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.

Modelling the control of bovine brucellosis in India.

Brucellosis imposes substantial impacts on livestock production and public health worldwide. A stochastic, age-structured model incorporating herd demographics was developed describing within- and between-herd transmission of Brucella abortus in dairy cattle herds. The model was fitted to data from a cross-sectional study conducted in Punjab State of India and used to evaluate the effectiveness of control strategies under consideration. Based on model results, stakeholder acceptance and constraints regarding vaccine supply, vaccination of replacement calves in large farms should be prioritized. Test and removal applied at early stages of the control programme where seroprevalence is high would not constitute an effective or acceptable use of resources because significant numbers of animals would be 'removed' (culled or not used for breeding) based on false positive results. To achieve sustained reductions in brucellosis, policymakers must commit to maintaining vaccination in the long term, which may eventually reduce frequency of infection in the livestock reservoir to a low enough level for elimination to be a realistic objective. This work provides key strategic insights into the control of brucellosis in India, which has the largest cattle population globally, and a general modelling framework for evaluating control strategies in endemic settings.

Exploring smart phone based colorimetric technology for on-site quantitative determination of adulterant (neutralizer) in milk.

There is a great concern regarding the safety of milk not only for human health but also for its economic consequences. The portable sensing devices, which can collect and analyze data within food supply chains at critical control points are lacking. Smart phones have now emerged as an integral part of each home, lab, farm and factory. It is having a provision of digital camera and computation; with widespread applicability including food analysis. The use of soda as a milk neutralizer is a usual practice but has a detrimental human health impact. This investigation explored an easy, economic, fast, repeatable, and field applicable Smartphone-based sensing technology, which was standardized and in-house validated for the quantitative determination of neutralizer in milk samples. The method had simple steps of spot-test response and digital image evaluation with the Red Green Blue process. The linearity of the method was shown by analytical curves ranging from 0.125% (1250 ppm) to 1% (10,000 ppm) that were characterized by R2 > 0.99. The limit of detection of 0.11% demonstrated the sensitivity of the method which was found better than the existing wet chemical spot test. Comparison with the existing spectroscopic method revealed no statistically significant difference between the observations using paired t-test at a confidence level of 95%.

Seroprevalence and risk factor investigation for the exposure of Toxoplasma gondii among veterinary personnel in Punjab, India.

Toxoplasma gondii, a globally important food borne zoonotic parasite, infects most of the warm-blooded animals as well as people. Veterinarians and para vets are considered at risk of T. gondii exposure. We determined the seroprevalence of T. gondii in veterinary personnel and investigated the associated risk factors in Punjab, India. Two hundred and five blood samples collected from veterinary personnel were tested for the presence of Toxoplasma IgG and IgM antibodies using ELISA. The apparent and true seroprevalence of T. gondii with 95% confidence interval (CI) were estimated. Information about participant demographics, and possible routes of exposure was collected using a self-completed questionnaire at the time of blood collection. For risk factor investigation, a veterinary person was considered Toxoplasma seropositive using a combination of tests in parallel, i.e. if it was positive in either IgG or IgM ELISA. A mixed effects logistic regression model was constructed to evaluate the association of demography, occupational and non-occupational factors with Toxoplasma seropositive status. The apparent and estimated true seroprevalence of T. gondii antibodies using Toxoplasma IgG ELISA was 8.78% (95% CI 5.63-13.45%) and 7.36% (95% CI 4.04-12.29%), respectively. The apparent and estimated true seroprevalence using Toxoplasma IgM ELISA was 0.49% (95% CI inestimable - 2.71%) and 0.51% (95% CI inestimable - 2.83%), respectively. After adjusting other variables in the final model, consuming mutton and owning a cat were associated with large odds of being Toxoplasma seropositive. In this study occupational exposure does not seem to play an important role for the exposure of T. gondii in veterinary personnel in Punjab state of India. The seroprevalence of toxoplasmosis in veterinary personnel is comparatively low in occupationally exposed veterinary personnel in Punjab, India.

Comparative analysis of Methicillin-Resistant Staphylococcus aureus (MRSA) and Borderline Oxacillin Resistant Staphylococcus aureus (BORSA) in community and food of animal origin.

