Microbial dynamics and climatic interactions in pig sheds: Insights into airborne microbes and particulate matter concentrations.

Emissions of airborne pollutants from livestock buildings affect indoor air quality, the health and well-being of farmers, animals and the environment. This study aimed to evaluate the microbial count within pig sheds and its relationship with meteorological variables (temperature, relative humidity and air velocity) and particulate matter (PM10 and PM2.5) and microbial diversity. Sampling was conducted both inside and outside of two pig sheds over three seasons (summer, rainy and winter), with regular monitoring at fortnightly intervals. Results showed that the bacterial and fungal counts ranged from 0.07 to 3.98 x 103 cfu/m3 inside the sheds and 0.01 to 1.82 x 103 cfu/m3 outside. Seasonal variations were observed, with higher concentrations of particulate matter detected during the winter season, followed by summer. Climatic variables such as temperature, air velocity and relative humidity demonstrated significant impacts on the abundance of Enterobacteriaceae and fungi, while air velocity specifically influenced the presence of mesophilic bacteria and staphylococci. Importantly, no significant disparities were found between microbial counts and particulate matter levels. Staphylococcaceae emerged as the predominant bacterial family, while Aspergillus and Cladosporium spp. were the dominant fungal species within the pig sheds. The average levels of airborne bacteria and fungi in pig sheds were found to be within the recommended range, which can be attributed to the loose housing design and lower animal population on the farms.

Situation Analysis and Recommendations for the Biosecurity Status of Dairy Farms in Punjab, India: A Cross-Sectional Survey.

Farm biosecurity is an important herd management strategy to assure infection prevention and animal health. The present study aimed to evaluate the implementation of biosecurity measures on dairy farms in Punjab, India, using the standardized Biocheck.UGent™ scoring system. Convenience sampling was used to select 94 dairy farms, comprising both cattle and buffaloes, with a mean herd size of 74.8 animals. The study found that the mean external and internal biosecurity scores for the selected dairy farms were 45.4% and 43.7%, respectively. Among the subcategories, the highest external biosecurity score was observed for 'vermin control and other animals' (63%), and the highest internal biosecurity score was observed for 'adult cattle management' (76.6%). Whereas the lowest score for external biosecurity was observed for 'purchase and reproduction' (30.6%), and the lowest score for internal biosecurity was observed for 'health management' (33.6%). The overall mean biosecurity score of the present study was 44.8%, which was lower than the overall mean global score of 52%. The correlation analysis indicated a positive correlation between herd size and overall biosecurity scores, indicating that larger farms had, on average, higher biosecurity scores. Based on these results, it can be concluded that the implementation of biosecurity measures on dairy farms in Punjab needs improvement, especially in the subcategories of 'purchase and reproduction' and 'health management'. Further research to identify the factors influencing the implementation of biosecurity measures on dairy farms in the region is recommended.

Assessing and Prioritizing Zoonotic Diseases in Punjab, India: A One Health Approach.

Zoonotic diseases have a significant impact on both human and animal health globally. The present study was planned to prioritize the zoonoses in Punjab state of India. To develop a zoonotic disease prioritization scoring system, a comprehensive approach has been taken, including literature review, key person interviews with animal health experts (n = 12) and medical professionals (n = 7), and nine focus group discussions (FGDs) with veterinary academicians, medical professionals, and field veterinary doctors. The scoring system comprises of seven major criteria, each assigned a weightage score (ws): prevalence/incidence of the disease (ws = 0.20), severity of illnesses in humans (ws = 0.18), epidemic potential (ws = 0.16), socio-economic burden (ws = 0.16), availability of effective control and prevention measures (ws = 0.15), inter-sectoral collaborations (ws = 0.1), and bioterrorism potential (ws = 0.05). The finalized scoring system, accompanied by a list of 15 selected zoonotic diseases, was implemented among a group of 23 professionals engaged in zoonoses research (n = 7), animal health (n = 10), and medical health (n = 6) to determine their prioritization. The zoonotic diseases prioritized for the Punjab (India) included, Brucellosis (0.70) > Rabies (0.69) > Anthrax (0.64) > Leptospirosis (0.62) = Toxoplasmosis (0.62) = Highly Pathogenic Avian Influenza (HPAI) (0.62) > Bovine tuberculosis (0.61) > Q fever (0.60) > Cysticercosis (0.59) > Listeriosis (0.58) > Crimean-Congo haemorrhagic fever (CCHF) (0.57) > Japanese encephalitis (0.56) = Echinococcosis (0.56) > Dermatophytosis (0.53) > and Scrub typhus (0.48), respectively. Higher priority is suggested for endemic zoonoses (e.g., brucellosis and rabies) as compared to those with epidemic potential (e.g., CCHF, HPAI etc.) in Punjab. Results of the current study will help in the development of targeted control and prevention strategies for zoonotic diseases in Punjab and other geographical regions facing similar challenges.

