Antibiogram, Genomic and Phylogeny of Stool and Seafood Isolates from Some Cholera-Prone Coastal Communities in Rivers State, Nigeria

Cholera is a significant public health threat across the globe, especially in coastal regions with poor water supply. This study was carried out to determine the antibiogram, genomic, and phylogeny of stool and seafood isolates from some cholera-prone coastal communities in Rivers State, Nigeria. A total of 400 stool samples and 42 different seafood were aseptically collected and examined using standard microbiology and molecular techniques. An antibiogram of isolates from seafood and stool samples was assayed. Genes for virulence, resistance, and relatedness of bacteria identified were also determined. The isolates from the stool and seafood were examined for susceptibility to some selected antibiotics. The findings showed the prevalence rate of cholera in the communities as follows: 16% in Kaa, 30% in Andoni, 4% in Ogu/Bolo, and 10% in Abua/Odual. The isolates from stool were susceptible to Ciprofloxacin and Gentamycin with a susceptibility rate of 94.12% each while 100% resistance was recorded against Amoxicillin-clavulanic acid, 94.12% against Amikacin and 88.24% against Colistin. For the sea foods, the isolates were susceptible to gentamycin and ciprofloxacin with a susceptibility rate of 91.43% and 82.86% respectively. Resistance was also recorded against Colistin (88.57%) and Azithromycin (82.86%). Testing the isolates for the presence of 16SrRNA genes showed that all were positive with 1500 bp 16SrRNA gene band size. TEM, OXA, SHV, and CTX-M resistant genes were detected whereas the virulence genes were TDH and AcrB. The phylogenetic analysis revealed isolates from seafood to be Aeromonas dhakensis, Vibrio parahaemolyticus, Vibrio azureus, and Providencia rettgeri, while in stool samples they were Enterobacter sichuanensis, Enterobacter hormaechei, Providencia sneebia, and Proteus vulgaris. Providencia stuartii, Proteus mirabilis, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae were common isolates from both seafood and stool samples. This study has shown that not all reported cases of cholera are caused by Vibrio cholerae. Therefore, attention should be paid to other water-borne bacteria in every outbreak, especially in coastal communities.

Surveillance of multidrug resistant uropathogenic bacteria in hospitalized patients in Indian

Objective:To record surveillance,antibiotic resistance of uropathogens of hospitalized patients over a period of 18 months.Methods:Urine samples from wards and cabins were used for isolating urinary tract infection(UTI)-causing bacteria that were cultured on suitable selective media and identified by biochemical tests;and their antibiograms were ascertained by Kirby-Bauer's disc diffusion method,in each 6-month interval of the study period,using 18 antibiotics of five different classes.Results:From wards and cabins,1 245 samples were collected,from which 996 strains of bacteria belonging to 11 species were isolated,during April 2011 to September2012.Two Gram-positive,Staphylococcus aureus(S.aureus)and Enterococcus faecalis(E.faecalis),and nine Gram-negative bacteria,Acinetobacter baumannii,Citrobactcr sp.,Escherichia coli,Enterobacter aerogenes,Klebsiella pneumoniae.Klebsiella oxytoca,Proteus mirabilis,Proteus vulgaris and Pseudomonas aeruginosa were isolated.Both S.aureus and E.faecalis were vancomycin resistant,and resistant-strains of all pathogens increased in each 6-month period of study.Particularly,all Gram-negatives were resistant to nitrofurantoin and co-trimoxazole,the most preferred antibiotics of empiric therapy for UTI.Conclusions:Antibiograms of 11 UTI-causing bacteria recorded in this study indicated moderately higher numbers of strains resistant to each antibiotic studied,generating the fear of precipitating fervent episodes in public health particularly with bacteria,Acinetobacter baumannii,Escherichia coli,Klebsiella pneumoniae and S.aureus.Moreover,vancomycin resistance in strains of S.aureus and E.faecalis is a matter of concern.

