API 20E Research Articles

ISOLATION AND IDENTIFICATION OF BRENNERIA NIGRIFLUENS AS CAUSAL AGENT OF BARK CANKER DISEASE ON WALNUT IN IRAQ

Bark canker disease has recently found in Sulaimani province. Disease incidence reached to 17.2% in Tawella. Bacterial isolates were identified by their morphological and biochemical characteristics using the API 20E system. Fifty-one and half percent of the isolates were identified as Brenneria nigrifluens and 36.3% as Pantoea spp. B. nigrifluens isolates formed circular colonies with entire margins and creamy color on nutrient agar. Biochemical tests classify B. nigrifluens isolates into seven groups. Vitek GN system further used to confirm the identification. The isolates produced necrotic lesion of different size on artificial inoculated walnut branches, but no symptoms on detached leaves. Isolates 22, 28 and 31 induced typical symptoms on two years old seedlings. B. nigrifluens successfully re-isolated from the inoculated seedlings. All B. nigrifluens isolates were resistant to erythromycin and cephalexin, most of them (16/17) were resistant to ampicillin, (13/17) resistant to vancomycin, (12/17) resistant to rifampin and amikacin; while (10/17) were moderately resistant to penicillin; (7/17) were moderately susceptible to streptomycin and (6/17) to gentamicin. Chloramphenicol, tobramycin, and tetracycline showed high efficiency in bacterial growth inhibition. Minimum inhibitory and minimum bactericidal concentrations of five chemicals against 17 bacterial isolates showed high efficiency of Kocide in killing 94.1% and inhibiting of 100% of the isolates at 0.31g/L In vitro, followed by Nordox which kill 70.6% and inhibit 76.4% of the isolates at 0.65 and 0.32 g/L respectively. In vivo studies confirm high efficacy of Kocide in disease control and restriction of vertical and horizontal extension of the cankers followed by Nordox.

Characterization of probiotic Bacillus subtilis isolated from Nile tilapia (Oreochromis niloticus) digestive tract and evaluation its positive impact on health and immunity.

In the present study, a probiotic, Bacillus subtilis was isolated from the digestive tract of freshwater Nile tilapia collected from production pounds of the Central Laboratory of Aquaculture Research, Egypt. The isolated strain of Bacillus subtilis was biochemically identified using API 20E and 16S rRNA. The Biosafety. assay confirmed that the isolate was not pathogenic to Nile tilapia fish. Isolated strain was able to survive at increasing concentrations of bile. Isolated strain was sensitive to most used antibiotics. The Isolated strain showed antagonistic activity against bacterial fish pathogens, including Aeromonas hydrophila and pseudomonas aeruginosa. Isolated strain did not show any hemolytic activity against human blood in the blood agar medium. To evaluate the positive impact of the supplementation of an isolated strain of Bacillus subtilis on hematological, biochemical and immunological parameter of Nile tilapia. A total of 120 fish (20 g) were randomly divided into 2 groups (control and treatment with 3 replicates). The control groups were fed the basal diet. Treatment groups were supplemented with B. subtilis 1X109 cfu/g. Results showed that Nile tilapia fed with diets containing B. subtilis for 7 weeks had a significant increase in Hb%, RBCs count, HCT values and WBCs count. Also, there is a significant improvement in liver function test (ALT, AST, Total protein and Albumin). Also, there is a significant increase in immunological parameter (NBT, Lysozyme). Moreover, fish fed with B.subtilis supplemented diets showed improved survival rates after A. hydrophila infection. Keywords:- Bacillus subtilis , Nile tilapia , API 20E, Positive impact.

Salmonella Carriage among Patients in Fako Division, Cameroon: A Cross-sectional Study of Its Prevalence and Associated Risk Factors