The aim of this study was to illustrate the relative pervasiveness of Borderline Oxacillin Resistant Staphylococcus aureus (BORSA) and Methicillin-Resistant Staphylococcus aureus (MRSA) in community and food of animal origin and their relationship with other genetic determinants. Staphylococcus aureus isolates were subjected to E-test using the antibiotics: oxacillin, ceftriaxone, cotrimoxazole, vancomycin, genotypic tests for the genes mecA, vanA, blaZ, pvl gene and SCCmec typing. The prevalence of S. aureus (MRSA) in the food of animal origin and community settings was 21% (1.8% MRSA) and 21.9% (7.4% MRSA), respectively. SCCmec type V was prevalent among the food of animal origin, while SCCmec type IVa among the community isolates. The likelihood of MRSA presence among community isolates was three times more than in isolates from chicken and milk samples. Likewise, the likelihood of detecting pvl positive MRSA (pvl+MRSA) isolates was 4-fold higher in the community setting than in the food of animal origin. The mecA negative BORSA (mecA-BORSA) was a frequently observed phenotype among S. aureus isolates. Also, co-detection of pvl and cotrimoxazol resistance was reported in this study although there was no noteworthy correlation of cotrimoxazol resistance with the type of sample. Isolates from milk and community settings exhibit higher minimum inhibitory concentration to vancomycin (Vancomycin MIC creep, 2-4 µg/mL). SIGNIFICANCE: Current study provides the information on the statistical relationship between the genetic determinants of S. aureus with respect to sample type, and additionally the correlation that exists between the pvl and MRSA, pvl and cotrimoxazol resistance, vancomycin MIC and MRSA/Methicillin-Susceptible S. aureus (MSSA).

The Extent and Structure of Peri-urban Smallholder Dairy Farming in Five Cities in India.

Livestock keeping is common in many cities in India, driven by the demand for animal-source foods, particularly perishable milk. We selected five cities from different regions of the country and conducted a census in 34 randomly selected peri-urban villages to identify and describe all smallholder dairy farms. In total 1,690 smallholder dairy farms were identified, keeping on average 2.2 milking cows and 0.7 milking buffaloes. In Bhubaneswar, the proportion of cows milking was only 50%, but in other cities it was 63-73%. In two of the five cities, more than 90% of the farmers stated that dairy production was their main source of income, while <50% in the other cities reported this. In one of the cities, only 36% of the households kept milk for themselves. Market channels varied considerably; in one city about 90% of farms sold milk to traders, in another, 90% sold to the dairy cooperative, and in another around 90% sold directly to consumers. In conclusion, peri-urban dairy systems in India are important but also varying between different cities, with only one city, Bengaluru, having a well-developed cooperative system, and the northeastern poorer region being more dependent on traders. Further studies may be needed to elucidate the importance and to design appropriate developmental interventions.

Pesticide Residues in Peri-Urban Bovine Milk from India and Risk Assessment: A Multicenter Study.

Pesticides residue poses serious concerns to human health. The present study was carried out to determine the pesticide residues of peri-urban bovine milk (n = 1183) from five different sites (Bangalore, Bhubaneswar, Guwahati, Ludhiana and Udaipur) in India and dietary exposure risk assessment to adults and children. Pesticide residues were estimated using gas chromatography with flame thermionic and electron capture detectors followed by confirmation on gas chromatography-mass spectrometer. The results noticed the contamination of milk with hexachlorocyclohexane (HCH), dichloro-diphenyl trichloroethane (DDT), endosulfan, cypermethrin, cyhalothrin, permethrin, chlorpyrifos, ethion and profenophos pesticides. The residue levels in some of the milk samples were observed to be higher than the respective maximum residue limits (MRLs) for pesticide. Milk samples contamination was found highest in Bhubaneswar (11.2%) followed by Bangalore (9.3%), Ludhiana (6.9%), Udaipur (6.4%) and Guwahati (6.3%). The dietary risk assessment of pesticides under two scenarios i.e. lower-bound scenario (LB) and upper-bound (UB) revealed that daily intake of pesticides was substantially below the prescribed acceptable daily intake except for fipronil in children at UB. The non-cancer risk by estimation of hazard index (HI) was found to be below the target value of one in adults at all five sites in India. However, for children at the UB level, the HI for lindane, DDT and ethion exceeded the value of one in Ludhiana and Udaipur. Cancer risk for adults was found to be in the recommended range of United States environment protection agency (USEPA), while it exceeded the USEPA values for children.

