Prevalence of extended spectrum β-lactamase, AmpC β-lactamase and metallo β-lactamase mediated resistance in Escherichia coli from diagnostic and tertiary healthcare centers in south Bangalore, India

Background: The increasing reports on multidrug resistant Escherichia coli has become a potential threat to global health. Here, we present a cross-sectional study to characterize extended spectrum β-lactamase, AmpC β-lactamase and metallo β-lactamase producing E. coli isolated from different human clinical samples.Methods: A total of 300 clinical Gram negative bacterial isolates were collected and re-characterized for the identification of E. coli following standard microbiological techniques. The antimicrobial susceptibility of E. coli isolates was initially screened by Kirby-Bauer disk diffusion and MIC methods. The resistant isolates were confirmed to be ESBL, AmpC and MBL producers by their respective phenotypic confirmatory tests of combined disc method.Results: We identified 203 (68%) E. coli and 97 (32%) Non-E. coli isolates. The highest recovery of E. coli was from urine samples 72 (35%). Combined disc method using ceftazidime/ceftazidime+clavulanic acid and cefotaxime/cefotaxime+clavulanic acid confirmed 156 (79%) and 144 (73%) E. coli as ESBL producers, respectively. Thirty-four (34%) and 16 (27%) resistant E. coli isolates were confirmed to be AmpC and MBL producers, likewise.Conclusions: Increased prevalence of ESBL, AmpC and MBL producing E. coli were observed. Beta-lactamase mediated resistance appears to be prime mechanism in the multidrug resistant E. coli. Thus, early detection of beta lactamase producing E. coli is necessary to avoid treatment failure and prevent the spread of MDR

Understanding Antibiotic Usage on Small-Scale Dairy Farms in the Indian States of Assam and Haryana Using a Mixed-Methods Approach-Outcomes and Challenges

The use and misuse of antibiotics in both humans and animals contributes to the global emergence of antimicrobial resistant (AMR) bacteria, a threat to public health and infection control. Currently, India is the world's leading milk producer but antibiotic usage within the dairy sector is poorly regulated. Little data exists reflecting how antibiotics are used on dairy farms, especially on small-scale dairy farms in India. To address this lack of data, a study was carried out on 491 small-scale dairy farms in two Indian states, Assam and Haryana, using a mixed method approach where farmers were interviewed, farms inspected for the presence of antibiotics and milk samples taken to determine antibiotic usage. Usage of antibiotics on farms appeared low only 10% (95% CI 8-13%) of farmers surveyed confirmed using antibiotics in their dairy herds during the last 12 months. Of the farms surveyed, only 8% (6-11%) had milk samples positive for antibiotic residues, namely from the novobiocin, macrolides, and sulphonamide classes of antibiotics. Of the farmers surveyed, only 2% (0.8-3%) had heard of the term "withdrawal period" and 53% (40-65%) failed to describe the term "antibiotic". While this study clearly highlights a lack of understanding of antibiotics among small-scale dairy farmers, a potential factor in the emergence of AMR bacteria, it also shows that antibiotic usage on these farms is low and that the possible role these farmers play in AMR emergence may be overestimated

Evaluation of an In-house Indirect ELISA for Differential Detection of IgM and IgG anti-Brucella Antibodies in Human Brucellosis

Brucellosis caused by various species of the genus Brucella is one of the most important zoonotic diseases of global importance with veterinary, public health, and economic concerns. The study aimed to standardize IgM and IgG-based iELISA to detect anti-Brucella antibodies for serodiagnosis of acute and chronic human brucellosis. The test was standardized using 1:320 dilution of smooth lipopolysaccharide (sLPS) antigen from B. abortus S99 strain, 1:80 serum dilution, 1:4000 anti-human IgM and IgG conjugates, respectively for both IgM and IgG iELISA. The cut-off using 50 each brucellosis positive and negative human sera panel samples was set at ≥ 42 for both IgM and IgG iELISA. A total of 700 human sera samples were evaluated (137 veterinary doctors, 157 artificial inseminators, and 406 veterinary assistants). Overall, the study detected 8.3%, 8.1%, 8%, and 6.1% positivity by in-house IgG iELISA, RBPT, IgM iELISA, and SAT tests, respectively. Considering commercial iELISA kit as a gold standard, the sensitivities of IgM and IgG iELISA were 90% and 97.9%, respectively, whereas, specificities were >99%. The study established >98% specificity and >90% sensitivity for differential detection of immunoglobulin classes in the standardized iELISA. The developed assay outperformed the other evaluated tests with a shorter assay time and can be implemented in both endemic and non-endemic regions for surveillance and diagnosis of human brucellosis

