The occurrence of antibiotic residues in milk constitutes a potential risk to the health of consumers. The present study describes the optimisation and validation of a high-performance liquid ...chromatographic (HPLC) method for the simultaneous determination of sulphadiazine (SDZ), sulphamethoxazole (SMX), oxytetracycline (OTC), doxycycline (DOX), tetracycline (TC), enrofloxacin (ENRO) and chloramphenicol (CLP) residues in bovine milk using colchicine (COL) as internal standard. The determination of these antimicrobials was carried out on C
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analytical column using high-performance liquid chromatographic-diode array detection (HPLC-DAD). The extraction method involving deproteinisation of the milk sample followed by a solid-phase extraction (SPE) clean-up of antibiotic residues has been developed. The method was validated according to the European Commission Decision 2002/657/EC and applied for the analysis of antibiotic residues in 21 raw milk samples collected from Ludhiana, Punjab, India. The recoveries for the studied antibiotics ranged from 83.3–111.8% with relative standard deviations between 3.5 and 16.2%. The limits of quantification for these antibiotics, with the exception of chloramphenicol, were below the maximum residue limits (MRLs), making the method suitable for performing routine analysis.
Q Fever is a zoonotic disease of significant animal and public health concern, caused by Coxiella burnetii (C. burnetii), an obligate intracellular bacterium. This study was done to evaluate the ...diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of three diagnostic methods to diagnose C. burnetii infection in cattle and buffaloes in Punjab, India: an indirect ELISA method applied in serum samples and a trans-Polymerase Chain Reaction (trans-PCR) technique applied in milk samples and genital swabs, using a Bayesian latent class analysis. Conditional independence was assumed between the tests, given (i) the different biological principle of ELISA and trans-PCR and (ii) the fact that the trans-PCR was performed on different tissues. The ELISA method in the serum samples showed the highest DSe of 0.97 (95% Probability Intervals (PIs): 0.93; 0.99) compared to the trans-PCR method applied in milk samples 0.76 (0.63; 0.87) and genital swabs 0.73 (0.58; 0.85). The DSps of all tests were high, with trans-PCR in genital swabs recording the highest DSp of 0.99 (0.98; 1), while the DSp of trans-PCR in milk samples and ELISA in serum samples were 0.97 (0.95; 0.99) and 0.95 (0.93; 0.97) respectively. The study results show that none of the applied tests are perfect, therefore, a testing regimen based on the diagnostic characteristic of the tests may be considered for diagnosis of C. burnetii.
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.
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.
In the present investigation, groundwater samples (n = 508) collected from northern, central, and southern Punjab were analyzed for the detection of potentially hazardous metals (Pb, Cd, Ni, Cr) and ...As using atomic absorption spectrometer. Lead was observed as a predominant contaminant with 167 (32.8%) samples exceeding the permissible limit, followed by As, with 19 (3.7%) samples above the limit set by WHO. Spatial variation for the occurrence of metals showed statistically significant levels of Ni and Cr in groundwater from central Punjab, and As from northern Punjab. Assessment of health risks to these pollutants revealed As as a noxious pollutant, with hazard indices and carcinogenic risks significantly higher than the recommended limits in the three regions; and hence, posing a serious threat to the study population. Also, the risk values were higher for children than adults, indicative of their higher susceptibility to the ill-effects of these pollutants.
Taenia solium (T. solium) cysticercosis remains a neglected zoonotic disease in India. The current study was planned to estimate the prevalence of T. solium porcine cysticercosis in the Punjab state ...of India, to compare this prevalence with the disease prevalence in pigs reared outside Punjab and to assess the distribution of the parasite in pig carcasses.
Two slaughter shops were selected in each of the 22 districts of Punjab. Pigs slaughtered on the day/s of inspection were post-mortem inspected to identify the presence of T. solium cysts. Estimated true prevalence was estimated by taking into account the diagnostic sensitivity (38%) and specificity (100%) of post-mortem inspection using the Rogan-Gladen estimator. Positive carcasses were purchased and brought to the laboratory to assess the tissue distribution of T. solium cysts and to conduct PCR targeting large subunit rRNA gene, internal transcribed spacer 1 gene, ITS1 gene and Cytochrome oxidase I gene. The selected PCR products were submitted for sequencing and phylogenetic analyses were performed.
We contacted 71 shop owners to achieve a sample of 44 shops for the study. We inspected 642 pigs reared in Punjab and 450 imported from other states at these slaughter shops. In addition, we sampled 40 pigs from an abattoir located in the state capital. Of the 642 pigs reared in Punjab, 9 had T. solium cysts with an apparent prevalence of 1·40% (95% CI: 0·74%, 2·64%) and the estimated true prevalence of 3.69% (95% CI: 1·95%, 6·95%). Pigs imported from outside the state had a significantly higher prevalence (odds ratio: 2·58; 95% CI: 1·12, 5·98; p-value: 0·026) as 15 of the 450 imported pigs were positive (apparent prevalence: 3.33%; 95% CI: 2.03%, 5.43%; estimated true prevalence: 8.77%; 95% CI: 5.34%, 14.28%). None of samples was positive from the pigs sampled at the abattoir in the state capital. The PCR confirmed T. solium cysts from all the 24 positive samples. We counted a median of 897 (range 526-1964) cysts per infected pig from the 19 infected pig carcasses inspected. The phylogenetic tree based on the alignment of partial cytochrome oxidase 1 sequences indicated all positive samples to be clustered with the T. solium Asian genotype. The analysis did not indicate the presence of T. asiatica in the slaughter pigs.
