Treatable Traits in Chronic Respiratory Disease: A Comprehensive Review.

Chronic respiratory diseases are major contributors to the global burden of disease. While understanding of these diseases has improved, treatment guidelines have continued to rely on severity and exacerbation-based approaches. A new personalised approach, termed the "treatable traits" approach, has been suggested to address the limitations of the existing treatment strategies. We aim to systematically review the current evidence regarding treatable traits in chronic respiratory diseases and to identify gaps in the current literature. We searched the PubMed and Embase databases and included studies on treatable traits and chronic respiratory diseases. We then extracted information on prevalence, prognostic implications, treatment options and benefits from these studies. A total of 58 papers was included for review. The traits identified were grouped into five broad themes: physiological, biochemical, psychosocial, microbiological, and comorbidity traits. Studies have shown advantages of the treatable traits paradigm in the clinical setting. However, few randomised controlled trials have been conducted. Findings from our review suggest that multidisciplinary management with therapies targeted at treatable traits has the potential to be efficacious when added to the best practices currently implemented. This paradigm has the potential to improve the holistic care of chronic respiratory diseases.

The infant pharyngeal microbiomes: origin, impact and manipulation.

PURPOSE OF REVIEW: There has been an exponential increase in research into infant microbiome evolution, and it appears that pharyngeal microbiota are associated with clinical phenotypes (e.g. infection and asthma). Although broad consensus views are emerging, significant challenges and uncertainties remain. RECENT FINDINGS: Infant pharyngeal microbiome research is limited by low biomass, high temporal diversity and lack of agreed standards for sampling, DNA sequencing and taxonomic reporting. Analysis of amplicon sequence variants and improved cost and availability of whole-genome sequencing are promising options for improving taxonomic resolution of such studies. Infant respiratory microbiomes arise, at least in part, from maternal flora (e.g. the respiratory tract and breastmilk), and are associated with environmental and clinical factors (e.g. mode of feeding and delivery, siblings, daycare attendance, birth season and antibiotic usage). Interventional research to modify the infant pharyngeal microbiota has recently been reported, using dietary supplements. SUMMARY: Further work is needed to improve characterization of the infant pharyngeal microbiomes, including routes of bacterial acquisition, role of environmental factors and associations with disease phenotypes. Methodological standards are desirable to facilitate more reproducible, comparable research. Improved understanding may enable manipulation of infant pharyngeal microbiota to improve clinical outcomes.

Antibacterial activity against porcine respiratory bacterial pathogens and in vitro biocompatibility of essential oils.

Bacterial respiratory infections affecting pigs such as pneumonia, pleuropneumonia, and pleurisy, are a major health concern in the swine industry and are associated with important economic losses. This study aimed to investigate the antibacterial activities of essential oils against major swine respiratory pathogens with a view to developing a potential alternative to antibiotics. Their synergistic interactions with the bacteriocin nisin was also examined. Lastly, we assessed the in vitro biocompatibility of the most efficient essential oils using a pig tracheal epithelial cell line. Of the nine essential oils tested, those from cinnamon, thyme, and winter savory were the most active against Streptococcus suis, Actinobacillus pleuropneumoniae, Actinobacillus suis, Bordetella bronchiseptica, Haemophilus parasuis, and Pasteurella multocida, with minimum inhibitory concentrations and minimum bactericidal concentrations ranging from 0.01 to 0.156% (v/v). The main component found in cinnamon, thyme, and winter savory oils were cinnamaldehyde, thymol, and carvacrol, respectively. Treating pre-formed S. suis and A. pleuropneumoniae biofilms with thyme or winter savory oils significantly decreased biofilm viability. We also observed a synergistic growth inhibition of S. suis with mixtures of nisin and essential oils from thyme and winter savory. Concentrations of nisin and cinnamon, thyme and winter savory essential oils that were effective against bacterial pathogens had no effect on the viability of pig tracheal epithelial cells. The present study brought evidence that essential oils are potential antimicrobial agents against bacteria associated with porcine respiratory infections.

Associations between management practices and within-pen prevalence of calf diarrhea and respiratory disease on dairy farms using automated milk feeders.

