Rapid and visual detection of Shiga-toxigenic Escherichia coli (STEC) in carabeef meat harnessing loop-mediated isothermal amplification (LAMP).

Shiga toxigenic E. coli are important foodborne zoonotic pathogens. The present study was envisaged to standardize loop-mediated isothermal amplification assays targeting stx1 and stx2 genes for rapid and visual detection of STEC and compare its sensitivity with PCR. The study also assessed the effect of short enrichment on the detection limit of LAMP and PCR. The developed LAMP assays were found to be highly specific. Analytical sensitivity of LAMP was 94 fg/µLand 25.8 fg/µL for stx-1 and stx-2 while LOD of 5 CFU/g of carabeef was measured after 6-12 h enrichment. The study highlights the importance of short (6-12 h) enrichment for improving the sensitivity of LAMP. The entire detection protocol could be performed within 9 h yielding results on the same day. The developed LAMP assays proved to be a handy and cost-effective alternative for screening STEC contamination in meat.

Draft Genome Sequence of a Pasteurella multocida Strain Isolated from a Spotted Deer (Axis axis) in India.

Here, we report the genome sequence of a Pasteurella multocida strain isolated from the heart blood of a spotted deer (Bareilly, India). The 2.44-Mbp genome has 2,227 coding sequences, with a G+C content of 40.7%.

In vivo therapeutic efficacy of Curcuma longa extract loaded ethosomes on wound healing.

Since ancient times, medicinal plants are widely accepted to promote the health and wellness of animals and mankind. The medicinal plant-based therapies have limitations of delayed onset of action, inconsistent absorption, low bioavailability, oxidation, and poor solubility. The encapsulation studies suggested improved efficacy. Therefore, the present study attempts to evaluate the efficacy of Curcuma longa extracts encapsulated in Ethosome on wound healing model compared to crude extract. The Curcuma longa extract swere prepared by cold percolation method and total curcuminoid content was determined by Reverse phase-HPLC. Three Ethosomal suspensions (ETS1, ETS2, and ETS3) were prepared and characterized for particle distribution, morphology, and absorption spectrum by Zetasizer, Scanning Electron Microscopy, and FTIR respectively. The Ethosomal suspension with the highest entrapment efficiency was applied topically at a varying concentrations (0.25, 0.5, and 1 g/cm2) on the surgically created wounds in rats. The efficacy of wound healing was evaluated by clinical observation, macroscopic evaluation of granulation tissue, colour digital image processing, and histology. The methanolic extract of Curcuma longa showed better antibacterial potential than ethanolic and aqueous. The total Curcuminoid content in the Curcuma longa rhizome was 4.03%. The size, PDI, zeta potential, and viscosity of Ethosomal suspension ranged from 34.8 to 371 nm, 0.236-1.178, 15.6-36.8mV, and 0.8460-0.8510, respectively. The ETS3 was found the most optimum combination with the highest entrapment efficiency and the topical application at a dose rate of 0.5 g/cm2 and 1.0 g/cm2 resulted in comparable wound contracture, pain score, histopathological score as compared to control groups.It was concluded that the Curcuma longa encapsulation in Ethosome resulted in improved wound appearance, granulation tissue score, and appearance with a shortened period of wound resolution at the cellular level as compared to crude extract.

Molecular epidemiological studies on avian pathogenic Escherichia coli associated with septicemia in chickens in India.

