Chlamydia psittaci is a zoonotic pathogen mainly transmitted by psittacine birds and poultry. The low shedding rate of the pathogen in the apparently healthy birds and human clinical cases may result in false-negative results. In the present study, a droplet digital PCR (ddPCR) assay was developed and compared with optimized quantitative PCR (qPCR) for the detection of C. psittaci from the clinical samples. The ddPCR assay was found to be comparatively more sensitive than the qPCR, wherein the limit of detection (LOD) of ddPCR was upto 2.4 copies of the DNA template, whereas, the qPCR could detect upto 38 copies of the DNA template in the reaction mixture. Overall, the developed ddPCR assay was found to be robust, specific, and could reliably quantify up to 17.8 copies of the DNA template. Finally, the applicability of the developed ddPCR assay was tested by screening the field samples (n = 124), comprising lung tissues from dead poultry and feral birds; pooled faecal samples from the free-living birds, commercial and backyard poultry farms; pharyngeal and cloacal swabs collected from the duck farms. Of these, a total of seven samples were found to be positive by the ddPCR, whereas, three samples could be detected as positive using the qPCR. The developed ddPCR could serve as a reliable screening tool, particularly in those clinical samples wherein the shedding of C. psittaci is substantially very low.

Background: Cationic antimicrobial peptides (AMPs) have attracted considerable attention to source antimicrobial as well as antibiofilm technology solutions. In vitro and in vivo antimicrobial efficacies of three short-chain AMPs, Indolidicin, Cecropin A (1–7) – Melittin (CAMA), lactoferricin (17–30), and their combination were explored for the first time against multi-drug resistant enteroaggregative Escherichia coli (MDR-EAEC). Methods and materials: Minimum Inhibitory Concentrations (MICs) and Minimum Bactericidal Concentrations (MBCs) of AMPs were determined by broth micro-dilution technique. The AMPs were tested for its stability at temperatures (70 °C and 90 °C), proteases (Proteinase-K and trypsin) and physiological concentration of cationic salts (NaCl and MgCl2); safety in sheep RBCs and cell lines (RAW 264.7 and HEp-2); adverse effect on beneficial flora and membrane permeability. In vitro dose- and time-dependent time kill assay was also carried out. Later, in vivo studies of AMPs (alone and in combination) employing Galleria mellonella (survival assay, bacterial burden, haemocyte density, melanisation assay, cytotoxicity assay and histopathological examination) was carried out. Results: All the three AMPs were stable (except for Indolicidin with trypsin), tested safe at lower MIC and no adverse effect against beneficial flora was observed. AMPs alone and in combination, exhibited bacterial membrane permeability. In vitro time-kill assay revealed concentration-cum-time dependent clearance of MDR-EAEC in the AMP-treated groups, while, in the in vivo G. mellonella experiment, the infected group treated with AMPs (alone and in combination) revealed an increased survival rate, immunomodulatory effect, reduced MDR-EAEC counts and were tested safe to the larval cells which concurred with histopathological observations. Conclusion: The AMPs seem to be promising candidates for further investigations using appropriate animal models for MDR-EAEC strains.

