Effect of antibiotic treatment on Mycoplasma hyopneumoniae detection and infectious potential.

Mycoplasma hyopneumoniae (M. hyopneumoniae) causes significant economic losses in the swine industry. Antibiotics with activity against Mycoplasma spp. are employed for disease mitigation and pathogen elimination. However, veterinarians are often challenged with the detection of M. hyopneumoniae by PCR after antibiotic treatment, thus raising the question whether the bacterium is still infectious. The objective of this study was to evaluate the effect of tulathromycin treatment on M. hyopneumoniae detection and infectious potential during the acute and chronic phases of infection. For each infection phase, one age-matched naïve gilt was placed in contact with one M. hyopneumoniae infected gilt that was either treated with tulathromycin, treated and vaccinated, or non-treated, for 14 days. Four replicates per treatment group were performed for each infection phase. A numerical reduction in relative bacterial load was observed in acutely treated gilts compared to non-treated gilts. The rate at which naïve gilts became infected with M. hyopneumoniae was numerically reduced when co-housed with treated, acutely infected gilts compared to those housed with non-treated, infected gilts. During the chronic infection phase, M. hyopneumoniae was detected by PCR in more than 50 % of treated infected gilts and persisted for up to three months post-treatment. Transmission was not detected in all treatment groups however, the possibility that the pathogen was infectious could not be completely ruled out. Further research focused on assessing M. hyopneumoniae detection and viability post-treatment is necessary to guide control and elimination efforts.

Antimicrobial treatment of Mycoplasma hyopneumoniae infections.

Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary agent of enzootic pneumonia, a chronic and economically important respiratory disease of pigs. Control and prevention of M. hyopneumoniae infections can be accomplished by optimization of management and housing conditions, and by vaccination. The present paper summarizes the current knowledge on the main characteristics and efficacy of antimicrobials used for the treatment of clinical M. hyopneumoniae infections, the in vitro and in vivo activities of these antimicrobials and the reported resistance mechanisms against some. Potentially active antimicrobials against M. hyopneumoniae include tetracyclines, macrolides, lincosamides, pleuromutilins, amphenicols, aminoglycosides, aminocyclitols and fluoroquinolones. Antimicrobial treatment can be administered either orally or parenterally. Based on the overall results of efficacy studies performed under experimental and/or field conditions, the majority of agents belonging to these antimicrobial classes improved clinical parameters (clinical signs, lung lesions) and reduced performance losses due to M. hyopneumoniae infection. Antimicrobials may, however, not be able to prevent infection or to eradicate the bacterium from the respiratory tract. The decision to medicate should, therefore, be considered carefully. M. hyopneumoniae shows an intrinsic resistance against ß-lactam antibiotics, sulfonamides and trimethoprim. A few reports have shown acquired antimicrobial resistance against some antibiotics, along with associated resistance mechanisms. The results of antimicrobial susceptibility testing are difficult to interpret in terms of treatment outcome, as no clinical breakpoints have been defined for M. hyopneumoniae.

Antimicrobial susceptibility and genetic profile of Mycoplasma hyopneumoniae isolates from Brazil.

Mycoplasma hyopneumoniae is the etiologic agent of porcine enzootic pneumonia, responsible for major production losses worldwide. The bacteria have a limited metabolism and need to obtain molecules from the growth environment, which causes multiple difficulties for in vitro culture. These limitations have a negative influence on the ability to carry out research for the development of the rational use of antimicrobials and vaccines. The objective of this investigation was to evaluate the genetic profile and in vitro susceptibility of field isolates of M. hyopneumoniae to different antimicrobials. All 16 isolates obtained from the samples presented 100% of identity in the partial sequence of 16S rRNA gene when compared to M. hyopneumoniae. A dendrogram was created using the PCR results of the genes related to pathogenicity, and the isolates were distributed into four clusters, suggesting genetic variability among four different isolates circulating on the same farm. The minimum inhibitory concentration of the isolates was higher for the antimicrobials tylosin (< 0.001-16 mg/L) and spiramycin (< 0.001-16 mg/L) than for enrofloxacin (< 0.001-0.125 mg/L) and tiamulin (< 0.001-0.125 mg/L). Our results demonstrate the genetic variability among M. hyopneumoniae isolates from pigs of the same farm, with differences in their susceptibility to antimicrobial agents.

Cathepsin L promotes secretory IgA response by participating in antigen presentation pathways during Mycoplasma Hyopneumoniae infection.