Bacterial septicemia causes huge economic losses in the poultry industry and there is no systematic research available in India on the connection of various pathogens associated with septicemia. The present molecular epidemiological study was conducted to investigate the association of different bacterial and immunosuppressive viral pathogens in septicemia suspected chickens. A total of 443 chicken carcasses with septicemic conditions from 71 different flocks were included in this study. Heart blood swabs were subjected to bacterial culture for Salmonella spp., Pasteurella multocida, Escherichia coli, and Gallibacterium anatis. Of these 51 flocks tested for E. coli, 49 (96.1%) flocks were found positive. Among flocks tested for Salmonella spp., 2 flocks were found positive. All tested flocks were found negative for G. anatis and P. multocida as well as air sac swabs tested negative for Mycoplasma spp. Bacterial cultural examination revealed that majority of septicemic chickens were found to be infected with E. coli and these E. coli isolates showed the highest resistance to vancomycin (60%), followed by erythromycin (50%) and cefotaxime (38%) and maximum sensitivity to cefotaxime and clavulanic acid combinations (81.5%), followed by chloramphenicol (69.6%) and ertapenem (67.2%). Among the 5 avian pathogenic E. coli (APEC) virulence genes were detected in 36 flocks and highest frequency of iss (100%), followed by ompT or iutA (97.2%), hly (61.1%) and iroN (47.2%) genes. On polymerase chain reaction (PCR) screening, 10.5, 4.5, 52.2, 19.4, 9.0, 4.5, 20.1 and 19.4% of the flocks were positive for G. anatis, Ornithobacterium rhinotracheale, APEC, Salmonella spp., Mycoplasma gallisepticum, Mycoplasma synoviae, chicken infectious anemia virus and Marek's disease virus, respectively. To our knowledge, the present study is first on the etiology of septicemia in chicken flocks in India. The present study infers that the majority of septicemic deaths in broiler chickens less than 8 weeks have been connected with APEC and majority of E. coli isolates are multidrug resistance, suggesting the need for surveillance and intervention to curb the inadvertent use of antibiotics. Although, incidence of G. anatis association with septicemia was reported, still requires a rigorous epidemiological study to determine the actual prevalence. However, more detailed studies encompassing vast geographical area with large sample size and long duration of the studies are necessary to provide a clear picture of the interaction of different pathogens causing septicemia in chicken.

A comparative genomics approach for identifying genetic factors in Escherichia coli isolates associated with bovine diseases.

AIMS: E. coli are ubiquitously present bacterial pathogens that cause septicaemia, diarrhoea and other clinical illness in farm animals. Many pathogen factors can be associated with disease conditions. Currently, studies inferring E. coli genetic factors associated with infection in bovines are limited. Hence, the present study envisaged to determine the pathogen genetic factors associated with bovine disease conditions. METHOD AND RESULTS: The comparative genomic analysis involved genome sequence data of 135 diseased and 145 healthy bovine origin E. coli strains. Phylogroups A and C, as well as pathotypes ExPEC and EPEC, were found to have a strong connection with bovine disease strains. STEC strains, including EHEC, seem to play a less important role in bovine disease. Sequence types (STs) predominant among strains from diarrhoeal origin were ST 301 (CC 165) and ST 342. Correlation of core genome phylogeny with accessory gene-based clustering, phylogroups and pathotypes indicated lineage-specific virulence factors mostly associated with disease conditions. CONCLUSIONS: Comparative genomic analysis was applied to infer genetic factors significant in bovine disease origin E. coli strains. Isolates from bovine disease origin were enriched for the phylogroups A and C, and for the pathotypes ExPEC and EPEC. However, there was minimal evidence of STEC involvement. The study also indicated predominant genetic lineages and virulence genes (pap, sfa and afa) associated with disease origin strains. SIGNIFICANCE AND IMPACT OF STUDY: The study revealed significant pathotypes, phylogroups, serotypes and sequence types associated with bovine disease conditions. These identified genetic factors can be applied for disease diagnosis, implementing vaccines and therapeutic measures. In addition, E. coli isolates from the bovine species revealed a complex pattern of disease epidemiology.

Antimicrobial resistance pattern of extended-spectrum ß-lactamase-producing Escherichia coli isolated from fecal samples of piglets and pig farm workers of selected organized farms of India.

BACKGROUND AND AIM: Extended-spectrum ß-lactamase (ESBL)-producing Escherichia coli are gradually increasing worldwide and carry a serious public threat. This study aimed to determine the antimicrobial resistance pattern of ESBL-producing E. coli isolated from fecal samples of piglets and pig farm workers. MATERIALS AND METHODS: Fecal samples from <3-month-old piglets (n=156) and farm workers (n=21) were processed for the isolation of ESBL-producing E. coli in MacConkey agar added with 1 µg/mL of cefotaxime. E. coli (piglets=124; farm workers=21) were tested for ESBL production by combined disk method and ESBL E-strip test. Each of the ESBL-positive isolate was subjected to antibiotic susceptibility testing. The ESBL-producing E. coli were further processed for genotypic confirmation to CTX-M gene. RESULTS: A total of 55 (44.4%, 55/124) and nine (42.9%, 9/21) ESBL-producing E. coli were isolated from piglets and farm workers, respectively. Antibiotic susceptibility testing of the ESBL-positive E. coli isolates from piglets and farm workers showed 100% resistance to ceftazidime, cefotaxime, cefotaxime/clavulanic acid, ceftazidime/clavulanic acid, and cefpodoxime. A proportion of 100% (55/55) and 88.9% (8/9) ESBL-positive E. coli were multidrug resistance (MDR) in piglets and farm workers, respectively. On genotypic screening of the ESBL E. coli isolated from piglets (n=55), 15 were positive for the bla CTX-M gene and of the nine ESBL E. coli from farm workers, none were positive for the bla CTX-M gene. CONCLUSION: Although there was no significant difference in isolation of ESBL-producing E. coli between piglets and farm workers, the ESBL-positive E. coli from piglets showed relatively higher MDR than farm workers.

