Multiple Antimicrobial Resistance Index Research Articles

Prevalence and antimicrobial susceptibility of Streptococcus equi isolated from horses in Santa Catarina state, Southern Brazil.

The objective of this study was to determine the prevalence of Streptococcus equi (S. equi subsp equi and S. equi subsp zooepidemicus) in the state of Santa Catarina and evaluate the antimicrobial susceptibility of the isolates. For this, 420 nasal swab samples were collected from randomly selected horses. Isolation and phenotypic characterization of the bacteria were performed by sowing on 5% sheep blood agar, followed by analysis of morphotinctorial characteristics and biochemical analysis. To differentiate the main beta-hemolytic Streptococcus in horses, the fermentation profiles of the sugar's lactose, maltose, sorbitol, and trehalose were used, which were confirmed at the subspecies level by the PCR technique. The antimicrobial susceptibility panel was defined by the disk diffusion method, testing 13 antimicrobials from ten different classes, all regularly used in equine medical clinics, followed by the calculation of the multiple antimicrobial resistance index. Ten strains of S. equi were isolated, with a prevalence of 2.38% (10/420). Of the total positive samples, 3% (3/10) were confirmed as belonging to S. equi subsp equi and 70% (7/10) were confirmed as belonging to S. zooepidemicus. Multidrug resistance was observed in 60% (6/10) of isolates. The antimicrobial with the greatest resistance was clindamycin with 70% (7/10), followed by beta-lactams, with 40% (4/10) resistance to penicillin and 30% (3/10) to ceftiofur. The isolates were 100% (10/10) sensitive to gentamicin, chloramphenicol, levofloxacin, and vancomycin. This was the first study carried out in the state, and based on these data, it can be said that Santa Catarina has a low prevalence of S. equi and the presence of multi-resistant strains of S. equi was confirmed in the equine herd in Santa Catarina.

Antimicrobial susceptibility profile of bacteria of the genus Aeromonas and Escherichia coli isolated from samples of yellow hake (Cynoscion acoupa).

The aim of this study was to evaluate the antimicrobial susceptibility profile of Aeromonas sp., and Escherichia coli isolated from samples of yellow hake (Cynoscion acoupa). We analyzed 53 Aeromonas spp. and four E. coli isolates. We observed increased resistance of E. coli to levofloxacin and sulfa-trimethoprim as well as resistance of Aeromonas spp. to ampicillin, amoxicillin-clavulanate, cefuroxime, and cefotaxime. The multiple antimicrobial resistance(MAR) index indicated multidrug resistance in 90.54% (n=48) of Aeromonas spp. isolates and in 50% (n=2) of E. coli isolates. One strain of Aeromonas spp. was resistant to all 11tested antimicrobials (MAR index = 1.00). In vitro,piperacillin + tazobactam was the most effective antimicrobial for E. coli,and cefepime and levofloxacin were the most effective antimicrobials for Aeromonas spp. Therefore, in case of illnesses caused by these microorganisms, these antimicrobials should be used. The multidrug resistance of Aeromonas spp. and E. coli in this study is elevated. This is worrisome considering the increase in bacteria resistant to multiple drugs, reducing the options for successful clinical antimicrobial use.

Multi-drug resistant (MDR) Gram-negative pathogenic bacteria isolated from poultry in the Noakhali region of Bangladesh.

Rapidly increasing antibiotic-resistant bacterial strains in Bangladesh's food and farm animals stem from the excessive and inappropriate use of antibiotics. To assess the prevalence of multi-drug resistant (MDR) Gram-negative bacteria in poultry chicks, we sought to isolate and identify strains carrying antimicrobial resistance genes. Isolation and identification involved biochemical tests, 16S rRNA sequencing, and PCR screening of species-specific genes. MDR patterns were evaluated using CLSI guidelines with seventeen antibiotics across twelve classes. Targeted gene sequences were amplified for the detection of Extended-spectrum β-Lactamase (ESBL), carbapenem, tetracycline, sulfonamide, and colistin resistance genes. Common isolates, such as Escherichia coli, Klebsiella pneumoniae, Proteus penneri, and Enterobacter hormaechei, exhibited average Multiple Antimicrobial Resistance (MAR) indices of 0.66, 0.76, 0.8, 0.84, and 0.81, 0.76, 0.84, 0.41 for broiler and layer chicken, respectively. Providencia stuartii and Salmonella enterica, exclusive to broiler samples, had MAR indices of 0.82 and 0.84, respectively. Additional isolates Morganella morganii, Aeromonas spp., and Wohlfahrtiimonas chitiniclastica were found in layers (Average MAR indices: 0.73, 0.71, and 0.91). Notably, M. morganii, E. hormaechei and W. chitiniclastica were identified for the first time in Bangladeshi poultry chicken, although their evolution is yet to be understood. In this study, Pan-drug resistance was observed in one P. stuartii (broiler) and one Aeromonas spp. (layer) with a MAR index 1, while all isolates exhibited MAR indices >0.2, indicating MDR. Antimicrobial resistance (AMR) gene screening identified blaTEM, blaSHV, tetA, and sul1 in a majority of the MDR strains. Interestingly, E. coli (lactose positive and negative) and E. hormaechei were exclusively found to possess the tetB gene. In addition, E. coli (lactose negative), Klebsiella pneumoniae, Enterobacter hormaechei, M. morganii, and P. stuartii were observed to carry the colistin-resistant mcr-1 gene, whereas sul2 was detected in E. coli (lactose positive and negative), E. hormaechei, P. stuartii, and P. penneri. These findings emphasize the health risk of our consumers of both broiler and layer chickens as they have turned into a potent reservoir of various AMR gene carrying MDR and Pan-drug resistant bacteria.