Data on management practices used with automated milk feeders (AMF) are needed to identify factors associated with calf health in these systems. The objectives of this observational, longitudinal, cross-sectional study were to estimate the prevalence of calf diarrhea (CD) and bovine respiratory disease (BRD), and to identify factors associated with prevalence of these diseases at the pen level on dairy farms feeding milk to group-housed calves with AMF. Seventeen dairy farms with AMF in Ontario, Canada, were visited 4 times, seasonally, over 1 yr. The clinical health of all calves (n = 1,488) in pens (n = 35) with AMF was scored to identify the number of calves with CD and BRD. Data on calf, feeder, and pen management practices were analyzed using generalized linear mixed regression models for each disease. Overall calf-level prevalence of CD and BRD were 23 and 17%, respectively. Median (interquartile range, IQR) within-pen prevalence of CD and BRD were 17% (7 to 37%) and 11% (0 to 28%), respectively. Median age (IQR) for diarrheic calves was 25 d (16 to 42 d), and for calves with BRD was 43 d (29 to 60 d). Factors associated with lower within-pen prevalence of CD were the administration of vitamin E and selenium at birth [odds ratio (OR) = 0.56; 95% confidence interval (CI): 0.32 to 0.99], feeding of probiotics (OR = 0.44, 95% CI: 0.22 to 0.93), and adding fresh bedding every 2 to 3 d (OR = 0.43; 95% CI: 0.24 to 0.76) compared with every 7 or more days. In contrast, sharing air with older cattle (>9 mo old) was associated with increased within-pen prevalence of CD (OR = 4.54, 95% CI: 1.88 to 10.52). Additionally, total bacteria counts ≥100,000 cfu/mL in milk samples taken from the AMF mixing jar were associated with increased within-pen prevalence of CD during the summer visit (OR = 3.34; 95% CI: 1.31 to 8.54). Increased total solids in milk or milk replacer (OR = 0.48, 95% CI: 0.27 to 0.85) and feeding whole milk versus milk replacer (OR = 0.29, 95% CI: 0.11 to 0.75) were associated with lower within-pen prevalence of BRD. Factors associated with greater within-pen prevalence of BRD were sharing air with weaned cattle up to 8 mo old (OR = 3.21, 95% CI: 1.26 to 8.16), and greater depth of the wet bedding pack. The use of maternity pens for reasons other than just calving was associated with increased prevalence of both CD and BRD (OR = 1.85, 95% CI: 1.03 to 3.33; OR = 2.61, 95% CI: 1.21 to 5.58, respectively). These results suggest that isolation from older animals and frequent cleaning of the feeder and pen may help to reduce disease prevalence in group-housed calves fed with an AMF.

Treatment history and antimicrobial susceptibility results for Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni isolates from bovine respiratory disease cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2013 to 2015.

Bovine respiratory disease is the most costly disease facing the cattle industry. Increasing resistance to antimicrobial treatment has been presented as a significant contributing factor, often through summarized susceptibility testing data. We assessed the relationship between previous antimicrobial treatment and antimicrobial susceptibility results from isolates of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni cultured from bovine respiratory cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2013 to 2015. Antimicrobial susceptibility data from 1,251 bacterial isolates were included for analysis. More bacterial isolates from cattle that received antimicrobial treatment showed resistance compared to isolates from untreated cattle, and the percentage of resistant isolates increased as the number of antimicrobial treatments increased. Resistance to enrofloxacin, spectinomycin, tilmicosin, and tulathromycin was present in >75% of M. haemolytica isolates from cattle that had received 3 or more antimicrobial treatments; resistance to each of those 4 antimicrobials was present in ≤10% of M. haemolytica isolates from untreated cattle. Similar but less dramatic trends were apparent for isolates of P. multocida and H. somni. The percentage of multi-drug resistant bacterial isolates also increased with the number of treatments. Results of our study suggest that previous antimicrobial treatment may have a profound effect on antimicrobial susceptibility testing. Summarized susceptibility results from diagnostic laboratories should not be used to make generalized statements regarding trends in antimicrobial resistance without providing context regarding antimicrobial treatment history.

Recreational Waterborne Illnesses: Recognition, Treatment, and Prevention.

