Cysticercus. Purification of PCR products lead to clear products of 340 bp for US Nagar isolate; 338bp for Nainital isolate; 319 bp for Bageshwar isolate; 330 bp for Almora isolate; 339 bp for Rampur isolate for Echinococcus. For Cysticercus, clear products of 322 bp for US Nagar isolate; 418 bp for Bageshwar isolate and 354 bp for Nainital isolate were obtained. Phylogenetic analysis of the isolates show that Echinococcus isolates from the present study are 100% similar to the isolates from South Korea, Iran, Italy, Israel, Turkey and Iraq (G1 isolate). The T. solium isolates share 97.92% resemblance with Mexico isolate (TsChb4), 98.02% with Bangalore isolate, 98.59% with Meghalaya isolate (Ts-MI) and, 97.99% with Uttar Pradesh isolate (TsInd D). The Bageshwar isolate of T. solium is shown to be the root of the phylogenetic tree with other isolates being the branches. This is the first report of prevalence of G1 isolate of Echinococcus in buffaloes. Asian isolate of T. solium is found in the pigs of Uttarakhand, thus explaining the significance of this study in understanding the zoonotic potential of both the parasites.
  • ThesisItemOpen Access
    Epidemiological studies on rabies through meta-analysis
    (G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Chauhan, Ram Swaroop; Upadhyay, A.K.
    In the present study, meta-analysis on rabies in India by the use of a random-effect model was done to estimate the prevalence of the disease in India. The data is obtained from the peer-reviewed articles and publications on this disease during 2010-2020. The data which is used in the present study includes the studies in which the samples are completely random. The Meta-analysis for the epidemiology and sero-prevalence of rabies was done on a total of 32 studies. Further subgroup analysis was done like analysis for species, geographical regions, and diagnostic tests.The total sample size for prevalence estimation in humans are 49828 and sero-prevalence estimation in dogs by ELISA and RFFIT are 1856 and 689 respectively. Pooled prevalence of published papers using random-effect model for rabies in humans was estimated 65% (95%CI: 40%-86%) and in dogs by ELISA and RFFIT was estimated 53%(95%CI: 33%-73%) and72%(54%CI: 95%- 86%) respectively.Publication bias for rabies in humans through regression test revealed significant publication bias (z = 0.6947, p> 0.05). Sero-prevalence of rabies in dogs by ELISA by rank correlation test showed non-significant (Kendall’s tau = 0.1111, p> 0.05) and regression test revealed significant publication bias (z= 0.2142, p>0.05). For seroprevalence of rabies in dogs by RFFIT the rank correlation test showed non-significant (Kendall’s tau = 0.4667, p > 0.05)and regression test revealed significant publication bias (z= 0.3222, p>0.05). The majority of bite victims were between the ages of 0-20 (21.49%) followed by(20.30%) the ages of 21-40. In the studies that were mentioned, males were disproportionately more likely (71.87%) to have been bitten by a dog than females (28.13%). The majority of victims suffer animal bites on their extremities. Maximum dog bites were recorded in the evening (62.9%). According to a survey, 15% of dogs in the nation were vaccinated. With the help of definite and precise clinical history with epidemiological rates, relation between associated factors may help in the identification of the highest disease burden that helps us to improve our knowledge to develop a plan of action for effective control and prevention measure
  • ThesisItemOpen Access
    Prevalence and characterization of non-typhoidal Salmonella isolates obtained from retail fish meat shops
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Sana Parveen; Sana Parveen; Sana Parveen; Maansi; Maansi; Maansi