Cathepsin L (CTSL) has been proved to help contain leishmaniasis and mycoplasma infection in mice by supporting cellular immune responses, but the regulatory functions of CTSL on mucosal immune responses haven't been tested and remain undefined. Here, we investigated the effects of CTSL on SIgA responses and invariant chain (Ii) degradations in the co-cultured swine dendritic cells (DCs) and B cells system in vitro. When the cells system were transfected with vector CTSL-GFP or incubated with recombinant CTSL (rCTSL) before they were infected with Mycoplasma hyopneumoniae (M.hp), SIgA significantly increased and Ii chain was degraded into smaller intermediates, while SIgA decreased when CTSL was knockdown or inhibited with E64. To confirm the SIgA responses promoted by CTSL contribute to the resistance to mycoplasma pneumonia, pigs injected with rCTSL before they were challenged with M.hp, showed milder clinical symptoms and histopathological damage of lungs, less mycoplasma burden together with higher secretion of SIgA, percentages of CD4+ T cells and level of MHC II molecules comparing with the group without rCTSL. Collectively, these results suggested that rCTSL could provide effective protection for piglets against mycoplasma pneumonia by enhancing M.hp-specific mucosal immune responses through its role in antigen presentation by processing the invariant chain.

Macrolide-Resistance Selection in Tibetan Pigs with a High Load of Mycoplasma hyopneumoniae.

Currently, tylosin tartrate is the first-line treatment for Mycoplasma hyopneumoniae infections in China. However, the efficacy of tylosin tartrate and resistance to this treatment in M. hyopneumoniae infections of Tibetan pigs are unknown. In this study, we examined the prevalence of M. hyopneumoniae infection in Tibetan pigs at three intensive farms in Tibet, China. In addition, we investigated the efficacy of tylosin tartrate treatment for porcine enzootic pneumonia by monitoring M. hyopneumoniae DNA eradication dynamics and macrolide resistance (MR). Eighty-two of 450 (18.2%) Tibetan pigs tested positive for only M. hyopneumoniae, and most of these animals (85.1%) had symptoms and signs of pneumonia. The elimination of M. hyopneumoniae DNA was substantially faster in Tibetan pigs with a lower pretreatment M. hyopneumoniae load, and the total eradication rate was 97.4% (75/77). Two Tibetan pigs tested positive for M. hyopneumoniae that contained macrolide resistance-determining mutations in the 23S rRNA gene. Our results indicate that the pretreatment M. hyopneumoniae load may be an effective predictor of macrolide treatment efficacy (and possibly that of other antimicrobial agents) and MR. Moreover, our results suggest that danofloxacin mesylate can be used as an alternative drug for the treatment of macrolide-resistant M. hyopneumoniae infection acquired during intensive farming.

Efficacy of in-feed medication with chlortetracycline in a farrow-to-finish herd against a clinical outbreak of respiratory disease in fattening pigs.

The efficacy of chlortetracycline (CTC) in-feed medication to treat pigs with clinical respiratory disease was investigated in a farrow-to-finish pig herd infected with Mycoplasma hyopneumoniae, and with clinical respiratory disease in growing pigs. In total, 533 pigs were included. The animals were vaccinated against M hyopneumoniae and porcine circovirus type 2 at weaning. At onset of clinical respiratory disease, they were randomly allocated to one of the following treatment groups: chlortetracycline 1 (CTC1) (two consecutive weeks, 500 ppm), chlortetracycline 2 (CTC2) (two non-consecutive weeks, with a non-medicated week interval in between, 500 ppm) or tylosin (T) (three consecutive weeks, 100 ppm). Performance (daily weight gain, feed conversion ratio), pneumonia lesions at slaughter and clinical parameters (respiratory disease score) were assessed. Only numeric differences in favour of the CTC2 group were obtained for the performance and the clinical parameters. The prevalence of pneumonia lesions was 20.5, 13.1 and 23.0 per cent (P<0.05) for the CTC1, CTC2 and T groups, respectively. The study demonstrated that CTC, when administered at onset of clinical respiratory disease via the feed at a dose of 500 ppm during two alternative weeks, was able to decrease the prevalence of pneumonia lesions, and numerically reduce performance losses and clinical signs.

Efficacy of florfenicol injection in the treatment of Mycoplasma hyopneumoniae induced respiratory disease in pigs.

