S.V.S. Malik

Effect of nisin and its combination with sodium chloride on the survival of Listeria monocytogenes added to raw buffalo meat mince

Antilisterial activity of nisin (Nisaplin), alone at concentrations of 400 and 800 IU/g and in combination with 2% sodium chloride was incorporated in raw buffalo meat mince. Samples of the raw meat mince were inoculated with 10(3) colony forming units (cfu)/g of L. monocytogenes and stored at 4°C for 16 days and at 37°C for 36 h. Initial estimates of pH, extract release volume, mesophilic and psychrophilic counts were found to be 5.74, 48 ml, 3.5×10(5) and 1.0×10(5) cfu/g of meat, respectively. The growth of L. monocytogenes in the treated groups was significantly (P<0.05) inhibited compared to the control group. The degree of inhibition increased with increasing concentration of nisin and decreasing storage temperature. Addition of 2% sodium chloride in combination with nisin increased the efficacy of nisin at both storage temperatures. The pH in the treated groups remained significantly lower (P<0.01) than in the control groups at both 4 and 37°C.

Listeria monocytogenes in spontaneous abortions in humans and its detection by multiplex PCR

Aim: To assess the extent of Listeria monocytogenes in causation of human spontaneous abortions by isolation methods and PCR analysis for the presence of virulence‐associated genes.

Isolation of Listeria monocytogenes and anti-listeriolysin O detection in sheep and goats

Studies were undertaken to compare the detection of anti-listeriolysin O antibodies (ALLO) and isolation of Listeria monocytogenes from meat and milk samples of sheep and goats. Out of 201 samples (87 of milk and 114 of meat) tested, 17.64% yielded Listeria species isolates. L. monocytogenes was isolated from 6.66 and 1.56% of meat (60) and (64) milk samples of goats, respectively, and 7.4% of sheep meat (54) samples. All the samples of ewes’ milk (23) were negative for L. monocytogenes. Seropositivity for ALLO was observed in 41.13 and 33.76% of goats and sheep, respectively. The culture positivity for L. monocytogenes and detection of ALLO did not show any agreement (k<0.202). # 2000 Elsevier Science B.V. All rights reserved.

Epidemiology and risk management of listeriosis in India

Listeria monocytogenes is a foodborne pathogen that can cause serious invasive illness, mainly in certain well-defined high-risk groups, including elderly and immunocompromised patients, pregnant women, newborns and infants. In India, this pathogen has been isolated from humans, animals and foods. The incidence of Listeria is generally comparable to those reported elsewhere in the world. In humans, maternal/neonatal listeriosis is the most common clinical form reported. Among animal populations, spontaneous abortions, subclinical mastitis, meningoencephalitis and endometritis were the commonest forms reported. The disease largely remains undiagnosed and under reported. From reported analyses of a variety of foods for Listeria, milk and milk products, meat and meat products, seafood and vegetables have been reported to be contaminated in India. The legal framework for microbiological safety of foods against microbes including L. monocytogenes is summarised. The epidemiological studies would help in understanding of the sources of infection and persistence and their risk assessment, routes of transmission, clinical forms and allow for better management of the infection.

Isolation of Coxiella burnetii from bovines with history of reproductive disorders in India and phylogenetic inference based on the partial sequencing of IS1111 element

In the present study, a total of 158 blood samples from 148 bovines and 10 dogs having a history of reproductive disorders were screened for Coxiella burnetii by trans-PCR method. In case of bovines, 6.08% (9/148) blood samples comprised of 4.54% (4/88) cattle and 8.33% (5/60) buffaloes turned out to be positive for C. burnetii DNA while all the samples from dogs (10) were found negative. Of the 9 PCR-positive bovine blood samples, the organism could be isolated only from 3 cases of buffaloes by chick embryo inoculation method. Further, to predict the homology and genetic diversity, the recovered C. burnetii isolates designated as Y1, Y3 and Y7 were partially sequenced for IS1111 gene. On phylogenetic analysis, Y3 and Y7 isolates clustered to a common node away from Y1 isolate. This study may enlighten the nature of circulating C. burnetii isolates in different parts of the world. To the best of our knowledge, this appears to be the first report describing phylogenic analysis of C. burnetii isolates based on IS1111 gene sequence.

Presence of a widely disseminated Listeria monocytogenes serotype 4b clone in India.

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

A multiplex PCR for detection of Listeria monocytogenes and its lineages.

A novel multiplex PCR assay was developed to identify genus Listeria, and discriminate Listeria monocytogenes and its major lineages (LI, LII, LIII). This assay is a rapid and inexpensive subtyping method for screening and characterization of L. monocytogenes.