Review of antibiotic use and resistance in food animal production in WHO South-East Asia Region.

Antimicrobial resistance is an emerging global threat to public health. The resistant bacteria in food animals can be transferred to humans through the food chain. Limited information on antimicrobial usage and resistance in food animals is available in Southeast Asia due to inadequate monitoring or surveillance systems. A literature review was conducted on antimicrobial use and resistance in food animal production in Southeast Asia for the period 2011-2020, to assess the scope and extent of antibiotic use and resistance. The countries included in the study were Bangladesh, Bhutan, Democratic People's Republic of Korea, India, Indonesia, Maldives, Myanmar, Nepal, Sri Lanka, Thailand and Timor-Leste. The information was categorised by country, production type and findings regarding antibiotic use and resistance. A total of 108 publications were included in the review. Results showed widespread use of critically and highly important antibiotics in livestock, poultry and aquacultured fish and their products. To curb the growing threat of antibiotic resistance, Southeast Asian countries need to strengthen surveillance and regulatory controls of antimicrobial use in food animal production through "One Health" approach.

Can Improved Farm Biosecurity Reduce the Need for Antimicrobials in Food Animals? A Scoping Review.

Limited and judicious antimicrobial usage (AMU) is considered the key to saving the success of human and veterinary medicine in treating infections. With the limited alternatives for antimicrobials, farm biosecurity (and herd management) is considered a promising tool to mitigate the non-judicious AMU and to maintain animal health, production, and welfare. The present scoping review aims to analyse the effect of farm biosecurity on AMU in livestock systems and formulate recommendations. Peer-reviewed manuscripts published between 2001-2022 were analyzed using the PRISMA framework using PubMed, Scopus, and Science Direct databases. After applying the inclusion criteria, 27 studies were found to assess the effect of farm biosecurity (or management practices) on AMU at the herd/farm level in quantitative/semi-quantitative terms. These studies were carried out in 16 countries, of which 74.1% (20/27) were from 11 European countries. The highest number of studies were from pig farms [51.8% (14/27)], followed by poultry (chicken) farms [25.9% (7/27)], cattle farms [11.1% (3/27)], and a single study from a turkey farm. Two studies include both pig and poultry farms. Most of the studies were cross-sectional [70.4% (19/27)], seven were longitudinal, and one was a case-control study. Complex interactions were observed among factors influencing AMU, such as biosecurity measures, farm characteristics, farmers' attitudes, availability of animal health services, stewardship, etc. A positive association between farm biosecurity and reduction in AMU was observed in 51.8% (14/27) of the studies, and 18.5% (5/27) showed that improvement in farm management practices was associated with a reduction in AMU. Two studies highlighted that coaching and awareness among farmers might lead to a decrease in AMU. A single study on economic assessment concluded biosecurity practices as a cost-effective way to reduce AMU. On the other hand, five studies showed an uncertain or spurious association between farm biosecurity and AMU. We recommend the reinforcement of the concept of farm biosecurity, especially in lower- and middle-income countries (LMICs). Further, there is a need to strengthen the evidence on the association between farm biosecurity and AMU in region- and species-specific farm settings.

