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The ladybird beetles of Scymninae, a subfamily of Coccinellidae are efficient biocontrol agents. Many earlier studies document their protective role associated with many crops. This subfamily is represented by 5 tribes, 15 genera and 138 species in the Indian subcontinent. However there is no scientific record of taxonomic description of the subfamily Scymninae within the Coccinellidae family of beetles (Coccinellidae: Coleoptera) in Haryana, India. In the present study, seven species from three genera and two tribes of sub-family Scymninae i.e. Nephus (Bipunctatus) bipunctatus (Kugelann, 1794), Nephus regularis (Sicard), Scymnus (Pullus) coccivora Ramakrishna Ayyar, 1925, Scymnus (Pullus) latemaculatus Motschulsky, Scymnus (Scymnus) nubilus Mulsant, Scymnus (Pullus) posticalis Sicard and Stethorus aptus Kapur were identified. It included the generation of keys to the tribes, genera, subgenera and species found during the study. It provided detailed taxonomy of the identified species based on various morphological characteristics such as coxal lines, antennae, mandibles, and male and female genitalia. This study marks the first taxonomic exploration of species within the Scymninae subfamily in Haryana, India. It will significantly contribute to understanding the biodiversity of beetles in the region and will lay the groundwork for further research and conservation initiatives. [ABSTRACT FROM AUTHOR]

Background: The purpose of this analysis is to evaluate the antimicrobial susceptibility of eight drugs effective against Helicobacter pylori (H. pylori) strains and the genetic diversity of H. pylori virulence genes to foresee clinical outcomes in North India. Materials and Methods: Fifty‐eight H. pylori strains isolated from patients suffering from various gastrointestinal (GI) diseases were included in the study. MICs of various antibiotics were determined by the agar dilution method. The chi‐squared test and Fisher exact test were used to determine the p‐value, which was considered significant at p‐value ≤ 0.05. RStudio 4.0 was used to for the data visualization. Results: The prevalence of drug resistance was found to be: cefixime (CFM) (41.3%), furazolidone (FZD) (34.4%), amoxicillin (AMX) (20.7%), levofloxacin (LVFX) (70.7%), metronidazole (MTZ) (39.6%), tetracycline (TET) (20.7%), clarithromycin (CLA) (17.2%), and rifabutin (RIF) (17.2%). Out of 58 H. pylori strains, 3 were pan susceptible. There were H. pylori strains with single‐drug resistance (21.8%, 12/55), dual resistance (30.9%, 17/55), triple resistance (20%, 11/55), and multidrug resistance (27.3%, 15/55). The resistance rate in MTZ, CLA and RIF were found to be significantly higher in females as compared to males (p = 0.005, p = 0.002, and p = 0.02), respectively. The resistance to TET exhibited significantly higher levels in gastritis compared to GERD, DU, and other disease groups (p = 0.04) respectively. Conclusion: TET, AMX, CLA, and RIF were found to be more effective antibiotics against H. pylori infections, whereas more studies are required to provide evidence on increasing resistance rate of LVFX. [ABSTRACT FROM AUTHOR]

Enteroaggregative Escherichia coli (EAEC) is an emerging enteric pathogen that causes acute and chronic diarrhea in developed and industrialized countries in children. EAEC colonizes the human intestine and this ability to form colonies and biofilm is an important step in pathogenesis. Here, we investigated the relationship between known or putative 22 EAEC virulence genes and biofilm formation in isolates derived from acute diarrhea and healthy children and their aggregative adherence (AA) pattern with Hep-2 cell lines. A total of 138 EAEC isolates were recovered from 1210 stool samples from children (age < 10 years) suffering from acute diarrhea and 33 EAEC strains isolated from 550 healthy children (control group) of different Anganwadi centers in Chandigarh region were included. Polymerase chain reaction using the primer pair pCVD432 identified E. coli isolates as EAEC. A total of 22 virulence-related genes have been identified using M-PCR chain reactions. The crystal violet method was used for the quantitative biofilm assay. Aggregative adherence assay was also studied using HEp-2 cell lines. Of 138 EAEC isolates from the acute diarrheal group, 121 (87.6%) EAEC isolates produced biofilm. In our findings, typical EAEC (62%) isolates were strong biofilm producers (37.5%) in the diarrheal group. Among adhesive variants, agg4A (39.6%) and aggA (21.6%) were the most common and were statistically significant (p = 0.01 and p = 0.03 respectively). We reported that the aggR gene along with the typical AA pattern was present in 71.4% of the EAEC strains in the diarrheal group, whereas it was present in 44% of the control group. Other aggR non-dependent genes like ORF3 and eilA may also lead to biofilm formation. In conclusion, there is significant heterogeneity in putative virulence genes of EAEC isolates from children and biofilm formation is associated with the combination of many genes. [ABSTRACT FROM AUTHOR]

Pasteurella multocida is widely distributed in all pig-rearing countries, affecting the economic viability and profitability of pig production. The present research highlights the molecular characterization and pathology of untypeable capsular serotypes of P. multocida in slaughtered pigs from prominent pig-rearing states of India. The prevalence of Pasteurellosis was 27.17% by Pasteurella multocida specific Pasteurella multocida specific PCR (PM-PCR). assay, while isolation rate was 7.62%. The microscopic lesions of bronchopneumonia, tonsillitis, and the presence of bacterial antigens in immunohistochemistry confirmed P. multocida with pathologies. In capsular typing, the majority of the isolates were untypeable with prevalence of 52.15% and 43.58% in molecular and microbiological methods, respectively. All the isolates showed the uniform distribution of virulence genes such as exbB, nanB, sodC, plpB, and oma87 (100%), while the variations were observed in ptfA, hasR, ptfA, pfhA, hsf-1, and plpE genes. The untypeable isolates showed higher prevalence of hsf-1 gene as compared to others. The untypeable serotypes showed a higher degree of resistance to ampicillin, amoxicillin, and penicillin antibiotics. The mouse pathogenicity testing of untypeable capsular isolates confirmed its pathogenic potential. The higher frequency of pathogenic untypeable isolates with antibiotic resistance profile might pose a serious threat to the pigs, and therefore, preventive measures should be adopted for effective control. [ABSTRACT FROM AUTHOR]

Aeromonas hydrophila, a natural inhabitant of aquatic environments is known to become pathogenic in fish under favourable conditions. It has also been reported to cause infections in humans and animals. The prevalence, virulence-associated genes, and antimicrobial resistance of 57 A. hydrophila isolates from freshwater fish farms in Andhra Pradesh, India, were studied. The isolates showed resistance to trimethoprim (35.9%), co-trimoxazole (21.1%), oxytetracycline (19.3%), doxycycline (17.5%), and enrofloxacin (8.93%), but were completely sensitive to ciprofloxacin. The majority of the A. hydrophila isolates were strong (33%), moderate (50%), and weak (17%) biofilm formers, playing roles in virulence and colonization in hosts. The virulence gene distribution in representative A. hydrophila isolates was very high for enolase (enol, 96.4%), followed by flagellin (fla, 94.7%), cytotonic enterotoxins (alt, 91.2%), cytotoxic enterotoxins (act, 84.2%), lipase (lip, 77.1%), serine protease (ahp, 54.3%), aerolysin (aerA, 50.8%), and elastase (ahyB, 43.8%), respectively. 80% of the isolates carried a maximum of four virulence genes, whereas none carried the DNAase (exu) or haemolysin (hly) genes. Hemolytic assays on blood agar demonstrated that 70% of the isolates were β-haemolytic. The findings of the study will help to create awareness among aquaculturists of the potential risk of the spread of isolates and the virulence-associated genes across other sectors and the need to adopt best management practices in aquaculture. [ABSTRACT FROM AUTHOR]

