Olaquindox resistance in the coliform flora of pigs and their environment: an ecological study

Faecal samples were taken weekly from a chosen pen on each of four commercial pig farms, two of which used olaquindox as a feed additive. Coliform bacteria were isolated from the samples and the incidence and level of resistance to olaquindox and to four therapeutic antibiotics determined. The coliforms isolated were biotyped to follow the emergence of drug-resistant sub-populations. The results showed that the coliform flora was complex and that a turnover of biotypes was associated with changes in the occupancy of the pen and, possibly, diet. This turnover led to large fluctuations in both incidence and level of resistance to olaquindox and the antibiotics. Olaquindox resistance was not specifically linked with resistance to any therapeutic antibiotic, and the possible origin of olaquindox-resistant strains is discussed in relation to the biotypes.     

Monitoring for the development of antimicrobial resistance during the use of olaquindox as a feed additive on commercial pig farms

Since 1982, when olaquindox was introduced as a pig-feed additive in the UK, about 12 commercial farms in Suffolk have been monitored annually to check for the possible emergence of resistance to olaquindox and chloramphenicol among the coliform flora of the pigs and their environment. In spite of the sampling variability and the impossibility of controlling the use of feed additives and management on the farms, the overall results obtained were consistent and, it is suggested, the method is widely applicable. A steady, albeit low, increasing incidence and level of resistance to olaquindox was recorded (1982–1984) on farms using it and, to a lesser degree, on neighbouring farms that did not. No significant increase in the level of chloramphenicol resistance was observed. Genetical studies on a selection of olaquindox-resistant isolates suggested that the genes determining resistance were likely to be borne on the chromosome.     

Monitoring for the development of antimicrobial resistance during the use of olaquindox as a feed additive on commercial pig farms

Since 1982, when olaquindox was introduced as a pig-feed additive in the UK, about 12 commercial farms in Suffolk have been monitored annually to check for the possible emergence of resistance to olaquindox and chloramphenicol among the coliform flora of the pigs and their environment. In spite of the sampling variability and the impossibility of controlling the use of feed additives and management on the farms, the overall results obtained were consistent and, it is suggested, the method is widely applicable. A steady, albeit low, increasing incidence and level of resistance to olaquindox was recorded (1982-1984) on farms using it and, to a lesser degree, on neighbouring farms that did not. No significant increase in the level of chloramphenicol resistance was observed. Genetical studies on a selection of olaquindox-resistant isolates suggested that the genes determining resistance were likely to be borne on the chromosome.     

Incidence of multiple antibiotic resistant Escherichia coli in the Bhavani River

Of 107 Escherichia coli strains isolated from the water, sediment and fish of the Bhavani River, all of which are considered potential causes of human enteric disease, 62% were resistant to more than four antibiotics. Levels of resistance to bacitracin, penicillin, and novobiocin were generally high whereas those to polymyxin-B and chloramphenicol were much lower. A high incidence of multiple antibiotic resistant E. coli was noted in all samples and the multiple antibiotic resistance index of the strains showed that 95% of the strains originated either from man or cattle.     

Serotypes and biotypes and antibiotic susceptibility of Haemophilus influenzae encountered in a clinical laboratory in Taiwan.

Forty-four serologically and biochemically typable Haemophilus influenzae isolates from clinical specimens in Taiwan were subjected to analysis in their relationship with source of isolation and age distribution. It was found that all isolates from blood and cerebrospinal fluid were serotype b, biotype I, and all were in children less than 4 years of age. Serotypes b and e, biotypes I and III were encountered to have the highest incidence of infection caused by H. influenzae in this area. All H. influenzae isolates were further tested for susceptibility to several selected antibiotics. All strains of this organism were susceptible to erythromycin and chloramphenicol. All but two strains were susceptible to tetracycline, whereas more strains were resistant to carbenicillin, gentamycin, keflin, and penicillin. Thirty-four percent strains were found to be resistant to ampicillin and all were beta-lactamase producer. No direct correlation between ampicillin resistance and serotypes or biotypes was recognized.     