Background: The present investigation of Coxiella burnetii infection in cattle and farm workers on an organized cattle dairy farm, which appears to be the first of its kind in India, was undertaken to assess the status of this largely neglected and masked zoonosis. Methods: A total of 665 samples comprising of serum (n = 224), milk (n = 217) and vaginal swabs (n = 224) collected from milch animals (n = 224) with a history of reproductive disorders were screened. Besides these, ticks (n = 114); animal feed (n = 4) and environmental samples (n = 13) as well as serum (n = 19) of farm workers were also collected. The animal sera and milk samples as well as human sera were tested for antibodies against C. burnetii by commercial ELISA kit, whereas, all the collected samples were subjected to trans-PCR targeting the IS1111 gene of C. burnetii. Results: A high positivity for coxiellosis was detected in sera (29.91%) and milk (26.73%) samples of dairy cattle as well as sera from human contacts (84.21%) by ELISA. The trans-PCR detected the pathogen in 12.94% sera, 14.73% vaginal swabs and 5.53% milk samples of cattle, and in one soil sample, however, the sera of the farm workers and tick were tested negative. Conclusions: The high positivity for coxiellosis among cattle and farm workers highlight the need to undertake extensive epidemiological studies to unravel the trends of C. burnetii infection in India. (C) 2018 The Authors. Published by Elsevier Limited on behalf of King Saud Bin Abdulaziz University for Health Sciences. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Enteroaggregative Escherichia coli (EAEC) is an emerging foodborne pathogen worldwide, often responsible for persistent diarrheoa in the infants and young animals. In present study, a total of 1519 diarrhoeal fecal samples from human infants (n=890) and young domestic animals (n=629) from eight different states of India were investigated. The recovered E. coli isolates by cultural method were further confirmed as EAEC by PCR targeting chromosomal associated genes (aaiA, astA, pilS, ecp,irp2, pic, fimA) and plasmid-borne genes (cvd432, aggR, aafA, aggA, agg3A) as well as by gold standard HEp-2 cell adherence assay. A total of 197 EAEC isolates were recovered from diarrhoeal cases of human infants (n=95) and young animals (n=102) which were further subjected to Pulse Field Gel Electrophoresis (PFGE) to address their genetic diversity and to the antimicrobial susceptibility test to know their susceptibility towards antibiotics used in routine practice. Overall, a highly diverse PFGE profile was observed for most of the EAEC test isolates. However, irrespective of place of isolation, sharing or circulation of identical clones of EAEC between different species of young animals including human infants was evident. The antibiogram profile revealed an alarming multidrug resistance (MDR) profile among diarrhoeal EAEC isolates of human & animal origin. The infant origin EAEC isolates were more resistant to Beta-lactam, Third generation Cephalosporins and Fluroquinolones, while the animal origin EAEC isolates were resistant to Beta-lactam, Tetracyclines and Sulphonamides. However, all the EAEC isolates of infant and young animals were found sensitive to Imipenem drug. 6. ABST. No. PS-1-11 Isolation, Identification and Antibiotic Sensitivity profile of Salmonella Enteritidis recovered from local poultry meat shops Diksha Gourkhede, Kaushik Satyaprakash, Bhoomika Sirsant, Jay Prakash Yadav, Richa Pathak, Deepa Ujjawl, D. B. Rawool, S. V. S. Malik and S. B. Barbuddhe Division of Veterinary Public Health, ICAR-IVRI, Izatnagar, Bareilly, U.P-243122 National Research Centre on Meat, Chengicherala, Hyderabad, Telengana500092 deepak.rawool@yahoo.com Abstract Non-typhoidal Salmonellae (NTS) are leading cause of foodborne infections and the emergence of antimicrobial resistance among them is a global concern. In the present study, isolation and molecular identification of S. Enteritidis was attempted from poultry faecal droppings, caecal and meat samples. The identified S. EnteritidisNon-typhoidal Salmonellae (NTS) are leading cause of foodborne infections and the emergence of antimicrobial resistance among them is a global concern. In the present study, isolation and molecular identification of S. Enteritidis was attempted from poultry faecal droppings, caecal and meat samples. The identified S. Enteritidis isolates were then subjected to antibiotic sensitivity studies as per CLSI 2017 guidelines. A total of 72 samples comprising of faecal droppings (n=23), caecum (25) and meat (n=24) samples were collected aseptically from local poultry meat shops of Bareilly and were processed immediately for isolation of Salmonella by standard conventional method. All the presumptive Salmonella colonies on XLD medium were processed for recommended biochemical tests as well as by PCR targeting genus specific inv A gene. Of the 72 samples, 13 were found to be positive for Salmonella genus by both, PCR and biochemical tests. These 13 isolates were then subjected to identification of S. Enteritidis serovar by serotype specific conventional PCR targeting sdf gene. Of the 13 genus confirmed Salmonella isolates, 9 S. Enteritidis isolates amplified sdf gene. The antibiotic sensitivity studies of the identified S. Enteritidis isolates revealed an alarming multidrug resistance (MDR), as of the 9 S. Enteritidis isolates, 7 were resistant to three or more classes of antibiotics namely β lactam group, aminoglycosides, tetracyclines, fluoroquinolones, sulfonamides and glycopeptides. These MDR S. Enteritidis isolates were further processed to determine the MIC range. The MIC range observed were 10-30 μg/ml, 10-240 μg/ml, >240 μg/ml, >240 μg/ml and >240 μg/ml for ciprofloxacin, tetracycline, ampicillin, vancomycin and sulphomethoxazole, respectively. 