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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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ThesisItem Open 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; MaansiThe 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.ThesisItem Open 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.ThesisItem Open 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; MaansiSalmonellosis, 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.ThesisItem Open Access Burden assessment and characterization of thermophilic Campylobacter in broiler chickens(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Dhayananth, B.; Upadhyay, A.K.Campylobacter is considered to be a major pathogen in terms of food safety concern since it causes gastrointestinal problems worldwide. As these organisms have an intricate epidemiological cycle and the sources of contamination are varied in nature, necessary steps should be taken for controlling the organism below the level of risk for humans. The present research was organized in such a manner to screen and detect thermophilic Campylobacters from a total of seven farms of poultry (Haldwani, Pantnagar, Lalkuan, Kiccha, Jawaharnagar, Shanthipuri farm 1, and Shanthipuri farm 2) and certain retail meat shops (pantnagar) located in the state of Uttarakhand. Totally 645 samples which includes 15 poultry meat samples, 446 poultry fecal samples and 184 environmental samples were processed according to their standard protocol for accomplishing the objectives of the study undertaken. Bacteria showing characteristic morphological and staining characteristics were confirmed biochemically. 16S rRNA gene (816 bp) was targeted for genus identification while lpxA gene was targeted for species differentiation of C. jejuni (331 bp) and C. coli (391 bp). Through simplex PCR 73 isolates were confirmed to be belonging to Campylobacter genus with an overall prevalence of 11.32%. Maximum recovery was found at lalkuan farm (33.77%) followed by Haldwani farm (16.13%), Pantnagar farm (11.93%), Shanthipuri farm1 (8.79%), Shanthipuri farm2 (6.76%), Pantnagar meat shops (6.67%) while least prevalence in Jawaharnagar farm (5.1%). No Campylobacter isolates were recovered from Kiccha. Prevalence of C. jejuni (52.05%) species was observed to be more than C. Coli (47.95%). All the 73 isolates were screened for the presence of ten virulence genes among which cadF, flaA and dnaJ showed 100% prevalence followed by cdtB(87.67%), cgtB(45.68%), pldA(31.51%) and wlaN(16.44%). None of the isoates were harbouring ciaB, iam, virB. Only 40 isolates sustained out of 73 isolates which were further processed for Kirby Bauer disc diffusion assay in which antibiotic sensitivity testing of Cefoxitin had highest resistance as 95% followed by Ciprofloxacin 80%, Nalidixicacid25%, Cefatoxime-22.5%, Ampicillin-20%, Tetracycline-12.5%, Clindamycin-7.5%, Erythromycin-5%. The antibiotics Gatifloxacin, Streptomycin were susceptible to all the isolates screened but some intermediate resistance was found in Sulfisaxazole and Levofloxacin. The isolates which developed resistance phenotypically were checked for their respective Antimicrobial Resistance Genes (ARGs). β-lactam resistance gene blaOXA-61 was prevalent in 32 isolates (86.49%), cmeB was found in 24 isolates (72.73%) , tet (O) was detected in 3 isolates (60%), ermB was detected in 1 isolate (50%). The resistance towards cephalosporin and fluoroquinolone group of antibiotics is increasing representing the over usage of antibiotics in poultry farms which should be taken care off and necessary measures to be undertaken to reduce its usage to avoid AMR among humans.ThesisItem Open Access Virulence characterization and antimicrobial resistance among non-typhoidal Salmonella isolates obtained from commercial broiler farms(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-02) Hussain, Sheeba; MaansiThe objective of this study was to determine the prevalence, virulence genes and antimicrobial resistance of non-typhoidal Salmonella isolated from commercial broiler farms. A total of 721 samples were collected from 8 commercial broiler farms, Ramnagar (n=1), Kiccha (n=1), Jawaharnagar (n=1), Shantipuri (n=1), Pantnagar (n=2) and Haldwani (n=2) representing two districts (Udham singh Nagar and Nainital) of Uttarakhand state, India. The samples (n=721) consist of poultry faeces (n=310), litter (n=127), water (n=127), feed (n=124) and manure (n=33). The overall prevalence of non-typhoidal Salmonella (NTS) was found to be 10.68% (n= 77). The highest prevalence of Salmonella was observed in water samples (19/127,14.96%) followed by poultry litter (18/127, 14.17%), poultry feed (12/124, 9.68%), poultry feces (27/310, 8.71%) and manure samples (1/33, 3.03%). According to the location, the highest prevalence was observed in Haldwani Farm 1 (27.19% ;31/144) followed by Shantipuri Farm (27.14% ,19/70), Haldwani Farm 2 (24.18%, 22/91), Pantnagar Poultry Farm-2 (3.92%, 4/102) and Pantnagar Farm-1 (0.98%, 1/102). Three different serotypes viz; Salmonella Kentucky (50.65%; 39/77), Salmonella Virchow (28.57%; 22/77) and Salmonella Typhimurium (3.89%, 3/77) were identified. Thirteen isolates (16.88%) were reported to be non typable. All 77 isolates were screened for the presence of 8 virulence genes by using PCR. The sopB gene harbored by 59 (76.62%) isolates followed by sipA 44 (57.14%), spvC 48 (62.34%), stn 47 (61.03%), mgtC 34, (44.16%), sopE1 25 (32.47%), gipA 16 (20.78%) and fliC 4 (5.19%) isolates. All 77 isolates (100%) were multidrug resistant (MDR). Among the 12 antimicrobials screened, highest resistance was observed for Erythromycin (100%, 77/77) and Nalidixic acid (100%, 77/77) followed by Tetracycline (97.40%, 75/77), Sulphafurazole (93.51%, 72/77), Cefotaxime (81.81%, 63/77), Cefazolin (80.52%, 62/77), Gatifloxacin (76.62%,59/77), Ciprofloxacin (71.43%, 55/77), Ampicillin (67.53%, 52/77), Levofloxacin (64.94%, 50/77), Cefoxitin (28.57%, 22/77) and Streptomycin (14.29%, 11/77). Pan-resistance was observed in six isolates comprising of 3 S. Kentucky, 1 S. Virchow and 2 Non typable Salmonella isolates. Fourty eight (62.34%) Salmonella isolates were identified as co-resistant to ciprofloxacin and cefotaxime. The most common resistance patterns against the 12 antimicrobials tested were observed as follows: AMP, E, LE, CIP, CTX, GAT, SF, TE, CZ, NA (17/77; 22.07%), AMP, CX, E, LE, CIP, CTX, GAT, SF, TE, CZ, NA (11/77; 14.29%), E, CTX, SF, TE, CZ, NA (6/77 ; 7.79%) and E, CTX, SF, TE, NA (6/77 ; 7.79%). All phenotypically resistant Salmonella isolates were assessed for 12 corresponding antimicrobial resistance genes. Out of 11 streptomycin resistant Salmonella isolates aadA1 and strA genes were carried by equal number of isolates, 7(63.64%) while aadA2 and strB genes were found in 8 (72.73%) and 2(18.18%) respectively. Only one isolate carried all the four tested resistant gene whereas one of the resistant isolates did not carry any of them. Sulphafurazole resistance encoding genes sul1 and sul2 were found in 25% (18/72) and 2.59% (2/72) isolates, respectively. Among the β- lactam genes, blaTEM gene was the predominant 22.22% (14/63) followed by blaCMY (1.59%, 1/63). Out of 3 tetracycline resistant genes tetA was detected in 29.33% (22/75) isolates, whereas none of the isolates carried tetB and tetG genes. Risk factors analysis revealed that the distance of manure heap from the poultry farms and the size of the flock were found to be significantly associated while size of the flock was negatively associated with the prevalence of Salmonella in the farms. Overall, our study revealed a high prevalence of multi drug resistant Salmonella in the broiler farms of the region. Co-resistant isolates (ciprofloxacin and cefotaxime) may pose a therapeutic threat and also highlights the irrational use of antimicrobials in the poultry farms in India.ThesisItem Open Access Epidemiological studies on physical, chemical, psychological and zoonotic hazards among veterinarians(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Parmar, Tanuja; Upadhyay, A.K.This study reports physical, chemical, zoonotic and psychological hazards relevant to Indian veterinarians as obtained by a self administered questionnaire. Out of total 392 respondents, 5.1% (20) reported no injury, 47.5% (186) respondents had 1-5 injuries, 