    Non-Typhoidal salmonellosis stands among the major food illnesses. Outbreaks from contaminated fishes have been witnessed worldwide. This study was performed to assess the presence of Non -Typhoidal Salmonella in fishes and associated environmental samples of 22 retail fish meat shops of 4 locations of Uttarakhand. For this, a total of 368 samples (fish meat swabs(n=46), gill swabs(n=23), skin swabs(n=25), intestine swabs(n=23), hand swabs(n=44), knife swabs(n=27), rinsing water(n=22), fish water(n=41), floor swabs(n=22), chopping board swabs(n=24), utensil swabs(n=24) and container swabs(47) were collected. The overall occurrence of Non-Typhoidal Salmonella was found to be 4.35% (16/368). The highest prevalence of Salmonella was observed in rinsing water (18.18%,4/22) sample followed by knife swabs ( 11.11%, 3/27) , chopping board swabs (8.33%, 2/24) , meat swabs (6.52%, 3/46) , floor swab (4.54%,1/22) , gill and intestine swab (4.34%, 1/23), fishwater (2.43%,1/41). Geographically, the highest prevalence was observed in Lalkuan (6.52%, 3/46) followed by Haldwani (5.55%, 10/180), Kiccha (2.12%, 1/47) and Pantnagar (2.08%, 2/96). Of 16 Salmonella isolates, confirmed using ompC (204 bp) gene, 14 were revealed as S.Typhimurium (87.5%, 14/16) while 2 did not exhibit the typh gene (401bp) amplicon size. All the 16 isolates screened for the presence of virulence genes using PCR commonly harbored sipA gene (87.5%, 14/16) followed by stn (75%, 12/16), sopB ( 68.75%, 11/16), sopE1 ( 56.25%, 9/16) mgtC ( 43.75%, 7/16) and spvC and gipA ( 12.5%, 2/16) genes each. Highest resistance was observed against Tetracycline and Ampicillin (93.75%, 15/16), Nalidixic acid (50%, 8/16), Ciprofloxacin (37.5%, 6/16) Ofloxacin, Cefotaxime and Sulfisoxazole ( 25%, 4/16) each, Chloramphenicol (12.5%, 2/16) and Streptomycin ( 6.25%, 1/16). Ten Salmonella isolates were multi drug resistant (MDR). Ten different antimicrobial resistance patterns were observed. Of these, only one pattern (TE, AMP, NA, CTX , OF, SF) was found common in 2 Salmonella isolates belonging to knife and meat swab sample. All phenotypically resistant and intermediate resistant isolates were screened for 6 corresponding antimicrobial resistance genes. The most commonly occurring resistance gene was gyrA (92.30%, 12 /13), blaTEM (53.33%, 8/15), aadA1 and strA (50%, 2/4), sul1 (30.76 %, 4/13) while tetA was not found in any of the isolates. Overall, our study detected occurrence of Non-Typhoidal Salmonella in the fish retail meat shops. Resistance to critically important flouroquinolones and highly important Cephalosporin and Tetracycline antibiotic detected in Salmonella isolates is a serious threat to public health which highlights the indiscriminate use of antimicrobials.
  • ThesisItemOpen Access
    Whole genome sequencing & bioinformatics of Campylobacter isolated from animals
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-11) Roma; Upadhyay, A.K.
    Campylobacter species are one of the leading cause of bacterial foodborne zoonoses. Worldwide, Pathogenic Campylobacter species are responsible for causing over 400–500 million infections cases each year. These pathogenic Campylobacter species are grouped into major human enteric pathogens (C. jejuni, C. jejuni subsp. jejuni (Cjj), C. jejuni subsp. doyley (Cjd), C. coli and C. fetus) minor pathogens (C. concisus, C. upsaliensis, C. lari and C. hyointestinalis) and major veterinary pathogens (C. fetus subsp. venerealis (Cfv) and C. Fetus subsp. fetus (Cff)). A total of 498 samples consisting of poultry caeca (120), litter (40), poultry rectal swabs (24), poultry skin (24), water samples (38),faeces of pigs (30), meat swabs (32), clinical dog faecal ssample (20), goat (32) and lab animals faeces (92) were screened for the presence of C. jejuni and C. coli using conventional isolation and identification procedures. The overall occurence of Campylobacter was 8.83 %. The samples collected were screened for campylobacters. Highest isolation rate was observed from lab animals (19.56 %, 18/92) followed by poultry caeca (13.33%,16/120), pig faeces (10%,3/30), meat swabs (9.37 %, 3/32) ,poultry faeces (7.14%, 3/42) and litter (2.5%, 1/40). Since loads of virulence genes are responsible for bacterial pathogenecity, resilence, stress response and environmental persistence. The individual detection of every gene by techniques like PCR is a tedious