Prevalence and antibiogram study of Salmonella and Staphylococcus aureus in poultry meat

Objective:To evaluate the presence and antibiogram pattern of Salmonella and Staphylococcus aureus(S.aureus)in retail poultry meat products.Methods:Foodborne pathogens(Salmonella and S.aureus)were isolated from poultry meat and confirmed with the help of biochemical and immunological test.Antibiogram of the isolates were examined by following CLS1 methods.Results:A total number of 209 poultry meat samples were collected and studied in this study.Out of which,5.26%were found contaminated with Salmonella while 18.18%were found contaminated with S.aureus.All the Salmonella and S.aureus isolates were found resistant to at least one antibiotic.About 72.72%of the Salmonella isolates showed resistance to tetracycline,while S.aureus isolates were also found highly resistant to tetracycline equal to 44.73%.One of the Salmonella isolates showed multi-drug resistance to almost six antibiotics out of nine antibiotics used in the study.Multidrug resistant S.aureus isolates were also found in the study.Conclusions:The study confirmed the presence of Salmonella and S.aureus in retail poultry meat.It is a potential threat to consumer health.To reduce the risk of contamination,good hygiene practices are necessary from processing to storage.

Prevalence and antimicrobial resistance of Salmonella spp.in raw retail frozen imported freshwater fish to Eastern Province of Saudi Arabia

Objective:To determine the proportion of imported frozen fish contaminated with Salmonella among retail food stores and supermarkets in the Eastern Province of Saudi Arabia.Methods:A total of 223 frozen freshwater fish purchased from different supermarkets and grocery stores were analyzed for the presence of foodborne pathogen Salmonella.The isolation of Salmonella was determined and confirmed by using the methods of US Food and Drug Administration's Bacteriological Analytical Manual.CHROMagar Salmonella plus,biochemical tests and API 20E strips.Antimicrobial susceptibilities of Salmonella isolates were determined by the disk diffusion method on Muller-Hinton agar,as described by Kirby-Bauer.in accordance with the guidelines of the Clinical and Laboratory Standards Institute.Results:Out of the total 223 fish samples(20 of catfish,18 of carfu,20 of mirgal,25 of milkfish,35 of mackerel,75 of tilapia,and 30 of rohu),89(39.9%)were tested positive for Salmonella.The prevalence of positive samples were reported for the freshwater fish of pangas(60.0%,n=12),carfu(27.7%,n=5),mirgal(35.0%,n=7),milkfish(52.0%,n=13),mackerel(31.4%,n=11),tilapia imported from Thailand(64.0%,n=16),tilapia imported from India(28.0%,n=14),rohu imported from Thailand(26.6%,n=4)and rohu imported from Myanmar(46.6%,n=7).A total of 140 isolates of Salmonella spp.were yielded from at least seven different types of frozen freshwater fish imported from 5 different countries and were tested for their susceptibility to 16 selected antimicrobial agents.The highest antibiotic resistance was observed to tetracycline(90.71%)followed by ampicillin(70%)and amoxicillin-clavulanic acid(45%).Conclusions:The obtained results of this study shows that these raw retail imported frozen freshwater fish are contaminated with potentially pathogenic Salmonella spp.And the study recommend and suggest that there is a need for adequate consumer measures.

Characterization, prevalence and antibiogram study of Staphylococcus aureus in poultry

Objective: To reveal the presence of methicillin resistant Staphylococcus aureus(S. aureus)(MRSA) in poultry samples and to determine the antibiogram pattern against five antibiotics.Methods: Samples from different poultry farm of Chittagong city, Bangladesh were examined for S. aureus by different biochemical tests and confirmed as MRSA by identifying the presence of mec A gene using PCR. Antibiotic resistance pattern in S. aureus was determined by antibiotic disk diffusion method.Results: In this study, a total of 60 samples(30 from nasal swabs and 30 from cloacal swabs) were used, of which 54 were confirmed as S. aureus by different biochemical tests. Among these, 12 were confirmed as MRSA by detecting mec A gene using PCR.During antibiogram study, both nasal and cloacal samples showed the highest resistance against penicillin-G and the lowest resistance was observed against neomycin.Conclusions: Based on the present study, it can be said that different antibiotics are used extensively in poultry that leads to MRSA and is alarming for human health.