MexAB-OprM Efflux Pump of Pseudomonas aeruginosa Offers Resistance to Carvacrol: A Herbal Antimicrobial Agent.

Carvacrol is a herbal antimicrobial agent with in vitro activity against several bacterial pathogens. However, multidrug resistant strains of Pseudomonas aeruginosa are resistant to herbal antimicrobial compounds including carvacrol. Resistance of P. aeruginosa to carvacrol is not well studied. This study was aimed to identify the gene(s) associated with carvacrol resistance, thus to understand its mechanisms in P. aeruginosa. A herbal drug resistant strain was isolated from a hospital environment. Carvacrol sensitive mutant was generated using transposon mutagenesis. The inactivated gene in the mutant was identified as mexA, which is part of the mexAB-oprM operon. Inactivation of the mexA gene resulted in a >31-fold reduction in MIC of carvacrol, whereas a >80-fold reduction was observed in the presence of drug efflux inhibitor phenylalanine-arginine ß-naphthylamide (PAßN). The parental herbal-resistant strain was completely killed within 3 h of incubation in the presence of carvacrol and PAßN. The mexA inactivation did not affect the resistance to other herbal compounds used. The results demonstrate that resistance to carvacrol in P. aeruginosa is mediated by the MexAB-OprM efflux pump.

Development and evaluation of isothermal amplification assay for the rapid and sensitive detection of Clostridium perfringens from chevon.

Loop-mediated isothermal amplification (LAMP) is a promising, simple, rapid and sensitive molecular detection method. In the present study, LAMP assay was developed for detecting Clostridium perfringens in chevon. Primers were designed to detect the cpa gene of C. perfringens. A panel of 19 bacterial strains, including 3 C. perfringens and 16 other strains, were included in this study to standardize and evaluate the LAMP assay. No false positive amplification was observed indicating 100% specificity of the assay. The detection limit of LAMP and conventional PCR in the DNA extracted from pure C. perfringens was 0.34 pg and 3.4 pg, respectively. This revealed that LAMP assay is 10 times more sensitive than conventional PCR. The sensitivity of the LAMP assay for the detection of C. perfringens in raw chevon was found to be 1.2 × 102 CFU/g after 6-h enrichment and 1.2 × 105 CFU/g without enrichment in artificial spiking studies. Improved C. perfringens detection of 12 CFU/g within 12 h was obtained proving that LAMP assay is significantly faster than traditional methods that take >2 d. The developed LAMP assay also detected the targeted organism in clinical and environmental samples with the sensitivity and specificity of 97% and 84%, respectively with Kappa agreement of 0.824 respects to PCR assay. This method shows immense potential for routine diagnosis and monitoring of C. perfringens in food, environment and clinical samples. This is the first report in which the LAMP assay was optimized for the detection of C. perfringens in chevon.

Selection and characterization of probiotic lactic acid bacteria and its impact on growth, nutrient digestibility, health and antioxidant status in weaned piglets.