Introduction: Burkholderia cepacia complex (Bcc) clonal complex (CC) 31, the predominant lineage causing devastating outbreaks globally, has been a growing concern of infections in non-cystic fibrosis (NCF) patients in India. B. cenocepacia is very challenging to treat owing to its virulence determinants and antibiotic resistance. Improving the management of these infections requires a better knowledge of their resistance patterns and mechanisms. Methods: Whole-genome sequences of 35 CC31 isolates obtained from patient samples, were analyzed against available 210 CC31 genomes in the NCBI database to glean details of resistance, virulence, mobile elements, and phylogenetic markers to study genomic diversity and evolution of CC31 lineage in India. Results: Genomic analysis revealed that 35 isolates belonging to CC31 were categorized into 11 sequence types (ST), of which five STs were reported exclusively from India. Phylogenetic analysis classified 245 CC31 isolates into eight distinct clades (I-VIII) and unveiled that NCF isolates are evolving independently from the global cystic fibrosis (CF) isolates forming a distinct clade. The detection rate of seven classes of antibiotic-related genes in 35 isolates was 35 (100%) for tetracyclines, aminoglycosides, and fluoroquinolones; 26 (74.2%) for sulphonamides and phenicols; 7 (20%) for beta-lactamases; and 1 (2.8%) for trimethoprim resistance genes. Additionally, 3 (8.5%) NCF isolates were resistant to disinfecting agents and antiseptics. Antimicrobial susceptibility testing revealed that majority of NCF isolates were resistant to chloramphenicol (77%) and levofloxacin (34%). NCF isolates have a comparable number of virulence genes to CF isolates. A well-studied pathogenicity island of B. cenocepacia, GI11 is present in ST628 and ST709 isolates from the Indian Bcc population. In contrast, genomic island GI15 (highly similar to the island found in B. pseudomallei strain EY1) is exclusively reported in ST839 and ST824 isolates from two different locations in India. Horizontal acquisition of lytic phage ST79 of pathogenic B. pseudomallei is demonstrated in ST628 isolates Bcc1463, Bcc29163, and BccR4654 amongst CC31 lineage. Discussion: The study reveals a high diversity of CC31 lineages among B. cenocepacia isolates from India. The extensive information from this study will facilitate the development of rapid diagnostic and novel therapeutic approaches to manage B. cenocepacia infections. [ABSTRACT FROM AUTHOR]

Streptococcus suis is an emerging bacterial pathogen of huge economic impact to the swine industry worldwide. The information regarding the carrier status of S. suis in the slaughtered pigs along with its genetic characterization is not available in Indian pig population, which needs to be addressed for the therapeutic and preventive measures. In the present study, 563 palatine tonsils of apparently healthy slaughtered pigs were probed for the prevalence, and genetic characterization of S. suis and prevalence were found to be 15.45% and 32.68% by bacteriological and molecular methods, respectively. In 87 isolates recovered, 6 cps-types were detected showing the predominance of serotype 7 (24.13%) and 5 (18.39%), whereas 11 cps-types were detected in tonsillar DNA involving cps-types 9 (28.26%) and 7 (14.13%) as the major serotypes with arcA+/sly+/epf+/mrp− being the prevalent genotype. The histopathological changes with the immunodetection of S. suis antigen confirmed its persistence in asymptomatic carriers. Of 87 bacterial isolates, 7 isolates (serotypes 7 & 2) were pathogenic to Swiss albino mice showing the classical lesions of meningitis and septicemia. The presence of virulent serotypes of S. suis in healthy slaughtered pigs suggests a great health risk to the people engaged in piggery operations and in-contact pigs. [ABSTRACT FROM AUTHOR]

Background: Non-typhoidal Salmonella (NTS) is one of the four critical worldwide reasons for diarrhoeal infections and causes a noted zoonotic infection termed non-typhoidal salmonellosis. Non-typhoidal salmonellosis generally causes self-limited gastroenteritis, whereas in immunocompromised conditions can result in invasive infections. Aim and Objectives: To detect the likely NTS serovars causing non-typhoidal Salmonellosis and their virulence genes. Material and Methods: This is a prospective cross-sectional research work carried out at the Enteric Diseases Division, Central Research Lab, Kasturba Medical College, Manipal from January 2016 - June 2018. Stool samples were collected from patients with diarrhoea admitted to the tertiary care centre in Udupi district, India. Stool samples were processed according to the World Health Organisation laboratory protocol. Non-typhoidal Salmonella isolated were subjected to PCR for the detection of virulence genes. Results: Of the 1599 diarrheal samples processed, 55 NTS were isolated with a prevalence of 3.43 %. invA gene was existent in 83.6% of the isolates, spvCgene in 61.8%, stn in 100% sopB in 96.4% and sefA in 5.45%. Conclusion: The presence of virulence genes among NTS increases the complications of non-typhoidal salmonellosis. Routine antimicrobial susceptibility tests for salmonellae should be carried out and then reported to prevent the associated morbidity and mortality. [ABSTRACT FROM AUTHOR]

Tilletia indica is a quarantine fungal pathogen that poses a serious biosecurity threat to wheat-exporting countries. Acquiring genetic data for the pathogenicity characters of T. indica is still a challenge for wheat breeders and geneticists. In the current study, double digest restriction-site associated-DNA genotyping by sequencing was carried out for 39 T. indica isolates collected from different locations in India. The generated libraries upon sequencing were with 3,346,759 raw reads on average, and 151 x 2 nucleotides read length. The obtained bases per read ranged from 87 Mb in Ti 25 to 1,708 Mb in Ti 39, with 505 Mb on average per read. Trait association mapping was performed using 41,473 SNPs, infection phenotyping data, population structure, and Kinship matrix, to find single nucleotide polymorphisms (SNPs) linked to virulence genes. Population structure analysis divided the T. indica population in India into three subpopulations with genetic mixing in each subpopulation. However, the division was not in accordance with the degree of virulence. Trait association mapping revealed the presence of 13 SNPs associated with virulence. Using sequences analysis tools, one gene (g4132) near a significant SNP was predicted to be an effector, and its relative expression was assessed and found upregulated upon infection. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Typhi and Salmonella Paratyphi are causative agents of enteric fever. Salmonella Typhi persists as a biofilm on gallstones. Hence, we studied the biofilm formation, antibiogram, and virulence genes of S. enterica serovars. Antibiogram of S. enterica serovars from human blood and stool samples were studied by Kirby–Bauer disk diffusion method and biofilm by microtiter plate method. We studied the minimum inhibitory concentration of the isolates by Vitek-2 semiautomated system. Polymerase chain reaction was done to detect invA and spvC genes. Of the 55 isolates studied, 36 (65.45%) were Salmonella Typhi, 13 (23.63%) were Salmonella Paratyphi A, 2 (3.64%) were Salmonella Typhimurium, and 4 (7.28%) were Salmonella spp. Resistance to ciprofloxacin and nalidixic acid were found to be 81.8% and 92.7%, respectively. Chloramphenicol and cotrimoxazole-susceptible strains were 98.18%. One each of Salmonella Typhi, Salmonella Paratyphi A, and S. enterica isolates formed weak biofilm at 28°C. However, at 37°C eight Salmonella Typhi produced weak biofilm in the presence of bile. One Salmonella Paratyphi A and two Salmonella spp. formed weak biofilm in the absence of bile. All the isolates had the invA gene. Salmonella Typhimurium had invA and spvC genes. Bile may contribute to biofilm formation and persistence of the Salmonella Typhi on gallstones, which may lead to carrier state. Changing antibiotic susceptibility pattern of Salmonella serovars is observed in our geographic area. The presence of invA and spvC genes indicate the ability of invasiveness and intracellular survival. [ABSTRACT FROM AUTHOR]