Antimicrobial resistance pattern of methicillin-resistant Staphylococcus aureus in the food industry

There is increasing concern about the impact on public health of methicillin-resistant Staphylococcus aureus (MRSA) associated with animal food products. MRSA remains a serious problem because of the high incidence and multidrug resistance of the strains, even for strains isolated from foods, food environments and food handlers. The objectives of this study are: (i) to evaluate the susceptibility of S. aureus strains isolated from food, food handlers and food-processing environments to 14 antibiotics currently used in veterinary and human therapy; (ii) to assess the presence of the mecA gene. A total of 1007 samples were collected from food, food handlers, and environments and were analyzed for the presence of S. aureus. S. aureus was present in 165 of the 1007 samples. A total of 157 isolates were methicillin-susceptible S. aureus (MSSA) and 8 isolates were MRSA. In particular, out of 8 MRSA strains detected, 4 strains harboured the mecA gene. All MRSA strains were resistant to at least one of the tested antibiotics and 6 strains demonstrated multi-resistance. Considering the high level of resistances in S. aureus and the isolation of MRSA strains, the surveillance of antimicrobial resistance and the spreading of this pathogen is of crucial importance in the food production chain. These data are useful in improving background data on antimicrobial resistance of S. aureus isolated from food, processing environments and food handlers, supporting the prudent use of antibiotics and the development of international control programs.     

Phenotypical characteristics of Shiga-like toxin Escherichia coli isolated from sheep dairy products

AIMS: To analyse phenotypical characteristics of Shiga toxin-producing Escherichia coli (STEC) strains from ovine origin. METHODS AND RESULTS: A total of 13 STEC strains (eight O157 and five non-O157) isolated from sheep dairy products were used in this study. Biochemical traits, motility, haemolytic activity, resistance to tellurite-cefixime, maximum growth temperature and antibiotic resistance were determined. The STEC strains were grouped into nine biochemical and physiological biotypes (five for the O157 and four for the non-O157 strains). All STEC strains showed resistance to bacitracin, cloxacilin, penicillin and tylosin. CONCLUSIONS: Different biotypes and antibiotic resistance patterns of STEC isolated from sheep dairy products were observed. SIGNIFICANCE AND IMPACT OF THE STUDY: This work will be a contribution to the better characterization of STEC isolated from sheep dairy products, which have, to date, been scarcely studied, and to the better understanding of the risks associated with its consumption.     

Antibiotic growth promoter olaquindox increases pathogen susceptibility in fish by inducing gut microbiota dysbiosis

Low dose antibiotics have been used as growth promoters in livestock and fish. The use of antibiotics has been associated with reduced pathogen infections in livestock. In contrast, antibiotic growth promoter has been suspected of leading to disease outbreaks in aquaculture. However, this phenomenon is circumstantial and has not been confirmed in experimental conditions. In this study,we showed that antibiotic olaquindox increased the susceptibility of zebrafish to A. hydrophila infection. Olaquindox led to profound alterations in the intestinal microbiota of zebrafish, with a drastic bloom of Enterobacter and diminishing of Cetobacterium. Moreover, the innate immune responses of zebrafish were compromised by olaquindox(P0.05). Transfer of microbiota to GF zebrafish indicated that while the immuo-suppression effect of olaquindox is a combined effect mediated by both OLA-altered micro biota and direct action of the antibiotic(P0.05), the increased pathogen susceptibility was driven by the OLA-altered microbiota and was not dependent on direct antibiotic effect. Taken together, these data indicate that low level of OLA induced gut microbiota dysbiosis in zebrafish, which led to increased pathogen susceptibility.     

Antimicrobial resistance of bacteria isolated from slaughtered and retail chickens in South Africa

Animal feed is increasingly being supplemented with antibiotics to decrease the risk of epidemics in animal husbandry. This practice could lead to the selection for antibiotic resistant micro-organisms. The aim of this study was to determine the level of antibiotic resistant bacteria present on retail and abattoir chicken. Staphylococci, Enterobacteriaceae, Salmonella and isolates from total aerobic plate count were tested for resistance to vancomycin, streptomycin, methicillin, tetracycline and gentamicin using the disc diffusion susceptibility test; resistance to penicillin was determined using oxacillin. Results from the antibiotic code profile indicated that many of the bacterial strains were displaying multiple antibiotic resistance (MAR). A larger proportion of resistance to most antibiotics, except for vancomycin, was displayed by the abattoir samples, therefore suggesting that the incidence of MAR pathogenic bacteria was also higher in the abattoir samples. This resistance spectrum of abattoir samples is a result of farmers adding low doses of antibiotics to livestock feed to improve feeding efficiency so that the animals need less food to reach marketable weight. The lower incidence of MAR pathogenic bacteria in the retail samples is a result of resistance genes being lost due to lack of selective pressure, or to the fact that the resistant flora are being replaced by more sensitive flora during processing. The use of subtherapeutic levels of antibiotics for prophylaxis and as growth promoters remains a concern as the laws of evolution dictate that microbes will eventually develop resistance to practically any antibiotic. Selective pressure exerted by widespread antimicrobial use is therefore the driving force in the development of antibiotic resistance. This study indicated that a large proportion of the bacterial flora on fresh chicken is resistant to a variety of antibiotics, and that resultant food-related infections will be more difficult to treat.     