32.1% (126) encountered 5-10 injuries and 15.3% (60) veterinarians had more than 10 injuries during last 5 years. Animal related injuries like bite (31.8%), scratch (65.1%), kick (62.8%), horn wound (14%), needle prick (89.2%), injuries due to falling / lifting animals / moving heavy equipments (61.3%) and fracture (3.8%) were commonly reported physical injuries Total no. of veterinarians involved in taking radiographic examinations were 19.1% (75/392). Most of the veterinarians taking x-rays were academic veterinarians. Hundred eighty seven (47.7%) veterinarians were using antineoplastic agents to treat animals and out of those 2.1% (4) veterinarian accidently injected drugs to themselves. Thirty two (8%) veterinarian, experienced adverse effect due to disinfectant and 0.7% (3) veterinarians reported adverse effects due to pesticides. The types of allergy sustained by veterinarians were sneezing, eye/ nose/ throat infection, cough, skin irritation and latex gloves allergy. Ringworm (13.5%) and fungal infection (26.5%) were most common zoonotic infection among veterinarians. Low level stress reported among 45% participants, moderate among 34% and high level of stress among 21% participants. The 43.9% veterinarians used gloves, 39.8% were wearing apron and only 1% respondents were using goggles. 36.7% participants did not use any protective gear. Routine deworming within every 6 months followed by 43.6% of the respondents, 31.1% in the last 1 year and 17.1% had done it once in the last two years. Percentage of the veterinarians did not practice routine deworming was 8.2%. Majority of veterinarians 73% (286) rarely go for medical checkup or only when required. The awareness levels among the veterinary health professionals was near optimal but the need was felt to implement efforts aimed at addressing deterrence of occupational hazards by developing and executing improved safe handling practices and safety measures.ThesisItem Open Access Isolation, identification and antimicrobial susceptibility of public health significant organisms present in organic agricultural farms of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-07) Rautela, Richa; MaansiStudies are lacking in India regarding the presence of public health significant organisms in the environment and plant samples at organic farms. The present work was carried out on two highly significant public health organisms i.e. Salmonella and Listeria in five organic agricultural farms located at Kotabagh, Dhamola, Ramnagar and two locations of Pantnagar in Uttarakhand state, India. A total 500 samples, comprising 350 environmental samples viz; soil (n=227), manure (n= 66) and water (n=57) besides, 150 plant samples viz; rhizosphere (n=50), roots (n=50) and leaves/grains (n=50) were processed for isolation of Salmonella and Listeria. Standard protocols were used for isolation of both organisms. The morphological and biochemical characterization was attempted. Genus specific PCR targeting invA (284 bp) gene for Salmonella and prs (370 bp) gene for Listeria was used for molecular confirmation. A total of 11 Salmonella isolates were recovered with an overall prevalence of 2.2% while Listeria could not be isolated representing 0% prevalence. Amidst environmental sources maximum detection (8.7%) was observed from water. Three isolates each were isolated from manure (4.5%) and soil (1.32%).Highest prevalence was noticed at Ramnagar (6.4%) followed by Pantnagar-2 (3.7%) and Pantnagar-1(2.2%). Any isolate could not be detected from Kotabagh and Dhamola. Out of 11 Salmonella isolated, 7 belonged to Group C1; one was confirmed to be Salmonella Gabon and 3 Salmonella Miyazaki. Kirby-Bauer disc diffusion assay was performed for antibiotic sensitivity on all the 11 Salmonella isolates. Most frequent resistance was observed for cefotaxime (72.72%) followed by ciprofloxacin (45.45%), cefazolin (36.36%) and cefoxitin (27.27%). Nalidixic acid, streptomycin, sulphafurazole and tetracycline, each exhibited 18.18% resistance. Only 1 isolate showed resistance against gatifloxacin (9.09%). All the isolates showed susceptibility towards amoxicillin and levofloxacin. Salmonella isolates showing phenotypic resistance