Methicillin-resistant staphylococcus aureus (MRSA) sequence type 28 (ST 28) and spa type t021 is a CC30, prototype of ST-30, Community Associated-MRSA (CA-MRSA) (lukS-lukF +). It is a multi-drug resistant strain harbouring staphylococcal endotoxins, haemolysins, ureolysin, serine protease, and antimicrobial resistance genes. In this study, we report the draft genome sequence of this MRSA isolated from the most commonly used food fish, ribbon fish (Trichiurus lepturus). The total number of assembled paired-end high-quality reads was 7,731,542 with a total length of 2.8Mb of 2797 predicted genes. The unique ST28/t021 CA-MRSA in fish is the first report from India, and in addition to antibiotic resistance is known to co-harbour virulence genes, haemolysins, aureolysins and endotoxins. Comprehensive comparative genomic analysis of CA-MRSA strain7 can help further understand their diversity, genetic structure, diversity and a high degree of virulence to aid in fisheries management. [ABSTRACT FROM AUTHOR]

Objective: This study was designed to discover the dissemination of virulence genes in Methicillin-resistant Staphylococcus aureus from clinical, community and environmental settings. Results: This study includes 1165 isolates collected from hospital, community and environmental settings. Among them sixty three were confirmed as MRSA with varied SCCmec types viz; type I, type II, type III, type IV, type V, type VI, type VII, type VIII and type XII. The virulence gene such as sea (n = 54), seb (n = 21), eta (n = 27), etb (n = 2), cna (n = 24), ica (n = 2) and tst (n = 30) was also revealed from this study. The study underscores coexistence of resistance cassette and virulence genes among clinical and environment isolates which is first of its kind from this part of the world. [ABSTRACT FROM AUTHOR]

Objective: Burkholderia pseudomallei, the etiological cause of melioidosis, is a soil saprophyte endemic in South-East Asia, where it constitutes a public health concern of high-priority. Melioidosis cases are sporadically identified in nonendemic areas, usually associated with travelers or import of goods from endemic regions. Due to extensive intercontinental traveling and the anticipated climate change-associated alterations of the soil bacterial flora, there is an increasing concern for inadvertent establishment of novel endemic areas, which may expand the global burden of melioidosis. Rapid diagnosis, isolation and characterization of B. pseudomallei isolates is therefore of utmost importance particularly in non-endemic locations., Data Description: We report the genome sequences of two novel clinical isolates (MWH2021 and MST2022) of B. pseudomallei identified in distinct acute cases of melioidosis diagnosed in two individuals arriving to Israel from India and Thailand, respectively. The data includes preliminary genetic analysis of the genomes determining their phylogenetic classification in rapport to the genomes of 131 B. pseudomallei strains documented in the NCBI database. Inspection of the genomic data revealed the presence or absence of loci encoding for several documented virulence determinants involved in the molecular pathogenesis of melioidosis. Virulence analysis in murine models of acute or chronic melioidosis established that both strains belong to the highly virulent class of B. pseudomalleii., (© 2024. The Author(s).)

Background: Despite high vaccination coverage using acellular (ACV) and whole-cell pertussis (WCV) vaccines, the resurgence of pertussis is observed globally. Genetic divergence in circulating strains of Bordetella pertussis has been reported as one of the contributing factors for the resurgence of the disease. Our current knowledge of B. pertussis genetic evolution in circulating strains is mostly based on studies conducted in countries using ACVs targeting only a few antigens used in the production of ACVs. To better understand the adaptation to vaccine-induced selection pressure, it will be essential to study B. pertussis populations in developing countries which are using WCVs. India is a significant user and global supplier of WCVs. We report here comparative genome analyses of vaccine and clinical isolates reported from India. Whole-genome sequences obtained from vaccine strains: WCV (J445, J446, J447 and J448), ACV (BP165) were compared with Tohama-I reference strain and recently reported clinical isolates from India (BPD1, BPD2). Core genome-based phylogenetic analysis was also performed using 166 isolates reported from countries using ACV. Results: Whole-genome analysis of vaccine and clinical isolates reported from India revealed high genetic similarity and conserved genome among strains. Phylogenetic analysis showed that clinical and vaccine strains share genetic closeness with reference strain Tohama-I. The allelic profile of vaccine strains (J445:ptxP1/ptxA2/prn1/fim2–1/fim3–1; J446: ptxP2/ptxA4/prn7/fim2–2/fim3–1; J447 and J448: ptxP1/ptxA1/ prn1/fim2–1/fim3–1), which matched entirely with clinical isolates (BPD1:ptxP1/ptxA1/prn1/fim2–1 and BPD2: ptxP1/ptxA1/prn1/fim2–1) reported from India. Multi-locus sequence typing (MLST) demonstrated the presence of dominant sequence types ST2 and primitive ST1 in vaccine strains which will allow better coverage against circulating strains of B. pertussis. Conclusions: The study provides a detailed characterization of vaccine and clinical strains reported from India, which will further facilitate epidemiological studies on genetic shifts in countries which are using WCVs in their immunization programs. [ABSTRACT FROM AUTHOR]

Salmonella has emerged as one of the most important food-borne pathogens for humans as well as animals and the ability of biofilm formation by these bacteria has further aided their survival in unfavorable environment. Characterization of these biofilm producing bacteria isolated from pigs and chicken may lead to formulation of strategies for prevention and control of Salmonella infections. Therefore, the present study was conducted to isolate Salmonella from pigs and poultry of Mizoram, determine their biofilm producing ability by phenotypic and genotypic methods along with their virulence and antimicrobial resistance properties. A total of 15 Salmonella spp. (pig=9, poultry=6) was isolated from 100 faecal samples from pigs and 50 cloacal swabs from poultry and biofilm producing ability of the isolates was determined by microtiter plate assay. A total of 10 (66.67%) isolates were found to be biofilm producer. All the biofilm producing bacterial isolates were investigated for antimicrobial sensitivity and distribution of selected biofilm associated genes (csgA, csgD and adrA), virulence genes (invA, stn and sefA) and antimicrobial resistance (AMR) genes (blaTEM, blaSHV and blaCTX-M). The most prevalent resistance was found against ceftazidime (80%), ceftriaxone (80%), cefixime (70%), cefotaxime (70%), gentamicin (70%), cotrimoxazole (60%) and ampicillin (60%). A total of 7 (70%) isolates were resistant to at least three different classes of antimicrobial agents and considered as multidrug resistant. All the isolates were positive for adrA (100%) but negative for csgA and csgD genes. The most frequent virulence gene was invA (100%) and stn (100%). Among the AMR genes, blaTEM (60%) was found to be the major AMR determinants. Moreover, a total of 7 Salmonella isolates were positive for at least one of t biofilm associated genes, virulence genes and AMR genes. [ABSTRACT FROM AUTHOR]