Introduction: This study was aimed at evaluating the prevalence and the risk factors of Salmonellosis in patients who were consulted in some medical facilities in Fako Division of Cameroon.
 Methods: A cross-sectional study was carried out from November 2017 to November 2018 in three hospitals in Fako division of Cameroon; Tiko District Hospital, Mutengene Medical Center and Buea Regional Hospital. A total of 510 individuals presenting with symptoms of Salmonellosis were administered comprehensive questionnaire. Salmonella enterica strains were cultured from stool and identified using API 20E. Data was entered into Excel and imported into STATA v.12 for Windows, for statistical analysis. Odd ratios were calculated to determine the risk factors associated with Salmonellosis.
 Results: Fifty Salmonella enterica strains were isolated giving a prevalence of 9.8%. Univariate analysis showed the following risk factors for Salmonellosis: area of residence; suburban p=0.037, OR=5.7 95% CI (1.1-30.03) and rural p=0.077, OR=2.3 95% CI (0.91-5.76), overcrowding (2 persons in a room) p=0.047, OR=2.3 95% CI (1.01-5.41); drinking tap-water, p=0.032 OR=0.38(.16-.092); left-over drugs from the pharmacy, p=0.906 OR= 1.07(0.32-3.55) as being relatively significant risk factors.
 Conclusion: The prevalence was found to be higher among the very young and older people greater than 45 years. The risk factors identified in this study are: age, area of residence; overcrowding; consuming locally prepared yoghurt or Kosam; eating out or auto-medication by taking leftover drugs. These findings highlight the need of reinforcement of hygiene promotion especially in infants and overpopulated communities, educate on proper prescription and usage of drugs, in addition to the intensification of environmental interventions.

Distribution of blaCTX\u2009\u2212\u2009M, blaTEM, blaSHV and blaOXA genes in Extended-spectrum-\u03b2-lactamase-producing Clinical isolates: A three-year multi-center study from Lahore, Pakistan

BackgroundFrequency of extended-spectrum-β-lactamase-producing clinical isolates is increasing worldwide. This is a multi-center study which was aimed to check the frequency of third-generation cephalosporin resistance and distribution of the key genetic determinants of Extended-spectrum-β-lactamase-producing Clinical isolates in Pakistan.MethodsA total of 2372 samples were processed in three tertiary care hospitals and one diagnostic research center of Lahore, Pakistan during Aug-2014 to Sep-2017. Analytical profile index (API 20-E) was used for biochemical characterization of isolates. Antibiotic susceptibility testing (AST) and third generation cephalosporin resistant (3GC-R) isolates were subjected to: double disc synergism test (DDST), combination disc test (CDST) and epsilometric test (E-test) for confirmation of ESBL-production. PCR amplification of isolates with plasmid and genomic DNA was performed. Amplicon sequences were checked for gene-variants and statistical analyses were performed to check the significance of data.ResultsA total of 497/995 (50%) isolates including Escherichia coli 65% (n = 321), Klebsiella spp. 25% (n = 124) and Pseudomonas. 5% (n = 24), Enterobacter spp. 4% (n = 20) and Acinetobacter spp. 2% (n = 8) were screened as third generation cephalosporin resistant (3GC-R). Urine 56% (n = 278) followed by pus 20% (n = 99) and wound swab 6% (n = 29) were frequent sources. Incidence of ESBL-producers detected by combination disc test was 79% (n = 392). PCR revealed blaCTX − M (76%) gene followed by blaOXA (52%), blaTEM (28%) and blaSHV (21%) were most prevalent among ESBL-producers detected by CDST. blaCTX − M − 1(65%), blaOXA (78%) and blaTEM (57%) genes were carried on plasmids. Amplicon sequencing revealed blaCTX − M − 15 (75%), blaOXA − 1 (49%) and blaTEM − 1B (34%) and 21 (n = 28) isolates carried three genes in them.ConclusionPrevalence of ESBL-producing isolates has increased 1.13 folds during study years. Isolates had high prevalence of ESBL-encoding blaCTXM − 15 gene and narrow spectrum blaOXA − 1 and blaTEM − 1B were also prevalent.

English

The objective of this study was to evaluate the emergence of beta-lactam resistance related to extended spectrum beta-lactamase (ESBL) encoding genes (TEM, SHV, CTX-M-1 and OXA-48) in Escherichia coli from Brazzaville. In the period between January 2016 and May 2017, 89 strains in E. coli were isolated from hospitalized patients, outpatients and domestic sewage. The E. coli strains were identified by the API 20E system. An antibiogram was performed on isolated trains by the disk diffusion method. The ESBL phenotype was detected using the synergistic technique according to CA-SFM (ESBL). Genes were detected using PCR and characterized by sequencing. An overall prevalence of 48.31 (43/89) and rates of 74.42, 23.26, 9.30, and 6.97% for blaTEM genes blaCTX-M-1, blaSHV and blaOXA-48 were detected. 25.58% were community strains and 74.42% hospital. The majority were isolated urine (81.40%) and the urology department was more represented. Except for imipenem, colistin strains of ESBL showed high resistance to other antibiotics than non-yielding ones (p <0.05). This high prevalence of detected ESBL, the high level of resistance to antibiotics raises fears of a high risk of multidrug-resistant bacteria and call on the authorities for a policy of monitoring resistance. Key words: Escherichia coli, extended spectrum beta-lactamase (ESBL), carbapenemases, Brazzaville.