Antimicrobial Resistance and Plasmid Profiles of <i>Salmonella enterica</i>Serovars from Different Sources in Lagos, Nigeria

Introduction: Salmonella enterica Serovars remains one of the leading pathogens that cause diarrhoea and bloodstream infections in developing countries. The emergence of multidrug resistant (MDR) Salmonella has become a serious problem globally. This study investigated the antibiotic resistance and plasmid profiles of Salmonella isolates from different sources. Methods: Seventy-three samples comprised of clinical (30), hand swab (15), food (10) and water (18) were analyzed bacteriologically. Salmonella isolates were identified and characterized by standard procedures. Isolates were subjected to antimicrobial susceptibility testing and were further screened for plasmid DNA by standard methods. Results: A total of 27 Salmonella isolates made up of 5 (18.5%) S. typhi, 6 (22.2%) S. enteritidis, 9 (33.3) S. typhimurium, 5 (18.5%) S. cholerasuis, and 1 (3.7%) each of S.arizonae and S. vichow were obtained in this study. All the isolates developed resistance to three or more antibiotics evaluated. Four distinct resistance profiles: TetAmpCol, TetAmpColCot, TetAmpColCip and TetAmpColCotCip were recorded with 63% of the isolates exhibiting resistance profile TetAmpColCot. Specifically 23 of 27 (85.2%) of the isolates harboured plasmid DNA comprised of 12 distinct plasmid profiles of different sizes ranging from 3.2 kb to 30.2kb. Salmonella isolates of the same species from different sources differed in plasmid profile. Plasmid profile was found to show good discriminatory capability compared to antibiotics resistance profile. Conclusion: This study revealed that both resistance antibiogram and plasmid profile are still viable epidemiological tools for tracing the source of Salmonella isolates. A need for prudent use of antibiotics is suggested.

Urinary Tract Infections Identified in the Preoperative and Their Sensitivities to Antibiotics

Introduction: Urinary tract infection is a public health problem. Patients who have to undergo urology surgery are exposed to infectious complications if there is an untreated urinary tract infection. The objective of our study is to identify all the germs responsible for urinary tract infections discovered in the preoperative assessment of hospitalized patients with their susceptibility and antibiotic resistance profiles. Patients and Methods: This is a retrospective and descriptive study of 124 cases of cytobacteriological examination of urine collected over 2 years from January 1st, 2016 to December 31st, 2017, performed in the urology department of the University Hospital of Bocar S. Sall of Kati. The parameters studied were: age, sex, reason for admission, the diagnosis chosen, the result of the cytobacteriological examination of urine and the antibiogram. Results: We collected 124 cases of cytobacteriological examination of urine during this period. The average age of our patients was 58 years old with extremes ranging from 3 years to 84 years. The sex ratio was 6 in favor of men. 86 patients, or 69%, were admitted for micturition disorders. In 53.2%, the diagnosis was BPH followed by urolithiasis 22.6%. Cytobacteriological examination of urine identified 55 cases of urinary infection, 44%. In 34 cases or 60%, the germ responsible was an E. coli. Two germs were sensitive only to one antibiotic. Conclusion: This study made it possible to have an idea about the antibiotic susceptibility of germs involved in urinary tract infections. Awareness-raising for the proper use of antibiotics needs to be strengthened to minimize the risk of increasing antibiotic resistance.

A Retrospective Cross Sectional Study of Blood Culture Results in a Tertiary Hospital, Ekiti, Nigeria

The current study was conducted to determine the epidemiology and antibiotic sensitivity pattern of bacteria isolated from blood of septicemic patients for improved antibiotic therapy. A three-year descriptive study was done at Microbiology Laboratory, Ekiti State University Teaching Hospital, Ado Ekiti, from April 2012 to April 2015. Information compiled from patients’ records includes age, sex, isolated organisms and antibiotic susceptibility patterns. Three hundred and thirteen blood cultures were collected from neonatology and pediatrics wards, Out Patients’ Department (OPD) and from other adult patients. Forty one culture plates yielded mono microbial growth (no polymicrobial growth), giving an incidence of 13.1% positive blood culture (N = 41/313). There were 58.4% Gram negative bacilli and 41.6% Gram positive cocci in the microbial growth. Bacteria isolated were Staphylococcus aureus 34% (14/41), Klebsiella species 22% (9/41), Enterococci 17% (7/41), Proteus species 12% (5/41), Escherichia coli 7% (3/41) and Streptococcal pneumoniae 7% (3/41). There was a (35%) higher occurrence of septicemia in neonates than in any other age groups in the hospital. Bacterial sensitivity to 13 antibiotic agents was determined by antibiotics disc diffusion using modified Kirby Bauer’s method. Gram-positive organisms showed a higher antibiotic sensitivity ranging from 14% - 100% than the Gram-negative bacteria (11% - 80%). Staphylococcus aureus and Klebsiella species are the most prevalent organisms. The third generation Cephalosporins (Ceftriaxone) and Floroquinolone (Levofloxacin, Ofloxacin) have proved reliable for management of these blood infections.