Brucellosis in India: results of a collaborative workshop to define One Health priorities.

Brucellosis is an important zoonosis worldwide. In livestock, it frequently causes chronic disease with reproductive failures that contribute to production losses, and in humans, it causes an often-chronic febrile illness that is frequently underdiagnosed in many low- and middle-income countries, including India. India has one of the largest ruminant populations in the world, and brucellosis is endemic in the country in both humans and animals. In November 2017, the International Livestock Research Institute invited experts from government, national research institutes, universities, and different international organizations to a one-day meeting to set priorities towards a "One Health" control strategy for brucellosis in India. Using a risk prioritization exercise followed by discussions, the meeting agreed on the following priorities: collaboration (transboundary and transdisciplinary); collection of more epidemiological evidence in humans, cattle, and in small ruminants (which have been neglected in past research); Economic impact studies, including cost effectiveness of control programmes; livestock vaccination, including national facilities for securing vaccines for the cattle population; management of infected animals (with the ban on bovine slaughter, alternatives such as sanctuaries must be explored); laboratory capacities and diagnostics (quality must be assured and better rapid tests developed); and increased awareness, making farmers, health workers, and the general public more aware of risks of brucellosis and zoonoses in general. Overall, the meeting participants agreed that brucellosis control will be challenging in India, but with collaboration to address the priority areas listed here, it could be possible.

Prevalence, molecular detection and risk factors investigation for the occurrence of Toxoplasma gondii in slaughter pigs in North India.

BACKGROUND: Toxoplasma gondii, an important food borne zoonotic parasite, infects almost all warm-blooded animals including pigs. People primarily become infected with T. gondii via consuming meat of infected animals. Status of T. gondii is largely unknown in pigs in India including northern regions. We, therefore, determined the prevalence of T. gondii infection in pigs from North India. RESULTS: DNA of T. gondii was detected in 6.7% (54/810) of the tested slaughter pigs. Highest prevalence was observed in pigs from Punjab (8.2%) followed by Chandigarh (5.3%) and Uttarakhand (4.8%). Phylogenetic analysis revealed that the isolates from pigs had 96-100% nucleotide identity with Type I RH strain (AF179871), 96-99.7% with VEG type III strain (LN714499) and 67-72% with type II ME 49 strain (XM002370240). However, low level of polymorphism in the targeted B1 gene did not allow the determination of the clonal lineages of the isolates. Antibodies against T. gondii was reported in 48.3% (73/151) of the sera obtained from pigs slaughtered at Chandigarh abattoir, and scavenging by pigs was a significant risk factor. CONCLUSION: Prevalence of T. gondii DNA was low in pigs in North India, however, presence of the parasite warrants food safety concerns. Further studies are required to identify the clonal lineage of T. gondii circulating in pigs reared in North India. Pig farmers should be educated about the hygienic management practices.

Prevalence, multidrug resistance and molecular typing of methicillin-resistant Staphylococcus aureus (MRSA) in retail meat from Punjab, India.

OBJECTIVES: This study reports the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in retail meat from Punjab, India. METHODS: Classical microbiological methods were applied to isolate and identify S. aureus isolates. Isolates also underwent Etest. PCR and sequencing were used to identify and characterise antimicrobial resistance genes. MLST, SCCmec and spa typing were performed. RESULTS: A total of 408 meat and 101 swab samples were processed for S. aureus isolation. Phenotypic resistance was highest to penicillin (90.97%), followed by ciprofloxacin (61.80%), tetracycline (45.14%) and erythromycin (11.11%). Isolates from chicken samples showed significantly higher MICs for tetracycline than chevon and pork samples and significantly higher MICs for trimethoprim/sulfamethoxazole and gentamicin than chevon and swab samples (P<0.05). No isolates were phenotypically resistant to vancomycin (MICs of 0.5-2µg/mL). Most isolates (52.78%, 95% CI 44.63-60.93%) were multidrug-resistant and carried resistance genes to penicillin (blaZ), oxacillin (mecA), gentamicin (aacA-aphD), erythromycin (ermB, ermC) and tetracycline (tetK, tetL, tetM). MRSA was only found in chicken samples (2.72%; 4/147). Seven S. aureus (5.07%) were borderline oxacillin-resistant (MIC range 4-8µg/mL). All MRSA were SCCmecV-pvl+-t442, among which three isolates were ST5. Their genotype was mecA+, blaZ+, aacA-aphD+, tetK+, ermC+/-. Among the erythromycin-resistant isolates, 25% were MRSA, of which 12.5% isolates expressed an inducible macrolide-lincosamide-streptogramin B (iMLSB) phenotype. CONCLUSION: These data confirm the presence of ST5-t442-MRSA-SCCmecV-pvl+ and iMLSB MRSA in meat samples, indicating a potential role of meat in the dissemination of multidrug-resistant S. aureus strains and successful MRSA lineages in Punjab.