Illness after recreational water activities can be caused by a variety of agents, including bacteria, viruses, parasites, algae, and even chlorine gas. These illnesses are more common in summer. Waterborne illnesses are underreported because most recreational activity occurs in unsupervised venues or on private property, and participants tend to disperse before illness occurs. Symptoms of waterborne illness are primarily gastrointestinal, but upper respiratory and skin manifestations also occur. Gastrointestinal symptoms are usually self-limited, and supportive treatment may be all that is necessary. However, some infections can cause significant morbidity and mortality. Cryptosporidium and Giardia intestinalis are the most common cause of gastrointestinal illness and have partial chlorine resistance. Respiratory infections are typically mild and self-limited. However, if legionnaires' disease develops and is unrecognized, mortality may be as high as 10%. Cellulitis caused by Vibrio vulnificus can result in serious illness, amputation, and death. Early and appropriate antibiotic treatment is important. Chronically ill and immunocompromised persons are at high risk of infection and should be counseled accordingly.

EFFICACY AND SAFETY OF BAN HUANG ORAL LIQUID FOR TREATING BOVINE RESPIRATORY DISEASES.

BACKGROUND: Ban Huang oral liquid was developed as a veterinary compound preparation by the Lanzhou Institute of Husbandry and Pharmaceutical Sciences of the Chinese Academy of Agricultural Sciences (CAAS). The purpose of this study was to determine whether the oral liquid preparation of traditional Chinese medicine, Ban Huang, is safe and effective for treating respiratory diseases in cattle. MATERIALS AND METHODS: Acute oral toxicity experiments were conducted in Wistar rats and Kunming mice via oral administration. The minimum inhibitory concentration of the drug against Mycoplasma bovis in vitro with the double dilution method was 500 mg/mL, indicating good sensitivity. The results of laboratory pathogen testing, analysis of clinical symptoms, and analysis of pathological anatomy were combined to diagnose bovine respiratory diseases in 147 Simmental cattle caused by mixed infections of M. bovis, bovine respiratory syncytial virus, bovine parainfluenza virus type 3, and Mannheimia haemolytica. These cattle were randomly divided into three groups: drug treatment group 1 (treated via Tilmicosin injection), drug treatment group 2 (treated with Shuang Huang Lian oral liquid combined with Tilmicosin injection), and drug treatment group 3 (treated with Ban Huang oral liquid combined with Tilmicosin injection). Treatment effects were observed within 7 days. RESULTS: The results showed no toxicity and a maximum tolerated dose greater than 20 g/kg BW. For the 87 cattle in drug-treatment group, the cure rate was 90.80%, whereas the response rate was 94.25%. The cure rate of drug treatment group was increased by 14.13% in comparison with that of drug control group 1 and by 7.47% in comparison with that of drug control group 2 (both P < 0.05). CONCLUSION: This study demonstrates that Ban Huang oral liquid is a safe and effective treatment for bovine respiratory diseases, especially for mixed infection caused by M. bovis, bacteria, and viruses.

Vitamin A deficiency is associated with gastrointestinal and respiratory morbidity in school-age children.

Infection is an important cause of morbidity throughout childhood. Poor micronutrient status is a risk factor for infection-related morbidity in young children, but it is not clear whether these associations persist during school-age years. We examined the relation between blood concentrations of micronutrient status biomarkers and risk of gastrointestinal and respiratory morbidity in a prospective study of 2774 children aged 5-12 y from public schools in Bogotá, Colombia. Retinol, zinc, ferritin, mean corpuscular volume, hemoglobin, erythrocyte folate, and vitamin B-12 concentrations were measured in blood at enrollment into the cohort. Children were followed for 1 academic year for incidence of morbidity, including diarrhea with vomiting, cough with fever, earache or ear discharge with fever, and doctor visits. Compared with adequate vitamin A status (≥30.0 µg/dL), vitamin A deficiency (<10.0 µg/dL) was associated with increased risk of diarrhea with vomiting [unadjusted incidence rate ratio (IRR): 2.17; 95% CI: 0.95, 4.96; P-trend = 0.03] and cough with fever (unadjusted IRR: 2.36; 95% CI: 1.30, 4.31; P-trend = 0.05). After adjustment for several sociodemographic characteristics and hemoglobin concentrations, every 10 µg/dL plasma retinol was associated with 18% fewer days of diarrhea with vomiting (P < 0.001), 10% fewer days of cough with fever (P < 0.001), and 6% fewer doctor visits (P = 0.01). Every 1 g/dL of hemoglobin was related to 17% fewer days with ear infection symptoms (P < 0.001) and 5% fewer doctor visits (P = 0.009) after controlling for sociodemographic factors and retinol concentrations. Zinc, ferritin, mean corpuscular volume, erythrocyte folate, and vitamin B-12 status were not associated with morbidity or doctor visits. Vitamin A and hemoglobin concentrations were inversely related to rates of morbidity in school-age children. Whether vitamin A supplementation reduces the risk or severity of infection in children over 5 y of age needs to be determined.

