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.

Genotypic characterization of Listeria monocytogenes isolated from humans in India.

Listeria monocytogenes is a foodborne pathogen associated with severe diseases in humans and animals. The genotypic analysis of 17 L. monocytogenes isolates recovered from humans in India during 2006-2009 using multiplex serotyping PCR allowing serovar predictions, conventional serology and by pulsed field gel electrophoresis (PFGE) is presented. The isolates were recovered from patients exhibiting various clinical conditions. A multiplex-PCR based serotyping assay revealed 88·24% (15/17) of the strains belonging to the serovar group 4b, 4d, 4e and 11·76% (2/17) to the serovar group 1/2b, 3b. Conventional serology indicated that 13 (76·47%) L. monocytogenes isolates to be of serotype 4b, 2 (11·76%) serotype 4d, and 2 (11·76%) serotype 1/2b. Ten ApaI and nine AscI pulsotypes were recognized among the 17 human isolates. PFGE analysis allowed discrimination among isolates of the same serotype and among isolates from the same sampling areas or those isolated from different areas. Thus, PFGE together with multiplex-PCR serotyping allows rapid discrimination of L. monocytogenes strains. In addition, the predominance of L. monocytogenes serotype 4b is of concern, as this serotype has been most frequently associated with human listeriosis outbreaks.

PCR analysis of Pasteurella multocida isolates from an outbreak of pasteurellosis in Indian pigs.

An outbreak of pasteurellosis with high mortality was recorded in indigenous pigs in India. The presence of Pasteurella multocida in samples collected from dead pigs was detected by smear examination and isolation, and later by P. multocida specific polymerase chain reaction (PM-PCR). P. multocida was detected in all the samples collected from dead pigs, with nine strains ultimately isolated. All the isolates were positive by PM-PCR. Six isolates showed CAPA and three were of CAPD capsular types. All the isolates were negative for toxigenic gene (toxA). The isolates were sensitive to oxytetracycline, doxycycline, gentamycin, erythromycin, ampicillin, amoxycillin, chloramphenicol and enrofloxacin and resistant to sulphadiazine and cloxacillin. The PCR assays used in this study have been shown to be useful diagnostic tools for P. multocida detection and characterization.

Evaluation of indirect and avidin-biotin enzyme linked immunosorbent assays for detection of anti-listeriolysin O antibodies in bovine milk samples.

Listeria monocytogenes is a foodborne pathogen that causes a wide spectrum of diseases in humans and animals. Enzyme linked immunosorbent assays (ELISA) [indirect and avidin-biotin (A-B)] for detecting L. monocytogenes antibodies in bovine milk samples (n = 2060) were standardized and evaluated by comparison with bacteriological examination. The tests were standardized by checker board titration. Highly purified listeriolysin O (LLO) was used as an antigen. Receiver operating characteristic (ROC) analysis was performed to decide the cut-off values. The ROC analysis revealed the sensitivities of indirect and A-B ELISA as 100% and specificities as 97.1 and 99.9% respectively. Listeria monocytogenes was isolated from 105 (5.1%) milk samples collected from 52 farms. Anti-LLO IgG antibodies were detected from 137 and 112 milk samples when tested by indirect and A-B ELISA respectively. Of the 52 farms screened, 28 (53.8%) yielded one or more isolates of L. monocytogenes and 33 (63.5%) of the farms had one or more animals simultaneously positive by one or both the assays for anti-LLO antibodies.

Genotypic characterization of Listeria spp. isolated from fresh water fish.

A total of 200 samples (muscles and viscera, 100 of each) of fresh water fish, walking catfish (Clarias batrachus) were screened for Listeria spp. All the samples were subjected to a two-step enrichment followed by plating on selective media. Confirmation of the isolates was on the basis of biochemical characters, haemolysis on blood agar and Christie, Atkins, Munch Petersen test. A total of 39 isolates of Listeria spp. were recovered. Of these 26 (67%), 8 (21%), 3 (8%) and 2 (5%) were Listeria monocytogenes, Listeria seeligeri, Listeria grayi and Listeria welshimeri, respectively. The isolates were subjected to a PCR assay for detection of the virulence-associated genes individually or together. The plcA, actA, hlyA and iap genes were detected in six strains, three genes (actA, hlyA and iap) in nine strains, the plcA, hlyA and iap in our strain, the hlyA and iap were in three strains, actA and hlyA in four strains, plcA and hlyA in our strain and hlyA in two strains. The hlyA and iap were also detected in L. seeligeri.

Isolation of pathogenic Listeria monocytogenes in faeces of wild animals in captivity.

The isolation of pathogenic Listeria spp. in faecal samples of captive wild animals was studied. Isolation of the pathogen was attempted from the samples by selective enrichment in University of Vermont Medium and plating onto Dominguez-Rodriguez isolation agar, PALCAM agar and modified McBride Listeria agar. Pathogenicity of the isolates was tested by Christie, Atkins, Munch Petersen test, phosphotidylinositol-specific phospholipase C assay, mice inoculation test and chick embryo bioassay. Listeria monocytogenes was isolated from eight (16%) of 50 faecal samples from six different mammals and one bird. Out of eight isolates, one isolate from jackal proved to be pathogenic by all the pathogenicity testing assays. PCR amplification of virulence genes suggested that the isolate was potentially pathogenic.

Evaluation of herbal coccidiostat 'Coxynil' in broiler.

Anticoccidial efficacy of "Coxynil" a polyherbal preparation was tested against Eimeria tenella in broilers. Body weight of birds challenged with E. tenella in Coxynil treated groups was higher as compared to Coxynil untreated. Oocyst out put, lesion score, HI titres against New Castle disease virus were significantly higher in Coxynil supplemented groups in comparison to Coxynil un-supplemented groups. Examination of ceaca of the birds, revealed that the Coxynil interfered with life cycle of coccidia. The typical second generation schizonts were absent in ceacal section of Coxynil treated groups. The results indicate that Coxynil is effective herbal coccidiostat.