7. ABST. No. PS-1-13 Does sanitizing/hygienic interventions improves the microbiological quality of street foods? Anukampa, D.K. Singh, K.N. Bhilegaonkar, Ashok Kumar, D. Bardhan, Vinodh Kumar, O.R., Shagufta Bi., Sivakumar, M., Pruthvishree, B.S., Karthikeyan, R. and Z.B. Dubal Division of Veterinary Public Health, ICARIndian Veterinary Research Institute, Izatnagar, Bareilly243 122, U.P., India. drzunjar@yahoo.co.in Abstract A total of 1020 foods of animal origin and associated environmental samples were collected aseptically before and after intervention from 18 local food vendors of Bareilly (U.P) to assess the efficacy of an intervention to improve the microbial quality of street foods. Aerobic plate count (APC), enumeration of Staphylococcus aureus, E. coli, sulphite reducing Clostridia, presence of Salmonella spp. and Listeria monocytogenes of the samples was performed. The pre and post-intervention microbiological quality of food samples were assessed and the mean log reduction of APC was observed for swabs of hand (HS), plate (PS), table (TS) and cloth (CS) were within the range of 2.56-5.61, 1.96–5.31, 2.56–6.32 and 2.66–6.77 log10 cfu/cm. The E. coli were in the range of 1.12–3.78, 1.22–3.72, 1.69-4.16 and 0-4.33 log10 cfu/cm and S. aureus in the range of 1.30-3.74, 1.47-2.30, 1.36–2.55 and 1.2–2.86 log10 cfu/cm for HS, PS, TS and CS samples, respectively. Percent positivity in samples was also reduced significantly from 48.58% to 38.12% for E. coli, and 44.66% to 34.85% for S. aureus. The cooked food samples showed low microbial count before and after intervention strategies whereas the raw foods and processed/ready-to-eat foods showed a different trend in which 88.33% & 73.33% samples were within the acceptable limits before intervention and the microbial load was reduced drastically and within acceptable limits after an intervention. BeforeA total of 1020 foods of animal origin and associated environmental samples were collected aseptically before and after intervention from 18 local food vendors of Bareilly (U.P) to assess the efficacy of an intervention to improve the microbial quality of street foods. Aerobic plate count (APC), enumeration of Staphylococcus aureus, E. coli, sulphite reducing Clostridia, presence of Salmonella spp. and Listeria monocytogenes of the samples was performed. The pre and post-intervention microbiological quality of food samples were assessed and the mean log reduction of APC was observed for swabs of hand (HS), plate (PS), table (TS) and cloth (CS) were within the range of 2.56-5.61, 1.96–5.31, 2.56–6.32 and 2.66–6.77 log10 cfu/cm. The E. coli were in the range of 1.12–3.78, 1.22–3.72, 1.69-4.16 and 0-4.33 log10 cfu/cm and S. aureus in the range of 1.30-3.74, 1.47-2.30, 1.36–2.55 and 1.2–2.86 log10 cfu/cm for HS, PS, TS and CS samples, respectively. Percent positivity in samples was also reduced significantly from 48.58% to 38.12% for E. coli, and 44.66% to 34.85% for S. aureus. The cooked food samples showed low microbial count before and after intervention strategies whereas the raw foods and processed/ready-to-eat foods showed a different trend in which 88.33% & 73.33% samples were within the acceptable limits before intervention and the microbial load was reduced drastically and within acceptable limits after an intervention. Before intervention the number of samples positive for S. aureus and number of samples within acceptable limits for raw foods, processed/ready-to-eat foods, cooked foods was 45.33%, 45.23%, 21.05% and 66.67%, 75.0%, 91.66%, respectively. After the intervention, there was a reduction in pathogenic counts for processed/ready-to-eat foods, raw foods and cooked foods. Interestingly, before intervention 82.89% of food samples were within the acceptable limits which increased to 90.78% after intervention. The cooked samples were less frequently contaminated with E. coli and S. aureus, while most of the raw food samples showed the presence of E. coli and S. aureus. The samples like chicken gravy, cooked chicken, omelet, boiled egg & boiled milk were negative for S. aureus. Few samples of raw egg, omelet, salad, chutney and meat samples were positive for SRC and Salmonella, while improved bacterial quality was noticed after an intervention. Interestingly, L. monocytogenes could not be recovered from any of the samples. It was observed that the sanitizing or hygienic interventions improve the microbiological quality of street foods. 