Oxidative stress, gene expression and histopathology of cultured gilthead sea bream (Sparus aurata) naturally co-infected with Ergasilus sieboldi and Vibrio alginolyticus

BackgroundParasitic and bacterial co-infections have been associated with increasing fish mortalities and severe economic losses in aquaculture through the past three decades. The aim of this study was to evaluate the oxidative stress, histopathology, and immune gene expression profile of gilthead sea bream (Sparus aurata) co-infected with Ergasilus sieboldi and Vibrio alginolyticus.ResultsVibrio alginolyticus and Ergasilus sieboldi were identified using 16 S rRNA and 28 S rRNA sequencing, respectively. The collagenase virulence gene was found in all Vibrio alginolyticus isolates, and the multiple antimicrobial resistance index ranged from 0.286 to 0.857. Oxidant-antioxidant parameters in the gills, skin, and muscles of naturally infected fish revealed increased lipid peroxidation levels and a decrease in catalase and glutathione antioxidant activities. Moreover, naturally co-infected gilthead sea bream exhibited substantial up-regulation of il-1β, tnf-α, and cyp1a1. Ergasilus sieboldi encircled gill lamellae with its second antennae, exhibited severe gill architectural deformation with extensive eosinophilic granular cell infiltration. Vibrio alginolyticus infection caused skin and muscle necrosis in gilthead sea bream.ConclusionThis study described some details about the gill, skin and muscle tissue defense mechanisms of gilthead sea bream against Ergasilus sieboldi and Vibrio alginolyticus co-infections. The prevalence of co-infections was 100%, and no resistant fish were detected. These co-infections imbalance the health status of the fish by hampering the oxidant-antioxidant mechanisms and proinflammatory/inflammatory immune genes to a more detrimental side. Our results suggest that simultaneous screening for bacterial and parasitic pathogens should be considered.

MOLECULAR IDENTIFICATION AND ANTIBIOGRAM OF METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS FROM WOUNDS OF BOTH IN- AND OUT-PATIENTS AT UNIVERSITY OF BENIN TEACHING HOSPITAL (UBTH), BENIN CITY, NIGERIA

Methicillin-resistant Staphylococcus aureus (MRSA) stands as a prominent bacterial pathogen associated with various human infections, characterized by its remarkable ability to rapidly develop resistance to multiple antibiotics. The study aimed to characterize MRSA isolated from wounds among hospital patients. A total of eighty-seven unique S. aureus isolates were obtained from wound cultures collected from patients at the University of Benin Teaching Hospital (UBTH) in Benin City, Nigeria. These samples underwent assessment for methicillin resistance through conventional cultural methods, and detection of methicillin resistance (mecA) and S. aureus specific (nuc) genes using PCR amplification. The MRSA isolates underwent assessment for antimicrobial resistance through disc diffusion testing and for virulence factors using phenotypic testing procedures. An overall prevalence of 34/87 (39.1%) MRSA isolates was recorded. Based on the source of the isolate, the distribution of MRSA isolates from in-patients was 22/56 (39.3%) and out-patients was 12/31 (39.7%). Total Panton valentine leucocidin (PVL) screening revealed 11/34 (32.4%). All the isolates were resistant to penicillin 34/34 (100%) and cefepime 34/34 (100%). Lower level of resistance was demonstrated towards ciprofloxacin 3/34 (8.8%). Overall, every isolate displayed resistance to at least three antibiotics, resulting in a multiple antimicrobial resistance index of at least 0.33. The total virulence factors observed were beta-hemolytic activity 22/34 (64.7%), protease activity 14/34 (41.2%), and lipase activity 19/34 (55.9%). This study emphasized the significance of regularly monitoring MRSA prevalence within both the hospital and the local community. Therefore, the need for better intervention and continuous enlightenment on risks associated with the abuse...