Monitoring of antimicrobial usage among adult bovines in dairy herds of Punjab, India: A quantitative analysis of pattern and frequency.

The present study aimed to evaluate the antimicrobial usage (AMU) pattern in dairy herds of Punjab, India. The on-farm quantification of AMU in adult bovine animals by the manual collection of empty drug containers ("bin method") along with the records of the treatment was carried out in 38 dairy farms involving 1010 adult bovines for 1 year from July 2020 to June 2021. The farm owners were asked to record the antibiotic treatments as well as to deposit empty antibiotic packaging/vials into the provided bins placed at the farms. A total of 14 different antibiotic agents in 265 commercial antibiotic products were administered to the dairy herds during the study. A total of 179 (67.55%) administered products contained antimicrobials of "critical importance" as per the World Health Organization (WHO). Mastitis (54.72%), followed by the treatment of fever (19.62%), reproductive problems (15.47%), and diarrhea (3.40%) accounted for the majority of drugs administered in the herds during the study period. The most commonly used antibiotics were enrofloxacin (89.47% herds; 21.51% products), followed by ceftriaxone (50% herds; 12.83% products), amoxicillin (50% herds; 12.83% products), oxytetracycline (55.26% herds; 11.70% products), and procaine penicillin (47.37% herds; 12.83% products). The highest quantity of AMU [in terms of antimicrobial drug use rate (ADUR)] was observed for ceftiofur, followed by ceftriaxone, procaine benzyl penicillin ceftizoxime, enrofloxacin, cefoperazone, amoxicillin and ampicillin. A total of 125 (47.17%) products contained "highest priority critically important antimicrobials" (HPCIA) and 54 (20.37%) products contained "high priority critically important antimicrobials". In terms of overall number of animal daily doses (nADD), the highest priority critically important antimicrobials (HPCIA) of the WHO such as third-generation cephalosporins and quinolones, respectively accounted for 44.64 and 22.35% of the total antibiotic use in the herds. The bin method offers an alternative to monitoring AMU as a more accessible approach for recording the actual consumption of antimicrobials. The present study, to the best of our knowledge, is the first of its kind to provide an overview of the qualitative and quantitative estimate of AMU among adult bovines from India.

Characterization of chicken eggs associated Escherichia coli and Staphylococcus aureus for biofilm production and antimicrobial resistance traits.

The present study assessed the prevalence, virulence characteristics, antimicrobial resistance and biofilm-forming ability of E. coli and S. aureus recovered from egg samples in Ludhiana, Punjab. A total of 393 samples from hatcheries (n = 238), retail shops (n = 94), and households (n = 61) were collected. The prevalence of E. coli was observed as 11.70% and 9.16% for S. aureus. A total of 41.30% of E. coli isolates were positive for aggR gene and 52.17% were for fimA gene; while 36.11% of the S. aureus isolates were positive for coa gene. A high proportion of E. coli (76.10%) and S. aureus (69.44%) isolates were resistant toward ≥3 tested antibiotic classes. A total of 39.13% of E. coli isolates were moderate biofilm former, whereas the majority of the S. aureus (41.67%) were weak biofilm former. No significant difference regarding biofilm formation was observed between MDR and non-MDR isolates of E. coli and S. aureus. Biofilm genes viz., fimC and crl were reported in 43.47% and 80.43% of E. coli isolates, respectively; while icaA and icaD genes were reported in 58.34% and 47.22% of S. aureus isolates, respectively. A strong metabolic activity among 52.17% of E. coli and 41.66% of S. aureus isolates was observed using XTT assay. The present study highlights the need for applied food safety measures across the egg production chain of the region to prevent the development of MDR strains and biofilms.

Qualitative Study on Antimicrobial Usage and Resistance in the Dairy Chain: A Situation Analysis and Solutions by Stakeholders from Punjab, India.