Knowledge, attitude and practices relating to zoonotic diseases among livestock farmers in Punjab, India.

Zoonotic diseases cause significant health and economic impact in developing countries such as India. Many zoonotic diseases are prevalent in the livestock and as an occupational zoonosis in the livestock farmers in India. Lack of knowledge on the disease transmission, prevention and control measures is a potential high risk for the occurrence of zoonotic diseases in the livestock and its keepers in India. We conducted this study to understand knowledge, attitude and practices of livestock farmers regarding zoonoses. Five villages from each of the 22 districts of the state were conveniently selected (n = 110). Farmers available at village community sites were enrolled in the study and requested to complete a custom designed questionnaire (n = 558). In addition, livestock farmers attending basic livestock husbandry training were also surveyed (n = 301). Data from questionnaires was used to create three index variables: (a) knowledge score; (b) attitude score and (c) practice score. Association between demographic and other explanatory variables with knowledge score was evaluated using linear regression analyses. Similarly, the association between knowledge and attitude score with practice score was evaluated. Of the 859 participants, 685 (80%) livestock farmers had heard the term 'zoonoses' but only 345 (40%), 264 (31%) and 214 (25%) farmers were aware of the zoonotic nature of tuberculosis, Japanese encephalitis and taeniosis, respectively. For practices, 23% farmers reported consumption of raw milk and only 10% and 8% livestock farmers ever got their animals tested for brucellosis and tuberculosis, respectively. The low level of education and being a cattle farmer were negatively associated with the farmer's knowledge on zoonotic diseases. The attitude score was positively associated with the practice score of the participants. The results indicate need for educating the livestock farmers particularly those with a low level of education to reduce the health and economic impact of zoonotic diseases in India.

Estimation of the health and economic burden of human brucellosis in India.

High seroprevalence estimates of brucellosis in livestock species and humans in India suggest that the disease is a significant public health concern in the country. We developed an economic model and conducted an assessment of the impact of human brucellosis in India to describe the current situation to help formulate prevention and control strategies. Economic losses of human brucellosis were calculated based on the official records and the data from epidemiological surveys conducted in India. These data were used to estimate the disability adjusted life years (DALYs) due to human brucellosis. Uncertainty was incorporated in the model by applying probability distributions for certain input parameters. The analyses were further supplemented by conducting sensitivity analyses to determine which parameters had the biggest influence on the outcome of economic losses. The annual median losses due to human brucellosis were estimated to be Rs 627.5 million (uncertainty interval [95% UI] Rs 534.8-741.2 million; US $ 10.46 million) with a loss of Rs 442.3 million (95% UI 371.0-516.0; US $ 7.37 million) among adults and Rs 185.0 million (95% UI 124.0-255.0; US $ 3.08 million) among children. Human brucellosis in India caused a loss of 177 601 (95% UI 152 695-214 764) DALYs at the rate of 0.15 (95% UI 0.13-0.17) DALYs per thousand persons per year. The DALYs were found to be 0.29 (95% UI 0.08-0.70) per thousand persons per year in occupational and 0.13 (95% UI 0.06-0.18) in non-occupational adult population. This is the first systematic analysis of the health impact of human brucellosis in India and of indirect/production losses occurring due to human brucellosis anywhere in the world. The results indicate that brucellosis causes considerable economic losses and has a reasonable health impact, particularly among occupational groups. Intervention policies need to be strengthened to reduce the socio-economic impact of human brucellosis in India.