Bacteria Recovered from Cultured Gilt-Head Seabream (Sparus aurata) and their Antimicrobial Susceptibilities

Objective: Gilt-head seabream (Sparus aurata) is an important species for Turkish aquaculture and bacterial diseases are oneof the limiting factors for the production of this species. The aim of this study is the identification of the bacterial disease agents in cultured gilt-head seabream in Turkey and the determination of their antibacterial susceptibilities. Materials and Methods: In this study, 27 diseased gilt-head seabream samples between 3-130 g, showing various clinical symptoms, were examined bacteriologically. Bacterial isolates recovered from moribund fish samples were identified by using standard bacteriological methods and API 20E rapid identification strips. Antibiotic susceptibility tests of 10 different substances against these isolates were determined by using Kirby-Bauer disc diffusion method.Results: Vibrio anguillarum, V. scophthalmi, V. logei, V. harveyi, Pseudomonas anguilliseptica, P. stutzeri, Staphylococcus sp. and Micrococcus luteus isolates were isolated from diseased fish samples as pure and mixed infection. It was detected that API 20E may give erroneous results for fish pathogenic bacteria since they are not included in the database of the kit. Generally, enrofloxacin, florphenicol and oxytetracycline were found to be the most effective antibiotics against bacterial isolates.Conclusion: This study is the first report for V. scophthalmi, V. logei, V. harveyi, P. stutzeri and M. luteus infections of gilt-head seabream in Turkey.

THE PREVALENCE AND PHENOTYPIC CHARACTERIZATION OF EXTENDED-SPECTRUM β-LACTAMASES-PRODUCING ESCHERICHIA COLI STRAINS ISOLATES RECOVERED FROM TERTIARY HOSPITALS IN SANA'A CITY, YEMEN

The emergence of ESBL producing Escherichia coli in clinical isolates is posing a serious threat for treating nosocomial infections. The aim of the study was to determine the frequency of extended spectrum β-lactamase (ESBL) producing Escherichia coli isolated from clinical specimens in several teaching and general hospitals in Sana'a city, Yemen, and to compare the phenotypic methods used for the characterization of ESBL producing strains. This cross sectional observational study was conducted from 1st July to 28th of August 2017, at the Department of Medical microbiology, Faculty of Medicine and Health Sciences, University of Sciences and Technology in Sana'a city, Yemen. A total number of 3500 various clinical samples were analyzed during the study period. Escherichia coli were identified using API 20E system and ESBL detection was carried out using double-disk synergy test (DDST) and CLSI confirmatory test. Escherichia coli were isolated from 100 samples, out of which 63 (63%) were ESBL producers and 37 (37%) were non-ESBL producers. The gender distribution of ESBL producing Escherichia coli was 35/45 (77.8%) in males and 28/55 (50.9%) in females. Highest frequency of ESBL producing Escherichia coli was detected in sputum (100%), wounds (83.3%) and urine (65.7%) samples. Comparison of DDST and CLSI confirmatory test showed that 65 (65%) isolates were characterized by DDST and 63 (63%) using CLSI confirmatory test. All ESBL-positive isolates were susceptible to imipenem, indicating that this agent is the best drug for treating serious infections caused by ESBL-producing E. coli. In conclusion, the present study shows moderately high frequency of ESBL producing Escherichia coli among patients suffering from sepsis in tertiary hospitals in Sana'a city. DDST was found to be less efficient in ESBL detection as compared to CLSI confirmatory test.

Expression of dnak, groES and cps genes in irradiated Klebsiella pneumoniae strains isolated from UTI Egyptian patients

T his investigati on presents a brief study for monitoring the correlation between the expression of heat-shock protein (HSP) genes; dnak, and groES as well as the capsular polysaccharide (caps) gene and antimicrobial susceptibility of extensively drug resistant (XDR) Klebsiella pneumoniae in response to a low dose of Gamma (γ) radiation. The XDR K. pneumoniae strains, that were obtained from (UTI) Egyptian patients were identified using the API 20E strip automated system. Their antimicrobial susceptibilities were monitored using standard disc diffusion method in parallel with their minimum inhibition concentrations (MICs) towards standard antibiotics using a micro-titer method, before and after exposure to γ-radiation at a dose of 24.4 Gray (Gy). The expression of dnak, groES, and cps genes was detected by qRT-PCR under the same conditions. The results demonstrate increasing antibiotic resistance after exposure to γ-radiation. Also, a doubling in the MIC of the tested strains towards CAZ(ceftazidime) was observed. These results were confirmed by an elevation in the expression of dnak, groES, and cps genes in response to γ-radiation. These preliminary results need more confirmaions, because they assume difficulties in curing immunocompromised patients infected with XDR K. pneumoniae by the currently available antibiotics.