Polygenes and modifier genes for tetracycline and penicillin resistance in Neisseria gonorrhoeae

The genetic basis for spontaneous resistance to tetracyline (Tet) and penicillin (Pen) in Neisseria gonorrhoeae was investigated. Tet and pen are polygenes which confer small but distinct levels of resistance to Tet and Pen, respectively. Mtr is a multiple-drug resistance polygene which increases resistance to Tet and Pen (as well as to other unrelated antibiotics). Tem is a modifier gene affecting resistance toTet and Pen. Pem is a modifier gene for Pen resistance. The following gene combinations code for resistance to five antibiotics: tet, mtr and tem for Tet; pen, mtr, pem and tem for Pen; tet, tem and mtr for doxycycline; pen and pem for ampicillin; pen, pem and mtr for nafcillin.     

Presence of various Acinetobacter species in urinary tract infections

Species classification of 39 strains of Acinetobacter isolated from urinary tract infections (UTI) was performed by API 20NE tests. On the whole 24 biotypes were differentiated which formed 4 species and one biotype had no defined species classification. Acinetobacter baumani was most numerously represented. Seventy seven percent of strains were isolated from urine samples from women. About 90% of Acinetobacter strains were isolated as single pathogens. Sensitivity of these strains to 23 antibiotics and chemotherapeutics using ATB-UR and disc diffusion technique was evaluated. Of all cephalosporins tested ceftazidime was the most active. All strains tested were sensitive to imepenem. The high percentage of strains was sensitive to quinolones of IIIrd generation. The high percentage of strains was resistant to nitrofurantoin.     

三种抗生素对B型和Q型烟粉虱内共生菌的去除效果比较研究

利用烟粉虱Bemisia tabaci(Gennadius)内共生菌特异性引物,研究了内共生菌在B、Q型烟粉虱种群中的分布和感染率,同时评价了3种不同的抗生素利福平、氨苄青霉素和硫酸卡那霉素分别在3种不同的浓度下(100.0、50.0及25.0μg/mL)对烟粉虱内共生菌的去除效果。结果表明:B、Q型烟粉虱原生内共生细菌Portiera的带菌率均为100.0%;B、Q型烟粉虱次生内共生菌Hamiltonella的带菌率分别为91.7%和100.0%;B型烟粉虱次生内共生菌Rickettsia的带菌率为87.5%,Q型为0;其它次生内共生菌在B、Q型烟粉虱中均未检测到。利福平、氨苄青霉素和硫酸卡那霉素在3种不同的浓度下均不能去除B、Q型烟粉虱Portiera;利福平、氨苄青霉素在3种不同的浓度下均能完全去除B型烟粉虱Rickettsia,硫酸卡那霉素在不同浓度下去除Rickettsia的效果不同;3种抗生素去除Hamiltonella的能力受抗生素种类以及浓度的影响。同一抗生素在不同浓度下去除Hamiltonella的效果均是100.0μg/mL>50.0μg/mL>25.0μg/mL;不同浓度的抗生素去除Hamiltonella的效果均是利福平>氨苄青霉素>硫酸卡那霉素,各浓度与各抗生素之间的去除Hamiltonella的效果均具有显著性差异。     

Antimicrobial susceptibilities of Aeromonas spp. isolated from environmental sources

Aeromonas spp. are ubiquitous aquatic bacteria that cause serious infections in both poikilothermic and endothermic animals, including humans. Clinical isolates have shown an increasing incidence of antibiotic and antimicrobial drug resistance since the widespread use of antibiotics began. A total of 282 Aeromonas pure cultures were isolated from both urban and rural playa lakes in the vicinity of Lubbock, Texas, and several rivers in West Texas and New Mexico. Of these, at least 104 were subsequently confirmed to be independent isolates. The 104 isolates were identified by Biolog and belonged to 11 different species. The MICs of six metals, one metalloid, five antibiotics, and two antimicrobial drugs were determined. All aeromonads were sensitive to chromate, cobalt, copper, nickel, zinc, cefuroxime, kanamycin, nalidixic acid, ofloxacin, tetracycline, and sulfamethoxazole. Low incidences of trimethoprim resistance, mercury resistance, and arsenite resistance were found. Dual resistances were found in 5 of the 104 Aeromonas isolates. Greater numbers of resistant isolates were obtained from samples taken in March versus July 2002 and from sediment versus water. Plasmids were isolated from selected strains of the arsenite- and mercury-resistant organisms and were transformed into Escherichia coli XL1-Blue MRF'. Acquisition of the resistance phenotypes by the new host showed that these resistance genes were carried on the plasmids. Mercury resistance was found to be encoded on a conjugative plasmid. Despite the low incidence of resistant isolates, the six playa lakes and three rivers that were sampled in this study can be considered a reservoir for antimicrobial resistance genes.     