process. The next generation sequencing concept play an extremely important role in this context. Among next generation sequencing also, whole genome sequencing (WGS) and assembly is best method for the characterization of isolates as it can detect every gene of the organism coding for it’s identification, virulence, pathogenicity and antibiotic resistance.The study characterised the entire genome of the infectious isolates by analysis of WGS sequence data via use of both web based and command based tools. Virulence genes pattern was observed using web based analysis tools namely VFDB Virulence Factor Database (http://www.mgc.ac.cn/VFs) and wgMLST (Whole-genome MLST) which is pangenome based tool was used using core genome genes/loci and all acessory genes/loci to detect lineage – specific genes/loci. A total of 23 genes were detected by VFDB and 33 virulence genes were detected via SRST2. Only resistance to beta – lactam were found. Overall, our study detected only one antimicrobial resistance gene that is blaₒₓₐ -₄₈₉ commonly found in Campylobacter spp. indicating good antibiotic management in areas of Pantnagar, Rudrapur and Barielly
  • ThesisItemOpen Access
    Molecular genotyping of multidrug-resistant Salmonella typhimurium isolates using repetitive sequence-based PCR fingerprinting technique
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2020-08) Sivakumar, V.; Maansi
    Non-typhoidal salmonellae, responsible for salmonellosis in humans, are a threat to food safety impacting human health in the form of hospitalizations and in severe cases, fatalities. Such an impact also results in huge economic losses. Several food borne outbreaks resulting from zoonotic non- typhoidal Salmonella serovars have been reported, of which, S. Typhimurium has shown predominance. Epidemiological investigations of an outbreak requires accurate identification of the infection source and the transmission route for effective implementation of preventive measures against microbial food-borne pathogens. For this, techniques which can detect differences among the isolates at genomic level are applied. Therefore, in the present investigation, multidrug-resistant isolates of Salmonella Typhimurium were genotyped using repetitive sequence-based PCR (rep-PCR) fingerprinting. A total of 45 MDR S. Typhimurium isolates belonging to various sources and locations were procured from the Department of Veterinary Public Health and Epidemiology, CVASc, GBPUAT, Pantnagar. All the isolates were tested for purity and were confirmed on the basis of cultural, biochemical, serotyping and molecular (duplex PCR targeting genus specific ompC gene and serovar specific typh gene) assay. The isolates were subjected to ERIC, REP, BOX and GTG5-PCR analysis after DNA extraction and PCR confirmation. ERIC, REP, BOX and GTG5-PCR fingerprinting generated reproducible band patterns ranging from 3-10, 1-5, 6-14 and 2-10 separate bands, respectively. Cluster analysis revealed 9 types from ERIC-PCR, 7 types from REP-PCR, 16 types from BOXPCR and 9 types from GTG5-PCR. Hundred percent typeability was obtained with all genotyping techniques except REP-PCR, which differentiated 39 Typhimurium isolates only (86.6%). The isolates’ sharing similar band patterns is suggestive of a genetic similarity between them directing towards the multifactor involvement in the transmission of Salmonella. Faecal isolates and meat swab isolates sharing identical band patterns and grouped into a single cluster is indicative of a likely cross contamination. No unique pattern was observed in penta resistant (ACSSuT) profile isolates by our typing techniques. DI value of BOX-PCR was highest (0.940) followed by GTG5-PCR (0.862), REP-PCR (0.846) and ERIC-PCR (0.843). On comparing the techniques, the BOX-PCR exhibited good DI value, typeability and complex band pattern on gel in differentiating the isolates. Hence, to conclude, the BOX-PCR fingerprinting technique was found useful in the genotyping of isolates suitable enough to find its application in an epidemiological investigation during an outbreak.