Persistent Organic Pollutants Residues in Human Breast Milk from Bathinda and Ludhiana Districts of Punjab, India.

In the present study, persistent organic pollutants (POPs), including six congeners of polychlorinated biphenyls (PCBs) and organochlorine pesticides, were estimated in human breast milk samples collected from two districts of Punjab (India). The mean concentrations of POP residues were recorded higher in Bathinda district (PCBs: 33.7; DDTs: 519.2; HCHs: 46.6 ng g-1 lipid wt.) than Ludhiana (PCBs: 24.2; DDTs; 415.3; HCHs; 35.5 ng g-1 lipid wt.). Levels of PCBs and DDTs were observed higher in primiparas, whereas HCHs residues were seen more in multiparas. Risk analysis to POPs exposure through breast milk reflected that the daily intake for some infants was close to or above the tolerable daily intake limit for detrimental effects, which may raise a health concern. Comparative evaluation of present data indicated that DDT and HCHs residue levels in human breast milk from Punjab, India were among the lowest values reported for developing countries. The first-order kinetic reaction at a steady-state condition used to estimate the half-life of DDT and HCH suggested that DDT levels have declined from 18,211 to 490 ng g-1 lipid wt. with a half-life (Tdec1/2) of 3.25 years over a span of 15 years. Similarly, HCH levels have decreased from 8609 to 46.6 ng g-1 in this duration with Tdec1/2 of 2.25 years. Because some infants are still at risk, continuous monitoring of POPs in human milk is needed for surveillance and interpretation of time trends and for linkage to strict enforcement of agricultural regulations.

Risk factors for occupational Brucella infection in veterinary personnel in India.

Brucellosis is an important animal and human health issue in developing countries. This study aimed to identify risk factors associated with occupational Brucella infection in veterinary personnel in India. Blood samples were collected from 279 veterinary personnel working in the public sector. Sera were tested with rose bengal plate test (RBPT), standard tube agglutination test (STAT), and IgG and IgM ELISAs. Information about participant demographics, risk of exposure and infection control practices was collected using a self-completed questionnaire. The outcome measure of Brucella infection was created based on a positive RBPT or STAT test and a positive IgG ELISA test. Binomial logistic regression analyses were conducted to evaluate associations between explanatory variables and the outcome variable. Of the 279 participants, 61 (21.9%), 67 (24.0%), 55 (19.7%) and 150 (53.8%) were positive in RBPT, STAT, IgM and IgG ELISA, respectively. Compared to a trained veterinarian, veterinary pharmacists and animal handlers had greater odds of being test positive, suggesting that they were at greater risk of Brucella infection. Number of years spent working with animals was associated with greater odds of a person being positive for Brucella infection (p = .015). Counter-intuitively, those using personal protective equipment (PPE) for handling sick animals were found to be at greater risk, suggesting that either the use of PPE is inappropriate-making it ineffective-or that it is reverse association whereby those experiencing brucellosis symptoms start using PPE. Brucellosis is a common occupational zoonosis among veterinary personnel in India with the risk being higher in paraveterinary staff than veterinarians and in those who have been practicing for a longer period of time. Further investigations are required to clarify the effectiveness of PPE to reduce Brucella infection in veterinary personnel in India.

Exploring animal rabies endemicity to inform control programmes in Punjab, India.