8. ABST. No. PS-1-14 Efficacy of benzalkonium chloride, hydrogen peroxide with per acetic acid and chlorine on clean and dirty raw shelled eggs sanitation and egg safety Santosh Sajjan, Madhavaprasad C.B. and Prashant S. Bagalkote Department of Veterinary Public Health and Epidemiology Veterinary College, KVAFSU, Vinobanagar, Shivamogga, Karnataka. *Corresponding author: email: dr.santoshsajjan@gmail.com, Tel: +91-8861233410 dr.santoshsajjan@gmail.com, santoshsajjan@rediffmail.com Abstract A study was conducted to evaluate the efficacy of some commonly used sanitizers in the food processing industries viz. benzalkonium chloride, hydrogen peroxide with per acetic acid and chlorine water for surface sanitation of raw shelled eggs by artificially challenging with Salmonella Enteritidis and Listeria monocytogenes MTCC 1135under clean and dirty conditions by in-vitro trials. Minimum Inhibitory Concentration (MIC) of the selected sanitizers was determined according to the modified Rideal-Walkar test by tube dilution technique and capacity of sanitizers under clean and dirty conditions by Kelsey-Sykes test. The study revealed that, the MIC of benzalkonium chloride, hydrogen peroxide with per acetic acid and chlorine water was 25

Clostridium perfringens is one of the most important globally recognised gastroenteric pathogen in humans as well as animals. The present study was aimed to know the similarities/divergence among C. perfringens type A isolates of human and animal origin using the pulsed-field gel electrophoresis (PFGE) as a molecular tool. The enterotoxic isolates obtained by screening of human diarrhoeal cases (n = 130), diarrhoeal cases of pig (n = 52) and goat (n = 50), meat samples viz., pork (n = 59) and chevon (n = 57) were characterized by standard cultural and biochemical methods followed by PCR Assays. Accordingly, a total of 11 C. perfringens type A characterized isolates (16S rRNA(+), cpa(+), cpb(2)(+) and cpe(+)) recovered from human diarrhoeal cases (n = 3); diarrhoeal cases of pig (n = 2) and goat (n = 2); meat samples viz. pork (n = 2) and chevon (n = 2) were examined employing PFGE. The observed clustering pattern in PFGE analysis showed the relatedness between isolates from diarrhoeal goat and chevon (90-100%); diarrhoeal pig and pork (65-68%); moreover, isolates from human diarrhoeal cases were exhibiting lineage to cases from goat and pig diarrhoea as well pork and chevon by 62-68% relatedness. The outcome of the present study indicates the probable contamination of this pathogen to the human food chain through faeces from suspected food animals viz. goat and pig and their improperly cooked meat. (C) 2017 Faculty of Veterinary Medicine, Cairo University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Details about the members of the Indian Listeria Consortium are provided in the Supplementary Data.Emerging Microbes and Infections (2016) 5, e55; doi:10.1038/emi.2016.55; published online 8 June 2016

Background: Infectious diarrhoea particularly due to pathogenic bacteria is a major health problem in developing countries, including India. Despite significant reports of diarrhoeagenic Escherichia coli (DEC) pathotypes across the globe, there is paucity of studies which reveal their relatedness with respect to their isolation from different sources. This present study determines isolation and identification of DEC pathotypes from different sources, their genetic characterization and antibiogram sensitivity profiling. Methods & Materials: A total of 336 samples comprising of diarrhoeic stool samples from infants (n = 103), young animals (n = 106), foods and associated environmental sources (n = 127) were screened for E. coli. The identified E. coli were confirmed as DEC pathotypes by using PCR based assays. These isolates were further studied for their genetic diversity using Pulse Field Gel Electrophoresis (PFGE) subtyping tool and their antibiogram profile was determined against seven commonly used drugs. Results: Of the four DEC pathotypes investigated, Enteroaggregative E. coli (EAEC) was found to be the predominant pathogen with an isolation rate of 16.50% from infants, 17.92% from young animals and 10.24% from foods and environmental sources. These EAEC isolates, on further characterization revealed predominance of 'atypical' EAEC, with an isolation rate of 10.68% from infants, 15.09% from young animals and 10.24% from foods and associated environmental sources. On PFGE analysis, discrimination was also evident within DEC pathotypes, as only closely related EAEC isolates clustered together irrespective of their source of isolation. Further, higher antibiotic resistance pattern was observed among the isolated DEC pathotypes as almost 86.44% of isolates were found to be resistant against ≥3 tested drugs. Conclusion: EAEC pathotype in particular 'atypical' strains were found to be the predominant pathogen. On PFGE analysis, sharing and circulation of EAEC isolates between human and animal, including foods and associated environmental sources was evident. Besides this, an alarming antimicrobial resistance profile was observed for majority of the recovered DEC pathotype isolates.