Genetic diversity, virulence potential and antimicrobial susceptibility of Listeria monocytogenes recovered from different sources in India

Listeria monocytogenes isolates (n = 36) recovered from human and animal clinical cases and foods from different geographical regions of India were characterized using multiplex PCR-based serotyping, pulsed field gel electrophoresis (PFGE), in vitro and in vivo pathogenicity tests and antibiogram profiling. Multiplex PCR-based serotyping distributed L. monocytogenes isolates into 3 serogroups, of which 91.67% belonged to 4b, 4d, 4e serogroup, followed by 5.56% to 1/2a, 3a and 2.78% to 1/2b, 3b serogroups. PFGE analysis using ApaI and AscI restriction enzymes revealed 17 pulsotypes among 36 L. monocytogenes isolates with 6 major clusters having similar fingerprint profile within their cluster and 11 unique fingerprint profiles. Interestingly, PFGE analysis inferred that foods of animal origin could be a significant source of infection for spread of listeriosis among human populations. Furthermore, on comparison of in vitro and in vivo pathogenicity tests, an overall good correlation was observed between hemolytic titer assay and chick embryo inoculation test as most of the isolates with a hemolytic titer of ≥ 16 were found to be lethal to chick embryo. All the isolates were found to be susceptible to tested antimicrobials except for one animal isolate which showed resistance towards co-trimoxazole.

Genetic diversity and antibiogram profile of diarrhoeagenic Escherichia coli pathotypes isolated from human, animal, foods and associated environmental sources

Introduction Infectious diarrhoea particularly due to pathogenic bacteria is a major health problem in developing countries, including India. Despite significant reports of diarrhoeagenic Escherichia coli (DEC) pathotypes around the globe, studies which address genetic relatedness, antibiogram profile and their correlation with respect to their isolation from different sources are sparse. The present study determines isolation and identification of DEC pathotypes from different sources, their genetic characterisation, antibiogram profile and their correlation if any. Materials and methods A total of 336 samples comprising diarrhoeic stool samples from infants (n=103), young animal (n=106), foods (n=68) and associated environmental sources (n=59) were collected from Bareilly region of India. All the samples were screened by using standard microbiological methods for the detection of E. coli. The identified E. coli were then confirmed as DEC pathotypes using polymerase chain reaction–based assays. Those DEC pathotypes identified as Enteroaggregative E. coli (EAEC) were further confirmed using HEp-2 adherence assay. All the isolated DEC pathotypes were studied for their genetic diversity using pulsed-field gel electrophoresis (PFGE), and antimicrobial susceptibility testing was performed by using disc diffusion method as per Clinical Laboratory Standards Institute guidelines. Results and discussion Of the four DEC pathotypes investigated, EAEC was found to be the predominant pathogen with an isolation rate of 16.5% from infants, 17.9% from young animals, 16.2% from foods and 3.4% from the associated environmental sources. These EAEC isolates, on further characterisation, revealed predominance of ‘atypical’ EAEC, with an isolation rate of 10.7% from infants, 15.1% from young animals, 16.2% from foods, and 3.4% from the associated environmental sources. On PFGE analysis, discrimination was evident within DEC pathotypes as 52 unique pulsotypes were observed for 59 recovered DEC pathotypes. However, a few EAEC isolates were found to be clonal (clusters A, B, C, D, F, G, and H) irrespective of their source of isolation, suggests sharing and/or circulation among different sources. Further, a high antibiotic resistance pattern was observed among isolated DEC pathotypes as almost 86.4% of isolates were found to be resistant against ≥3 tested drugs.

Seroprevalence and molecular detection of coxiellosis among cattle and their human contacts in an organized dairy farm

BACKGROUND The present investigation of Coxiella burnetii infection in cattle and farm workers on an organized cattle dairy farm, which appears to be the first of its kind in India, was undertaken to assess the status of this largely neglected and masked zoonosis. METHODS A total of 665 samples comprising of serum (n=224), milk (n=217) and vaginal swabs (n=224) collected from milch animals (n=224) with a history of reproductive disorders were screened. Besides these, ticks (n=114); animal feed (n=4) and environmental samples (n=13) as well as serum (n=19) of farm workers were also collected. The animal sera and milk samples as well as human sera were tested for antibodies against C. burnetii by commercial ELISA kit, whereas, all the collected samples were subjected to trans-PCR targeting the IS1111 gene of C. burnetii. RESULTS A high positivity for coxiellosis was detected in sera (29.91%) and milk (26.73%) samples of dairy cattle as well as sera from human contacts (84.21%) by ELISA. The trans-PCR detected the pathogen in 12.94% sera, 14.73% vaginal swabs and 5.53% milk samples of cattle, and in one soil sample, however, the sera of the farm workers and tick were tested negative. CONCLUSIONS The high positivity for coxiellosis among cattle and farm workers highlight the need to undertake extensive epidemiological studies to unravel the trends of C. burnetii infection in India.