ISOLATION, CHARACTERIZATION AND ANTIBIOTIC SUSCEPTIBILITY PATTERN OF DIARRHEAGENIC ESCHERICHIA COLI (DEC) AMONG CHILDREN ATTENDING SOME SELECTED HOSPITALS WITHIN KADUNA METROPOLIS

Diarrhoea has been associated with significant morbidity and mortality, ranked the second cause of death in children aged 0 to 5 years. Bacteria, viruses, protozoa, and helminths have all been implicated in diarrhoea, however rotavirus and diarrhoeagenic E. coli (DEC) are the most common cause. This study aimed to determine the antibiotic susceptibility pattern of diarrheagenic Escherichia coli isolated from children 0-5 years attending selected hospitals in Kaduna metropolis. A total of 264 stool samples were collected from children attending four selected hospitals in Kaduna metropolis. Standard culture procedures and molecular techniques such as PCR and 16s rRNA were employed in isolation and characterization of diarrhoeagenic E. coli from the stool samples. The study established a prevalence of 24.2% for diarrheagenic Escherichia coli, and all the isolates demonstrated multiple antimicrobial resistance index (MARI) of 0.5 and above, and showed significant resistance against augmentin (100%), amoxacillin (100%), ampicillin-cloxacillin (ampiclox) (100%), erythromycin (100%), gentamycin (100%), cefoxitin (100%), cefuroxime (95%), ceftriazone (95%) and ceftaxidime (85%). The least (60%) resistance was observed against imipenem. The study concluded that antibiotics have not been very effective in the treatment of E. coli-related diarrhoea. The study, therefore, recommends the implementation of programmes geared towards good hygiene, good nutrition and good health

The Bacterial Profile and Antimicrobial Susceptibility Patterns of Urinary Tract Infection Patients at Pawe General Hospital, Northwest Ethiopia.

Urinary tract infection remains the most common infection widespread worldwide in both community and hospital settings. Rapidly increasing antibiotic resistance of uropathogens is resulting in limited treatment options. Thus, understanding the current uropathogens and their antimicrobial susceptibilities is essential for effective urinary tract infection treatment. The purpose of this study was to isolate, characterize, and determine the antimicrobial susceptibility patterns of bacterial pathogens associated with urinary tract infection at Pawe General Hospital in Northwest Ethiopia. A hospital-based cross-sectional study design was conducted from January to April, 2020, at Pawe General Hospital. Midstream urine specimens were collected from 141 individuals with suspected urinary tract infection for bacteriological identification and antimicrobial susceptibility testing. Among the 141 study participants, twenty-nine (20.6%) showed significant bacteriuria. Escherichia coli (42.6%) had the highest proportion of isolated uropathogen followed by Klebsiella spp. and Pseudomonas spp. each (10.7%); Proteus spp. (9.3%); coagulase negative staphylococci, Staphylococcus aureus, and Enterobacter spp. each (6.7%); Citrobacter spp. (4%); and Enterococcus faecalis and Streptococcus spp. each (1.3%). Outpatient isolates showed a resistance of 64% and 78.6% to amoxicillin-clavulanic acid and tetracycline, respectively. Inpatients showed 63.9% and 87.2% of resistance to cephalexin and tetracycline. It was also observed that all the isolates have a multiple antimicrobial resistance index greater than 0.20 except Citrobacter spp. (0.142) in inpatients. Even though in this locality, most isolates were sensitive to ceftriaxone, gentamicin, ciprofloxacin, nitrofurantoin, and norfloxacin, they are considered appropriate antimicrobials for empirical treatment of urinary tract bacterial infections. Periodic monitoring of etiology and drug susceptibility is highly recommended, along with health education on the transmission and causes of urinary tract infection.

Bovine mastitis in northeastern Brazil: Occurrence of emergent bacteria and their phenotypic and genotypic profile of antimicrobial resistance

The aim of this study was to identify emergent pathogens associated with bovine mastitis in northeastern Brazil and to characterize them for phenotypic and genotypic resistance to antimicrobials. A total of 321 milk samples from cows with subclinical mastitis were collected, and the isolates obtained in culture were identified using matrix-associated laser desorption-ionization - time of flight mass spectrometry. Phenotypic and genotypic antimicrobial resistance tests were performed. We identified 72 bacteria considered emergent in the study region: Enterococcus faecalis (26.3%; 19/72), Streptococcus agalactiae (22.2%; 16/72), Enterococcus faecium (20.0%; 15/72), Escherichia coli (6.9%; 5/72), 6.9% (5/72) Lactococcus garvieae (6.9%; 5/72), Acinetobacter baumannii (5.5%; 4/72), Bacillus subtilis (1.3%; 1/72), Kocuria marina (1.3%; 1/72), Macrococcus caseolyticus (1.3%; 1/72), Microbacterium resistens (1.3%; 1/72), Micrococcus luteus (1.3%; 1/72), Streptococcus dysgalactiae (1.3%; 1/72), Streptococcus hyovaginalis (1.3%; 1/72) and Streptococcus pluranimalium (1.3%; 1/72). The antibiogram revealed the following resistance profiles: ampicillin (77.7%; 56/72), cefoxitin (69.4%; 50/72), erythromycin (61.1%; 44/72), oxacillin (63.8%; 46/72), penicillin (79.1%; 57/72), tetracycline (63.8%; 46/72), gentamicin (25.0%; 18/72), and vancomycin (20.8%; 15/72). Of the isolates, 83.4% (60/72) showed multiple resistance to antimicrobials. The tetM gene was identified in 43.0% (31/72) of the isolates, followed by tetL (31.9%; 23/72), and blaZ (26.3%; 19/72). 83.4% (60/72) of the isolates presented a multiple antimicrobial resistance index higher than 0,2. Emergent bacteria with zoonotic and multiresistant potential occur in cows with mastitis in northeastern Brazil. It is necessary to monitor the occurrence of these and other bacteria in livestock environments and develop control strategies to prevent their spread.