Received: Revised: Accepted: 2013-08-19 2013-08-27 2013-08-28 The epidemiology of Q–fever with regard to the role of foods of animal origin in transmission of Coxiella burnetii and their risk assessment remains largely unknown in most parts of the world including India. In the present study, a total of 591 food samples of different animal origin including milk (518), meat (60) and shell eggs (13) were screened for the presence of the pathogen by trans–PCR assay targeting transposon like element IS1111 of C. burnetii. . The analysis revealed an overall positivity in 4.06% (24/591) food samples, all of which were of milk. The milk samples showed an overall positivity of 4.63% (24/518), with the detection of C. burnetii in 5.55% (23/414) bovine and 1.42% (1/70) ovine milk samples. However, all the samples of camel milk (n = 34), meat (n = 60) and shell eggs (n = 13) turned out to be negative for the pathogen. The detection of C. burnetii in the bovine and ovine milk indicates a potential health risk for domestic livestock as well as human beings, especially those who consume raw or unpasteurized milk. The study of screening foods of animal origin for occurrence of C. burnetii in India by highly sensitive and specific molecular diagnostic tools is largely lacking, and in this context, the present study appears to be the first of its kind in the country. It is recommended that milk should be consumed only after pasteurization and, dairy as well as other foods of animal origin should be included in surveillance and monitoring programmes for food–borne pathogens for the risk assessment of C. burnetii infection. All copyrights reserved to Nexus® academic publishers

The essential oil of basil (Ocimum basilicum L.) is used for flavoring foods, dental and oral products. Stability analysis was conducted on 30 accessions of basil germplasm (exotic as well as indigenous). These accessions were augmented from different countries namely, India (10), USA (10), USSR (7), Germany (1), Hungary (1) and Poland (1). These were evaluated under four diverse environments comprising two locations and two years (2004 and 2005). The mean performance and stability parameters of 30 accessions were analyzed under four environments for nine characters, namely, number of primary branches/plant, lamina length (cm), lamina width (cm), leaf-stem ratio, plant height (cm), fresh herbage yield/plant (g), dry herbage yield/plant (g), essential oil content (%) and essential oil yield/plant (ml). All 30 genotypes had squared deviation equivalent to zero (S2 di=0) showing consistent performance of the genotypes among all the environments. Essential oil is the economic product of basil. Five genotypes viz. EC388788, IC333332, IC336833, IC388891 and EC338773 were identified as desirable and stable in relation to essential oil yield per plant. Two genotypes (EC388788 and IC333332) were found superior for essential oil yield, fresh herbage yield and dry herbage yield/plant over the four environments. Hence, these genotypes could be recommended for their direct utilization across the environments.

Variability studies conducted in 30 Ocimum genotypes (20 exotic and 10 indigenous) in two environments during kharif (rainy season) 2004 and 2005 revealed highly significant variation for all the characters studied. More fresh herbage yield per plant was recorded in the genotypes namely IC333332, EC338785, EC128730 and EC388788. However, high essential oil yield per plant was extracted from genotypes IC110267 and EC388890. The pooled data analysed at phenotypic level revealed that fresh herbage yield per plant had highest positive direct effect with essential oil yield per plant and maximum direct negative effect in essential oil content. The essential oil yield per plant had the highest direct positive effect at phenotypic level on essential oil content followed by fresh herbage yield per plant whereas maximum negative effect was observed in number of flower-whorls per spike and days to seed maturity. The above genotypes performed well in respect of fresh herbage and essential oil yield per plant which could be considered for incorporation in breeding programme to develop an improved variety coupled with high yield potential to enhance the production and productivity.