  • ThesisItemOpen Access
    Studies on non-typhoidal Salmonella isolates obtained from indigenous and exotic layers of an organized poultry farm
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Nagpal, Aastha; Maansi
    The present study was undertaken to assess the prevalence, virulence characteristics and antimicrobial resistance at phenotypic and genotypic level of non-typhoidal Salmonella isolated from layer flocks of indigenous (Uttara fowl and Kadaknath) and exotic (RIR, White Leghorn and Australorp) breeds of an organized farm at Pantnagar. A total of 470 samples were collected from 5 layer breeds, Uttara fowl (n=220), Kadaknath (n=55), RIR (n=91), White Leghorn (n=55) and Australorp (n=49). The samples (n=470) comprised of poultry faeces (n=90), litter (n=75), feed (n=70), water (n=65), eggs (n=120) [egg surface (60) + egg content (60)] and caecal content (n=50) of dead birds.The overall prevalence of non-typhoidal Salmonella was 4.89% (23/470). Higher prevalence was observed in exotic breeds (8.21%, 16/195) than indigenous breeds (2.55%, 7/275). Among the different breeds, RIR showed the higher occurrence (17.58%, 16/91), followed by Uttara fowl (2.73%, 6/220) and Kadaknath (1.82%, 1/55). White Leghorn and Australorp did not reveal any presence of Salmonella. Of the total samples (n=470), the highest prevalence was observed in water samples (7/65, 10.77%), followed by poultry faeces (6/90, 6.67%), caecal content (3/50; 6.0%), litter (4/75, 5.33%), feed (2/70, 2.86%) and egg samples (1/120, 0.83%). Only one sample of egg surface rinse was positive (1/60, 1.67%) and none of the egg content samples (n=60) showed positive result. Serotyping revealed the presence of a single serovar viz. Salmonella Typhimurium (91.3%, 21/23), while two isolates (8.7%, 2/23) remained untypable. All 23 isolates were screened for the presence of 8 virulence genes by PCR. Majority of the isolates (22) carried sipA (95.65%) followed by sopB 17 (73.91%), sopE1 14 (60.87%), stn 13 (56.52%), fliC 11 (47.83%) and mgtC 7 (30.43%), spvC and gipA 3 each (13.04%). All 23 isolates when tested against 13 antimicrobials, showed highest resistance for Erythromycin 23 (100%) followed by Ampicillin 15 (65.22%), Nalidixic Acid 13 (56.52%), Ciprofloxacin 12 (52.17%), Cefazolin 9 (39.13%), Cefotaxime 8 (34.78%), Sulfisoxazole 7 (30.43%), Enrofloxacin 6 (26.09), Gatifloxacin 5 (21.74%), Cefoxitin and Tetracycline 4 each (17.39%), Levofloxacin 3 (13.04%) and Streptomycin 2 (8.70%). Phenotypic co-resistance against Ciprofloxacin and Cefotaxime was identified in 5 (5/23, 21.74%) isolates. Eighteen out of twenty-three isolates (78.26%) were multidrug resistant (MDR). Sixteen different antimicrobial resistance patterns were observed. Of these, common resistance patterns were CZ CTX E AMP, NA CIP LE GAT EX E AMP SF, S E, NA CIP CTX E AMP TE SF, E AMP, NA CIP CZ CX CTX E AMP and NA CIP CZ CX E AMP (all 2/23, 8.7%). The multiple antibiotic resistance (MAR) index was found in a range of 0.15-0.69. All phenotypically resistant (including intermediate) isolates were screened for 16 corresponding antimicrobial resistance genes. The most commonly occurring resistant gene was gyrA (23/23, 100%) followed by parC (22/23, 95.65%), aadA1 (2/4, 50%), sul1 (3/7, 42.86%), blaTEM (5/23, 21.74%) and qnrS (1/23, 4.35%) while blaPSE, blaCMY, aadA2, strA, strB, sul2, tetA, tetB, tetG and qnrA were not found in any of the isolates. None of the isolates exhibited Class1 integrons. Overall, our study detected low occurrence of NTS from the layer flocks of an organized farm. Co-resistance to clinically important antimicrobials, ciprofloxacin and cefotaxime observed in this study is a matter of concern that necessitates only deliberate use of antimicrobials in the poultry farms in the country.
  • ThesisItemOpen Access
    Isolation, characterization and prevalence of thermophilic Campylobacters in poultry farms and meat vendors using novel enrichment method
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Bisht, Piyush; Upadhyay, A.K.