The rising prevalence of antimicrobial resistance in animal foods and injudicious antibiotic use in the dairy sector pose significant threats to public health. Focus group discussions (FGDs) and key informant interviews (KIIs) were conducted with a strategic sample of four stakeholder groups (114 participants) associated with antibiotic usage in the dairy sector of Punjab. The FGDs were conducted among veterinarians (n = 56), para-veterinarians (n = 28), and KIIs were conducted among chemists (n = 18) and dairy quality managers (n = 12) during 2020-2021. FGDs and qualitative interviews of various stakeholders depict existing risk practices in the fields that may promote antimicrobial resistance. The present study revealed that widely prevalent quackery (treatment practices carried out by unauthorized persons without any recognized diploma/degree) and self-treatment by farmers, over-the-counter availability of antibiotics, low veterinarian per animal ratio, and lack of awareness among the society about the potential public health effects of antimicrobial resistance were the main risk factors for injudicious antibiotic use in the dairy sector. The present study involved a comprehensive approach targeting the stakeholders in the dairy sector and their reflections on judicious antimicrobial usage and antimicrobial resistance adapted to the dairy farming of Punjab. There is an urgent need for the advocation of policies that consider the specific challenges faced by the dairy sector to simultaneously improve access to veterinary services as well as strengthen antibiotic stewardship.

Prevalence, molecular characterization, and antimicrobial resistance profile of Clostridium perfringens from India: A scoping review.

Clostridium perfringens is one of the most important foodborne pathogens that causes histotoxic diseases and intestinal infections in both humans and animals. The present scoping review has been designed to analyze the literature published during 2000-2021 from India on the prevalence, molecular characterization, and antimicrobial resistance profile of C. perfringens isolates recovered from humans, animals, animal-based foods, and associated environmental samples. The peer-reviewed articles retrieved from four electronic databases (Google Scholar, PubMed, Science Direct, and Web of Science) were assessed using PRISMA-ScR guidelines. A total of 32 studies from India were selected on the basis of their relevance and inclusion criteria. The overall prevalence of C. perfringens among domestic animals having history of clinical symptoms and among healthy animals was found to be 65.8% (508/772) and 42.8% (493/1152), respectively. The pathogen was also detected in clinically affected wild animals (75%), healthy wild animals (35.4%), and captive birds (24.5%). The detection of C. perfringens among poultry having necrotic enteritis and among healthy birds was found to be 66.8% (321/480) and 25.6% (80/312), respectively. The detection of pathogen among animal-based foods (i.e., meat, milk, and fish and their products) and environmental samples depicted a prevalence of 20.8% (325/1562) and 30.2% (23/76), respectively. However, the prevalence of C. perfringens among humans having history of diarrhea and among healthy humans was found to be 25% (70/280) and 23.2% (36/155), respectively. The genotyping of C. perfringens isolates revealed that toxin type A was found to be the most prevalent genotype. Along with the alpha toxin gene (cpa), beta (cpb), epsilon (etx), iota (itx), enterotoxin (cpe), beta-2 toxin (cpb2), and NetB (netB) toxins were also detected in different combinations. Antimicrobial resistance profile of C. perfringens isolates recovered from different sources demonstrated that the highest resistance was detected against sulphonamides (76.8%) and tetracycline (41.3%) by phenotypic and genotypic detection methods, respectively. Comprehensive scientific studies covering different geographical areas at the human-animal-environment interface are crucial to generalize the real magnitude of C. perfringens-associated problem in India and for establishing a reliable database.

Biofilms as a microbial hazard in the food industry: A scoping review.