Antibiotic use, knowledge, and practices of milk vendors in India's informal dairy value chain

BackgroundMilk vendors play an important role in India's dairy value chain; however, their food safety practices are poorly understood. From a milk safety perspective, vendor behavior is significant because it has the potential to affect both consumer and producer behavior. This study describes the types of milk vendors in two Indian states, in an attempt to investigate vendors' hygienic knowledge and practices toward safety and antimicrobial resistance (AMR). MethodsA cross-sectional study was conducted in the states of Assam and Haryana, India. In selected villages, all the milk vendors identified at the time of visit were interviewed. A questionnaire was used to assess the knowledge and practices on antibiotics, milk safety and hygiene. The milk samples were tested for presence of antibiotic resistant bacteria using antibiotic susceptibility testing. ResultsIn total, 244 milk vendors were interviewed during the survey. Out of these, 146 (59.8%) of the vendors traded raw milk, while 40.2% traded pasteurized milk. Vendors were categorized depending on whom they supplied milk to. Five categories were identified: (a) those who sold at grocery shops; (b) those who sold on roadside (roadside vendors); (c) those who sold from door to door; (d) those who sold to sweet makers/tea stalls, and (e) those who sold from own home/other entity. The level of training among vendors on milk hygiene was non-existent and the knowledge related to antibiotics was low. Most of them [210/244 (86.07%)] agreed that boiled milk is always safer than raw milk but almost half [119 (48.77%)] of them admitted that sometimes they drink milk without boiling it. Most vendors believed that they could identify whether milk is safe or not for consumption just by its appearance and smell. Out of 124 milk samples collected from surveyed milk vendors and tested for the presence of antibiotic-resistant bacteria, 80 (64.52%) were tested positive. ConclusionThis study highlights the low levels of knowledge regarding food safety among milk vendors. It shows the predominance of informal milk vendors in the surveyed states and prevalence of AMR bacteria in milk traded by them. Training may be a beneficial strategy for addressing the issue

Molecular Characterization of Methicillin-Resistant Staphylococci from the Dairy Value Chain in Two Indian States

Bovine milk and milk products may contain pathogens, antimicrobial resistant bacteria, and antibiotic residues that could harm consumers. We analyzed 282 gram-positive isolates from milk samples from dairy farmers and vendors in Haryana and Assam, India, to assess the prevalence of methicillin-resistant staphylococci using microbiological tests, antibiotic susceptibility testing, and genotyping by PCR. The prevalence of genotypic methicillin resistance in isolates from raw milk samples was 5% [95% confidence interval, CI (3–8)], with 7% [CI (3–10)] in Haryana, in contrast to 2% [CI (0.2–6)] in Assam. The prevalence was the same in isolates from milk samples collected from farmers [5% (n = 6), CI (2–11)] and vendors [5% (n = 7), CI (2–10)]. Methicillin resistance was also observed in 15% of the isolates from pasteurized milk [(n = 3), CI (3–38)]. Two staphylococci harboring a novel mecC gene were identified for the first time in Indian dairy products. The only SCCmec type identified was Type V. The staphylococci with the mecA (n = 11) gene in raw milk were commonly resistant to oxacillin [92%, CI (59–100)] and cefoxitin [74%, CI (39–94)], while the isolates with mecC (n = 2) were resistant to oxacillin (100%) only. All the staphylococci with the mecA (n = 3) gene in pasteurized milk were resistant to both oxacillin and cefoxitin. Our results provided evidence that methicillin-resistant staphylococci occur in dairy products in India with potential public health implications. The state with more intensive dairy systems (Haryana) had higher levels of methicillin-resistant bacteria in milk