The present study was aimed to develop an effective probiotic lactic acid bacteria (LAB) from piglet feces and in vitro characterization of probiotic properties. To confirm host-species specificity of probiotics, the efficacy of isolated LAB on growth, nutrient utilization, health and antioxidant status was observed in early weaned piglets. A total of 30 LAB were isolated from feces of five healthy piglets (28d old). All isolates were Gram positive, cocco-bacilli and catalase negative. Out of thirty LAB isolates, twenty were shortlisted on the basis of their tolerance to pH (3.0, 4.0, 7.0 and 8.0) and bile salts (0.075, 0.15, 0.3 and 1.0%). Whereas, fourteen isolates were selected for further in vitro probiotic characterization due higher (P<0.05) cell surface hydrophobicity to toluene (>45 percent). These isolates fermented twenty-seven different carbohydrates but were negative for ONPG, citrate and malonate. Also enabled to synthesize amylase, protease, lipase and phytase. They were sensitive to penicillin, azithromycin, lincomycin, clindamycin, erythromycin, cephalothin and chloramphenicol and resistant to ciprofloxacin, ofloxacin, gatifloxacin, vancomycin and co-trimoxazole. Except three isolates, all showed antagonistic activity (>60% co-culture activity) against Escherichia coli, Salmonella Enteritidis, Salmonella serotype (ser.) Typhimurium, Staphylococcus intermedius, Staph. chromogenes, Proteus mirabillis, Areomonas veonii, Bordetella bronchioseptica and Klebsialla oxytoca. The isolate Lacp28 exhibited highest tolerance to acidic pH and bile salts (up to 0.3%), phytase activity, cell surface hydrophobicity, antagonistic activity and co-culture assay (>80% growth inhibition). Host specificity of Lacp28 was further confirmed by heavy in vitro adhesion to pig intestinal epithelium cells compared to chicken. Hence, Lacp28 was selected and identified by phylogenetic analysis of 16S rRNA as Pediococcus acidilactici strain FT28 with 100% similarity (GenBank accession nos. KU837245, KU837246 and KU837247). The Pediococcus acidilactici FT28 was selected as potential probiotic candidature for in vivo efficacy in weaned pigs. Thirty-six crossbred piglets (28d) were randomly distributed into three groups (four replicates of three each) namely, basal diet without probiotics (T0) or with Lactobacillus acidophilus NCDC15 (conventional dairy-specific probiotic; T1) or Pediococcus acidilactici FT28 (swine-specific probiotic; T2). At end of the experiment, six piglets of similar body weight were selected to conduct digestion trial for estimation of nutrient digestibility. Results of the study indicated that supplementation of both probiotics improved (P<0.001) FCR compared to control without significant effect in average daily gain and DM intake. However, the apparent digestibility of crude protein and ether extract was better (P<0.01) in pigs fed P. acidilactici FT28 compared control and L. acidophilus fed groups. The total WBC and RBC count, serum glucose, total protein, albumin and globulin concentration was higher (P<0.05) in P. acidilactici FT28 fed group with better (P<0.05) catalase and superoxide dismutase activity measured in erythrocyte. It is concluded that species-specific Pediococcus acidilactici FT28 isolated with potential in vitro probiotic properties and also hold probiotic candidature by showing the potential capabilities with higher nutrient digestibility, heamato-biochemical and antioxidant status compared to control and Lactobacillus acidophilus NCDC15.

Development and Evaluation of Simple Dot-Blot Assays for Rapid Detection of Staphylococcal Enterotoxin-A in Food.

The present study was aimed to develop and evaluate dot-blot assays for rapid detection of staphylococcal enterotoxin-A (SEA) in food. Dot blots were developed in two formats, indirect and sandwich utilizing mouse monoclonal anti-SEA and rabbit polyclonal anti-SEA antibodies. In indirect dot-blot format, recombinant SEA was directly coated on NCM dot-blot strip and detection was carried out by anti-SEA antibodies. In sandwich dot-blot format, SEA was trapped between anti-SEA capture and detection antibodies. Both the dot-blot assays exhibited a sensitivity of ~48 ng ml-1 when tested in different food matrices. The developed assays were highly specific as no cross-reactivity was detected with other classical staphylococcal enterotoxins, toxigenic bacteria and foodborne pathogens. Sensitivity and specificity of developed indirect and sandwich dot-blot assays with respect to PCR was found to be 100 and 99%, respectively. The results shows that the developed dot-blot assays can be used as rapid preliminary screening tests for detection of SEA in food or determining the toxigenic potential of staphylococci, especially in resource-limited settings.

Evaluation of complementary diagnostic tools for bovine tuberculosis detection in dairy herds from India.