Biofilms pose a serious public health hazard with a significant economic impact on the food industry. The present scoping review is designed to analyse the literature published during 2001-2020 on biofilm formation of microbes, their detection methods, and association with antimicrobial resistance (if any). The peer-reviewed articles retrieved from 04 electronic databases were assessed using PRISMA-ScR guidelines. From the 978 preliminary search results, a total of 88 publications were included in the study. On analysis, the commonly isolated pathogens were Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Escherichia coli, Bacillus spp., Vibrio spp., Campylobacter jejuni and Clostridium perfringens. The biofilm-forming ability of microbes was found to be influenced by various factors such as attachment surfaces, temperature, presence of other species, nutrient availability etc. A total of 18 studies characterized the biofilm-forming genes, particularly for S. aureus, Salmonella spp., and E. coli. In most studies, polystyrene plate and/or stainless-steel coupons were used for biofilm formation, and the detection was carried out by crystal violet assays and/or by plate counting method. The strain-specific significant differences in biofilm formation were observed in many studies, and few studies carried out analysis of multi-species biofilms. The association between biofilm formation and antimicrobial resistance was not clearly defined. Further, viable but non-culturable form of the foodborne pathogens is posing an unseen (by conventional cultivation techniques) but potent threat to the food safety. The present review recommends the need for carrying out systematic surveys and risk analysis of biofilms in food chain to highlight the evidence-based public health concerns, especially in regions where microbiological food hazards are quite prevalent.

Pharmacokinetic-Pharmacodynamic Study of Ampicillin-Cloxacillin Combination in Indian Thoroughbred Horses (Equus caballus) and Safety Evaluation of the Computed Dosage Regimen.

The pharmacokinetics of ampicillin-cloxacillin, given as single intravenously dose of 10 mg.kg-1 (5 mg.kg-1 of ampicillin plus 5 mg.kg-1 of cloxacillin) was examined in clinically presented Indian thoroughbred horses (n = 6) in order to design appropriate dosing strategies. Drug concentrations in plasma were determined by high performance liquid chromatography (HPLC) and pharmacokinetic parameters were derived by non-compartmental analysis using WinNonlin software. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of ampicillin-cloxacillin against quality control strains of Escherichia coli and Staphylococcus aureus, grown in Muller Hinton Broth, were determined by broth microdilution method. For ampicillin, area under plasma drug concentration time curve (AUC) was 15.2 ± 0.54 µg.h.ml-1, mean residence time (MRT) was 1.33 ± 0.06 h and clearance (Cl) was 0.33 ± 0.01 L.h-1.kg-1. For cloxacillin, AUC was 18.0 ± 0.9 µg.h.ml-1, MRT was 1.28 ± 0.02 h and Cl was 0.28 ± 0.01 L.h-1.kg-1. MIC of ampicillin-cloxacillin combination against E. coli and S. aureus was determined to be 0.4 µg.ml-1. PK-PD integration indicated that to maintain %T > MIC value 50% for bacteria with MIC ≤ 0.4 µg.ml-1, an appropriate intravenous dosage regimen of ampicillin-cloxacillin combination in horses would be 15 mg.kg-1 (i.e. 7.5 mg.kg-1 of ampicillin plus 7.5 mg.kg-1 of cloxacillin), to be repeated at 12 h intervals. Safety profile of the recommended regimen did not significantly alter any of the 16 biochemical or haematological parameters studied.

Association between aflatoxin M1 excretion in milk and indicators of rumen fermentation in bovines.

Aflatoxins and its metabolites negatively impact the ruminant health and production. The present cross-sectional study was aimed to determine the effect of aflatoxins on rumen fermentation by deducing the correlation between the aflatoxin M1 (AFM1) excretion in milk and indicators of rumen fermentation in bovines. The indicators of rumen fermentation were taken into account and correlated with AFM1 concentration in milk of 120 bovines (cattle (n = 82) and buffalo (n = 38)). The AFM1 in milk samples (n = 120) was quantified by ELISA kit. The correlation analysis revealed that with increase in excretion of AFM1 in milk, the pH (r = 0.38), methylene blue reduction time (MBRT) (r = 0.43), sedimentation activity time (SAT) (r = 0.31) and ammonia nitrogen content (r = 0.34) of rumen liquor increase, whereas the total volatile fatty acid (TVFA) content (r = - 0.25), total bacterial count (TBC) (r = - 0.43) and total protozoal count (TPC) (r = - 0.14) of rumen liquor decrease. The results of the present study suggest that the presence of aflatoxins in rumen could have negative effect on the process of rumen fermentation. Therefore, the prevention of primary entry point(s) of AFB1 through the feed of bovines is important for the animal health as well as public health.