Biotyping and resistotyping for epidemiological tracing of the fecal origin of Klebsiella pneumoniae in urinary infections

Twenty-four (82.7%) out of 29 patients suffering from hospital acquired urinary infections by Klebsiella pneumoniae had the same species in their faeces. Biotyping of 24 urinary and 219 fecal strains of K. pneumoniae resulted in 50 different biotypes - an average of four biotypes per fecal sample. Ten patients (34.4%) had the same biotype in urine and faeces without any correlation with previous vesical catheterization (p greater than 0.05). Using resistotyping to four chemical compounds selected among 34 tested substances (brilliant green, malachite green, potassium tellurite and mercuric chloride) 16 different resistotypes were found. Fourteen patients (58.3%) presented the same resistotype in urine and faeces but only in five patients was there correlation with simultaneous biotyping identity. Simultaneous occurrence of identical biotypes or resistotypes in faeces and urine occurred in only 54.2% of cases. However, there was a significant association between resistance ot mercuric and tellurite ions in fecal and urinary strains isolated from the same patient (p less than 0.001).     

Characterization of intermediate biotypes in atrazine-susceptible populations of Chenopodium polyspermum L. and Amaranthus bouchonii thell. in Hungary

Intermediate biotypes for atrazine herbicide resistance in Chenopodium polyspermum and Amaranthus bouchonii were characterized by a peculiar chlorophyll fluorescence induction curve. The intermediate biotypes were isolated from progenies of susceptible plants in maize grown in alternate years without atrazine. The lethal dose in seedling treatments was lower than that of the resistant plants but higher than for susceptible plants. Atrazine at 10 μM was near the I₅₀-value for in vivo nitrite reductase activity in both intermediate biotypes. The activity of nitrite reducttase in the intermediate biotypes was about 75% of that of susceptible biotypes. These characteristics of intermediate biotypes were maternally inherited in crosses.     

Uncultured soil bacteria are a reservoir of new antibiotic resistance genes

Antibiotic resistance genes are typically isolated by cloning from cultured bacteria or by polymerase chain reaction (PCR) amplification from environmental samples. These methods do not access the potential reservoir of undiscovered antibiotic resistance genes harboured by soil bacteria because most soil bacteria are not cultured readily, and PCR detection of antibiotic resistance genes depends on primers that are based on known genes. To explore this reservoir, we isolated DNA directly from soil samples, cloned the DNA and selected for clones that expressed antibiotic resistance in Escherichia coli. We constructed four libraries that collectively contain 4.1 gigabases of cloned soil DNA. From these and two previously reported libraries, we identified nine clones expressing resistance to aminoglycoside antibiotics and one expressing tetracycline resistance. Based on the predicted amino acid sequences of the resistance genes, the resistance mechanisms include efflux of tetracycline and inactivation of aminoglycoside antibiotics by phosphorylation and acetylation. With one exception, all the sequences are considerably different from previously reported sequences. The results indicate that soil bacteria are a reservoir of antibiotic resistance genes with greater genetic diversity than previously accounted for, and that the diversity can be surveyed by a culture-independent method.     

Prevalence and antibiotic resistance of <Emphasis Type="Italic">Escherichia coli</Emphasis> in tropical seafood

Summary The occurrence and antibiotic resistance of Escherichia coli in tropical seafood was studied. A 3-tube MPN method was used for determining the level of faecal contamination of fresh and processed seafood. Of the 188 samples tested which included finfish, shellfish, water and ice, 155 were positive for the presence of faecal coliforms following incubation at 44.5 °C. However, E. coli was isolated from only 47% of the samples positive for faecal coliforms. The antibiotic resistance of 116 strains isolated from seafood was tested using 14 different antibiotics including ampicillin, cephalothin, chloramphenicol, ciprofloxacin, gentamycin, nalidixic acid, streptomycin and vancomycin. Seven strains were resistant to more than five antibiotics of which one was resistant to eight antibiotics. The multiple drug resistant strains harboured plasmids of varying sizes. Antibiotic susceptibility studies revealed that seafood from India contains multiple antibiotic resistant strains of E. coli which may serve as a reservoir for antibiotic resistance genes in the aquatic environment. All the strains used in this study did not harbour any virulence genes commonly associated with pathogenic E. coli, when tested by polymerase chain reaction (PCR).     