    Campylobacters are Gram negative helical bacilli belonging to the family Campylobacteraceae and are of major public health significance as they are one of the leading cause of food borne gastroenteritis worldwide. The present study was carried out to find a novel technique for isolation of Campylobacter spp. and and determine the prevalence of thermophilic Campylobacter spp. in poultry and their living environment at different poultry farms and meat vendors located in Kumaon region of Uttarakhand state. Most of the Campylobacter culture media described in the literature are supplemented with sheep or horse blood, therefore, this study examined the use of goat blood as potential substitutes for sheep blood in blood-supplemented bacteriologic media commonly used in clinical microbiology laboratories. A total of 381 samples comprising of 156 poultry caeca, 86 poultry faeces, 25 goat faeces, 10 sheep faeces, 24 meat swabs and 80 environmental samples viz.; water (n=32) and litter (n=48) were collected from three (n=3) poultry farms and meat shops (n=4) were analyzed, of which 49 samples showed characteristic colonies, either having a spreading or watery nature. They were typical Gram negative spiral rods and had characteristic cork screw motility. All the 49 Campylobacter isolates were confirmed using biochemical and molecular assays. In latex agglutination test, all the isolates produced characteristic agglutination. Genus-specific PCR amplification of 16SrRNA gene yielded expected product of 816 bp in all the isolates. In multiplex PCR assay conducted targeting lpxA gene for the identification of C. jejuni (331 bp) and C. coli (391bp) was used. According to research, the mean of the viable count of bacteria obtained from media supplemented with Goat blood was 0.921x108 c.f.u./ml and sheep blood was 0.936x108 c.f.u./ml. The Chi square analysis (ϗ2) was performed which proves the result to be non-significant showing similarities between the results obtained by both the media. Hence, the data indicates that goat blood can also be used as alternative and all the studies were carried on goat blood. The prevalence rate of thermophilic campylobacters sample was found to be 12.79% (11/86) in poultry faeces, 21.15% (33/156) in poultry caeca, 12.5% (3/24) in meat swabs and 4.16% (2/48) in litter from poultry farms. No thermophilic Campylobacter was isolated from goat faeces, sheep faeces and water samples from the poultry farms. Farmwise, the highest prevalence in poultry farms was detected in Anandpur (11.7%) followed by Haldwani (7.5%), and Pantnagar (5.3%). Among the chicken meat shops, highest prevalence was detected in Pantnagar (24%) followed by Haldwani (23.3%), Lalkuan (15%) and Nainital (6.6%). Out of the total of 49 thermophilic Campylobacter isolated, comprising of 36 C. jejuni (67.34%) and 13 C. coli (32.65%). As poultry serve as important source of Campylobacter, awareness among the poultry farmers and chicken meat retailers should be created to prevent the further transmission of this zoonotic agent.
  • ThesisItemOpen Access
    Molecular characterization of non-typhoidal Salmonella serovars isolated from commercial broiler farms and retail chicken meat shops with reference to virulence and antimicrobial resistance
    (G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-06) Saini, Shubham; Maansi
    Salmonellosis, primarily caused by Salmonella Typhimurium and S. Enteritidis, is deemed as one of the most common and most important food-borne zoonoses worldwide. The present study was undertaken to determine the prevalence, virulence genes and antimicrobial resistance of non-typhoidal Salmonella isolates obtained from commercial broiler farms and retail chicken meat shops. A total number of 556 samples were collected from five commercial broiler farms and thirty-two retail chicken meat shops situated in/around towns/cities of the Udham Singh Nagar and Nainital districts of Uttarakhand state. The samples comprised of poultry faeces (n=121), litter (n=56), drinking water (n=38) and feed (n=38) from the broiler farms (n=253), and chicken meat swabs (n=133), cutting surface/chopping board swabs (n=46), hand swabs of meat cutters/handlers (n=46), knife swabs (n=46) and meat rinsing water (n=32) from retail chicken meat shops (n=303). The overall prevalence of 5.57% (n=31) was