Emergency preparedness for public health threats, surveillance, modelling & forecasting.

In the interconnected world, safeguarding global health security is vital for maintaining public health and economic upliftment of any nation. Emergency preparedness is considered as the key to control the emerging public health challenges at both national as well as international levels. Further, the predictive information systems based on routine surveillance, disease modelling and forecasting play a pivotal role in both policy building and community participation to detect, prevent and respond to potential health threats. Therefore, reliable and timely forecasts of these untoward events could mobilize swift and effective public health responses and mitigation efforts. The present review focuses on the various aspects of emergency preparedness with special emphasis on public health surveillance, epidemiological modelling and capacity building approaches. Global coordination and capacity building, funding and commitment at the national and international levels, under the One Health framework, are crucial in combating global public health threats in a holistic manner.

Knowledge, Attitude, and Practices (KAP) Survey among Veterinarians, and Risk Factors Relating to Antimicrobial Use and Treatment Failure in Dairy Herds of India.

The indiscriminate usage of antimicrobials in the animal health sector contributes immensely to antimicrobial resistance (AMR). The present study aims to assess the antimicrobial usage pattern and risk factors for AMR in animal husbandry sector of India. A cross-sectional survey about Knowledge, Attitude, and Practices (KAP) among veterinarians was carried out using a questionnaire comprising of 52 parameters associated with antibiotic use and the emergence of AMR in dairy herds. Respondents' KAP scores were estimated to rank their level of knowledge, attitude, and practice. Furthermore, risk factors associated with treatment failure were analyzed by univariable and multivariable analyses. Out of a total of 466 respondents, the majority had average knowledge (69.5%), neutral attitude (93.2%), and moderate practice (51.3%) scores toward judicious antibiotic usage. Veterinarians reported mastitis (88.0%), reproductive disorders (76.6%), and hemoprotozoan infections (49.6%) as the top three disease conditions that require antibiotic usage. Most of the veterinarians (90.6%) resorted to their "own experience" as the main criteria for antibiotic choice. The use of the highest priority critically important antimicrobials (HPCIA) listed by the World Health Organization (WHO) in animals, particularly quinolones (76.8%) and third-generation cephalosporins (47.8%), has been reported. On multivariable regression analysis of the risk factors, the lack of cooperation of the dairy farmers in the completion of a prescribed antibiotic course by the veterinarian and the demand for antibiotic use even in conditions not requiring antibiotic use were found to be significantly associated with the outcome variable "treatment failure" having respective odds of 1.8 (95%CI: 1.1-3.0) and 3.6 (95%CI: 2.3-5.8) (p < 0.05). The average KAP score of veterinarians, poor farm management practices, lack of awareness among farmers on prudent antibiotic use, and lack of antibiotic stewardship are the significant factors that need attention to combat the rising AMR in veterinary sector in India.

Assessment of aflatoxin contamination in dairy animal concentrate feed from Punjab, India.