Antimicrobial Resistance among Enterobacteriaceae Found in Chicken and Cow Droppings and Their Public Health Importance

Introduction: The recent surge in the number of antimicrobial resistant cases from hospitals and communities has created a need to study the points and sources of exposure to certain bacteria and determine their susceptibility to commonly used antibiotics. This study aimed at identifying and screening for drug-resistant Enterobacteriaceae isolated from chicken droppings and cow dungs in Onitsha, Anambra state, in the South-Eastern part of Nigeria. Methods: This is a cross-sectional descriptive study which included 50 chickens and 50 cow dung samples collected from five poultry houses and cow ranches respectively using sterile swab sticks. The samples were transported to the laboratory and processed following standard microbiological protocols. Isolates in the samples were recovered using MacConkey Agar, Eosin Methylene Blue Agar and Salmonella-Shigella Agar following standard microbiological procedures and then identified/characterized biochemically using commercial API 20E identification kits following the standard manufacturer’s protocol. Isolates were subjected to antibiotic susceptibility testing on Muller Hinton Agar using Kirby Bauer double-disc diffusion technique. The multiple antibiotics resistance index was determined as well. Isolates with reduced susceptibility to Ceftazidime were screened for extended spectrum beta-lactamase, AmpC- and metallo-beta-lactamase-production using Rosco Diagnostic kit. Results: Sixty-two (100%) Gram-negative bacteria were isolated from a total of 100 samples collected from both sites, out of which 43 (69.4%) are Enterobacteriaceae. A total of 30/43 (69.8%) Enterobacteriaceae including K. pneumoniae, S. enteritica, S. odorifera, E. coli, K. intermediate, P. stuartii, E. aerogenes, P. penneri, P. mirabilis and C. braakii were recovered from chicken droppings, whereas 13/43 (30.2%) Enterobacteriaceae including K. pneumoniae, S. enteritica, S. odorifera, E. coli, K. intermediate, P. stuartii, E. aerogenes, P. penneri, P. mirabilis and C. braakii were recovered from cow dungs. Two (12.5%) different isolates demonstrated metallo-beta-lactamase and cephalosporinase (AmpC) production. The isolates were susceptible to six antibiotics tested except Augmentin and Nitrofurantoin where the resistance is 100% and 85% respectively while Ceftriaxone and Ofloxacin had the best antibacterial activity against the isolates from both sites. Conclusion: The bacteria of public health importance isolated from these sites and their antibiogram profile have shown the need for proper monitoring and management of animal wastes in order to mitigate the threat to human health in the spirit of One Health as well as contribute to the fight against antibiotic resistance.

Molecular characteristics,antibiogram and prevalence of multi-drug resistant Staphylococcus aureus (MDRSA) isolated from milk obtained from culled dairy cows and from cows with acute clinical mastitis