High burden of multidrug resistant bacteria detected in Little Akaki River

Spread of antimicrobial-resistant bacteria between humans and animals occurs when the environment is contaminated with animal and human wastes. A total of 30 samples were collected from the Akaki river to identify antimicrobial-resistant bacteria. Bacterial enumeration and characterization was done by spreading serially diluted water samples on MacConkey agar. Sixty four bacterial isolates were identified and susceptibility tested using VITEK 2. The most frequently identified bacteria were Providencia alcalifaciens 10 (15.6%), Kluyvera cryocrescens 9 (14.1%) and Citrobacter freundii 7(10.9%), respectively. Multiple drug resistant bacteria were identified, constituting 17 (28%) of the 64 identified isolates. Multiple antimicrobial resistance (MAR) index of the six sites laid in the range 0.13–0.27, being the highest score located downstream of all the sampling sites. Species MAR index varied from 0.12 to 0.40. Out of 64 isolates, 54 (84.4%) of them were resistant to Ampicillin. On the contrary, most of the isolates were sensitive to Amikacin and meropenem. In conclusion, our findings indicated E.coli count was above the WHO permissible levels. The predominant isolates were P. alcalifaciens, and C. freundii. The MAR index of major isolates was greater than two, implying the study sites were exposed to high-risk sources of human or animal contamination.

Antimicrobial Resistance of Aeromonas Species Isolated from Cultured Indian Major Carp, Labeo rohita: Possible Public Health Concern

In India, Labeo rohita is widely cultured and consumed freshwater fish. Aeromonads are etiological agents of major bacterial fish diseases like furunculosis, haemorrhagic septicaemia, skin ulcers, fin/tail rot and dropsy, causing significant economic losses in carp culture. Aeromonas species are widely distributed in aquatic environment which is considered as important vehicle of Aeromonas infections to fish and humans. Some of the Aeromonas spp. causes gastroenteritis, septicaemia, peritonitis, meningitis and eye infections in humans. In the present study Aeromonas species were isolated from diseased freshwater fish Labeo rohita collected from two districts viz., West Godavari and SPSR Nellore of Andhra Pradesh, India. A Total of 12 Aeromonas spp. were isolated and identified by biochemical tests. A. veronii bv. veronii (35%) was dominant when compared to other Aeromonas spp. Further, Antimicrobial resistance and multiple Antimicrobial resistance (MAR) of all Aeromonas spp. were tested against 17 antibiotics being frequently used for human diseases. The Antimicrobial resistance of all the 12 Aeromonas spp. have shown significantly high (p<0.05) resistance (100%) to ampicillin, amoxyclave and oxytetracycline except A. cavernicola when compared to other antibiotics. The MAR index of Aeromonas spp. ranged from 0.18-0.76, which indicates origination of isolated Aeromonas spp. from high risk sources of contamination. A. hydrophila, A. veronii bv. sobria, A. veronii bv. veronii, A. schubertii and A. jandaei isolated in this study were found to be pathogenic to humans also. The results revealed the pathogenic potential of Aeromonas infections in freshwater fish culture and emerging threats to public health.

English

Chicken meat, which could be a healthy and nutritionally food, is regrettably incriminated as a source of Salmonella typhimurium which has a great ability to cause human salmonellosis. This study aimed to assess the prevalence of multidrug-resistant S. typhimurium in chicken meat and the effects of essential oils on its viability. A total of 300 chicken meat and its products samples were streaked on XLD agar plates, which was followed by identification of the isolates based on biochemical and serological tests. Ten isolates were serotyped as S. typhimurium then assayed for susceptibility to 14 antimicrobials by the single diffusion method. Eight isolates (80%) showed multiple antimicrobial resistance (MAR) for 3 or more antimicrobials with MAR index of 0.4857 in average. Serotyped S. typhimurium strain with the highest antimicrobial resistance, confirmed by 16s RNA sequencing, was selected for studying the effects of thyme, oregano, and lemon essential oils with concentrations of 0.5, and 1% on its viability after inoculating into chicken fillets by intensity of 3.0×106 and on sensory traits of chicken fillets on 2nd, 4th, 6th, and 8th days of inoculation during cold storage (4ºC). All results showed a significant reduction of S. typhimurium counts with highest inhibition obtained using 1% lemon essential oil. The sensory properties of treated chicken fillets were improved by all used essential oils, compared to the control samples after 6th day, and 8th day of the storage period. The samples treated with 0.5, and 1% lemon essential oil revealed the highest improvement of sensory attributes. This study proved that the majority of S. typhimurium existing in chicken meat are Multidrug-resistant and have no negative effect on sensory traits, hence, posing a public health hazard. Natural essential oils have, also, great antimicrobial effect on S. typhimurium, thus it could replace chemical antimicrobials.