AIM: A cross-sectional study was undertaken to know the herd prevalence and evaluate the single intradermal tuberculin testing (SITT), culture isolation, and polymerase chain reaction (PCR) analysis for the diagnosis of bovine tuberculosis (TB). MATERIALS AND METHODS: A total of 541 cows of three dairy farms of Bareilly and Mukteshwar were screened by SITT followed by collection of pre-scapular lymph node (PSLN) aspirates (71), milk (54), and blood (71) samples from reactor animals. These clinical samples were processed for culture isolation and direct PCR-based identification and species differentiation. RESULTS: Out of 541 cows screened by SITT, 71 (13.12%) animals were found positive. Mycobacteria were isolated from 3 (4.22%) PSLN aspirate but not from any cultured milk and blood samples. 28 (39.43%) PSLN aspirate and 5 (9.25%) milk samples were positive for Mycobacterium TB (MTB) complex (MTC) by PCR amplification for the IS6110 insertion sequence; however, blood samples were found negative. For species differentiation, multiplex-PCR using 12.7 kb primers was conducted. Out of 28 PSLN aspirate, Mycobacterium bovis was detected in 18 (64.28%) and MTB in 8 (28.57%), whereas 2 aspirate samples (7.14%) were positive for both the species. All the five milk positive samples were positive for M. bovis. CONCLUSION: Direct detection of bovine TB by a molecular-based method in dairy animals after preliminary screening was appeared to be more sensitive and specific compared to the conventional method (i.e., culture isolation). Its application in form of serial testing methodology for the routine diagnosis and thereafter, culling of infected stock may be suggested for the control programs in dairy herds. The PSLN aspirate was found to be the most suitable specimen for culture isolation and PCR-based detection of Mycobacterium spp. among live infected animals.

Axone, an ethnic probiotic containing food, reduces age of sexual maturity and increases poultry production.

Axone (Akhuni) is a homemade synbiotic (Nagamese fermented soybean product) served as side dish in North Eastern India. In this study, effects of Axone feeding on growth, weight gain, sexual maturity and egg production on Vanaraja birds (a strain of poultry bird developed at PDP Hyderabad for villages and backyard poultry) were evaluated. Axone incorporation in commercial poultry feed at the rate of 5% (W/W) significantly improved growth rate (weight gain) both in male (p 0.001) and female (p 0.05) chicks, reduced age by 13 days at first egg laying (p 0.01), increased egg production (p ≤ 0.001) and improved egg weight (p ≤ 0.01). Microbiological analysis of Axone sample revealed that the major bacteria in Axone samples were Bacillus coagulans, well known for their probiotic value.

Evaluation of vaccine candidate potential of deltaaroA, deltahtrA and deltaaroAdeltahtrA mutants of Salmonella enterica subspecies enterica serovar Abortusequi in guinea pigs.

Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi), a host adapted Salmonella causes abortions, still births and foal mortality in equids. Though known since more than 100 years, it is still a problem in many of the developing countries including India. There is dearth of really good vaccine affording immunity lasting at least for one full gestation. In search of a potential vaccine candidate, three defined deletion mutants (deltaaroA, deltahtrA and deltaaroAdeltahtrA) of S. Abortusequi were tested in guinea pig model for attenuation, safety, immunogenicity, humoral immune response, protective efficacy and persistence in host. The deltahtrA and deltaaroAdeltahtrA mutants were found to be safe on oral inoculation in doses as high as 4.2 x 10(9) cfu/animal. Also through subcutaneous inoculation deltaaroAdeltahtrA mutant did not induce any abortion in pregnant guinea pigs. All the three mutants did not induce any illness or death in 1-2 week-old baby guinea pigs except deltahtrA mutant which caused mortality on intraperitoneal inoculation. Inoculation with mutants protected against challenge and increased breeding efficiency of guinea pigs. After >4.5 months of mutant inoculation, guinea pigs were protected against abortifacient dose of wild type S. Abortusequi and mother guinea pigs also conferred resistance to their babies to the similar challenge. Early humoral immune response of S. Abortusequi mutants was characteristic. Faecal excretion of deltaaroA and htrA mutants was detected up to 45 days of inoculation in guinea pigs while deltaaroAdeltahtrA mutant could not be detected after 21 days of inoculation. The results indicated that the double deletion mutant (deltaaroAdeltahtrA) was the most effective and safe candidate for vaccination against S. Abortusequi through mucosal route of inoculation.

Effect of pH and salinity on pathogenicity of Flavobacterium columnare and Myxobacterium sp. in Indian cat fish, Clarias batrachus (Linn.) and Heteropneustes fossilis (Bloch.).