Previous studies estimate that one-third of the annual global burden of rabies (~20,000 cases) occurs in India. Elimination of canine rabies is essential to reduce this burden. Surveillance of animal cases can assess both the risk to humans and the efficacy of control strategies. The objective of this study was to describe the spatial and temporal occurrence of reported confirmed cases of rabies in animals in Punjab, India, from 2004 to 2014. We analysed passive surveillance data on 556 samples submitted from 2004 to 2014 to GADVASU, Ludhiana, Punjab, India. Regression and time series analyses were conducted to understand seasonal and long-term variation of cases and identify cross-correlation of monthly cases between species. Spatio-temporal analyses assessed spatial autocorrelation of date of reporting, mean geographic centres of disease occurrence and clustering of cases using Kulldorff's space-time permutation statistic. The annual number of submissions and proportion of confirmed cases were consistent throughout 2004-2014. Most submissions (320; 57.6%) were confirmed rabies cases, including dogs (40.6%), buffalo (29.7%) and cattle (23.1%). Regression analysis of monthly cases in dogs showed seasonal variation with significant increases in cases in March and August. Monthly case numbers in buffalo decreased over time. Long-term temporal trend was not detected in dog and cattle cases. Time-series models identified significant cross-correlation between dog and buffalo cases, suggesting that buffalo cases were spillover events from dogs. Significant spatio-temporal variation or clusters of cases were not detected. These results indicate that rabies cases in animals-and therefore, the potential for exposure to humans-were temporally and spatially stable during 2004-2014 in Punjab, India. The endemic nature of rabies transmission in this region demands a coordinated, sustained control programme. This study provides baseline information for assessing the efficacy of rabies control measures and developing seasonally targeted dog vaccination and rabies awareness strategies.

Molecular characterization of antibiotic-resistant Staphylococcus aureus from livestock (bovine and swine).

AIM: The aim of this study was to figure the prevalence, phenotypic and genotypic antibiotic resistance (AR) pattern of Staphylococcus aureus isolated from bovine and swine nares. MATERIALS AND METHODS: Colonies with typical morphology on Baird-Parker agar supplemented with egg-yolk tellurite emulsion were selected and biochemically/genotypically identified as S. aureus. These strains were further subjected to epsilometer test for their sensitivity to various clinically important antibiotics and antibiotic susceptibility testing for amoxicillin/clavulanic acid, and double-disk diffusion testing was performed by the standard disc diffusion method following CLSI guidelines. S. aureus strains were also tested for the presence of AR genes, viz., blaZ, mecA, aacA-aphD, erm (ermA, ermB, ermC), tet (efflux genes tetK and tetL, tetM and tetO of the ribosomal protection family), and vanA. RESULTS: The nasal cavities of 17 out of 47 randomly selected bovine and 20 out of 28 randomly selected swine were positive for S. aureus, representing the prevalence of 36.2% (95% confidence interval [CI]: 22.5-49.9) and 71.4% (95% CI: 54.7-88.1), respectively. Most of the S. aureus strains showed higher resistance to penicillin (94.6%, minimal inhibitory concentration [MIC] ≥1.5 µg/ml) followed by ciprofloxacin (56.7%, MIC ≥32 µg/ml) and tetracycline (18.9%, MIC ≥32 µg/ml). About 10-15% of the strains were resistant to gentamicin (MIC 16 µg/ml) and oxacillin (MIC 6-8 µg/ml). None of the strains were resistant to vancomycin (MIC 0.25-1.5 µg/ml). In this study, 32.4% strains were resistant to three or more than three antibiotics and prevalence of this multi-drug resistant S. aureus was 45% (95%CI: 26.6-63.4) and 17.6% (95%CI: 6.7-28.5) in swine and bovine nasal samples, respectively. Four strains from pigs were borderline oxacillin-resistant S. aureus MIC 6-8 µg/ml, but none were mecA positive. Two of these strains were ß-lactamase hyperproducers. Among the resistance genes blaZ, tetK, tetL, tetM, ermB, and aacA-aphD were found. CONCLUSION: Our results demonstrated the absence of mecA and pvl gene, but the presence of multi-drug resistant S. aureus in the nares of healthy animals which has a potential to spread in a community.

The Prevalence of Extended-Spectrum Beta-Lactamase-Producing Multidrug-Resistant Escherichia Coli in Poultry Chickens and Variation According to Farming Practices in Punjab, India.