Occurrence of Virulence and Antimicrobial Resistance Determinants in Vibrio harveyi Isolated from Marine Food Fish Cultured in Korea.

Vibrio harveyi is a significant cause of infection in both marine animals and humans. It has been reported frequently in seafood-borne infections worldwide. This study was conducted to determine the potential health impact of the V. harveyi isolated from marine food fish cultured in Korea concerning their virulence and antimicrobial resistance. A total of 49 V. harveyi samples were isolated by biochemical tests and multiplex PCR. Phenotypic detection of virulence factors resulted DNase activity (81.63%), hemolysis (α = 75.51% and β = 12.25), gelatinase activity (71.43%), protease production (71.43%), phospholipase activity (65.31%), and lipase production (34.69%). Virulence genes, including VPI, tlh, tdh, toxR, VAC, and ctxAB, were detected in 57.14%, 44.90%, 36.73%, 22.45%, 12.24%, and 8.16% of the isolates, respectively. Resistance to ampicillin (77.55%), oxacillin (69.39%), nalidixic acid (53.06%), amoxicillin (46.94%), oxytetracycline (46.94%), colistin sulfate (34.69%), fosfomycin (34.69%), chloramphenicol (32.65%), streptomycin (32.65%), cephalothin (28.57%), oxytetracycline (26.53%), ceftriaxone (20.41%), erythromycin (14.29%), and cefoxitin (12.24%) was detected in disc diffusion assay. Most of the isolates were classified as multidrug resistant as they scored multiple antimicrobial resistance index ≥0.2. Furthermore, antimicrobial resistance genes tetB, qnrA, intI1 (Class 1 integron integrase), aac(6')-Ib, blaSHV, blaCTX-M, strA-strB, tetA, aphAI-IAB, qnrC, qnrS, and blaTEM were found in 81.63%, 67.35%, 61.22%, 46.94%, 44.90%, 44.90%, 36.73%, 18.37%, 10.20%, 10.20%, 8.16% and 6.12% of the isolates, respectively. In conclusion, the development of antimicrobial resistance among V. harveyi will ultimately reduce the efficacy of antimicrobials used for treating and can favor the development of more virulent V. harveyi strains.

Antimicrobial-resistant in Escherichia coli isolated from different effluent locations within Ahmadu Bello University, Zaria, Nigeria

The safety of municipal water is increasingly becoming of concern globally. Agricultural activities, industrial and residential effluents and community waste are ways through which water sources are contaminated and resistant bacteria can be spread via effluents to municipal water. The study aimed to isolate and determine the distribution of antimicrobial drug-resistant Escherichia coli from different points of the University sewer system in April 2018. A total of 48 samples were collected twice weekly from the six randomly selected inspection chamber sites out of the 14 identified sites. The selected sites of the sewer were located in some hostels, markets and health service areas within the ABU. main campus. The samples were processed by culturing on an EMB agar plate followed by biochemical characterization using conventional biochemical tests and Microbact 12E. An antimicrobial sensitivity test was also carried out using 13 different antibiotic discs. The results obtained revealed that the Community market had an isolation rate of 4(50%), while Sickbay had 3(37.5%) and Danfodiyo hostel with 2(25%). Multiple antimicrobial resistance index (MARI) was found to be 0.31 from four isolates (36%) of E. coli of which 3(75%) were sampled from Sickbay and 1(25%) from ABU Dam. Also, five isolates (45%) had MARI of 0.23, of which 2(40%) were sampled from Danfodiyo hostel, 1(20%) from Ribadu hostel and 2(40%) from Community market. The E. coli isolates were more resistant to Ampicillin, tetracycline and cephalothin. Other bacteria isolated were Klebsiella ozaenae, Hapnea alvei and Morganella morganii all with MARI of 0.31. There is a need for public health awareness on the effect of discharging antibiotic-resistant E. coli contaminated effluent into the environment and water bodies. Hence, the public health significance of recycling such water for domestic usage and agricultural purpose.

Rethinking Manure Application: Increase in Multidrug-Resistant Enterococcus spp. in Agricultural Soil Following Chicken Litter Application.