No evidence for early fitness penalty in glyphosate‐resistant biotypes of Conyza canadensis: Common garden experiments in the absence of glyphosate

Strong selection from herbicides has led to the rapid evolution of herbicide‐resistant weeds, greatly complicating weed management efforts worldwide. In particular, overreliance on glyphosate, the active ingredient in RoundUp^®, has spurred the evolution of resistance to this herbicide in ≥40 species. Previously, we reported that Conyza canadensis (horseweed) has evolved extreme resistance to glyphosate, surviving at 40× the original 1× effective dosage. Here, we tested for underlying fitness effects of glyphosate resistance to better understand whether resistance could persist indefinitely in this self‐pollinating, annual weed. We sampled seeds from a single maternal plant (“biotype”) at each of 26 horseweed populations in Iowa, representing nine susceptible biotypes (S), eight with low‐level resistance (LR), and nine with extreme resistance (ER). In 2016 and 2017, we compared early growth rates and bolting dates of these biotypes in common garden experiments at two sites near Ames, Iowa. Nested ANOVAs showed that, as a group, ER biotypes attained similar or larger rosette size after 6 weeks compared to S or LR biotypes, which were similar to each other in size. Also, ER biotypes bolted 1–2 weeks earlier than S or LR biotypes. These fitness‐related traits also varied among biotypes within the same resistance category, and time to bolting was inversely correlated with rosette size across all biotypes. Disease symptoms affected 40% of all plants in 2016 and 78% in 2017, so we did not attempt to measure lifetime fecundity. In both years, the frequency of disease symptoms was greatest in S biotypes and similar in LR versus ER biotypes. Overall, our findings indicate there are no early growth penalty and possibly no lifetime fitness penalty associated with glyphosate resistance, including extremely strong resistance. We conclude that glyphosate resistance is likely to persist in horseweed populations, with or without continued selection pressure from exposure to glyphosate.     

Antimicrobial resistance of Salmonella spp. from pigs at slaughter in Spain in 1993 and 2001

AIM: To compare the incidence of antimicrobial resistance among Salmonella serotypes isolated in a pig slaughterhouse in Zaragoza (Spain) during 1993 and 2001. METHODS AND RESULTS: A total of 168 isolates representing 10 serotypes were examined by disc diffusion method using 17 antibiotics. Data showed that the majority of the strains were resistant to streptomycin (97%), sulfadiazine (93.4%) and tetracycline (83.3%). A large proportion of the collection was multidrug resistant (MDR, resistance to four or more antibiotics) with a greater incidence in 2001. The findings imply an increasing incidence of MDR amongst S. Typhimurium, and all S. Typhimurium-definitive phage type (DT) 104 isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulphonamide and tetracycline (R-ACSSuT). This resistance phenotype had spread among other phage and serotypes. Salmonella Ohio was also a MDR serotype and this is not a serotype normally associated with drug resistance. CONCLUSIONS: A large proportion of the strains were MDR and this showed that pork products could be a potential vehicle of MDR Salmonella food-borne infections. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings may have significant public health consequences and could contribute to the development of useful practices aimed at limiting the transmission of MDR Salmonella serotypes through the food chain.     

Genetic analysis of drug resistance in Neisseria gonorrhoeae: production of increased resistance by the combination of two antibiotic resistance loci.

The studies reported here demonstrate that increased resistance of Neisseria gonorrhoeae to penicillin, tetracycline, and chloramphenicol results from the combined effect of two resistance loci. As shown by experiments with deoxyribonucleic acid from transformants carrying only a single resistance locus, transformants with an incresed level of resistance to penicillin result from the combination of a penicillin-specific locus, pen, and a multiple resistance locus, mtr. Similarly, transformants with an increased level of resistance to tetracycline result from the combination of mtr and a tetracycline-specific locus, tet. Transformants with an increased level of resistance to chloramphenicol result from the combination of mtr and a chloramphenicol-specific locus, cml. Deoxyribonucleic acid dilution experiments established that only a single dose of each of the two required resistance loci is necessary to give higher-level resistance. Higher-level-resistant transformants were not obtained when a double dose of one resistance locus or a combination of loci pairs other than mtr and pen, mtr and tet, or mtr and cml was introduced into a recipient. Combinations of the mtr and tet genes resulted in increased resistance to semisynthetic tetracyclines. The presence of the mtr and pen genes resulted in increased resistance to penicillinase-stable penicillins.