recorded, which can be distributed as 4.35% (n=11) for the broiler farms, and 6.60% (n=21) for the chicken meat shops. The highest prevalence of Salmonella isolated from broiler farms was observed in water samples (02/38, 5.26%) and poultry faeces (06/121, 4.96%) followed by poultry litter (02/56, 3.57%) and poultry feed (01/38, 2.63%). Whereas in case of chicken meat shops, the highest prevalence of Salmonella was observed in chicken meat samples (13/133, 9.77%) followed by chopping board (03/46, 6.52), knife (02/46, 4.35%), rinsing water (01/32, 3.12%) and hand swabs of butchers (01/46, 2.17%). Geographically, the highest prevalence among broiler farms was recorded in the farm of Shantipuri (6.25%, 03/48), followed by Haldwani and Kiccha farms (5.00%, 03/60 and 02/40 respectively), followed by Kashipur farm (3.70%, 02/54) and Ramnagar farm (1.96%, 01/51). Whereas in case of chicken meat shops, a significantly higher prevalence of Salmonella was recorded from Pantnagar (8.33%, 05/60), followed by Rudrapur (7.84%, 04/51), Lalkuan (7.32%, 03/41), Haldwani (7.14%, 04/56), Kashipur (6.12%, 03/49) and Kiccha (2.17%, 1/46). Serotyping of thirty-one isolates revealed the presence of only one type of serotype viz; Salmonella Typhimurium (93.55%; 29/31), while two (6.45%; 2/31) isolates were found to be untypable (one each from poultry faeces and chicken meat). All the thirty-one isolates were screened for the presence of eight virulence genes (sipA, sopE1, fliC, mgtC, spvC, stn, sopB and gipA) by PCR. The most commonly encountered virulence gene was sipA, amplified in 100% (31) isolates, followed by mgtC 93.55% (29), sopB 87.10% (27), fliC 70.97% (22), gipA 64.52% (20), stn 64.52% (20), spvC 51.61% (16) and sopE1 16.13% (5) isolates. All 31 Salmonella isolates (100%) were multidrug resistant (MDR). Complete resistance of all the isolates was observed against Nalidixic acid and Erythromycin (100% each) followed by higher resistance against Ampicillin (87.09%), Ciprofloxacin (83.87%), Tetracycline (77.42%), Cefotaxime (74.19%), Gatifloxacin (70.97%), Cefazolin (67.74%) along with Sulphafurazole (67.74%), Streptomycin (58.06%), Levofloxacin (51.61%), Cefoxitin (38.71%) and Chloramphenicol (32.26%). Phenotypic co-resistance against Ciprofloxacin and Cefotaxime was identified in 21 (21/31, 67.74%) isolates. Three panresistant isolates, all 3 being S. Typhimurium were also identified. The most commonly expressed resistance patterns were AMP CX E LE CIP CTX GAT SF TE CZ NA S C (3/31; 22.07%), followed by AMP E LE CIP CTX GAT TE CZ NA S (2/31; 6.45%) and AMP CX E CIP CTX SF TE CZ NA S (2/31; 6.45%). All phenotypically resistant Salmonella isolates were screened for the presence of 16 corresponding antimicrobial resistance genes. Among the 28 intermediate and resistant Salmonella isolates, aadA1 gene was carried by all 28 (100%) isolates tested, while aadA2 gene was present in 11 (39.28%) isolates, strA gene in 17 (60.71%) isolates and strB gene was not detected in any of the isolates. Out of three Tetracycline resistant genes, tetA was present in 60% (15/25) isolates, whereas none of the isolates carried tetB and tetG genes. Sulphafurazole resistance encoding sul1 was harbored by 79.17% (19/24) isolates tested, while sul2 gene was not amplified in any of the isolates. Amongst the _-lactam resistance genes, blaTEM gene was detected in 100% (27/27) of the isolates, while blaPSE and blaCMY were found to be absent in all the isolates. Amid the fluoroquinolone resistance genes, gyrA and parC genes were found to be present in 93.55% (29/31) and 64.52% (20/31) of the isolates respectively, whereas none of the isolate revealed the presence of qnrA or qnrS genes. In the multiplex PCR performed among all MDR isolates for the detection of mcr-1 to mcr-5 genes coding resistance for colistin, no isolate exhibited the presence of any of the five genes targeted. Overall our study detected high prevalence of multi drug-resistant Salmonella in the broiler farms and chicken meat shops. Higher resistance to ‘critically important’ (fluoroquinolones) and ‘highly important’ (cephalosporins and tetracycline) antibiotics detected in Salmonella isolates of poultry origin is a serious public health threat which stresses on the need to curb the practice of irrational use of antimicrobials in the poultry sector in India.