Aflatoxins are one of the major environmental contaminants in animal feed and pose a potential threat to human health due to their secretion in the milk of lactating animals. The present study was conducted with the objectives to determine the occurrence of aflatoxins (B1, B2, G1, and G2) in dairy animal concentrate feed and to evaluate the effect of season, spatial variation, and dairy farm size on the levels of aflatoxins contamination. A total of 189 dairy animal concentrate feed samples were tested for aflatoxins with enzyme-linked immunosorbent assay (ELISA) as screening and high-performance liquid chromatography with fluorescence detection (HPLC-FLD) as confirmatory techniques. Of the total, 59% feed samples were found positive for aflatoxins, while 44% samples were detected with total aflatoxins levels higher than the tolerance limit established by the Food and Drug Administration (FDA) and 58% samples were found with aflatoxins B1 (AFB1) levels above the European Commission (EC) legal limit. AFB1 levels in dairy animal concentrate feed were found significantly higher during rainy (41.6 µg kg-1) and winter (35.9 µg kg-1) seasons as compared to the summer season (25.5 µg kg-1). The theoretical extrapolation of the AFB1 carry-over from animal feed to milk (aflatoxins M1) in different seasons may lead to 50-100% contamination of milk at levels above the EC tolerance limit. The incidence and levels of aflatoxins especially AFB1 in animal feed, not only pose a direct effect on animals but may also pose a concern for food safety in relation to the occurrence of aflatoxins M1 in milk. Therefore, continuous surveillance of aflatoxins in dairy animal feeds is required to reduce animal and consequently human exposure.

Epidemiology of brucellosis in cattle and dairy farmers of rural Ludhiana, Punjab.

Brucellosis is a zoonotic disease imposing significant impacts on livestock production and public health worldwide. India is the world's leading milk producer and Punjab is the state which produces the most cattle and buffalo milk per capita. The aim of this study was to investigate the epidemiology of bovine brucellosis to provide evidence for control of the disease in Punjab State, India. A cross-sectional study of dairy farms was conducted in humans and livestock in rural Ludhiana district using a multi-stage sampling strategy. The study suggests that brucellosis is endemic at high levels in cattle and buffalo in the study area with 15.1% of large ruminants testing seropositive and approximately a third of dairy farms having at least one animal test seropositive. In total, 9.7% of those in direct contact with livestock tested seropositive for Brucella spp. Persons that assisted with calving and/or abortion within the last year on a farm with seronegative livestock and people which did not assist with calving/abortion had 0.35 (95% CI: 0.17 to 7.1) and 0.21 (0.09 to 0.46) times the odds of testing seropositive compared to persons assisting with calving/abortion in a seropositive farm, respectively. The study demonstrated that persons in direct contact with cattle and buffalo in the study area have high risk of exposure to Brucella spp. Control of the disease in livestock is likely to result in benefits to both animal and public health sectors.

Potential risk factors associated with the occurrence of aflatoxin M1 in raw milk produced under different farm conditions.

The present study was carried out with the objectives to determine the occurrence of aflatoxin M1 (AFM1) in raw milk and identification of associated risk factors from three different types of animal farming systems. A total of 189 pooled raw milk samples were tested for AFM1 with enzyme linked immunosorbant assay (ELISA) as screening and high performance liquid chromatography with fluorescence detection (HPLC-FLD) as confirmatory techniques. AFM1 was found positive in 58% pooled milk samples with the mean level of 0.917 µg/L, while 50.8% and 36.5% samples were found above the legal limits set by European Commission (EC) and Food safety and standard authority of India (FSSAI), respectively for AFM1 in milk. Among all investigated risk factors, feeding of readymade concentrate feed and leftover household cereals, longer feed storage duration and feed storage quality were found significantly associated with presence of AFM1 in farm milk. High prevalence as well as levels of AFM1 in farm milk poses a health risk to the consumers especially children. Therefore, efforts are urgently required to create awareness among dairy farmers about aflatoxins contamination of feed and milk, to improve farm management practices and to control AFM1 level in milk below the legal limits.

Antibiotic residues and mycotoxins in raw milk in Punjab (India): a rising concern for food safety.