Objectives: To study the molecular characteristics, antibiogram and prevalence of multidrug resistant Staphylococcus aureus(S. aureus)(MDRSA) isolated from milk obtained from culled dairy cows and from cows with acute clinical mastitis.Methods: Bacteria were cultured from 188 quarter milk samples obtained from cows before culling(n=139) and from cows affected with acute mastitis(n=49) belonging to 10 dairy farms. The bacteria were identified using colony moiphology, Gram staining and biochemical characteristics. S. aureus isolates were then subjected to molecular characterization using PCR targeting 16 S rRNA and mecA gene to identify Methicillin resistant S. aureus(MRSA). The antibiogram of all isolates was performed using the Kirby-Bauer disk diffusion method against 10 commonly used antibiotics in dairy farms.Results: S. aureus was isolated from 19(13.7%) samples obtained from culled cows and 11(22.4%) samples obtained from cows with acute mastitis. In both culled cows and cows with acute mastitis, in vitro antibiogram revealed that 100% of S. aureus isolates were resistant to erythromycin, penicillin G, streptomycin, doxycyclin, and trimethoprim/sulpha. The prevalence of MRSA in milk of culled cows and cows with acute mastitis was 26.3% and 18.2%, respectively, with an overall prevalence of 3.7% among all samples. All MRSA isolates were completely resistant to all tested antibiotics. All MRSA isolates were positive for the presence of the mecA gene.Conclusions: MRSA carrying the mecA gene were isolated from mastitic milk from dairy cows in Jordan for the first time. MRSA may pose a potential health risk to the public, farm workers and veterinarians.

Quality Assessment of Surface and Drinking Water of Nakla Paurosova, Sherpur, Bangladesh

Water is one of the most vital elements of ecosystem and human being, but unfortunately nowadays, the pollution of surface and drinking water is an alarming problem. The present work deals with the assessment of physicochemical and bacteriological profile of several pond, jar and tube-well water samples to ensure its suitability for using and drinking. Total 30 samples were randomly selected and collected from Nakla Paurosova of Sherpur district by following the standard procedure. Bacteriological analysis was carried out by following the standard bacteriological methods. Most of the surface water sampling points were polluted by dumping of waste, cattle wash and were not suitable for drinking or other domestic purposes. Among three heavy metals, only Iron was detected in six tube-well water samples, one was also positive to arsenic, rest of the water bodies were negative to all of these metals. In case of most of the water bodies, different physicochemical properties were below standard limit. In pond water, the Total Viable Count (TVC) ranged from 2.7 × 107 cfu/100ml to 4.4 × 1015 cfu/100ml and Total Coliform Count (TCC) were 3.4 × 105 cfu/100ml to 4.8 × 1013 cfu/100ml, where the mean concentration of Heterotrophic Plate Count (HPC) was 2.4 × 105 cfu/100ml and 1.8 × 105 cfu/100ml in jar and tube-well water respectively. On the other hand, the Total Coliform Count of supply water was 33 cfu/100ml and tube-well water was 14 cfu/100ml. Fecal coliform was detected in all of the pond water samples, four jar and three tube-well water too. E. coli was present in all pond water samples, and also detected in 80% supply and 50% tube-well water also. Shigella spp. was found in two pond water and in one supply water, where tube-well water was free from it. Salmonella spp. was also detected in 30% of pond and 20% of supply water, whereas absent in tube-well water. 50% of pond, 40% of supply and 30% of tube-well water were contaminated with Vibrio spp. The total counts of these pathogenic bacteria exceeded the acceptable limit both surface and drinking water and also showed resistance against a broad range of commercially available antibiotics. Survey-based result revealed that, peoples of the study area who were using or drinking these water, were suffering from various water borne diseases. These kinds of water sources pose a major threat to public health. So, public awareness, proper treatment and precise management are needed prior to use and drink of these water.

Serotypes, Antibiogram and Genetic Relatedness of <i>Pseudomonas aeruginosa</i>Isolates from Urinary Tract Infections at Urology and Nephrology Center, Mansoura, Egypt

Background: Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that represents a major problem in many hospitals because of its increased resistance to antibiotics and the ability to cause nosocomial infections. The present study aimed to phenotype and genotype isolates of P. aeruginosa from inpatients with UTIs at Urology and Nephrology center, Mansoura, Egypt to study their relatedness. Methods: Thirty nine isolates of P. aeruginosa were phenotypically typed by determination of O-serotypes by slide agglutination technique and antimicrobial resistance patterns by disk-diffusion method. The genetic diversity of isolates was illustrated by performing RAPD-PCR using M13 primer. Results: Serotypes O11, O6 and O10 were the most prevalent. Isolates showed high resistance rates to antipseudmonal antibiotics with high incidence (51.3%) of multidrug resistance (MDR). Amikacin was the most effective. A significant correlation was found between O6, O10 and MDR. A relatively high polymorphism was demonstrated among P. aeruginosa isolates by using RAPD-M13 fingerprinting. Cross transmission was suggested by phenotypically and clonally identical isolates. Conclusion: The study demonstrates the role of combining both classical and molecular typing as a valuable mean to study the origin and cross transmission of P. aeruginosa in UTIs for better assessment of treatment and infection control.