Flavobacterium columnare (FC) and Myxobacterium sp. recorded persistently associated in fish hatchery and culture system of Himalayan and Sub - Himalayan regions were found to be pathogenic. The pH and salinity played a significant role on the pathogenicity of these potent pathogens in case of Clarias batrachus and Heteropneustes fossilis. LD50 value of FC was 10(4.5) CFU in both the fishes and those of Myxobacterium sp it was 10(6) CFU ml(-1) fish(-1). Fish challenged with F. columnare and Myxobacterium sp. (@ 0.2 ml fish(-1)) individually consisting 10(5-6) cfu ml(-1) exhibited explicit symptoms of columnaris disease and marked with ulceration and saddle back lesion on the dorsal side of body. Maximum reisolation of inoculated bacteria was recorded at pH 7.0 and 7.5 and at 0.0-0.5 (F. columnare) and 0.0-1.0% (Myxobacterium sp.) salinity. Foregoing results elucidated that F. columnare was more sensitive to salinity in comparison to Myxobacterium sp. and their pathogenicity significantly (p<0.05) depends on the salinity and pH that might be one of the physical factors to control their proliferation.

Plasmid profile and drug resistance pattern of zoonotic Salmonella isolates from Indian buffaloes.

BACKGROUND: Buffalo is the major source of animal protein in south-east Asia, including India; therefore, the presence of multiple drug resistance in Salmonella strains of buffalo meat and milk products is of immense public health concern. METHODOLOGY: Forty-six strains of Salmonella enterica subspecies enterica belonging to eight serovars (S. Anatum, 13; S. Weltevreden, 13; S. Rostock, 6; S. Typhimurium, 5; S. Gallinarum, 5; S. Stockholm, 1; S. Dublin, 1; and S. Orion, 2), isolated from buffalo meat and diseased buffaloes were studied for their antibiotic sensitivity and plasmid profile. RESULTS: All except six strains of Salmonella had one or more plasmids. Virulence plasmid of ~35MdA was present in 39 isolates while 19 strains had one to six additional plasmids with molecular weight ranging from 1 Mda > 35 Mda. A plasmid-free S. Anatum strain was resistant to seven drugs including fluoroquinolones, while strains having six to seven plasmids were resistant to fewer antimicrobial drugs. One S. Anatum isolate, resistant to 11 antibiotics, had only one plasmid. Eight serovars of Salmonella could be divided into 28 resistotypes on the basis of antimicrobial sensitivity assay. Most strains were resistant to streptomycin (84.8%) followed by kanamycin (58.7%), gentamicin (52.2%), ampicillin (50%) and oxytetracycline (50%). Few strains were resistant to cefotaxime (2.2%), amoxycillin (2.2%) and newer fluoroquinolones (6.5%). CONCLUSION: Multiple drug resistance was common among Salmonella isolates of buffalo origin, particularly against aminoglycosides, oxytetracycin, ampicillin and cephalexin. Presence of plasmids is not mandatory for occurrence of multiple drug resistance in S. enterica strains.

Drug resistance patterns of Salmonella isolates of equine origin from India.

INTRODUCTION: Salmonellosis is a zoonosis, and one of the most serious public health and animal health problems. METHODOLOGY: We studied 111 isolates of Salmonella belonging to 14 S. enterica subspecies enterica serovars namely S. Abortusequi (45), S. Weltevreden (1), S. Dumfries (2), S. Tshiongwe (1), S.I. 4,5,12:r,i:1,5 (12), S. Bovismorbificans (3), S. Drogana (8), S. Lagos (4), S. Kottbus (3), S. Richmond (1), S. Typhimurium (6), S. Newport (7), S. Paratyphi B var Java (17) and S. Saintpaul (5) isolated from equids in India. RESULTS: All strains studied were resistant to one or more antimicrobials. Strains were resistant to ampicillin (18, 16%), ampicillin+cloxacillin (6, 5%), cefotaxime (6, 5%), chloramphenicol (2, 2%), ciprofloxacin (9, 8%), gentamicin (27, 24%), kanamycin (37, 33%), nalidixic acid (10, 9%), furazolidone (97, 87%), streptomycin (33, 30%), sulphamethoxazole (91, 82%), tetracycline (48, 43%) and trimethoprim (5, 4.5%). Multiple-drug-resistance was detected in 84 (75.7%) isolates and was seen in isolates of all serovars except of S. Kottbus, a rare serovar in India. Salmonella isolates could be classified into 51 resistotypes but 47 (42.3%) isolates belonged to six major resistotypes. Resistotype 13 (resistant to furazolidone, sulphamethoxazole and tetracycline) was most common, followed by resistotype 19 (resistant to nalidixic acid, sulphamethoxazole and tetracycline), resistotype 28 (resistant to furazolidone, streptomycine, sulphamethoxazole and tetracycline) and resistotype 40 (resistant to furazolidone, gentamycin, kanamycin, streptomycine, sulphamethoxazole and tetracycline) including 11, 8, 8 and 7 strains of different serovars, respectively. CONCLUSIONS: This study revealed that antimicrobial drug resistance was common in Salmonella isolates from equids even towards those drugs not used in equids.