Antibiotic use, knowledge, and practices of milk vendors in India's informal dairy value chain

Background: Milk vendors play an important role in India's dairy value chain; however, their food safety practices are poorly understood. From a milk safety perspective, vendor behavior is significant because it has the potential to affect both consumer and producer behavior. This study describes the types of milk vendors in two Indian states, in an attempt to investigate vendors' hygienic knowledge and practices toward safety and antimicrobial resistance (AMR). Methods: A cross-sectional study was conducted in the states of Assam and Haryana, India. In selected villages, all the milk vendors identified at the time of visit were interviewed. A questionnaire was used to assess the knowledge and practices on antibiotics, milk safety and hygiene. The milk samples were tested for presence of antibiotic resistant bacteria using antibiotic susceptibility testing. Results: In total, 244 milk vendors were interviewed during the survey. Out of these, 146 (59.8%) of the vendors traded raw milk, while 40.2% traded pasteurized milk. Vendors were categorized depending on whom they supplied milk to. Five categories were identified: (a) those who sold at grocery shops; (b) those who sold on roadside (roadside vendors); (c) those who sold from door to door; (d) those who sold to sweet makers/tea stalls, and (e) those who sold from own home/other entity. The level of training among vendors on milk hygiene was non-existent and the knowledge related to antibiotics was low. Most of them [210/244 (86.07%)] agreed that boiled milk is always safer than raw milk but almost half [119 (48.77%)] of them admitted that sometimes they drink milk without boiling it. Most vendors believed that they could identify whether milk is safe or not for consumption just by its appearance and smell. Out of 124 milk samples collected from surveyed milk vendors and tested for the presence of antibiotic-resistant bacteria, 80 (64.52%) were tested positive. Conclusion: This study highlights the low levels of knowledge regarding food safety among milk vendors. It shows the predominance of informal milk vendors in the surveyed states and prevalence of AMR bacteria in milk traded by them. Training may be a beneficial strategy for addressing the issue

An Understanding of the Global Status of Major Bacterial Pathogens of Milk Concerning Bovine Mastitis: A Systematic Review and Meta-Analysis (Scientometrics)

In this study, the major mastitis pathogen prevalence in the cattle and buffalo of the world was estimated by a meta-analysis. Staphylococcus (S) species, Streptococcus (St) species, and Escherichia coli (Ec) prevalence studies reported during 1979–2019 were collected using online databases, and offline resources. A meta-analysis of these data was done with the meta package in R-Software. The Staphylococcus aureus was the major mastitis pathogen, mostly causing subclinical mastitis, Ec causing clinical mastitis and St causing subclinical and clinical mastitis. The pooled prevalence estimates of S, St, and Ec were 28%, 12%, and 11% in the world from 156, 129, and 92 studies, respectively. The S, St, and Ec prevalences were high in Latin America (51%), Oceania (25%), and Oceania (28%), respectively. Higher S, St, and Ec prevalences were observed by molecular methods, signifying high sensitivity and usefulness for future studies. Among bacterial species, S. aureus (25%) followed by coagulase-negative Staphylococcus species (20%), Escherichia coli (11%), St. agalactiae (9%), St. uberis (9%) were the important pathogens present in the milk of the world. We hypothesize that there is a urgent need to reduce mastitis pathogen prevalence by ensuring scientific farm management practices, proper feeding, therapeutic interventions to augment profits in dairying, and improving animal and human health