Occurrence of Typhoidal and Non-Typhoidal Salmonellae among Poultry Workers in the Southern, Nigeria

Typhoid fever and non-typhoidal salmonellosis remain endemic in most developing countries with large scale transmission through contaminated food and drinking water. Since the early 2000s, poultry has also been found to be a common food source for Salmonella enteritidis infections. A total of 500 blood, stool, cloaca swab and egg shell swab specimens were collected respectively from poultry, non-poultry workers and birds in Akwa Ibom State and screened for Salmonella species using standard cultural and serological techniques. The overall prevalence rate was 117 (23.4%). Salmonella species were distributed in all the three senatorial districts of the State, percentage distribution of Salmonella isolates was 42 (25%) for Uyo, 35 (20.8%) for Eket and 40 (23.8%) were isolated from Ikot Ekpene senatorial districts. Out of 372 human subjects screened 77 (20.7%) were positive and 38 (20.4%) were isolated from non-poultry workers while 39 (21.0%) were isolated from poultry Staff in the three senatorial districts. Out of 128 samples of poultry products analysed 40 (31.3%) were isolated. 23 (18%) were isolated from cloaca swabs while 17 (13.3%) were isolated from egg shell swabs. In this study, all the serological significant Typhoid fever cases were confirmed by blood culture. According to sero-grouping and source of sample collection 23.9% S. typhi was isolated from human followed by S. paratyphi A (20.5%) while S. enteritidis and S. gallinarum (19.7%). Among poultry products S. gallinarum, S. enteritidis, S, pullorum, S. paratyphi A, and S. typhimurium were isolated from bird Cloaca swab and Egg shell swab. Salmonella pullorum and S. typhi were not isolated from human and poultry respectively. High prevalence of Salmonella serovars such as S. gallinarium, S. typhi, S. Typhimurium and S. enteritidis with regular consumption of poultry and eggs without proper disinfection and appropriate boiling represent a serious public health risk in Akwa Ibom State. Demographic data obtained in this study showed that majority of the poultry workers across the three senatorial districts were within the ages of 21 to 30 (34%), 39% were none graduate while 30.4% were secondary school leavers. Fever was recorded as the highest clinical signs and symptoms followed by headache, abdominal cramps, diarrhoea and vomiting. Generally, all the isolates identified as Salmonella were tested for their susceptibility to antimicrobial agents. The results showed that Amoxacillin-Clavulanic acid was 100% susceptible to S. gallinarum, S. paratyphi A, 93% susceptible to S. typhi and 80% to S. typhimurium, S. enteritidis and S pullorum. Salmonella gallinarum was 100% sensitive to Ciprofloxacin and S. pullorum, whereas all isolates were 100% resistant to Oxacillin and Tylosin antibacterial agents used. The emergence of multiple drugs resistant Salmonella from human and poultry shows that the continuous use of drugs in poultry as growth promoters should be re-examined. Other factors as sources of Salmonella contamination in farms can be minimized by good hygienic practices and biosecurity measures.

Surveillance of Vibrio spp in Penaeus monodon Collected from Shrimp Pond of Satkhira, Bangladesh

Vibrio is the most common genera associated with crustaceans and often causing significant economic losses.Many Vibrio species are pathogenic to human and have been implicated in food borne diseases.The present study was conducted to identify Vibrio spp.from the tiger shrimp(Penaeus monodon)of shrimp pond at Satkhira,Bangladesh.A total number of 33 Vibrio species isolates were identified from 20 shrimp samples through a series of morphological,physiological and biochemical tests.The work reports the prevalence of Vibrio spp.in the pond environments and the existence of three Vibrio species such as V.alginolyticus,V.parahaemolyticus and V.harveyi were identified.In the study of antibiogram,all isolates were shown 100%sensitive to streptomycin,ciprofloxacin and chloramphenicol.Maximum 41%isolates were shown resistant to co-trimethaxozole whereas 30%and 24%resistant to azithromycin and novobiocin respectively.