The present study was envisaged with an aim to determine the occurrence of antibiotic residues; enrofloxacin, oxytetracycline, penicillin G, sulphamethoxazole and chloramphenicol as well as mycotoxins; aflatoxin M1 and ochratoxin A in raw milk samples collected from individual animals from dairy farms located in 9 districts of Punjab, India. A total of 168 raw milk samples were collected and analysed using commercially available competitive Enzyme linked immunosorbent assay kits. Out of these, 19 (11.3%) and 9 (5.4%) samples were found positive for antibiotic residues and mycotoxins, respectively. The milk samples were positive for enrofloxacin (4.8%), oxytetracycline (3.0%), penicillin G (3.0%) and sulphamethoxazole (0.6%) residues. The percentage of samples found above maximum residue limit established by Europen Union (EU)/Codex Alimentarius Commission (CAC) were 1.7%, 1.2% and 0.6% for enrofloxacin, oxytetracycline and penicillin G residues, respectively. Aflatoxin M1 was detected in 5.4% of monitored milk samples with 1.2% samples exceeded the established maximum levels of EU but were below the maximum levels established by CAC. The occurrence of antibiotic residues and mycotoxins in raw milk samples above maximum limits is a rising concern for food safety due to possibility of health risks to the consumers.

Listeria contamination in chevon and mutton from retail meat shops and slaughter house environment of Punjab, India.

Listeria monocytogenes is recognized as an emerging pathogen, with limited information or statistics on this pathogen. Moreover, poor hygienic practices in the unorganized meat sector favor its growth and survival. Therefore, the present study was carried out in Punjab, India to assess the prevalence of Listeria spp. in chevon and mutton samples, followed by its characterization. The prevalence of Listeria in chevon, mutton and swab samples collected from butchers' shops and slaughter houses was 1.82%, 3.21% and 6.66%, respectively. The 18 (2.67%) L. monocytogenes strains isolated from 786 samples carried one or more virulence-associated gene. Based on virulence genes, 6/18 isolates were potentially pathogenic. Serovar 4b (44.4%) was predominant, followed by serovars 1/2a (22.2%), 1/2b (22.2%) and 1/2c (11.1%). Detection of L. monocytogenes in meat and environment samples indicated poor hygiene, potential cross-contamination and risk of listeriosis to consumers and occupational workers. The findings of the study were also relevant to the fact that most of the slaughtering in Punjab, India is carried out in unorganized way, and this meat is not stored propery. Moreover, in the absence of a systematic foodborne disease surveillance and monitoring program, a database on foodborne diseases is absent, and needs to be established.

Pesticide residues in Indian raw honeys, an indicator of environmental pollution.

Honey has multifaceted beneficial properties, but polluted environment and unapproved apicultural practices have led to its contamination. In this study, QuEChERS method followed by chromatographic analysis by GC-µECD/FTD and GC-MS was validated and used for determination of 24 pesticides in 100 raw honey samples from various floral origins of Northern India. Matrix-matched calibrations showed that the method was selective and linear (r2 > 0.99) with detection limit < 9.1 ng g-1 for all the studied pesticides except for monocrotophos (21.3 ng g-1). The average recoveries at different fortification levels ranged from 86.0 to 107.7% with relative standard deviation < 20%. Pesticide residues were detected in 19.0% samples, and most prevalent compounds detected were dichlorvos in 6.0% samples followed by monocrotophos (5.0%), profenofos (5.0%), permethrin (4.0%), ethion (3.0%), and lindane (3.0%) with concentrations ranging from 58.8 to 225.5, from 96.0 to 430.1, from 14.6 to 43.2, from 27.8 to 39.6, from 25.6 to 28.0, and from 19.6 to 99.2 ng g-1, respectively. Honey samples originating from cotton, sunflower, and mustard crops (33.3%) that tested positive for pesticide residues were found to be significantly higher (p < 0.05) than the honey originating from natural and fruity vegetation (13.5%). Therefore, considering the contamination of environmental compartments due to extensive application of pesticides in the study area and their potential for subsequent transfer to honey by the expeditious bees, the results of present study proclaim that honey may be used as an indicator of environmental pollution. Further, estimated daily intakes of all contaminants were found to be at levels well below their acceptable daily intakes suggesting that consumption of honeys at current levels does not pose deleterious effects on human health. However, precautionary measures should always be taken considering the customary honey feeding in infants and cumulative effect of these chemicals in the foreseeable future.