The current study investigated the impact of chicken litter application on the abundance of multidrug-resistant Enterococcus spp. in agricultural soil. Soil samples were collected from five different strategic places on a sugarcane farm before and after manure application for four months. Chicken litter samples were also collected. Enterococci were enumerated using the Enterolert®/Quanti-Tray 2000® system and confirm and differentiated into species using real-time PCR. The antibiotic susceptibility profile of the isolates was determined using the disk diffusion method following the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. The overall mean bacterial count was significantly higher (p < 0.05) in manure-amended soil (3.87 × 107 MPN/g) than unamended soil (2.89 × 107 MPN/g). Eight hundred and thirty-five enterococci (680 from soil and 155 from litter) were isolated, with E. casseliflavus being the most prevalent species (469; 56.2%) and E. gallinarum being the least (16; 1.2%). Approximately 56% of all the isolates were resistant to at least one antibiotic tested, with the highest resistance observed against tetracycline (33%) and the lowest against chloramphenicol (0.1%); 17% of E. faecium were resistant to quinupristin-dalfopristin. Additionally, 27.9% (130/466) of the isolates were multidrug-resistant, with litter-amended soil harbouring more multidrug-resistant (MDR) isolates (67.7%; 88/130) than unamended soil (10.0%; 13/130). All isolates were susceptible to tigecycline, linezolid and gentamicin. About 7% of the isolates had a multiple antimicrobial resistance index > 0.2, indicative of high antibiotic exposure. Although organic fertilizers are regarded as eco-friendly compared to chemical fertilizers for improving soil fertility, the application of untreated animal manure could promote the accumulation of antibiotics and their residues and antibiotic-resistant bacteria in the soil, creating an environmental reservoir of antimicrobial resistance, with potential human and environmental health risks.

Virulotyping and genetic diversity of Aeromonas hydrophila isolated from Nile tilapia (Oreochromis niloticus) in aquaculture farms in Egypt

This study focused on the genotyping of Aeromonas hydrophila isolated from diseased Nile tilapia, virulence genes, and their relation with motile aeromonad septicemia (MAS) clinical signs, and the resistance profiles of the recovered isolates to the frequently used antimicrobials in aquaculture. We determined the prevalence of A. hydrophila and the associated mortality in fish in various farms. We also investigated the extent of homogeneity of isolates sampled from various aquaculture farms and fish organs and how these variables would drive A. hydrophila heterogeneity to help developing control strategies against this infection. Of the 41 (41%) A. hydrophila isolates recovered by culture from diseased fish in four aquaculture fish farms (faced 35–50% mortality rates) located at different geographical regions in Egypt, 39 (39%) were confirmed by 16S rDNA-PCR amplification. Five distinct patterns were found for the distribution of virulence genes among 41% of A. hydrophila isolates. Thirteen isolates (33.33%) were positive for aer gene; 9 (23.1%) harbored lip gene, 2 (5.13%) carried hlyA gene, and one isolate (2.56%) was positive for ast gene. A positive correlation was found between hlyA and ast, and between lip and aer genes (r = 0.6 and 0.5, respectively). All isolates were resistant to ampicillin and amoxicillin. The least resistance was noted for doxycycline and gentamicin (7.7%) followed by streptomycin (10.3%), trimethoprim/sulfamethoxazole (17.9%), chloramphenicol (25.6%), colistin (43.6%), and erythromycin (47.7%). Most isolates were susceptible to ciprofloxacin. Multiple antimicrobial resistance index values ranged from 0·2–0·6 among multidrug-resistant (MDR) isolates (87·2%). Enterobacterial repetitive intergenic consensus (ERIC)-PCR allowed differentiating A. hydrophila isolates into 3 ERIC clusters, each contained isolates belonging to different farms and organs. The fingerprinting profiles of isolates were diverse without any obvious regional or organ-specific isolates. Our results indicated that sampling localities and sampled organs differ, albeit none substantially, in terms of the respective isolates heterogeneity. In conclusion, the high prevalence of A. hydrophila in Nile tilapia and its diverse antimicrobial resistance and virulence profile suggest the increased risk of this infection and raise concerns about its public health importance. The prudent use of antibiotics in the fish industry is now becoming mandatory to avoid the great losses in aquaculture due to MDR A. hydrophila. Our study also indicated the importance of ERIC genotyping for the analysis of A. hydrophila diversity during future molecular epidemiology investigations.

Incidence of multiresistant antimicrobial Staphylococcus in the hands of Health Basic Unit professionals

Staphylococcus is an important etiologic agent of infections in hospital and healthcare settings. One of the means of proliferation of these agents is the contamination of the hands of professionals who perform health care. This research aimed to identify the possible incidence of multiresistant Staphylococcus aureus colonized in the hands of health professionals, before and after hand hygiene, and its correlation with the spread of infections in these environments. The research was carried out by biochemical tests in the laboratory, of which results indicated the colonization by Staphylococcus even after hand hygiene (88.3% positive and 11.7% negative); this allows the dissemination of the bacteria to other patients and environments. The samples were submitted to the multiple antimicrobial resistance index, and data showed the persistence of microorganisms resistant to the 15 antimicrobials used. So additional studies need to be done in order to suggest effective actions and awareness of health professionals about hand hygiene as an important preventive action in hospital and health care settings.