Prevalence and Study of the Clonality of Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Strains in Neonatology at the University Hospitals of Abidjan by the Pulsed Field Gel Electrophoresis and the Quantitative Antibiogram

Background: ESBL-producing strains of Klebsiella pneumoniae, one of the main causes of nosocomial and hospital-acquired infections, are commonly associated with therapeutic impasses. Surveillance of these multidrug-resistant pathogens is a crucial tool for controlling and preventing infections. This surveillance involves the use of appropriate molecular and phenotypic typing techniques. The choice of techniques is based on criteria such as discriminatory power, intra- and inter-laboratory reproducibility, epidemiological concordance, ease of use and cost. The aim of our study was to identify clusters of Extended-Spectrum Beta-Lactamase-producing Klebsiella pneumoniae (ESBL-K. pneumoniae) strains circulating in neonatology using quantitative antibiogram (QA) and Pulsed Field Gel Electrophoresis (PFGE). Materials and Methods: This cross-sectional study included 55 K. pneumoniae strains isolated from a total of 513 samples. These various samples are taken from newborns, healthcare personnel, and the environment. K. pneumoniae identification followed standard bacteriological procedures and was confirmed using the Vitek® 2 (bioMérieux). The detection of the ESBL phenotype was performed using the synergy test. QA and PFGE were used to identify clonal relationships between the various strains isolated. Concordance between these two methods was assessed by calculating Cohen’s KAPPA coefficient and Simpson’s diversity index. Results: Among the 55 K. pneumoniae strains included in this study, 58.2% (32/55) were found to be Extended-Spectrum Beta-Lactamase (ESBL) producers. Most of these strains were isolated from neonatal samples (blood samples and rectal swabs). The quantitative antibiogram method applied to 28 out of the 32 ESBL-producing strains revealed that the isolates were grouped into 5 clusters. Pulsed Field Gel Electrophoresis performed on a total of 16 ESBL-producing strains showed the existence of four profiles. A perfect concordance was observed between the two methods. Conclusion: The results of this study highlighted the existence of clonal strains of various origins within neonatology units.

Aeromonas veronii isolated from climbing perch(Anabas testudineus)suffering from epizootic ulcerative syndrome(EUS)

Motile Aeromonads are responsible for various diseases in fishes that cause significant economic losses in aquaculture.Aeromonas spp.are also frequently isolated from fish suffering from Epizootic Ulcerative Syndrome(EUS).The present study was conducted to isolate and identify A.veronii from Vietnam climbing perch(Anabas testudineus)suffering from EUS,evaluate the pathogenicity of the isolates,and find out antibiogram profile and susceptibility of the isolates to medicinal plant extracts.Vietnam climbing perch(A.testudineus)suffering from EUS was collected from fish farms located at Savar,Dhaka and bacteria were isolated from the external ulcerative lesion,and kidney and liver of the infected fish on nutrient agar plates.Preliminary phenotypic identification of the bacterial isolates was performed following morphological,physiological and biochemical characterization tests.The 16S rRNA gene sequencing was accomplished for further molecular identification of five selected isolates.To evaluate the pathogenicity of the isolates in vivo artificial infection challenge test was conducted through immersion method.Another in vivo pathogenicity test was done at three different temperatures(18◦C,28◦C,and 37◦C)to examine the effects of temperature on the pathogenicity of the isolates.Ten commercial antibiotics were used to find out the antibiogram profile of the isolates.Finally,susceptibility of the isolates was screened against 19 medicinal plants extracts.A total of 38 out of 46 isolates isolated from Vietnam climbing perch(A.testudineus)suffering from EUS were phenotypically identified as Aeromonas sp.Five randomly selected isolates were further confirmed as A.veronii by analyzing the 16S rRNA gene sequence homology.The pathogenicity of the isolates was found moderate to high(41%-80%mortality)in the artificially challenged fish.The highest and lowest mortality was obtained in the fish challenged with the isolates BFKA33 and BFKA12,respectively.The isolates were found resistant to several antibiotics,including amoxicillin,ampicillin,penicillin-G,and vancomycin.Nevertheless,extracts of three medicinal plant(Allium sativum,Syzygium aromaticum,and Tamarindus indica)demonstrated antibacterial activity to the isolates(BFKA12,BFKA15,BFKA18,BFKA29,and BFKA33).