Aquaculture is one of the fastest-growing agricultural sectors which serves as an invaluable source of animal protein. The presence of pathogenic microbes and antimicrobial resistance is a major concern in aquaculture. Aquaculture has been identified as a hotspot for resistant pathogens, but very less details are available on the origin of these pathogens. Here, to better understand the pathogenic incidence and their resistome and virulome, we have sequenced and analysed the genomes of Staphylococcus spp. isolates (n = 12) from shrimps obtained from aquaculture farms in Kerala, India. These isolates were typed as S. aureus (n = 4), S. haemolyticus (n = 3), S. epidermidis (n = 3), S. saprophyticus (n = 1) , and S. warneri (n = 1). Comparative genomic analysis of these isolates was done against reference and published Indian Staphylococcus spp. isolates to identify the strain origin and genetic relatedness. Most of the isolates had a very close identity with human isolates, indicating the human origin of these pathogens. Resistome analysis revealed, S. aureus harbouring anti-microbial resistance genes (ARGs) against methicillin, Norfloxacin, Gentamicin, etc. and S. haemolyticus harbouring ARG against methicillin, erythromycin, etc. S. aureus isolates presented quite an extensive virulome and their clustering with human-pathogens displays their potential for pathogenicity. We here, thus, call attention to monitoring the proper implementation of biosecurity measures in aquaculture farms to avoid entry and enrichment of such isolates. • Sequencing the genomes of staphylococcal isolates from shrimp aquaculture farms. • A comprehensive characterization of resistomes and virulome of the MRS isolates. • comparative analysis of sequenced isolates with strains from various sources – human, livestock and environment. [ABSTRACT FROM AUTHOR]

Purpose: To detect the presence of fimH and iss type 1, 2 and 3 genes in uropathogenic Escherichia coli (UPEC) isolates recovered from patients coming to the out patient department (OPD) of our hospital., Methods: E. coli isolates recovered from patients who had symptoms of urinary tract infection (UTI) were processed for the presence of fimH and iss genes. DNA was extracted using an in house method after which conventional PCR using forward and reverse primers targeting the four genes was carried out. The amplified products were electrophoresed and visualized in a gel documentation imager. Relevant demographic details of the patients were recorded on a pre-designed pro-forma and antimicrobial susceptibility testing of the isolates was done by disc diffusion method., Results: fimH was present in 87.5% of UPEC isolates whereas iss type 1 was seen in 7.3%, type 2 in 4.2% and iss type 3 in 71.9% isolates. Age of the patients ranged from 3 months to 82 ​yrs (mean 43.5 SD ​± ​18.20). UTI was more common in females (60.2%) as compared to males patients (39.8%). Dysuria (66.7%) was the most common symptom in the studied subjects and diabetes mellitus (42.6%) the most common co-morbidity. A total of 56.5% patients gave a history of prior antibiotic intake. The UPEC isolates were resistant to most of the antibiotics tested. However all the isolates were sensitive to polymyxin B and colistin. Fosfomycin resistance was seen in 9.5% of the UPEC isolates harbouring fimH gene., Conclusion: This is the first study that highlights the presence of iss type 3 gene in UPEC isolates along with the fimH and iss type 1 and 2 genes. The results of this study can serve as a stepping stone for future in depth research into the significance of the iss genes in causing UTI., Competing Interests: Conflict of interest None., (Copyright © 2023 Indian Association of Medical Microbiologists. Published by Elsevier B.V. All rights reserved.)

Genetic diversity of 13 isolates of Ralstonia solanacearum causing bacterial wilt in hot pepper and bell pepper (Capsicum annuum L.) from 6 states of India was assessed. All isolates of R. solanacearum belonged to biovar 3, race 1 and phylotype I. These isolates consisted of 4 distinct DNA types at 75% similarity coefficient using ERIC, BOX and REPPCRs techniques. Multilocus sequence analysis of hrpB, fliC and egl genes of 6 isolates of R. solanacearum along with 2 out group bacteria was done and they showed high level of variability within these three regions of the genome involving in pathogenicity. [ABSTRACT FROM AUTHOR]

This study reports the polyphasic identification, characterization of virulence potential, and antibiotic susceptibility of Aeromonas salmonicida subspecies salmonicida COFCAU_AS, isolated from an aquaculture system in India. The physiological, biochemical, 16s rRNA gene sequencing and PAAS PCR test identified the strain as Aeromonas salmonicida. The MIY PCR tests established the subspecies as ' salmonicida '. The in vitro tests showed the isolated bacterium as haemolytic with casein, lipid, starch, and gelatin hydrolysis activity, indicating its pathogenic attributes. It also showed the ability to produce slime and biofilm, and additionally, it possessed an A-layer surface protein. In vivo pathogenicity test was performed to determine the LD 50 dose of the bacterium in Labeo rohita fingerlings (14.42 ± 1.01 g), which was found to be 106.9 cells fish−1. The bacteria-challenged fingerlings showed skin lesions, erythema at the base of the fins, dropsy, and ulcer. Almost identical clinical signs and mortalities were observed when the same LD 50 dose was injected into other Indian major carp species, L. catla and Cirrhinus mrigala. Out of the twelve virulent genes screened, the presence of nine genes viz., aerA , act , ast , alt , hlyA , vapA, exsA , fstA , and lip were detected , whereas ascV, ascC , and ela genes were absent. The A. salmonicida subsp. salmonicida COFCAU_AS was resistant to antibiotics such as penicillin G, rifampicin, ampicillin, and vancomycin while highly sensitive to amoxiclav, nalidixic acid, chloramphenicol, ciprofloxacin, and tetracycline. In summary, we have isolated a virulent A. salmonicida subsp. salmonicida from a tropical aquaculture pond which can cause significant mortality and morbidity in Indian major carp species. • Isolation of A. salmonicida subsp. salmonicida strain COFCAU_AS form India. • In vitro and in vivo tests confirmed the virulent nature of COFCAU_AS. • COFCAU_AS possess 9 tested virulent genes: aerA , act , ast , alt , hlyA , vapA, exsA , fstA lip. • COFCAU_AS was resistant to penicillin G, rifampicin, ampicillin, and vancomycin. [ABSTRACT FROM AUTHOR]

The present study was undertaken with the prime objective of isolating and identifying Listeria spp. from various foods of animal origin sold at retail market outlets in the city of Navsari, Gujarat. Materials and Methods: Total 200 samples comprising of milk, milk products, meat, and fish (50 each) collected aseptically from local market which were subjected first to pre-enrichment in half strength Fraser broth followed by enrichment in full strength Fraser broth and subsequent plating on PALCAM agar. The growth with the typical colony characteristics were further identified up to species level on the basis of their morphological and biochemical characteristics. Cultures identified as Listeria monocytogenes were further subjected to in vitro pathogenicity tests and detection of different virulenceassociated genes viz. actA, hlyA, and iap using polymerase chain reaction. Results: Of the total 200 food samples of animal origin; 18 (9%) were found positive for Listeria spp. which were identified as Listeria seeligeri (6, 33.3%), Listeria innocua (5, 27.7%), Listeria welshimeri (4, 22.2%), and L. monocytogenes (3, 16.6%). The highest prevalence was observed in milk samples (8). Species wise, 6 isolates of L. seeligeri which included two each from cow milk, buffalo milk, and meat samples; 5 L. innocua isolates included four recovered from fish and one from meat sample; 4 L. welshimeri comprised of two isolates from ice cream and one each from buffalo milk and meat sample; and 3 isolates of L. monocytogenes recovered from milk (1 cow and 2 buffalo milk). All 3 L. monocytogenes isolates screened for the presence of virulence genes viz. actA, hlyA, and iap using the specific primers revealed the presence of all the genes suggesting the possibility of danger of foodborne listeriosis among raw milk consumers. Conclusion: Listeria spp. was isolated from 9% (18/200) of the animal origin food samples viz.; milk, milk products, meat, and fish with the highest prevalence in the milk samples. L. monocytogenes was isolated from 3 milk samples only. L. seeligeri was the predominant species isolated followed by L. innocua, L. welshimeri, and L. monocytogenes in this study. L. monocytogenes were found to carry virulence genes like actA, hly A, and iap genes suggesting the pathogenic potential of these isolates. [ABSTRACT FROM AUTHOR]