Occurrence of Multi-Resistant Staphylococcus aureus in Artisan Goat Coalho Cheese in Northeastern Brazil

Background: The artisanal goat coalho cheese is one of the products obtained that stand out in 11 the dairy goat farming of the Northeast of Brazil. Despite its importance, goat cheese is often 12 made under inadequate hygienic-sanitary conditions and usually uses raw goat's milk, 13 increasing the risk of product contamination. Among the pathogens carried by goat coalho 14 cheese, Staphylococcus aureus stands out, being responsible for cases of food poisoning and 15 persistent infections that are difficult to treat. This study aimed to evaluate the contamination, 16 genotypic and phenotypic resistance of Staphylococcus aureus isolated from artisanal coalho 17 cheese made with goat milk produced in the Northeast region of Brazil.Materials, Methods &amp; Results: This study analyzed only artisanal coalho cheeses made with raw goat's milk and purchased directly from farms. Twelve samples of artisanal coalho cheeses made with raw goat's milk were collected (1 sample per property) in 8 municipalities in the state of Pernambuco, Northeast region of Brazil. For microbiological analysis of enumeration of Colony Forming Units (CFU/g) of Staphylococcus spp. the methodology recommended by the International Organization for Standardization (2019) and recognized by the Ministry of Agriculture, Livestock and Supply was used. After enumeration, 5 colonies were selected per enumerated plate, a total of 180 Staphylococcus spp. was obtained. These were subjected to thermal extraction of genetic material to search for the nuc gene by Polymerase Chain Reaction, the isolates carrying the nuc gene were subjected to genotypic and phenotypic evaluation of antimicrobial resistance. After the phenotypic analysis, the Multiple Antimicrobial Resistance Index was evaluated. In all samples, Staphylococcus spp. and were considered unfit for consumption, with the lowest count being 9.4x103 CFU/g and the highest 6.4x106 CFU /g. Of the 180 isolates, 28.34% (51/180) were positive for the detection of the nuc gene. All resistance genes except mecA, mecC, and norB were detected. Of the 51 S. aureus isolates, 31.37% (16/51) were considered multi-resistant and presented a Multiple Antimicrobial Resistance Index above 0.2.Discussion: After microbiological analysis it was found that all samples of coalho cheese were out of standards and unfit for human consumption in accordance with Ordinance n° 146/1996 of the Ministry of Agriculture, Livestock and Supply of Brazil. Furthermore, the contamination of goat coalho cheeses is a risk to public health. During sample collection were found inadequate hygiene conditions in the environment used for cheese production. The presence of Staphylococcus aureus can be attributed to hygienic-sanitary failures in cheese production. From a health point of view, it is even more alarming when it comes to S. aureus carrying resistance genes. Although the 51 S. aureus isolates did not carry the mecA, mecC, norB genes and did not show phenotypic resistance to cefoxitin and oxacillin, all other genes were detected, indicating the circulation of S. aureus carrying the tet(L) genes, tet(M), tet-38, msrA, norA, and norC, which so far had not been reported in the production chain of goat coalho cheese in Brazil. Furthermore, the evaluation of the Multiple Antimicrobial Resistance Index identified the occurrence of multiple resistance to antimicrobials in 31.37% (16/51) of S. aureus at high risk to human health. The results obtained are quite worrying and serve as a warning to the scientific community and the Food Safety and Hygiene Inspection Services.Keywords: goat cheese, contamination, resistance, one health

Virulence Genes and Resistance Profile of Escherichia coli Isolated in Meat Meal Samples

Background: Feed is the main route of transmission of pathogenic microorganisms and is responsible for a large part of the cost of poultry production, so the inclusion of alternative foods in diets for monogastrics has been a constant. Among alternative foods most used in modern poultry farming are animal meal, however, when contaminated they constitute a route of transmission of several pathogenic agents, including Escherichia coli. In addition, there is a zoonotic potential, as poultry products are intended for human consumption. The objective of this research was to detect virulence genes, as well as to evaluate the resistance profile of Escherichia coli isolates from meat meal samples. Materials, Methods &amp; Results: A total of 40 Escherichia coli isolates were analyzed and the virulence genes surveyed iss, ompT, hlyF, iutA, and fimA identified by Polymerase Chain Reaction (PCR). The antimicrobial agents tested were: amoxycillin (30 μg), ceftiofur (30 μg), ciprofloxacin (5 μg), doxycycline (30 μg), florfenicol (30 μg), fosfomycin (200 μg), gentamicin (10 μg), norfloxacin (10 μg) and oxacillin (1 μg). It was possible to observe the occurrence of the iss resistance gene in 100% of Escherichia coli isolates, followed by hlyF (85%), fimA (75%), ompT (17.5%) and iutA (5%). Regarding the simultaneous detection for the genes, a greater association between the genes iss, hlyF and fimA (60%) was verified. All isolates showed resistance to oxacillin (100%), followed by doxycycline (25%), amoxicillin (22.5%), norfloxacin (17.5%), ceftiofur (15%), florfenicol (12.5%), fosfomycin (12.5%), ciprofloxacin (10%) and gentamycin (2.5%). In this study, a variation of the multiple antimicrobial resistance index (IRMA) was observed between 0.22 and 0.77. Discussion: The indiscriminate use of of antimicrobials as performance enhancers in production animals, may have contributed to the increase in antimicrobial resistance, with the occurrence of multiresistant Escherichia coli carrying virulence genes. Virulence genes present in Escherichia coli isolates are studied to understand the degree of influence they exert in the establishment of the disease, one of the most researched genes is the iss gene, involved in the processes that promote serum resistance. In this study, iss (100%) was present in all the isolates analyzed, although it is not the only mechanism used by these bacteria to reach internal organs and trigger an infection, this gene encodes an important mechanism associated with high levels of virulence. The second highest prevalence found was of the hlyF gene (85%), the high prevalence of hlyF suggests virulence potential, involved with the production of hemolysin and improvement of outer membrane vesicles associated with the release of toxins. The fimA gene (75%) was detected in a slightly lower percentage when compared to iss and hlyF. With the second lowest prevalence, the ompT gene (17.5%), is involved in a process that includes the proteolytic degradation of antimicrobial peptides and with the lowest prevalence the iutA gene (5%). Certain combinations of virulence genes make the strains easier to survive, adhere to, colonize and even the ability to develop septicemic conditions. Multiresistant E. coli strains, is a fact of concern for both animal and human health, since the presence of multiresistant strains, originating from poultry, can be transmitted from chicken carcasses. In this sense, the importance of sanitary control of the inputs used in animal feed is emphasized, as well as the prudent use of antimicrobials in animal production, with a view to producing a safe food, minimizing not only the economic losses, but also the risks to human health. Keywords: antimicrobial, bacterial resistance, colibacillosis, poultry.