Comparative efficacy of enrofloxacin and tulathromycin for treatment of preweaning respiratory disease in dairy heifers.

Preweaning respiratory disease continues to have a substantial effect on the current and future productivity of dairy replacement animals. Establishing an effective treatment plan for the preweaned calf may have a significant effect on well-being and lifetime productivity by limiting any early development of chronic disease. The primary objective of this study was to examine the efficacy of treatment with tulathromycin (TUL) or enrofloxacin (ENR) on the risk of re-treatment, with a secondary objective of investigating the effect of disease and subsequent treatment choice on average daily gain (ADG). A total of 1,141 Holstein heifers from 4 farms were observed and systematically scored for evidence of respiratory disease from birth through weaning or the time of death. At the time of diagnosis, calves were randomly and blindly allocated into 2 treatment groups. The overall incidence of respiratory disease was 60.9%. In the univariable analysis, the incidence of re-treatment between 7 and 10d of initial therapy for calves treated with ENR was greater than that in calves treated with TUL (27.6 vs. 21.2%). After adjusting for farm ID, clinical score at first treatment, and weight at first treatment, the odds of re-treatment were 1.5 times higher for calves treated with ENR than with TUL. The percentage of calves that required more than one re-treatment was higher for calves that received ENR compared with those that received TUL (9.3 vs. 4.1%). We observed no difference in ADG between calves treated with ENR or TUL, and no difference in ADG between calves that were treated for respiratory disease and those that were not treated for respiratory disease. Appropriate drug therapy for preweaning respiratory disease may have an important role in reducing the odds of re-treatment during the preweaning period.

Probiotics in the management of lung diseases.

The physiology and pathology of the respiratory and gastrointestinal tracts are closely related. This similarity between the two organs may underlie why dysfunction in one organ may induce illness in the other. For example, smoking is a major risk factor for COPD and IBD and increases the risk of developing Crohn's disease. Probiotics have been defined as "live microorganisms which, when administered in adequate amounts, confer health benefits on the host." In model systems probiotics regulate innate and inflammatory immune responses. Commonly used probiotics include lactic acid bacteria, particularly Lactobacillus, Bifidobacterium, and Saccharomyces, and these are often used as dietary supplements to provide a health benefit in gastrointestinal diseases including infections, inflammatory bowel disease, and colon cancer. In this respect, probiotics probably act as immunomodulatory agents and activators of host defence pathways which suggest that they could influence disease severity and incidence at sites distal to the gut. There is increasing evidence that orally delivered probiotics are able to regulate immune responses in the respiratory system. This review provides an overview of the possible role of probiotics and their mechanisms of action in the prevention and treatment of respiratory diseases.

Synergistic antibiotic activity of volatile compounds from the essential oil of Lippia sidoides and thymol.

Lippia sidoides Cham. (Verbenaceae) is used in the folk medicine as topical antiseptic in skin and mucous membranes and its therapeutic effect is attributed to the thymol presence. The objective of this work was to verify the chemical composition and antibiotic modifying activity of the essential oil extracted from the leaves of L. sidoides and its major component thymol. The essential oil was obtained by hydrodistillation and analyzed by GC/MS. The synergistic activity was evaluated using gaseous contact method. The essential oil was obtained (yield of 1.06%) and the GC/MS analysis identified the main constituents: thymol (84.9%) and p-cymene (5.33%). The antibiotic modifying activity was verified using the minimal inhibitory dose method and gaseous contact. It verified the interference of essential oil and thymol against all tested aminoglycosides. There were no statistical differences between the activity of the essential oil and thymol against Pseudomonas aeruginosa, indicating this to be the responsible composition for such activity. However, the oil was shown more effective when compared to the thymol against Staphylococcus aureus. The essential oil of L. sidoides and its major component thymol influence the activity of aminoglycosides and may be used as adjuvant in antibiotic therapy against respiratory tract bacterial pathogens.