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Not AvailableBrucellosis in swine is a contagious disease with greater zoonotic potential caused by Brucella suis. The study describes PAN India swine brucellosis sero-prevalence in 5431 stratified random serum samples collected during 2018–2019 from 26 out of 29 states and two out of seven union territories. The serum samples were tested for anti-Brucella antibodies by indirect ELISA and overall, 4.33% apparent prevalence (AP) was recorded. The AP is ≥ 10% in five states among 26 states, P ≥ 50% in four districts out of 117 districts screened and cent percent prevalence in two epi units out of 264 sampled. Significantly high seropositivity (p < 0.05) in male (6.08%) than female pigs (3.46%) and in ≥ 24-month-old pigs indicated older and male pigs as potential carriers of the disease. The study recorded endemicity of the swine brucellosis in few regions of India requiring periodical surveillance for control of the disease. Brucella testing of boars before breeding and awareness among farmers and veterinarians will aid in reduction of disease burden in the absence of vaccination policy.Not Availabl

Biocompatible Injectable Hydrogel with Potent Wound Healing and Antibacterial Properties

Two component injectable hydrogels that cross-link in situ have been used as noninvasive wound-filling devices, i.e., sealants. These materials carry a variety of functions at the wound sites, such as sealing leaks, ceasing unwanted bleeding, binding tissues together, and assisting in wound healing processes. However, commonly used sealants typically lack antibacterial properties. Since bacterial infection at the wound site is very common, bioadhesive materials with intrinsic antibacterial properties are urgently required. Herein, we report a biocompatible injectable hydrogel with inherent bioadhesive, antibacterial, and hemostatic capabilities suitable for wound sealing applications. The hydrogels were developed in situ from an antibacterial polymer, <i>N</i>-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC), and a bioadhesive polymer, polydextran aldehyde. The gels were shown to be active against both Gram-positive and Gram-negative bacteria, including drug-resistant ones such as methicillin-resistant <i>Staphylococcus aureus</i> (MRSA), vancomycin-resistant <i>Enterococcus faecium</i> (VRE), and β-lactam-resistant <i>Klebsiela pneumoniae</i>. Mechanistic studies revealed that the gels killed bacteria upon contact by disrupting the membrane integrity of the pathogen. Importantly, the gels were shown to be efficacious in preventing sepsis in a cecum ligation and puncture (CLP) model in mice. While only 12.5% of animals survived in the case of mice with punctured cecam but with no gel on the punctured area (control), 62.5% mice survived when the adhesive gel was applied to the punctured area. Furthermore, the gels were also shown to be effective in facilitating wound healing in rats and ceasing bleeding from a damaged liver in mice. Notably, the gel showed negligible toxicity toward human red blood cells (only 2–3% hemolysis) and no inflammation to the surrounding tissue upon subcutaneous implantation in mice, thus proving it as a safe and effective antibacterial sealant

Factors Influencing Antibiotic Prescribing Behavior and Understanding of Antimicrobial Resistance Among Veterinarians in Assam, India.

This study investigates factors influencing veterinarians' antibiotic prescribing behaviors and their understanding of antimicrobial resistance (AMR). The study used a telephone survey of 50 veterinarians conducted in five districts in Assam state, India. The survey sought information on the most prevalent animal diseases, veterinarians' awareness of potential preventive measures, including factors determining antimicrobial prescribing; the types of antimicrobials used for different health conditions in different species, and possible options to reduce antimicrobial use (AMU). The majority (86%) of respondents worked for the government, 98% reported having no written policy for the use of veterinary health products, and 58% have no on-site diagnostic facilities. Ceftriaxone, Enrofloxacin, and Oxytetracycline were the antibiotics (ABX) most frequently prescribed, by 76, 68, and 54% of veterinarians, respectively. These ABX were prescribed mainly for respiratory health problems and mastitis in cattle, and gastrointestinal infections in buffaloes, sheep, goat, and pigs. Severity of clinical symptoms, economic status of the livestock owner, and withdrawal period for ABX were ranked as very important factors for giving ABX. Less than two thirds (64%) were aware of the government ban for Colistin and only 2% were aware of a national plan for AMR. This study highlighted that ABX prescription is mostly based on tentative diagnosis given the lack of diagnostic facilities in most veterinary clinics. There is a need to enhance veterinary healthcare and to improve communication between policy makers and field veterinarians and, importantly, a need to disseminate clear prescribing guidelines on prudent AMU