This study investigated the efficacy of a single intramuscular injection of a new formulation of florfenicol to treat clinical respiratory disease following experimental Mycoplasma hyopneumoniae infection. M. hyopneumoniae-free piglets were allocated to three groups, namely, a treatment group (TG) and a positive control group (PCG), which were both inoculated endotracheally with a highly virulent isolate of M. hyopneumoniae, and a negative control group. At the onset of clinical disease, the TG received a single injection of florfenicol (30 mg/kg). All pigs were euthanased 4 weeks post-infection. Clinical symptoms were significantly reduced in the TG in comparison with the PCG. Average daily gain, feed conversion ratio, mortality and lung lesions were improved in the TG compared to the PCG, but the differences were not statistically significant.

Eradication of Mycoplasma hyopneumoniae from a swine finishing herd without total depopulation.

Using vaccination and medication, Mycoplasma hyopneumoniae (Mhyo) was eradicated from a finishing herd without total depopulation. Altogether 3243 feeder pigs originating from Mhyo-free herds were vaccinated once using an inactivated, adjuvanted vaccine before transporting them to a Mhyo-infected finishing herd. The Mhyo-infected groups of pigs were medicated with antimicrobial agents at the time of the arrival of the first groups of Mhyo-free, vaccinated feeder pigs. The groups were operated with an all-in-all-out method in rooms with separate ventilation and slurry disposal systems. Thereafter the farmer purchased only non-vaccinated feeder pigs originating from Mhyo-free sow herds. Serology gave no positive results for 5.5 years and it was concluded that the eradication programme had been successful in producing a Mhyo-free herd without total depopulation.

Genetic diversity of Mycoplasma hyopneumoniae isolates from conventional farrow-to-finish pig farms in Serbia.

Mycoplasma hyopneumoniae is a primary agent associated with mycoplasma pneumonia and the porcine respiratory disease complex (PRDC). Various reports have indicated that different strains of M. hyopneumoniae are circulating in the swine population. Lysates from lung swabs from naturally infected pigs of different ages were tested according to a new variable number of tandem repeats (VNTR) genetic typing method based on the polyserine repeat motif of the P146 lipoproteoadhesin, which can be applied directly on clinical material without isolation of M. hyopneumoniae. The aim was to determine the diversity of M. hyopneumoniae isolates from conventional farrow-to-finish pig farms located in different geographical areas of Serbia. PCR amplification was carried out using M. hyopneumoniae -specific designed, conserved primers (p146MH-L and p146MH-R) flanking the region encoding the repeat motif, followed by sequencing and cluster analysis. Five groups of M. hyopneumoniae with thirteen to twenty-four serine repeats were observed. Analysis of three samples from each farm indicated that the specific isolate is ubiquitous in pigs of different ages. Furthermore, seven clusters were observed within 27 tested samples. The results indicated a considerable diversity among M. hyopneumoniae field isolates in the swine population from conventional farrow-to-finish farms in Serbia and suggest close genetic relatedness of the corresponding isolates.

Evaluation of tulathromycin for the treatment of pneumonia following experimental infection of swine with Mycoplasma hyopneumoniae.

Tulathromycin was evaluated in the treatment of pneumonia in weaned pigs inoculated intranasally with Mycoplasma hyopneumoniae. Five days postchallenge, the pigs were randomized to treatment with a single IM administration of saline, a single IM administration of tulathromycin (2.5 mg/kg; day 0), or three IM administrations of enrofloxacin (5.0 mg/kg; days 0, 1, 2). Pigs were necropsied on day 12 or 13. Unchallenged controls remained healthy with no lung pathology. Compared with saline, coughing, mean lung lesion score, and proportional lung weight were significantly reduced and weight gain was significantly greater for tulathromycin-treated pigs (P < .05). Compared with enrofloxacin, there were no significant differences in proportional lung weight or weight gains, but coughing and lung lesion scores were greater for tulathromycin-treated pigs (P < .05). Tulathromycin was effective in the treatment of pneumonia following experimental infection with M. hyopneumoniae.

Efficacy of in-feed medication with tylosin for the treatment and control of Mycoplasma hyopneumoniae infections.