Antimicrobial Resistance Patterns and Molecular Characterization of &amp;lt;i&amp;gt;Klebsiella pneumoniae&amp;lt;/i&amp;gt; in Clinical Isolates at Mbarara Regional Referral Hospital

Background: Klebsiella pneumoniae is one of the most frequent opportunistic pathogens causing a range of infections and being resistant for beta-lactamases (ESBL) and Carbapenemases. Aim: The aim of the present study was to determine the antimicrobial resistance patterns and molecular characterization establishing the phenotypes and genotypes associated with drug resistance, an antibiogram of genotypically positive isolates for resistance of Klebsiella pneumoniae in clinical isolates at MRRH. Materials and Methods: A laboratory-based descriptive cross-sectional study that was conducted from September 2018 to May 2019 at MRRH. Klebsiella pneumoniae was identified by cultural and biochemical methods. Antibiotic sensitivity test was performed by modified Kirby-Bauer disc diffusion technique. ESBL production in Klebsiella pneumoniae was tested by double-disc synergy test, Carbapenemase production by MHT, Boronic Acid or EDTA test using Meropenem phenotypically and both resistance confirmed genotypically by Multiplex PCR. Results: Out of 1055 clinical isolates, 298 (28.2%) were found positive for Klebsiella.spp, 175 isolates were subcultured among which 22 (12.57%) were K. pneumoniae based on API 20E. Overall Sensitivity patterns of these Klebsiella pneumoniae isolates to Ceftriaxone, (Amoxicillin/Clavulanate), Gentamicin, Cefepime, Ciprofloxacin, Cefoxitin, Nitrofurantoin, Cefuroxime, piperacillin/tazobactam, Meropenem, Ceftazidime and cefotaxime were 72.7%, 63.7%, 54.5%, 45.5%, 31.8%, 31.8%, 27.3%, 27.3%, 22.7%, 22.7%, 18.2%, 9.1%, 9.1% respectively. ESBL producing K. pneumoniae was found at 68.18% (15/22) phenotypically. Genotypically; the ESBL genes were blaCTX-M (100%), blaSHV (80%) and blaTEM (100; 47%); 8/15 (73.3%) had CTX-M, SHV, TEM, 4/15 (26.67%) CTX-M, TEM, 3/15 (20.00%) CTX-M and SHV. Carbapenemase producing K. pneumoniae was found at 31.82% (7/22) phenotypically; 1/7 (14.28%) by MHT, 4/7 (57.14%) Boronic acid test and 2/7 (28.58%) EDTA test. Genotypically; 3/4 [(75%) 42.86%] had OXA-48, 1/4 [(25%) 14.28%] OXA-48 and KPC gene, 1/2 [(50%) 14.28%] KPC and VIM, 1/2 [(50%) 14.28%] KPC and KPC gene [(100%) 14.28%]. Conclusion/Recommendations: DDS to be used for ESBL production, MHT, Boronic Acid test and EDTA tests using Meropenem/or Imipenem for Carbapenemase-production routinely.

Prevalence, risk factors, and identification of Salmonella spp. in stray dogs of northwest Mexico

Salmonellosis has a worldwide relevance in aspects associated with public health, as only in 2009 were reported 93.8 million cases in humans. The objective of the study was to establish the prevalence, risk factors and bacteriological and molecular identification of Salmonella spp in stray dogs in urban, rural and coastal areas of Mexicali, a city in northwest Mexico. From May 2014 to February 2015, 385 dogs were tested. Sampling was performed by rectal swab and conventional bacteriological techniques were applied, for later implementation of the API 20E system and molecular identification by polymerase chain reaction (PCR). The data were analysed statistically by means of descriptive statistics and multiple logistic regression modelling. A prevalence of 6.27% was obtained in the dogs examined, the samples obtained were characterised to subspecies ( Salmonella enterica subspecies enterica and Salmonella enterica subspecie arizonae ). The geographical region with the highest prevalence in the study was the coast (10%), followed by the rural area (8.57%) and the urban area (5.8%), however, no significant statistical differences were detected. There was significant difference in the prevalence by age of dogs under one year ( P <0.05). The identification of Salmonella in dogs from northwest Mexico could correspond to serovars of zoonotic importance indicating a potential risk for the population. Key words : Salmonella spp, prevalence, stray dogs, public health.