Pasteurellosis in small ruminants affects the livelihood of small and marginal farmers of India. The present study was undertaken to understand the trends in gene carriage and antibiotic resistance pattern of Pasteurella multocida isolates recovered from small ruminants over a period of 10 years in India. A total of 88 P. multocida isolates of small ruminant origin were subjected to virulence gene profiling for 19 genes by PCR and antibiogram study employing 17 different antibiotics. Virulence genes like e xbB , exbD , tonB , oma87 , sodA , sodC , nanB and plpB (100% prevalence) and ptfA and hsf-2 (>90% prevalence) were found to be uniformly distributed among isolates. Unexpectedly, a very high prevalence (95.45%) of pfhA gene was observed in the present study. Dermonecrotoxin gene ( toxA) was observed in 48.9% of isolates with highest occurrence among serotype A isolates and interestingly, one of each isolate of serotype B and F were found to carry this gene. Antimicrobial susceptibility testing revealed 17.04% isolates to be multidrug resistant. Amongst all the antibiotics tested, most of the P. multocida isolates were found to be susceptible to enrofloxacin and chloramphenicol. This study highlights novel epidemiological information on frequency and occurrence of virulence genes among Indian isolates from small ruminants. [ABSTRACT FROM AUTHOR]

Abstract: Listeria monocytogenes isolated from Ganges water, human clinical and milk samples were characterized by antibiotic susceptibility, serotype identification, detection of virulence genes and ERIC- and REP-PCR fingerprint analyses. All isolates were uniformly resistant to ampicillin, except two isolates, and showed variable resistance to gentamicin, cotrimoxazole, ofloxacin, rifampicin and tetracycline. Of the 20 isolates found positive for pathogens, seven (four human and three water isolates) belong to serogroups 4b, 4d and 4e; six (one human and five water isolates) belong to serogroups 1/2c and 3c; four milk isolates belong to serogroups 1/2b and 3b; and three milk isolates belong to serogroups 1/2a and 3a. Two water isolates, all human isolates, except one (Pb1) lacking inlJ gene, and three milk isolates possess inlA, inlC, plcA, prfA, actA, hlyA and iap genes. The remaining water and milk isolates showed variable presence of inlJ, plcA, prfA, and iap genes. ERIC- and REP-PCR based analyses collectively indicated that isolates of human clinical samples belong to identical or similar clone and isolates of water and milk samples belong to different clones. Overall study demonstrates the prevalence of pathogenic L. monocytogenes species in the environmental and clinical samples. Most of the isolates were resistant to commonly used antibiotics. [Copyright &y& Elsevier]

Background & objectives: Limited information is available on shiga toxin producing Escherichia coli (STEC) in animals and birds from India. An outbreak of acute diarrhoea in poultry birds at Aizawl, Mizoram was investigated for detection and characterization of STEC and enteropathogenic E. coli (EPEC). Methods: E. coli was isolated and identified from rectal swabs, intestinal contents, heart blood and spleen of 19 poultry birds that died due to acute diarrhoea during the outbreak. Phenotypic characterization was done by standard bacteriological and biochemical techniques. All the isolates were serotyped based on their somatic antigens. Virulence genes (stx1, stx2, eaeA and hlyA) were detected by multiplex PCR assay. Results: A total of 42 E. coli isolates were obtained, of which 24 belonged to 3 serogroups (064, 089 and O91) and the remaining 18 were untypable (UT). Altogether, 14 (33.33%) isolates carried at least 1 virulence gene, of which 10 (23.81%) and 4 (9.52%) were recorded as STEC and EPEC, respectively. Of the 10 STEC isolates, one carried only stx2, one carried stx2 and hlyA, four carried stx1, stx2 and hlyA, two carried stx1, eaeA and hlyA genes and two carried stx1 and eaeA. Of the four EPEC isolates, two carried eaeA and hlyA, one carried only eaeA gene and 1 carried only hlyA gene. [ABSTRACT FROM AUTHOR]

Background & objectives: Bacillus cereus is one of the pathogens responsible for human diarrhoea, mainly due to consumption of contaminated food. The present study was undertaken to determine the occurrence of B. cereus among diarrhoeal patients and its phenotypic and genetic characteristics that determine the virulence and clonal features. Methods: Stool specimens were collected for two years from acute diarrhoeal patients attending the two referral hospitals in Kolkata. Presence of virulence genes in B. cereus was determined by PCR. Clonality was assessed by pulsed-field gel analysis (PFGE) by restriction digestion with SmaI and NotI enzymes. Enterotoxins were detected by haemolysin assay and using BCET-RPLA kit. Invasion assay was done on Hep-2 cell line. Antimicrobial susceptibility was tested by disc diffusion method. Results: B. cereus was identified in 54 (3.5%) of the 1536 diarrhoeal cases studied. Majority of the isolates were susceptible to many antibiotics but showed resistant to amoxyclav and cephalosporins. Six genes covering the two different enterotoxic complexes determining the pathogenicity of B. cereus have been characterized by PCR. The nhe genes were detected in a higher proportion than hbl. Except in two, clonal diversity was noticed among 21 B. cereus isolates. Haemolytic enterotoxin was detected in 76 per cent of the isolates. Majority of the isolates (67%) produced in vitro enterotoxin (BCET) confirming its involvement in the infection. Interpretation & conclusions: Though the presence of B. cereus was not high in patients with diarrhoea, several virulence factors confirm their association with diarrhoea. Distinct clonality was identified in majority of the isolates indicating their origin from different sources. [ABSTRACT FROM AUTHOR]

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. • APEC is most commonly associated with septicemia in chickens. • The frequencies of occurrence of different VAGs such as iroN, ompT, hly, iss and iutA genes in the APEC was 47.2, 97.2, 61.1, 100, and 97.2%, respectively. • The E. coli isolates showed the highest resistance to vancomycin (61%) and high sensitive to cefotaxime/clavulanic acid combination. • Gallibacterium anatis detected in septicemic chickens by PCR. [ABSTRACT FROM AUTHOR]