Ethnoveterinary application of Morinda citrifolia fruit puree on a commercial heifer rearing facility with endemic salmonellosis.

We have previously reported that Morinda citrifolia (noni) puree modulates neonatal calves developmental maturation of the innate and adaptive immune system. In this study, the effect of noni puree on respiratory and gastrointestinal (GI), health in preweaned dairy calves on a farm with endemic salmonellosis was examined. Two clinical trials were conducted whereby each trial evaluated one processing technique of noni puree. Trials 1 and 2 tested noni versions A and B, respectively. Puree analysis and trial methods were identical to each other, with the calf as the experimental unit. Calves were designated to 1 of 3 treatment groups in each trial and received either: 0, 15 or 30 mL every 12 hr of noni supplement for the first 3 weeks of life. Health scores, weaning age, weight gain from admission to weaning, and weaned by 6 weeks, were used as clinical endpoints for statistical analysis. In trial 1, calves supplemented with 15 mL noni puree of version A every 12 hr had a higher probability of being weaned by 6 weeks of age than control calves (P = 0.04). In trial 2, calves receiving 30 mL of version B every 12 hr had a 54.5% reduction in total medical treatments by 42 days of age when compared to controls (P = 0.02). There was a trend in reduced respiratory (61%), and GI (52%) medical treatments per calf when compared to controls (P = 0.06 and 0.08, respectively). There were no differences in weight gain or mortality for any treatment group in either trial.

Inhibitory activity of a standardized elderberry liquid extract against clinically-relevant human respiratory bacterial pathogens and influenza A and B viruses.

BACKGROUND: Black elderberries (Sambucus nigra L.) are well known as supportive agents against common cold and influenza. It is further known that bacterial super-infection during an influenza virus (IV) infection can lead to severe pneumonia. We have analyzed a standardized elderberry extract (Rubini, BerryPharma AG) for its antimicrobial and antiviral activity using the microtitre broth micro-dilution assay against three Gram-positive bacteria and one Gram-negative bacteria responsible for infections of the upper respiratory tract, as well as cell culture experiments for two different strains of influenza virus. METHODS: The antimicrobial activity of the elderberry extract was determined by bacterial growth experiments in liquid cultures using the extract at concentrations of 5%, 10%, 15% and 20%. The inhibitory effects were determined by plating the bacteria on agar plates. In addition, the inhibitory potential of the extract on the propagation of human pathogenic H5N1-type influenza A virus isolated from a patient and an influenza B virus strain was investigated using MTT and focus assays. RESULTS: For the first time, it was shown that a standardized elderberry liquid extract possesses antimicrobial activity against both Gram-positive bacteria of Streptococcus pyogenes and group C and G Streptococci, and the Gram-negative bacterium Branhamella catarrhalis in liquid cultures. The liquid extract also displays an inhibitory effect on the propagation of human pathogenic influenza viruses. CONCLUSION: Rubini elderberry liquid extract is active against human pathogenic bacteria as well as influenza viruses. The activities shown suggest that additional and alternative approaches to combat infections might be provided by this natural product.

Microbiological and mucociliary properties of the ethanol extract of Hymenocardia acida on selected respiratory clinical isolates.

The antimicrobial property of the ethanol leaf extract of Hymenocardia acida (H. acida) on some opportunistic respiratory pathogens was evaluated in this study. We also assessed the activity of the extract on tracheal mucociliary activity using murine tracheal mucus exudation and mucociliary motility in pigeons as experimental models. Phytochemical screening of the extract was done; and acute toxicity of the extract in mice was carried out using Lorke's method for estimation of its median lethal dose. Results show the presence of carbohydrates, saponins, tannins, flavonoids, alkaloids, resins, and balsams in the extract and the absence of anthraquinones, terpenes, and sterols. Results of the acute toxicity test showed that the extract was slightly toxic, with an estimated median lethal dose of 1,767.77 mg/kg body weight. At 50, 100, and 200 mg/kg body weight of H. acida, tracheal mucus exudation was increased by 14.29, 19.24, and 33.82%, respectively. The effect on mucociliary velocity was dose-dependent as 50, 100, and 200 mg/kg body weight of the extract led to increased ciliary activity by 7.69, 61.5, and 81.6%, respectively. The effects of the extract (200 mg/kg body weight) on mucus exudation and clearance were significant (p < .05) and higher than the effect of ammonium chloride. Although the extract did not inhibit the growth of C. albicans and K. pneumoniae, it exhibited moderate antimicrobial activity against Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, and Staphylococcus aureus. These findings show the mucociliary activity and antimicrobial properties of H. acida ethanol extract, and justify its use in the treatment of airway disorders.