Antibacterial Activity of Essential Oil of &amp;lt;i&amp;gt;Aeollanthus pubescens&amp;lt;/i&amp;gt; on Multidrug Resistant Strains of &amp;lt;i&amp;gt;Salmonella&amp;lt;/i&amp;gt; and &amp;lt;i&amp;gt;Escherichia coli&amp;lt;/i&amp;gt; Isolated from Laying Hens Farming in Benin

Infections due to Escherichia coli and Salmonella are of the major constraints for the laying hen’s industry as they cause mortality and serious economic losses. The use of conventional antibiotics to control bacterial has shown limits because it allows multidrug-resistance. The main objective of this study was to assess the antibacterial activity of essential oil from Aeollanthus pubescens on multidrug resistant strains of Salmonella and Escherichia coli isolated from laying hens faming in the department of Atlantique in Benin. Altogether, 11 strains of Salmonella and 16 strains of Escherichia coli have been isolated from 101 samples of different organs including liver, spleen, lung, feces and yolk according to standardized methods and their biochemical profile using API 20E gallery. Test of sensitivity was carried out on 11 antibiotics of six different families on identified strains in order to determine their resistance profile. A sensitivity test was carried out on multi-drug resistant strains with Aeollanthus pubescens essential oil to determine their sensitivity with regard to this essential oil. The results showed that the majority of Salmonella strains presented resistance to Tetracyclines (72.7%) and Sulfonamides (63.6%) and all Escherichia coli strains are resistant to Sulfonamides (100%) followed by Tetracycline (93.75%) and Ampicillin (75%). Aeollanthus pubescens essential oil was active on all the multi-drug resistant strains investigated with Minimal inhibitory concentration varying from 0.41 ± 0 mg/ml to 0.83 ± 0 mg/ml for Salmonella and from 0.41 ± 0 mg/ml to 1.66 ± 0 mg/ml for Escherichia coli (P Salmonella and the multi-drug resistant Escherichia coli investigated. Those results provide alternatives to control poultry bacterial pathologies in Republic of Benin. However, disease due to Escherichia coli and Salmonella must be taken more seriously and study on their resistance to antibiotic must be deepened as well.

Serological cross-reactivity of environmental Escherichia coli strains with Shigella-specific antisera

Many bacteria of clinical and environmental origin show evidence of sharing common surface antigens. The present study aimed for isolation of Escherichia coli strains that were serologically cross-reactive with Shigella species from freshwater ecosystems in Bangladesh by conventional cultural methods. Among twenty eight isolates, two isolates, termed 12(35) and 6(50) showed cross-reactivity with four polyvalent serogroup-specific Shigella antisera using slide agglutination assay. The isolates were identified and charcterized by cultural and biochemical properties and Western blot analysis. The isolates showed typical Escherichia coli cell morphology and cultural and biochemical properties and were identified as Escherichia coli by API 20E tests. Western blot analysis confirmed the isolates as cross-reactive with all the four group-specific Shigella antisera due to presence of immunogenic proteins and LPS. One of the isolates also showed cross-reactivity with multiple type-specific Shigella boydii antisera (monovalent) because of immunogenic proteins. Both the isolates were identified as nonpathogenic due to absence of virulence marker genes of diarrheagenic E. coli variants.This study revealed that a number of bacteria present in the environment could share important Shigella species surface antigens. Naturally occurring nonpathogenic environmental bacteria expressing surface antigens specific for certain types of Shigella could be a good choice for vaccine candidates against shigellosis.&#x0D; Bangladesh J Microbiol, Volume 33, Number 1-2, June-Dec 2016, pp 29-33

Detection of CTX-M genes from ��-lactam Resistance Proteus mirabilis associated with Urinary Tract Infection in Holy Karbala province, Iraq