The burden of antimicrobial resistance (AMR), particularly in India has been increased alarmingly. Methicillin-resistance in Staphylococcus aureus has been recognized as serious threat to the human especially if they are biofilm forming and equipped with virulence factors. In the present study monitoring of antibiotic resistant S. aureus was performed at three selected sites in Assam, India in August 2019 and February 2020. Ethnographic information was collected from the fish vendors in order to track and address potential sources of contamination. Twenty three potential methicillin-resistant S. aureus (MRSA) isolates were identified from the fish sold by the vendors and subjected to molecular characterization. The antimicrobial resistance profile of these MRSA isolates were regarded as multidrug-resistant (MDR) as they were resistant to ≥3 classes of antibiotics. The most prevalent resistance profile was; ampicillin-cefazolin-cefoxitin-gentamicin-norfloxacin-oxacillin-penicillin. Accessory gene regulators III (agr III) type MRSA (18/23, 78.26%) were found to be predominant compared to agr I type (5/23, 21.74%). Four isolates (17.39%) were observed to carry SCC mec -IV elements, which is a typical feature of community-associated MRSA (CA-MRSA). Two SCC mec -IV MRSA isolates were found to harbour panton-valentine-leucocidin (PVL) toxin genes and were resistant to macrolide in addition to beta-lactams. MLST and spa typing identified all MRSA as ST88 with spa type t2526. This is the first report from India on the incidence of ST88-SCC mec -IV (ST88-IV) MRSA in a fish market and its aquatic environs. The high prevalence of a single MLST clone, ST88, suggests that this lineage has a unique survival advantage in this environment. The study discusses the contribution of hospital wastewater in the dissemination of pathogenic MRSA clones to aquatic resources and then to humans through the food chain. • First report from India on the incidence of ST88-IV MRSA clone from market food fish. • Documented the prevalence of MRSA belonging to t2526; a very rare spa type. • Toxigenic potential of MRSA was described by the high prevalence rate of pvl , seb , seg and sei loci. • ica AD locus, clfB , fib and fnbB were the genes identified in MRSA to be associated with biofilm formation. • agr I and III were reported to be the prevalent allelic types. [ABSTRACT FROM AUTHOR]

The present study investigated the prevalence of Extended-Spectrum Beta Lactamase (ESBL) -producing E. coli and K. pneumoniae from the food fishes in retail markets in Assam, India. A total of 54 ESBL-producing E. coli and 12 K. pneumoniae isolates were recovered from 79 fish samples and were analyzed for antimicrobial resistance genes (ARGs) and virulence genes. E. coli isolates were categorized as multi drug resistant with resistance up to 12 different antibiotics with multiple antibiotic resistances (MAR) index ranging from 0.26 to 0.63. In E. coli, 100% resistance to cefotaxime along with 6% resistance to ceftazidime (third-generation cephalosporins) was observed. Moreover, 85% of the E. coli isolates were resistant to cefepime, a fourth-generation cephalosporin. K. pneumoniae showed resistance to 11 different antibiotics with MAR index value ranging from 0.21 to 0.57. All K. pneumoniae isolates showed 100% resistance to cefotaxime, 67% resistance to ceftazidime and 75% resistance to cefepime. Molecular characterization of ARGs revealed the presence of CTX-M group 1(CTX- M -15) in almost all E. coli isolates (98%, n = 53) and 100% in K. pneumoniae. A combination of uniplex and multiplex PCRs revealed fewer ARGs in E. coli isolates, with each isolate carrying 3 to 5 genes (tetA , dfrA1, sul1, sul2, qnrB, qnrS, aac(6ʹ)-Ib-cr). Majority of the E. coli were assigned to low-virulence phylogroup B1 and A while 8% of them belonged to pathogenic phylogroup D. 31 unique genetic profiles were identified for E. coli isolates by Pulsed-Field Gel Electrophoresis (PFGE) typing. K. pneumoniae isolates were highly diverse with 11 unique genetic profiles and a substantial ARG profile (bla TEM , bla SHV , bla OXA-1-like , tetA, strA, strB, dfrA1, sul1, sul2, qnrB, qnrS, aac(6ʹ)-Ib-cr, oqxA, oqxB). The frequency of ARGs ranged between 4 and 11. All K. pneumoniae isolates belonged to capsular serotype with wzi gene. Virulence gene iutA was prominent in all isolates while ybtS and kfu were confirmed in two isolates. Our findings raise concerns that fishes bought for consumption may serve as potential reservoirs of AMR genes and pose serious threat to public health. The study emphasizes the need for extensive surveillance of resistant strains in aquaculture and related settings, their in-depth analysis of population structure and transmission dynamics. • CTX-M group 1 (CTX- M -15), was the dominant Extended-Spectrum Beta Lactamase (ESBL) found in E. coli (98%) and K. pneumoniae (100%) recovered from food fishes in India. • The majority of E. coli were assigned to low-virulence phylogroup B1 (40%) and A (30%) while 8% of them belonged to pathogenic phylogroup D. • Each E. coli isolate carried 3 to 5 antimicrobial resistance genes (ARGs) (tetA , dfrA1, sul1, sul2, qnrB, qnrS, aac(6ʹ)-Ib-cr). • K. pneumoniae isolates were highly diverse with 11 unique PFGE profiles and a substantial ARG profile (bla TEM , bla SHV , bla OXA-1-like , tetA, strA, strB, dfrA1, sul1, sul2, qnrB, qnrS, aac(6ʹ)-Ib-cr, oqxA, oqxB). • Co-existence of ESBL genes with other ARGs such as quinolones resistance (qnrS and qnrB), tetracycline resistance (tetA), fluoroquinolones (oqxB and oqxA), sulphonamides-trimethoprim (sul2 and dfrA1) and streptomycin resistance genes (strA and strB). • K. pneumoniae belonged to capsular serotype (wzi gene) and harboured virulence genes iutA , ybtS and kfu. [ABSTRACT FROM AUTHOR]

Objective: To study the significance of enteroaggregative Escherichia coli (EAEC) as a pathogen causing acute diarrhea and a commensal in healthy nourished and malnourished children younger than five years of age in the Chandigarh region and to address possible traits of EAEC virulence genes, biofilm formation, phylogroups, and antibiotic resistance that would be correlated with diarrhea or carriage., Study Design: Stool samples were obtained from children with acute diarrhea (n = 548), as well as nourished (n = 550), and malnourished controls without diarrhea (n = 110). E coli isolates were confirmed as EAEC by pCVD432 polymerase chain reaction. Multiplex polymerase chain reactions were used to identify 22 virulence-related genes and phylogeny. Antibiotic susceptibility, adherence, and biofilm-forming potential also were studied., Results: Overall, 16.6% of children were malnourished. EAEC detection was greater among children with acute diarrhea (16%) than nourished (6%) and malnourished nondiarrheal controls (2.7%). We found an association of EAEC infections with age <2 years (P = .0001) in the diarrheal group. Adhesive variants adhesion fimbriae IV and adhesion fimbriae II were significantly associated with diarrhea. The aggR and aar genes showed a positive and negative association with the severity of disease (P = .0004 and P = .0003). A high degree of multidrug resistance was found (73.8%) in the diarrheal group. Most EAEC strains from the diarrheal group belonged to B2 and D phylogroups, whereas strains from non-diarrheal groups, which belonged to phylogroup B1., Conclusions: EAEC is a significant contributor to childhood diarrhea, its presence as a commensal, and the significance of the association of various virulence factors among the EAEC isolated from diarrheal and non-diarrheal stools. These data reinforce the importance of aggR and aar as positive and negative regulators and the contribution of AAF/II and AAF/IV fimbria for the pathobiology of EAEC., (Copyright © 2021 Elsevier Inc. All rights reserved.)