Efficacy of enrofloxacin, florfenicol and amoxicillin against Ornithobacterium rhinotracheale and Escherichia coli O2:K1 dual infection in turkeys following APV priming.

Experimental groups of 15 susceptible 3-week-old turkeys were inoculated oculonasally with avian metapneumovirus (APV) subtype A and susceptible Escherichia coli O2:K1 and Ornithobacterium rhinotracheale (ORT) bacteria, with a 3 days interval between viral and bacterial inoculation and approximately 8h between the two bacterial inoculations. The aims of the present study were to assess the efficacy of drinking-water administration of enrofloxacin for 3 and 5 days, amoxicillin for 5 days and florfenicol for 5 days for the treatment of the resulting respiratory disease, based on clinical and bacteriological examinations. Antimicrobial treatment started 1 day after dual bacterial inoculation. After infection, the birds were examined and scored for clinical signs daily, weighed at different times, and their tracheae swabbed daily. Five birds were euthanised and examined for macroscopic lesions at necropsy at 5 days post-bacterial inoculation (dpbi) and the remainder at 15dpbi. Samples of the turbinates, trachea, lungs, sinuses, air sacs, heart, pericardium and liver were collected for bacteriological examination. Recovery from respiratory disease caused by an APV/E. coli/ORT triple infection in 3-week-old turkey poults was overall most successful after enrofloxacin treatment, irrespective of treatment duration, followed by florfenicol treatment. Compared with the untreated group, clinical signs as well as ORT and E. coli multiplication in the respiratory tract were significantly reduced by both enrofloxacin treatments and the florfenicol treatment, with the enrofloxacin treatments showing significantly better reductions than the florfenicol treatment. Five-day treatment with amoxicillin, compared with the untreated group, did not cause a significant reduction in any of the aforementioned parameters.

[Ciprofloxacin and antibacterial therapy of respiratory tract infections].

Fluoroquinolones are present considered as an important independent group of chemotherapeutics within the class of quinolones, DNA-gyrase inhibitors characterized by high clinical efficacy and numerous indications, and clinical efficacy and humerous indications, and are known as a serious alternative to other highly efficient broad spectrum antibiotics. Significant clinical experience is accumulated with respect to ciprofloxacin, one of the first agents of the group up to date used clinically. In spite of the negative tendencies in development and distribution of antibiotic resistance, ciprofloxacin remains valid as an alternative drug in the treatment of many infections of various localization. Ciprofloxacin is one of the most useful agents in step-by-step or combined antibacterial therapy. Its us is substantiated by convincing microbiological, pharmacokintic and pharmacoeconomic reasons.

Persistence of Mycoplasma gallisepticum in chickens after treatment with enrofloxacin without development of resistance.

The ability of the avian pathogen Mycoplasma gallisepticum to persist despite fluoroquinolone treatment was investigated in chickens. Groups of specific pathogen free chickens were experimentally infected with M. gallisepticum and treated with enrofloxacin at increasing concentrations up to the therapeutic dose. When M. gallisepticum could no longer be re-isolated from chickens, birds were stressed by inoculation of infectious bronchitis virus or avian pneumovirus. Although M. gallisepticum could not be cultured from tracheal swabs collected on several consecutive sampling days after the end of the enrofloxacin treatments, the infection was not eradicated. Viral infections reactivated the mycoplasma infection. Mycoplasmas were isolated from tracheal rings cultured for several days, suggesting that M. gallisepticum persisted in the trachea despite the enrofloxacin treatment. The minimal inhibitory concentration (MIC) of enrofloxacin for most of the re-isolated mycoplasmas was the same as that of the strain with which the birds were inoculated. Furthermore, no mutation could be detected in the fluoroquinolone target genes. These results suggest that M. gallisepticum can persist in chickens without development of resistance despite several treatments with enrofloxacin.