BACKGROUND: Agricultural use of antimicrobials in subtherapeutic concentrations is increasing in response to the rising demand for food animal products worldwide. In India, the use of antimicrobials in food animal production is unregulated. Research suggests that many clinically important antimicrobials are used indiscriminately. This is the largest study to date in India that surveys poultry production to test for antimicrobial resistance and the occurrence of extended-spectrum ß-lactamases (ESBLs) modulated by farming and managerial practices. OBJECTIVES: Our goal was to survey poultry production for resistance to eleven clinically relevant antimicrobials and phenotypic occurrence of ESBLs as modulated by farming and managerial practices. METHODS: Eighteen poultry farms from Punjab were surveyed, and 1,556 Escherichia coli isolates from 530 birds were tested for susceptibility to 11 antimicrobials using the disk diffusion method and validated using VITEK 2 (bioMérieux, Marcy-L'Étoile, France). Samples from 510 of these birds were phenotypically tested for ESBL production using the combination disk method and confirmed using VITEK 2. Generalized linear mixed models were used to infer differences in resistance profiles associated with different farming practices and facility types. RESULTS: Resistance profiles were significantly different between broiler and layer farms. Broiler farms were 2.2 [ampicillin (AMP), p=0.017] to 23 [nalidixic acid (NX), p<0.001] times more likely to harbor resistant E. coli strains than layer farms. Adjusting for farm type (broiler vs. layer), the odds of resistance (although not statistically significant) to all antimicrobials except nitrofurantoin (NIT) were higher in independent facilities (IUs) as compared to contracted facilities (CFs). Increased prevalence of multidrug resistance (MDR; 94% compared to 60% in layers), including prevalence of ESBL-producing strains (87% compared to 42% in layers), was observed in broiler farms. CONCLUSIONS: Our findings suggest that unregulated use of clinically relevant antimicrobials in Indian broiler and layer farms may contribute to the emergence of resistance and support the need to curb the nontherapeutic use of medically important antimicrobials in food animal production. https://doi.org/10.1289/EHP292.

Molecular detection of Toxoplasma gondii in the slaughter sheep and goats from North India.

Toxoplasma gondii is an obligate intracellular parasite that infects almost all the warm blooded animals, including human beings. The disease usually remains asymptomatic but is a serious concern for pregnant women, developing foetus and immuno-compromised individuals. We collected 400 cardiac/skeletal muscle tissue samples from slaughter sheep (177) and goat (223) intended for human consumption from Punjab, Uttar Pradesh and Chandigarh states/union territory in North India. The samples were pepsin-HCl digested and DNA was extracted from all the digested samples. Nested-PCR was carried out to amplify 580bp and 531bp bands with external and internal sets of primers specific for B1 gene of T. gondii. Molecularly, six (1.5%) isolates of T. gondii were detected. In PCR, T. gondii DNA were detected from 1.69% and 1.34% of the sheep and goat samples, respectively. Three PCR amplified products were sequenced in both the directions and readable sequences were obtained. Due to a low level of polymorphism in the targeted B1 gene, the clonal lineages of different isolates could not be determined. The results indicate that T. gondii in slaughter sheep and goat presents a low food safety risk for public health in North India.

Seroepidemiology and molecular characterization of hepatitis E virus infection in swine and occupationally exposed workers in Punjab, India.

Hepatitis E virus (HEV) has two discrete epidemiological patterns: waterborne epidemics in developing countries only, caused by HEV genotype I, and sporadic zoonotic outbreaks in developing and developed countries caused by genotypes III and IV. This study was designed to investigate seroprevalence, molecular detection and the characterization of HEV by nested RT-PCR in swine as well as the occupational risk to exposed human population in Punjab state of north-western India. The occupational risk-exposed group comprised of swine farmers (organized - mixed feed feeders and unorganized - swill feeders), slaughterhouse workers, sewage workers and veterinary internes. During the study period, blood and faecal samples were collected from 320 swine and 360 humans with both high and low occupational exposure risks. The overall seroprevalence of swine HEV was 65.00%, with a significantly higher seropositivity in growing pigs (2-8 months of age). The prevalence of HEV RNA in swine faecal samples by nRT-PCR was 8.75% with a significantly higher detection in swill-fed pigs. With humans in the high occupational exposure risk population, significantly higher anti-HEV IgG seropositivity was observed (60.48%) as compared to control population (10.71%). Strong evidence of association between human anti-HEV IgG seropositivity and certain occupational exposure risk groups was observed (p < 0.05). This indicates that unorganized swine farmers, slaughterhouse workers and sewage workers have higher odds of HEV infection in this study region. Percentage of nucleotide similarity between swine and human HEV isolates was less than that found in countries with zoonotic HEV outbreaks. Molecular characterization revealed the circulation of G IV and G I genotypes among swine and human population in Punjab state, respectively.