High-risk hospital-associated multidrug-resistant (MDR) Enterococcus faecalis clonal complexes (CCs) such as CC2 and CC87 are enriched with virulence determinants that help to accumulate, colonize, and cause serious nosocomial infections. The aim of this study was to establish the epidemiology and clonal composition of 134 clinical E. faecalis isolates and to link molecular typing data with antimicrobial resistance and virulence determinants. All isolates were identified by conventional methods and confirmed by polymerase chain reaction (PCR) (16srRNA gene and ddl genes of E. faecalis / E. faecium) in 5-years. Disc diffusion test was performed on all strains. We screened all E. faecalis for aac(6′)-aph(2″) , van A, and van B resistance genes, and aggregation substance- asa1 , cytolysin- cylA , collagen-binding protein- ace , enterococcal surface protein- esp , gelatinase- gelE, and hyaluronidase- hyl virulence genes by PCR. Representative isolates of E. faecalis were characterized by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). Out of 539 patients with enterococcal infections, 134 (24.9%) had E. faecalis infections, 366 (67.9%) had E. faecium infections, and 39 (7.2%) had infections due to other enterococcal species. Of the 134 isolates, 79.1% and 61.9% isolates were high-level gentamicin resistant (HLGR) and MDR. In multivariate analysis, independent predictor for infection due to MDR E. faecalis strains was a surgical intervention (OR 2.41, 95% CI 1.17–4.96, P = 0·017). Overall, the observed rate of in-hospital mortality was 11.9%. The gelE, asa1, ace, cylA, esp and hyl genes were detected in 87.3%, 78.4%, 54.5%, 53.7%, 36.6% and 3.0%, respectively in E. faecalis isolates. The asaI, cylA, and gelE genes were significantly correlated with MDR E. faecalis. The PFGE analysis showed 28 clones with four major clones. MLST analysis revealed two sequence types-ST28 (CC87) and ST181 (CC2). This is the first Indian report on the emergence of the high-risk hospital-associated worldwide-disseminated ST28 (CC87) and ST181 (CC2), which have enriched with multiple virulence determinants and resistance to antibiotics, paticularly ampicillin. This report indicates serious health concern and calls for on-going surveillance, close monitoring, and improved infection control procedures to stop further spread of these isolates. Unlabelled Image • Aimes: To establish the epidemiology and clonal composition of 134 clinical E. faecalis isolates and to correlate antimicrobial resistance and virulence determinants. • A total of 539 enterococci, 134 (24.9%) were E. faecalis , 366 (67.9%) were E. faecium , and 39 (7.2%) were other enterococcal species. Of the 134 E. faecalis isolates, 79.1% and 61.9% isolates were HLGR and MDR. • The gelE, asa1, ace, cylA, esp and hyl genes were detected in 87.3, 78.4, 54.5, 53.7, 36.6 and 3.0% in E. faecalis isolates respectively. • This is the first report from India on the emergence of the high-risk hospital-associated worldwide-disseminated ST28 (CC87) and ST181 (CC2) enriched with multiple antibiotic resistance and virulence determinants. [ABSTRACT FROM AUTHOR]

Most environmental parameters have no consistent effect on the expression of bacterial genes responsible for their virulence. However, as fish are poikilothermic, the possibility of temperature variation having a pronounced effect on the expression of virulence-associated gene(s) of bacteria infecting the host needs to be investigated. In this study, the diversity of virulence genes in seven Aeromonas hydrophila isolates collected from diseased fish from different parts of India was characterized, and the effect of temperature variation on the extent of expression of their virulence was investigated. All bacterial isolates were screened for a total of nine bacterial virulent genes {aerolysin, hemolysin, cytoen, outer membrane protein TS (Omp TS), elastase, flagellin, lipase, β hemolysin and type 3 secretion system}, and the diversity in their presence or absence were marked at a particular in vitro condition. Three bacterial isolates (nos. 1, 7 and 2) were selected for further study, based on their ability to cause varied mortalities (20–100%) in Labeo rohita juveniles in intraperitoneal challenge study. Further, three isolates were injected intraperitoneally into L. rohita fingerlings at three different temperatures (i.e., 20, 28 and 37 °C) and at 6 h post-challenge, the kidney samples were collected to measure the levels of all nine bacterial virulence genes using semi-quantitative PCR. The maximum level of amplicons of virulence genes in all three A. hydrophila isolates was noticed at 28 °C as compared to 37 °C and 20 °C. It was also observed that haemolysin played a more prominent role in the expression of virulence, when compared to cytoen gene. Hence, it was concluded that water temperature does play a crucial role in governing virulence gene expression, and a temperature of 28 °C would be considered as suitable for looking into the pathogenicity of A. hydrophila for conducting any challenge study with this organism in tropical environment. • Studied A. hydrophila virulence related genes in in vitro and in vivo pathogenicity. • Investigated pathogenicity of A. hydrophila at varied water temperatures in L. rohita. • T3SS identified as a possible marker for identification of pathogenic isolates of A. hydrophila. [ABSTRACT FROM AUTHOR]

The religious mass gathering and bathing can pose a multitude of significant public health challenges and lead to severe alterations in the river microbial ecology. The Pandharpur Wari is an annual pilgrimage of Maharashtra, India, where millions of devotees carry the footprints of the saint-poets and pay their obeisance to Lord Vitthal on the 11th day of moon's waxing phase (Ashadi Ekadashi). As a part of the ritual, the engrossed devotees, walk over 250 km, take a first holy dip in a sacred river Indrayani at Alandi and secondly in Bhima River at Pandharpur. The MinION-based shotgun metagenomic approach was employed to examine the impact of spiritual mass bathing on environmental changes (concerning the river microbial community structure and functions); and public health aspects (in terms of changes in the pathogenic potential and antibiotic resistance). The analysis of bathing and post-bathing samples of both the rivers revealed alterations in the alpha and beta diversity, indicating significant spatiotemporal variations in the overall microbial structure and function. Furthermore, the analysis revealed up to 80% of differences in the abundance of virulence genes between the bathing and post bathing samples. We observed parallel increase of priority skin and enteric pathogens (ranging from 11% to 80%) such as Acinetobacter baumannii, Staphylococcus aureus , Streptococcus pyogenes, Mycobacterium tuberculosis , and Pseudomonas aeruginosa during the bathing event. Moreover, we observed a significant increase in the antibiotic resistance in the bathing samples of Bhima and Indrayani rivers respectively. Altogether, this is the first comprehensive metagenomic study unravelling the influence of religious mass-bathing on the riverine ecosystem. Image 1 • Study deciphers the impact of ritualistic mass bathing on the river microbial ecology. • Mass bathing resulted in alterations in the microbial community and functionality. • MinION sequencing revealed escalated ARGs in both the rivers during mass bathing. • Bathing samples showed a higher prevalence of priority pathogens and virulence trait. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus is an important shrimp & human pathogen and a special strain of it harbouring toxic plasmid can cause Acute Hepatopanreatic Necrosis Disease (AHPND) to shrimp. An attempt was made to isolate and further characterize this bacterium from shrimp farms of south east coast of India. From 381 shrimp farms sampled over a period of four years, 105 isolates were presumptively identified as V. parahaemolyticus. All the 105 isolates were found to be negative for PirA and PirB toxin, characteristic of AHPND V.parahaemolytiucs. The challenge experiments with representative isolates required higher number of bacteria to cause shrimp mortality. The isolates were negative for thermostable direct haemolysin (tdh), tdh related haemolysin (trh) and type III secretion system alpha and beta. The isolates were tested to be sensitive to majority of the antibiotics. This study thus indicates that V. parahaemolyticus isolates recovered from aquaculture ponds of Southeast coast of India are non-shrimp pathogenic and non-human pathogenic. Direct tissue samples (hepatopancreas and stomach) and Tryptic Soy Broth enriched bacteria population from these tissues were also found to be negative for PirA and PirB toxin, characteristic of AHPND. Based on this evidence, AHPND was not detected in any of the samples collected from these shrimp culturing areas. • Vibrio parahaemolyticus was found to be distributed in shrimp farms and other surrounding aquatic environments. • All the isolates were found to be non-virulent and non-pathogenic to shrimp and confirmed to belong non-AHPND group. • The isolates were found to be diverse and belong to at least 4 different phylogroups. • All the isolates were found to be sensitive to majority of the antibiotics tested. [ABSTRACT FROM AUTHOR]