Virulence, Resistance, and Genomic Fingerprint Traits of Vibrio cholerae Isolated from 12 Species of Aquatic Products in Shanghai, China.

Vibrio cholerae is a waterborne bacterium and can cause epidemic cholera disease worldwide. Continuous monitoring of V. cholerae contamination in aquatic products is imperative for assuring food safety. In this study, we determined virulence, antimicrobial susceptibility, heavy metal tolerance, and genomic fingerprints of 370 V. cholerae isolates recovered from 12 species of commonly consumed aquatic products collected from July to September of 2018 in Shanghai, China. Among the species, Leiocassis longirostris, Ictalurus punetaus, Ophiocephalus argus Cantor, and Pelteobagrus fulvidraco were for the first time detected for V. cholerae. Toxin genes ctxAB, tcpA, ace, and zot were absent from all the V. cholerae isolates. However, high occurrence of virulence-associated genes was detected, such as hapA (82.7%), hlyA (81.4%), rtxCABD (81.4%, 24.3%, 80.3%, and 80.8%, respectively), and tlh (80.5%). Approximately 62.2% of the 370 V. cholerae isolates exhibited resistance to streptomycin, followed by ampicillin (60.3%), rifampicin (53.8%), trimethoprim (38.4%), and sulfamethoxazole-trimethoprim (37.0%). Moreover, ∼57.6% of the isolates showed multidrug resistant phenotypes with 57 resistance profiles, which was significantly different among the 12 species (multiple antimicrobial resistance index, p < 0.001). Meanwhile, high incidence of tolerance to heavy metals Hg2+ (69.5%), Ni2+ (32.4%), and Cd2+ (30.8%) was observed among the isolates. The enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR)-based fingerprinting profiles classified the 370 V. cholerae isolates into 239 different ERIC-genotypes, which demonstrated diverse genomic variation among the isolates. Overall, the results in this study meet the increasing need of food safety risk assessment of aquatic products.

Antimicrobial Resistance Profiles of Diarrheagenic E. coli (DEC) and Salmonella Species Recovered from Diarrheal Patients in Selected Rural Communities of the Amathole District Municipality, Eastern Cape Province, South Africa.

PurposeThe emergence of multidrug-resistant bacteria remains as one of the major impediments towards the prevention and treatment of microbial infections and continues to be a serious threat to medicine. Henceforth, this study aimed at elucidating the antimicrobial resistance profiles of diarrheagenic E. coli (DEC) and Salmonella species recovered from diarrheal patients in selected rural communities of the Amathole District Municipality (ADM), Eastern Cape Province, South Africa (SA).MethodsThe antimicrobial resistance profiles of diarrheagenic E. coli (DEC) and Salmonella isolates were evaluated using antimicrobial susceptibility tests and the relevant antimicrobial resistance factors were elucidated by the Polymerase Chain Reaction technique.ResultsA sum of 324 diarrheagenic E. coli (DEC) and 62 Salmonella isolates were recovered from diarrheal stool specimens collected amongst diarrheal patients admitted in medical facilities/health-care centers within the ADM in the Eastern Cape Province, South Africa. Multiple antimicrobial resistance index mean values of 0.7 and 0.5 for DEC and Salmonella isolates, respectively, were observed in this study, indicating that these isolates were from sources where antimicrobials were frequently used. The antimicrobial resistance factors ampC, blaTEM, SulI and II, tet A and aadA were detected among antimicrobial-resistant DEC pathotypes and Salmonella isolates recovered in this study.ConclusionThe occurrence of the multiple antimicrobial-resistant DEC and Salmonella isolates with the relevant antimicrobial resistance factors in this study suggests a portentous human health threat associated with diarrhea and a major deterrent in medicine.