A total of 325 urine samples were collected from all ages and for both sexes from patients suffering from urinary tractinfections in the period from (15 February to 15 May, 2017) from Al Hussein Hospital in Karbala , The number of samples that gave a positive result for laboratory culture was 227 samples (84.69%) .The isolates were then identified according to the physiological and biochemical tests , It was found that the number of samples that were Gram negative was (59.9%) and the Proteus spp. obtained were (31.61%) of the total negative bacteria isolates. Proteus mirabilis isolates obtained from all isolates of Proteus spp. were 38 isolates formed ratio (88.37%) ,These isolates were initially diagnosed by culture them into the MacConkey agar and blood agar followed by a number of morphological and biochemical tests. The isolates were finally diagnosed using the API 20E system. Proteus mirabilis isolates obtained from urine samples of female were 27 isolates (71.05%) and 11 isolates (28.9%) from urine samples of male. The isolates were tested for 12 different types of ��-Lactam antibiotic to determine the most effective antibiotic toward these bacteria (Ampicillin, Piperacillin, Oxacillin, Cefazolin, Cephalothin , Cefoxitin , Ceftazidime , Ceftriaxone , Cefepime , Ertapenem , Imipenem and Aztreonam). The isolates of P. mirabilis showed a clear sensitivity to the Erytapenem , Imipenem , and Aztreonam , the nsensitivity ratio was 97.3% , 100% and 100% respectively. The polymerase chain reaction technique was also used to detect CTX-M gene which represents one of the three most frequent gene (TEM, SHV, CTX-M) in this bacteria The results of polymerase chain reaction showed that (17) isolates formed (44.74%) carrying the bla CTX-M gene. This study came to achieve the following objectives:

Monitoring of Pollution of Salmonella sp. in Raw Milk Using Virulence Gen Marker

Salmonella sp. is an enteropathogenic organism and it can spread through contaminated food which is rich in nutrition such as raw milk. The goal from this research is to find out the suitable pasteurization and time effect on the growth isolates Salmonella sp. that its invA presence is detected. Seventy-five (75) samples were taken in three locations of milk collection from groups of breeders. 10 ml of milk samples were grown in enrichment culture of modified Tryptic Soy Broth (mTSB) at 37oC for 12 hours. Cellular culture was then grown in differential selective medium of Salmonella Shigella Agar (SSA) and Chromocult Coliform Agar (CCA). From 75 milk samples produced 1392 colonies of typical Salmonella sp. pathogens. Then, isolates were selected using Triple Sugar Iron Agar (TSIA) and Urea Broth media, producing 3 suspected isolates of Salmonella sp which was known from colonies color. Results of confirmation using API 20E and 50 CHE produced two isolates of Salmonella spp. with %ID = 99.6% and one isolate of Salmonella typhi with %ID = 72.1%. All of the isolates were then detected for virulence factor using invA marker and all positive isolates have virulence factor. It indicates that raw milk contains polluted pathogenic bacteria.Keywords: raw milk, enteric pathogenic, virulence gen

Bacteriological quality of dry powder milk available in local markets of Bangladesh

Milk and milk powders are very rich in several nutrients and relished by everybody throughout the world. The present study was undertaken with the aim of investigating the bacteriological quality of locally available dry powder milk in Bangladesh. A total number of eight powder milk samples were collected from Gazipur city and its surrounding areas during the period from January 2017 to February 2017. The analysis comprised of enumeration of total viable bacterial count (TVBC), isolation of bacterial isolates and identification of pathogenic bacteria. Almost all the powder milk samples showed the total aerobic heterotrophic bacterial (TAHB) level above the standard acceptable range (&gt;104 CFU/g). Both gram positive and gram negative pathogenic bacteria viz. Staphylococcus aureus, Staphylococcus epidermidis, Bacillus, pseudomonas sp. and Enterobacter sp. were isolated from studied samples through morphological and biochemical characteristics. Pseudomonas sp. were confirmed by API 20E. These results highlighted the necessity to maintain appropriate sanitary and hygienic measures at each critical point in order to safeguard consumers from food borne pathogens.Asian J. Med. Biol. Res. September 2018, 4(3): 267-273

Analysis of bacteriological pollution and the detection of antibiotic resistance genes of prevailing bacteria emanating from pig farm seepage.