Salmonella is a major cause of morbidity and mortality in humans worldwide, and the infection with multidrug-resistant strains can cause severe diseases. Many coastal cities around the world discharge their wastewaters into the marine environment. These wastewaters contain a variety of pathogenic microorganisms that may have a role in the contamination of this ecosystem and have potential risks for public health. Using an environmental approach, the present study investigated the presence of Salmonella in sediment and water samples collected from Port Blair Bays. In this environmental approach, the provided information about the diversity of the Salmonella serovars, antibiotic resistance and the prevalence of virulence factors in Salmonella, especially from the coastal waters of Port Blair Bays. The occurrence of Salmonellae was significantly higher in water column samples (2.9%) than in those taken from the marine sediments (0.7%). Of the 133 positive Salmonella strains, 22 different serovars were identified. Salmonella enterica serovar Senftenberg was the predominant serovar, being represented by 54 isolates (42.5%), followed by serovar Typhimurium (19 isolates [15%]) and serovar Agona (12 isolates [9.4%]). The presence of virulence genes (filC, sitC, hilA, invA, sipC, hilD, hilC, invF, invE, invH, sipF, aadA, pare, gyrA, spaP and parC) and susceptibility studies with 10 selected antibiotics were also performed. The results of this study revealed that all Salmonella isolates were positive for targeted virulence genes and were resistant to at least one antibiotic. Antibiotic susceptibility studies revealed the presence of multidrug resistant Salmonella strains in coastal water, which usually from land base sources end up in the marine environment and may pose a significant risk on public health., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The emergence of extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae has been increasing rapidly across the world. The presence of virulence factors in ESBL producers further adds to the pathogenicity and severity of infection, which often complicate empirical therapy and sometimes result in treatment failures. In the present study, 227 non-repeated clinical isolates of K. pneumoniae obtained from different clinical specimens from a tertiary care hospital in India were analyzed to detect the genes responsible for ESBL production (blaTEM, blaCTX-M, and blaSHV), virulence (fimH-1, mrkD, entB, irp-1), and capsule production (K1-K2). Phenotypically identified 72 ESBL producing K. pneumoniae isolates were further subjected to PCR based genotypic analysis but only 20 were found to have at least one of the ESBL producing genes. blaTEM was the most predominant gene (100%), followed by blaSHV (90%), and blaCTX-M (85%). Similarly, the most common virulence genes were fimH-1 (70%), entB (65%), markD (55%), irp-1 (25%), K1 (25%), and K2 (20%). REP-PCR profile separated them into five major clusters (I-V), indicating the existing heterogeneity among the isolates. The resistance profile data obtained from the present study can serve as the information base to understand the infection pattern prevailing in the hospital and for physicians to recommend suitable antibiotics for the patients.

Objectives: This study aimed to identify the multiple drug resistance mechanisms and various virulence genes in high-level carbapenem-resistant Pseudomonas aeruginosa (CARPA) isolates collected from patients hospitalised in a tertiary care hospital in South India., Methods: A total of 156 CARPA isolates were included in the study. Multiplex PCR was optimised to detect carbapenemase and extended-spectrum β-lactamase (ESBL) genes. Semi-quantitative reverse transcription PCR (RT-PCR) was optimised to evaluate oprD deficiency. Efflux pump activity was determined by phenylalanine-arginine β-naphthylamide (PAβN) assay, and expression of mexA and mexC genes was evaluated by real-time quantitative PCR (qRT-PCR). Presence of the virulence genes exoS, algD, algU, lasR and rhlR were detected by qRT-PCR and plcH and lasB by RT-PCR., Results: Genes encoding carbapenemases and ESBLs were detected in 48.7% (bla VIM , 23.1%; bla NDM-1 , 17.3%; bla VIM +bla NDM-1 , 7.1%; and bla IMP , 1.3%) and 4% (bla VEB , 4.5%) of CARPA isolates, respectively. Loss of porin OprD was observed in nine isolates (5.8%), two of which harboured the bla VIM gene. Among efflux pump-positive isolates, mexA expression was significantly upregulated. algD expression was detected in 93% of CARPA isolates, followed by algU (89%), rhlR (84%), lasR (81%) and exoS (76%). The lasB and plcH genes were detected in 94% and 92% of isolates, respectively., Conclusions: Co-existence of multiple antimicrobial resistance mechanisms together with virulence genes in carbapenem-resistant isolates has become an alarming emerging threat. Continuous monitoring of multidrug-resistant pathogens is important for clinicians in order to determine therapeutic options against such infections., (Copyright © 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

Objective: The study was conducted to determine the association of the hlyD, papC and cnf-1 virulence genes with drug resistance in uropathogenic Escherichia coli (UPEC) isolated from cases of urinary tract infection (UTI)., Method: A total of 193 E. coli strains isolated from symptomatic cases of UTI in a tertiary care teaching hospital in Raichur, Northern Karnataka, India were included in the study. The antibiotic susceptibility pattern was determined by Kirby-Bauer's Disk Diffusion method, and the strains resistant to any of the third generation cephalosporins tested were further confirmed for extended-spectrum beta-lactamase (ESBL)-production by an E-strip test. Genotypic virulence markers, namely, hlyD, papC and cnf-1, were detected by the uniplex PCR method and the phylogenetic characterization was performed by a multiplex PCR assay., Results: The majority of the E. coli isolates belonged to the B2 phylogenetic group were significantly associated with ciprofloxacin-sensitivity and non-ESBL production (p<0.05). An increased prevalence of ciprofloxacin-sensitive strains over ciprofloxacin-resistant strains were observed among the UPEC isolates harboring the papC (72.9% vs. 40.2%; p<0.001), hlyD (43.7% vs. 21.6%; p<0.001) and cnf-1 (30.2% vs. 12.3%; p<0.05) genes. The presence of a multivirulent gene in the non-ESBL E. coli strains (44.5%) was significantly higher (p<0.05) than in the ESBL-producing strains (21%)., Conclusions: Among the UPEC isolates, the predominant B2 phylogenetic group was significantly associated with the ciprofloxacin-sensitive strains, as well as with the non-ESBL E. coli strains. The genotypic virulence markers of UPEC were associated with ciprofloxacin-sensitivity, and a significant number of the non-ESBL strains harbored multivirulent genes. The relationship between the presence of the virulence genes and ESBL production was complex and warrants further intensive studies., (Copyright © 2014 King Saud Bin Abdulaziz University for Health Sciences. Published by Elsevier Ltd. All rights reserved.)

We identified 281 Vibrio cholerae non-O1, non-O139 strains from patients with diarrhea in Kolkata, India. Cholera-like diarrhea was the major symptom (66.0%); some patients (20.3%) had severe dehydration. These strains lacked the ctxA gene but many had hlyA, rtxA, and rtxC genes. Pulsed-field gel electrophoresis showed no genetic link among strains.