were screened for the presence of β lactamase genes (blaTEM, blaCTX-M-9 and blaAmpC) using simplex PCR. Three S. Miyazaki serotypes possessed blaCTX-M-9 gene. None of the isolates exhibited blaTEM and blaAmpC genes. The study focuses on the presence of the public health significant Salmonella and Listeria in environmental and plant samples of the organic agricultural farms which could be significant source of food contamination.ThesisItem Open Access Molecular epidemiology of antimicrobial resistant non-typhoidal Salmonella isolated from retail chicken meat shops(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-05) Sharma, Jaishree; Deepak KumarThe objective of this study was to determine the prevalence, antimicrobial resistance and virulence factors of non-typhoidal Salmonella. A total of742 samples viz. meat swabs (n=188), poultry faeces (n=214), hand swabs (n=78), knife swabs (n=83), meat rinsing water (n=35), cutting surface swabs (n=31), chopping board swabs (n=41), utensil swabs (n=70) and litter (n=2) were collected from 39 retail chicken meat shops located at 6 different cities. The overall prevalence Non Typhoidal Salmonella (NTS) was found to be 9.43% (n= 70). The highest prevalence of Salmonella was observed in chicken meat (28/188, 14.89%), followed by knife (11/83, 13.25%), utensils (8/70, 11.43%), chopping board (3/41, 7.32%), rinsing water (3/35, 8.57%), poultry feces (15/214, 7.01%) and cutting slab surface samples (2/31, 6.45%). Geographically, the highest prevalence was observed in Lalkuan (20.99%, 17/81) followed by Pantnagar (15.60%, 17/109), Nainital(10.83%, 13/120), Rudrapur (9.78%, 9/92), Bareilly (5.83%, 7/120), Haldwani (5.66%, 6/106) and Kiccha (0.88%, 1/114). Three serotypes of Salmonella were identified, Salmonella Kentucky (74.29%; 52/70), S. Virchow (17.14%; 12/70) and S. Typhimurium (7.14%; 5/70). All 70 Salmonella isolates (100%) were multidrug resistant (MDR). Highest resistance was observed against Tetracycline (70/70, 100%) and Erythromycin (70/70, 100%) followed by Nalidixic Acid (69/70;98.57%), Ampicillin (67/70;95.71%), Ciprofloxacin (58/70;82.86%), Gatifloxacin (57/70;81.43%), Cefazolin (53/70;75.71%), Cefotaxime (36/70; 51.43%), Levofloxacin (35/70;50%), Sulfisoxazole (33/70;47.13%), Streptomycin (19/70;27.14%) and Cefoxitin (11/70; 15.71%). Twenty nine (29/70, 41.43%) Salmonella isolates were identified as coresistant to ciprofloxacin and cefotaxime. The most common resistance patterns were NA CIP GAT CZ E AMP TE (7/70, 10%), NA CIP LE GAT CZ CTX E AMP TE (6/70, 8.57%), S NA CIP LE GAT CZ CTX E AMP TE and NA CTX E AMP TE SF (both 5/70, 7.14%).These isolates were also resistant to 4 other antimicrobial agents (NA, E, AMP, TE). Salmonella isolates were screened for 12 antimicrobial resistance genes. Out of 47 resistant and intermediate streptomycin isolates, aadA1 and aadA2 were found to be 80.85% and 2.13% respectively, while strB was present in 6.38% isolates. Sulfisoxazole resistance genes sul1 and sul2 were detected in 82.35% (28/34) and 8.82% (3/34) isolates, respectively. Out of the β-lactam resistance genes, blaTEM was the most predominant (25.37%) followed by blaPSE and blaCMY (1.49%). The tetracycline resistance gene, tetA was detected in all Salmonella isolates (100%, 70/70). All 70 isolates were screened for the presence of virulence genes using PCR. A total of 66 isolates harbored sipA (94.29%) followed by mgtC (52/70; 74.29%), sopE1 (26/70; 37.14%), stn (24/70; 34.29%), sopB (9/70; 12.86%) genes, fliC gene (2/70; 2.86% each), spvC and gipA genes (1/70; 1.43%). Risk factor analysis revealed that none of the potential risk factors included were significantly associated with the Salmonella prevalence in the chicken meat shops. Overall, our study detected very high prevalence of multidrug resistant Salmonella in the chicken meat shop environments. Higher resistance to “critically important” (3rd and 4th generation fluoroquinolones) and “highly important” (1st generation cephalosporins and tetracycline) antibiotics detected in Salmonella isolates of poultry origin is a serious threat to public health which highlights the irrational use of antimicrobials in the poultry production in India.