Occurrence of Clostridium perfringens contamination in poultry feed ingredients:Isolation, identification and its antibiotic sensitivity pattern

This work has been undertaken to study the occurrence of Clostridium perfringens contamination in the poultry feed ingredients and find out its in-vitro antibiotic sensitivity pattern to various antimicrobial drugs. Two hundred and ninety-eight poultry feed ingredient samples received at Poultry Disease Diagnosis and Surveillance Laboratory, Namakkal, Tamil Nadu in South India were screened for the presence of C. perfringens. The organisms were isolated in Perfringens agar under anaerobic condition and subjected to standard biochemical tests for confirmation. In vitro antibiogram assay has been carried out to determine the sensitivity pattern of the isolates to various antimicrobial drugs. One hundred and one isolates of C. perfringens were obtained from a total of 298 poultry feed ingredient samples. Overall positivity of 33.89% could be made from the poultry feed ingredients. Highest level of C. perfringens contamination was detected in fish meal followed by bone meal, meat and bone meal and dry fish.Antibiogram assay indicated that the organisms are highly sensitive to gentamicin(100%), chlortetracycline(96.67%), gatifloxacin(93.33%), ciprofloxacin(86.67%), ofloxacin(86.67%) and lincomycin(86.67%). All the isolates were resistant to penicillin-G. Feed ingredients rich in animal proteins are the major source of C. perfringens contamination.

ClinicalandExperimentalStudyonTreatmentofAcuteRespiratoryTractInfectionwithShuanghuanglianAerosol（双黄连气雾剂）

Two hundred and two cases of acute respiratory tract infection ( ARI) were treated withShuanghuanglian (SHL) aerosol. In these cases, the majority were virus infection and 64% of them wascaused by the respiratory syncytial virus (RSV) . The virostatic and bacteriostatic test were done in vitro bythe cell culture method and it was shown that SHL could inhibit the RSV, para-influenza virus I - IV and 23kinds of pathogenic bacteria such as Staphylococcus aureus etc. The bacteriostatic effect was positively cor-related to the SHL concentration. Experimental study showed that SHL could enhance the NK cell activity,promote the production of a-interferon and raise the rate of lymphocyte transformation. The controlled obser-vation on SHL preparation with various dosage-forms revealed that the SHL aerosol in treating early ARlshowed better results than that of injections and oral liquid symptomatologically ( P < 0 . 05 ) . The total effec-tive rate was 96% .

Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater

Objective:To investigate plasmid DNA profiles and the antibiotic resistance of a total of 41 strains of Escherichia coli(E.coli)isolated from seawater and mussel collected from 15 different sampling stations in Sinop,Turkey.Methods:Most probable number technique was used for detection of E.coli.Antibiotic susceptibilities of the isolates were determined by the disc diffusion method.Plasmid DNA of the strains was extracted by the alkaline lyses procedure.Results:According to morphological and physiological properties,it was determined that the isolates belonged to E.coli species.Antibiotic susceptibility of the strains was determined against seven standard drugs using disc diffusion method.All isolates were resistant to bacitracin(100%),novobiocin(100%),ampicillin(12.5%),tetracycline(7.5%),ceftazidime(5%)and imipenem(2.5%),respectively,whereas the strains were susceptible to polymyxin B(100%).The multiple antibiotic resistance values for the strains were found in range from 0.28 to 0.57.In addition,plasmid DNA analyses results confirmed that 22 strains harbored a single or more than two plasmids sized approximately between 24.500 to 1.618 bp.The high-size plasmid(14.700 bp)was observed as common in 21 of all strains.Conclusions:As a result,our study indicated that the presence of antibiotic resistant E.coli strains in seawater and mussel might be potential risk for public health issue.