Management and disposal of pig farm seepage constitute a serious environmental challenge, and seepage discharge from agricultural waste‐water is considered to be one of the greatest contributors of organic substances, bacterial pathogens, and antibiotic resistance genes into the environment. The objectives of this study were to assess the level of bacteriological pollution and to identify the resident antibiotic‐resistant genes of culturable bacteria from a studied pig farm seepage. Enumeration of the viable bacterial cell of plated bacteria suspensions (10−1 to 10−8 cfu/mL) was performed; also, identification of pure bacterial colonies was done using an API 20E bacterial identification kit. CLSI guidelines for antimicrobial susceptibility testing were adopted to determine the antibiotic susceptibility/resistance of the cultured bacterial isolates. Identification of resident‐resistant genes was done using molecular biology procedures. The results on viable cells in seepage samples ranged from 4.30 × 102 to 1.29 × 109 cfu/mL. Pseudomonas luteola, Enterococcus vulneris, Salmonella choleraesuis spp arizonae, Escherichia coli, Enterobacter cloacae, Proteus mirabillis etc. were isolated from the pig farm soil samples. Almost all of the cultured isolates were resistant to Penicillin G, Vancomycin, Oxytetracycline, Spectinomycin, and Lincomycin. The most frequent resistant genes detected in the isolates were Van A, Van B, InuA, aph (3”)‐llla, blaTEM, Otr A, and Otr B. It was inferred from the study that Pig farm seepage has the ability to cause bacterial pollution that may negatively impact the natural environment, by introducing bacteria pathogens that harbor antibiotic‐resistant genes.

Analysis of adult damselfly fecal material aids in the estimation of antibiotic-resistant Enterobacterales contamination of the local environment.

Because damselflies are ubiquitously but focally present in natural environments and play a critical role as predators of other insect species, the fecal matter of damselflies may be useful for investigating antibiotic-resistant bacterial populations, including human pathogens, in local environments. We therefore examined the prevalence of antibiotic-resistant bacteria, including Enterobacterales, in fecal material from 383 damselflies (adults and larvae) collected from seven locations around Sapporo City, Japan, in 2016 and 2017. Fecal samples were plated on soybean casein digest (SCD) agar plates with and without antibiotics (SCD-A and SCD-w/o, respectively) to identify environmental bacteria and gut bacteria, respectively, and on MacConkey agar plates with antibiotics (MacConkey-A) to select for Gram-negative bacteria, including human pathogenic Enterobacterales species. The prevalence of colonies on each of the plates was compared, and representative colonies on MacConkey-A plates were identified to the species level using an API 20E kit and the MALDI Biotyper system. Overall, SCD-w/o plates showed a gut bacterial load of approximately 108 colony-forming units per adult damselfly or larva. There was a significant difference between the prevalence of colonies on the SCD-A and MacConkey-A plates, and a significantly increased prevalence of antibiotic-resistant bacteria on MacConkey-A plates was observed in samples collected from Shinoroshinkawa. Cluster analysis based on minimum inhibitory concentration values of 59 representative isolates from MacConkey-A agar plates revealed that samples from Shinoroshinkawa contained a higher prevalence of Enterobacterales than those from other sampling locations. Thus, fecal materials discharged by adult damselflies could be used in future studies as a simple tool for estimating antibiotic-resistant bacteria, including Enterobacterales species, in the local environment.

Comparative study on bacterial flora of the intestine in Persian sturgeon (Acipenser persicus) fingerlings reared in fiberglass tanks and earthen ponds

Stability of bacterial flora in fish intestine is very important, because the intestine is an important organ for outbreak of microbial infections and other fish diseases especially when the fish have not been vaccinated. The aim of this study was to determine the bacterial flora of intestine in sturgeon juvenile in earthen ponds. The results can be applied to prevent the diseases occurrence and health management. In this study, 90 individuals of Persian sturgeon ranged 3-5g in weight collected randomly by trawling in 3 earthen ponds as well as 15 individuals ranged 10-20g in weight by small net in 3 fiberglass tanks. After recording morphometric characteristics, the intestine fish and rearing water were cultured on TSA medium. Gram staining and complementary tests were conducted. In order to identify the bacterial species, various biochemical experiments and diagnostic kits of API 20E (special for gram - negative bacteria) were employed. According to morphometric measurements, in earthen ponds the mean weight and length of juveniles were 5.59 ± 3.18 and 11.4 ± 2.9, respectively. The results showed that in these ponds the mean facultative aerobic and anaerobic bacterial counts in the juvenile intestine were 5.59 ± 0.92 (log CFU.g-1) compared to 6.67 ± 0.34 (log CFU.ml-1) in rearing water. Furthermore, the following bacteria were found in the intestine of Persian sturgeon: Aeromonas sp., A. sobria, A. hydrophyla, Entrobacteriaceae and Corynebacterium, while the bacteria recovered from the rearing water were Aeromonas sp. and A. sobria. According to morphometric measurements, in the fiberglass tanks, the mean weight and length of juveniles were 12.13 ± 2.51 (g) and 15.11 ± 1.13 (cm), respectively. The results showed that in these tanks, the mean facultative aerobic and anaerobic bacterial counts in the fish intestine were 4.77 ± 0.04 (Log CFU.g-1) compared to 4.81 ± 1 (Log CFU/ml) in rearing water. Furthermore, the following bacteria were identified in the intestine of Persian sturgeon: Aeromonas sp., Micrococcus, Staphylococcus. The bacteria in the rearing water were A. sobria, Acinetobacter, Moraxella, Micrococcus and Staphylococcus.