Prevalence of vancomycin resistance and multiple drug resistance in enterococci in equids in North India.

INTRODUCTION: Vancomycin resistant and multi-drug-resistant enterococci are the major emerging pathogens in surgical, neonatal, and tertiary care units. METHODOLOGY: In this study, 267 enterococci from different clinical and non-clinical samples of equine origin were tested for their antimicrobial drug sensitivity against 19 antimicrobials using disc diffusion method. RESULTS: A total of 80.2% enterococci tested were resistant to vancomycin and 99.6% to multiple-drugs. There was a significant association between haemolytic potential and vancomycin resistance (chi(2), 0.00). Enterococci isolates from healthy equids were significantly (chi(2), 0.04) less resistant to vancomycin than the isolates from clinically sick animals. Besides vancomycin sensitivity, isolates were also tested for 18 more antimicrobial drugs; maximum numbers of isolates were sensitive to imipenem (75%) followed by tetracycline (60%), amoxicillin+clavulanic acid (54%), and minimum for cefdinir (4%). CONCLUSION: More than 80% strains of enterococci of equine origin were found resistant to vancomycin and 99.6% were multiple-drug resistant in Northern India. High prevalence of VRE and MDRE in healthy equids might be a potential danger for the health of persons in equine contact.

Infertility associated with sub clinical salmonellosis.

Subclinical infection of guinea pigs with isogenic wild type and aroA, htrA and aroA-htrA mutants of Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi) induced infertility, while mutants had little or no effect on conception rate in guinea pigs. Conception rate was significantly lower in guinea pigs inoculated with wild type (S-787) and aroA mutant of S. Abortusequi than those inoculated with intracellular survival deficient htrA or aroA-htrA mutants of S. Abortusequi. Chi-test analysis revealed that none of the three mutants could be attributed to low conception rate, but wild type Salmonella inoculation and chronic carriage of the pathogen were significant cause of low conception rate in guinea pigs. Role of S. Abortusequi in causation of infertility was proven from the experiment for the first time.

Prevalence of multidrug-resistant Salmonella on ready-to-eat betel leaves (Paan) and in water used for soaking betel leaves in North Indian cities.

The purpose of this study was to determine the prevalence of Salmonella in the water used by Paan vendors in 11 North Indian cities. The analysis of 776 water samples and 120 samples each of preprocessed Paan (from vendor stock) and ready-to-eat Paan collected from Bareilly revealed that four of the ready-to-eat Paan and 34 of the water samples contained multidrug-resistant Salmonella strains. The isolates belonged to five different serovars, i.e., Salmonella Newport (1), Salmonella Paratyphi B (1), Salmonella Teko (1), Salmonella Virchow (3), and Salmonella Saintpaul (32), and could also be classified into 18 different resistotypes. All of the isolates were sensitive to cotrimoxazole, and 97.27% of the isolates were sensitive to chloramphenicol, imipenem, ciprofloxacin, ceftriaxone, and neomycin. Multidrug resistance (against 5 to 18 antibiotics) was common, particularly for nalidixic acid (65.79%), cephalothin (68.42%), cefoperazone (57.89%), sulfamethizole (52.63%), furazolidone (65.79%), kanamycin (68.42%), doxycycline (50.00%), and cefotaxime (44.74%). Bacteriological analysis of 16 Salmonella-positive and 23 Salmonella-negative samples revealed that the presence of Salmonella in water samples had a negative correlation (r = -0.35) with coliform counts and a positive correlation (r = 0.55) with nonlactose fermenter counts. We determined that centrifugation was a rapid and cheap method for concentrating large samples of Salmonella. The presence of multidrug-resistant strains of zoonotic Salmonella on ready-to-eat Paan is a public health concern and may be one of the factors responsible for the hyperendemic status of salmonellosis in India.