Despite the underestimation of the prevalence due to missing mildly-infected carcasses, low participation and lack of representative sampling, the presence of heavily infected carcasses containing viable cysts, particularly those imported from outside the state, indicates that T. solium cysticercosis is an important food safety concern for pork consumers in Punjab, India. Measures should be taken to reduce the disease prevalence in pigs to reduce the disease burden in the public.
The dairy industry plays an important role in the economy and food security of India. A study of the dairy value chains was conducted in Punjab, India, to identify production constraints and ...biosecurity risks. Focus group discussions and key informant interviews were conducted during 2018-2019 with a total of 119 participants comprising veterinarians (41), paraveterinarians (15), veterinary academics (12), dairy farmers (46) and key informants (5). Input and output value chains were created, and potential risk nodes were identified that could facilitate the transmission of pathogens between animals, farms and villages. The majority of the participants were male (93%), middle-aged (68%) or worked in rural areas (75%). Most of the farmers self-cultivated their green fodder (82%), used the wheat straw from their own fields (60%) but purchased commercial feed (63%). Artificial insemination was used by 85% of farmers for cattle, but only 68% for buffaloes. Most of the farmers (76%) reported getting their animals vaccinated against foot-and-mouth disease and hemorrhagic septicemia. Animals were sold and purchased without any health certification and testing in most cases. Adoption of biosecurity measures by farmers and the use of personal protective equipment by veterinary personnel were very low. We recommend conducting epidemiological studies to further characterize the identified risk nodes, training of veterinary practitioners and farmers to ensure adequate biosecurity practices and the appropriate use of personal protective equipment.
The present study was conducted with the objectives to evaluate the prevalence of AFM1 in market milk and milk products from Ludhiana district of Punjab. A total of 408 samples comprising of milk ...(n = 109), butter (n = 97), cheese (n = 99) and curd (n = 103) were collected from seven tehsils of Ludhiana. The samples were tested for the estimation of aflatoxin M1 (AFM1) using ELISA technique; of which, 10% of the positive samples were confirmed by high performance liquid chromatography (HPLC). Further, the estimation of exposure and risk characterization of the consumer population of Ludhiana district to AFM1 from milk and milk products was done using deterministic and probabilistic methods. Overall, the mean levels (ppb) of AFM1 in milk, butter, cheese and curd were 0.314 ± 0.35, 0.100 ± 0.15, 0.321 ± 0.41 and 0.162 ± 0.22, respectively. The estimated daily intake (EDI: ng/kg body weight/day) of AFM1 among the consumers due to consumption of milk, butter, cheese and curd was 2.303, 0.020, 0.068 and 0.3287, respectively. Among the different income groups of consumers, the quantities of AFM1 consumed per day through milk and milk products were significantly higher in ‘high income group’. The risk of hepatocellular carcinoma (HCC) cases/year/105 individuals of different age groups showed that the value of HCC using deterministic approach was maximum (0.0106) in the age group ‘1–9 years’ and least (0.0020) in age group ‘21–60 years’. The Hazard Index (HI) value was found high for milk (11.54) and curd (1.64) as compared to butter (0.10) and cheese (0.34). The health risk assessment indicated that consumers, especially children, in the study area are at comparatively higher health risk to AFM1 owing to their low body weight and higher milk intake. The present study revealed the urgent need to control the occurrence of aflatoxins in milk through continuous monitoring and regular inspection to reduce the exposure among the consumer population.
•AFM1 presence was observed in market milk and milk products in Ludhiana, Punjab.•Risk characterization indicates AFM1 in milk may pose health risk.•The results warrants monitoring and control of aflatoxins in animal feed.
Q Fever is a zoonotic disease of significant animal and public health concern, caused by Coxiella burnetii (C. burnetii), an obligate intracellular bacterium. This study was done to evaluate the ...diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of three diagnostic methods to diagnose C. burnetii infection in cattle and buffaloes in Punjab, India: an indirect ELISA method applied in serum samples and a trans-Polymerase Chain Reaction (trans-PCR) technique applied in milk samples and genital swabs, using a Bayesian latent class analysis. Conditional independence was assumed between the tests, given (i) the different biological principle of ELISA and trans-PCR and (ii) the fact that the trans-PCR was performed on different tissues. The ELISA method in the serum samples showed the highest DSe of 0.97 (95% Probability Intervals (PIs): 0.93; 0.99) compared to the trans-PCR method applied in milk samples 0.76 (0.63; 0.87) and genital swabs 0.73 (0.58; 0.85). The DSps of all tests were high, with trans-PCR in genital swabs recording the highest DSp of 0.99 (0.98; 1), while the DSp of trans-PCR in milk samples and ELISA in serum samples were 0.97 (0.95; 0.99) and 0.95 (0.93; 0.97) respectively. The study results show that none of the applied tests are perfect, therefore, a testing regimen based on the diagnostic characteristic of the tests may be considered for diagnosis of C. burnetii.
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