IDENTIFICATION COMMON CAUSE OF NEONATAL SEPSIS BY ANALYTICAL PROFILE INDEX SYSTEM (API)

Background: Neonatal Sepsis is a bacterial infection of the blood in a neonate and an infant younger than 4 weeks of age. The analytical profile index or API is a classification of bacteria based on experiments, allowing fast identification. This system is developed for quick identification of clinically relevant bacteria. Objective: To identify the pattern of organisms in neonatal sepsis using API system in Baghdad City hospital, Al-Imamein Al-Kadhimein Medical City and Central Pediatrics Teaching Hospital. Methods: In this cross-sectional study, blood samples from 100 neonatal patients were inoculated into a blood culture bottle and incubated at 37 °C under aerobic conditions, subculture was done after 24 h of incubation, the growth was identified by phenotypic characteristics, gram's stain, and API system. Results: Positive blood cultures were detected in 82 (82%), according to API system the most prominent bacterial isolates from blood culture in neonates with early-onset sepsis were non-coagulase Staphylococcus (20.4%) Staphylococcus aureus (18.1 %), Acinetobacter bumanii (13.6%), Pseudomonas aeruginosa (11.36 %), Streptococcus agalactiae (11.3 %), while in late-onset sepsis the most common bacteria were Staphylococcus aureus (21.0%), non-coagulase Staphylococcus (13.1%), Citrobacter freundii and Pseudomonas aeruginosa (10.5 %) respectively. Conclusion: The API 20E may be useful for the identification of the bacterial species rarely described as pathogens in neonatal sepsis will help us to study the clinical burden resulting from the emergence of these species as causes for this neonatal infection. Keywords: Early-onset sepsis, late-onset sepsis, blood culture, API system Citation: Al-Mossawi HSM, Hassan JS, Al-bashier NM, Al-Omrani AAA. Identification common cause of neonatal sepsis by analytical profile index system (API). Iraqi JMS. 2018; 16(3): 327-334. doi: 10.22578/IJMS.16.3.12

Etiology of early onset neonatal sepsis in neonatal intensive care unit - Mansoura, Egypt.

This study was conducted to find out the bacterial causes of early onset neonatal sepsis and their susceptibility pattern for different classes of antibiotics in neonates admitted to neonatal intensive care unit (NICU) of Mansoura-Egypt. A descriptive cross-sectional study was conducted. All admitted newborns to our study were infants at <72 hours of age with clinical features of sepsis or product of an in-house delivery at Mansoura University Hospital with risk factors for neonatal sepsis. Identification of Gram-negative isolates was confirmed by API 20E kits (bioMerieux). Antibiotic susceptibility was performed by Kirby-Bauer disc diffusion method. Bacteria resistant to ≥3 antimicrobial classes were counted as multidrug resistant. One hundred eighty eight babies were admitted to our study. Positive blood culture was reported in 34.6% of newborn infants with prevalence of 4.02/1,000 live births. The most common isolated bacteria in early onset neonatal sepsis were coagulase negative staphylococci followed by Klebsiella pneumoniae and Serratia marcescens. The Gram-positive bacteria showed high resistance to ampicillin 93.9% while all the isolates were susceptible to vancomycin. The isolated Gram-negative bacteria were highly resistant to ampicillin [96.9%], amoxicillin-clavulanic acid [90.6%], cefotaxime [84.4%] and ceftazidime [84.4%]. Best sensitivity among all isolates was observed to imipenem. Multidrug resistance was observed among 45.5% of the Gram-positive and 68.8% of Gram-negative isolates. Appropriate identification of the source of infection and initiation of an effective management can reduce both mortality and morbidity associated with neonatal sepsis. The challenge is to initiate immediate empirical antibiotic therapy according to a strictly implemented updated antibiotic policy based on an individualized community established antimicrobial sensitivity pattern of microorganisms causing early onset neonatal sepsis.