The efficacy of in-feed medication with tylosin for the treatment of enzootic pneumonia was examined in an experimental Mycoplasma hyopneumoniae infection model. One group of 10 conventional M. hyopneumoniae-free pigs was inoculated intratracheally with a highly virulent field isolate of M. hyopneumoniae; a second group of 10 pigs was inoculated in the same way and after 12 days was given tylosin at 100 mg/kg feed for 21 days; a third group of 10 pigs was inoculated with sterile culture medium, and these pigs were not given tylosin. The pigs were examined daily for clinical signs and each pig was given a respiratory disease score. Thirty-three days after they had been infected the pigs were euthanased, the lung lesions were quantified and samples of lung were processed for immunofluorescence testing for M. hyopneumoniae. The mean (sd) respiratory disease and lung lesion scores were significantly higher (P<0.05) in both the infected groups than in the uninfected group. Between 23 and 33 days after infection the mean respiratory disease score of the pigs treated with tylosin was 0.54 (0.22), significantly (P<0.05) lower than that of the infected pigs which were left untreated, 1.54 (0.46); similarly, their average lung lesion score, 1.72 (1.20), was significantly lower than that of the untreated pigs, 5.27 (3.85).

Effectiveness of treatment with lincomycin hydrochloride and/or vaccination against Mycoplasma hyopneumoniae for controlling chronic respiratory disease in a herd of pigs.

A herd of pigs infected with Mycoplasma hyopneumoniae was used in a double-blind randomised trial to assess the effectiveness of three control strategies against chronic respiratory disease in growing-finishing pigs. One group of 61 pigs received 220 ppm lincomycin hydrochloride in the feed from day 71 to day 91, a second group was vaccinated against M. hyopneumoniae at four and 28 days of age, and a third group received both treatments; a fourth group was left untreated as a control. Throughout the nursery-finishing period (day 29 to slaughter) the average daily weight gain and feed conversion rate of all the treated groups were slightly better than in the controls, but there were no significant differences between them. There were no significant differences between the treated groups in terms of clinical signs, serology, pathology or mortality, which was very low throughout the trial.

Treatment of experimental enzootic pneumonia of the pig by norfloxacin or its 6-chloro analogue.

The 6-chloro analogue of norfloxacin (compound A) administered continuously in the feed at 400 ppm for 21 days markedly reduced the extent and activity of pneumonic lesions in pigs with pneumonia induced experimentally with an homogenate of pneumonic lung and broth cultures of Mycoplasma hyopneumoniae. Norfloxacin at 100 ppm or compound A at 200 ppm in the feed did not reduce the extent of lung lesions, although half the pigs treated with norfloxacin had lesions which appeared histologically to be healing. M hyopneumoniae was detected either by culture or immunofluorescence in the lungs of 60 per cent of the pigs treated with compound A at 400 ppm compared with all the pigs in the other groups. These results were related to the amount of drug in the lungs and body fluids during therapy. Only compound A at 400 ppm produced concentrations in the lungs and bronchial secretions exceeding the minimum inhibitory concentration against M hyopneumoniae. Mycoplasmacidal concentrations were not reached either in the lungs or bronchial secretions which might account partly for the frequent detection of M hyopneumoniae in the lungs after treatment. Drug resistance did not appear to be responsible for the persistence of M hyopneumoniae in vivo since the M hyopneumoniae isolates from the pigs after therapy were sensitive in vitro to both quinolones. As daily weight gain and feed-conversion efficiency improved in all groups of treated pigs compared with the controls, these effects were probably unrelated to the antimycoplasmal activities of the two quinolones.

Evaluation of tiamulin for treatment of mycoplasmal pneumonia in swine.

During 3 trials, using affected pigs of various ages, tiamulin was evaluated for treatment of experimentally induced mycoplasmal pneumonia. Pneumonia was induced in respiratory tract disease-free swine by intratracheal inoculation of a lung homogenate containing Mycoplasma hyopneumoniae. Eleven days after inoculation, when more than 20% of pigs were coughing, pigs were allotted to 3 or 4 groups (n = 8 pigs each) and were given regimens of no medication or 60 mg, 120 mg, or 180 mg of tiamulin/L of drinking water for 10 days. Twenty-one days after cessation of medication, pigs were euthanatized and then were necropsied. Results obtained from the 3 trials did not indicate significant difference among treatment groups in severity of macroscopic or microscopic lesions induced by M hyopneumoniae or in detection of M hyopneumoniae by use of immunofluorescent technique. Clinical evaluations, daily gain, and feed efficiency did not differ significantly among treatment groups. In this study, tiamulin administration did not have beneficial effects in swine with mycoplasmal pneumonia.

The synergistic activity of tiamulin and chlortetracycline: in-feed treatment of bacterially complicated enzootic pneumonia in fattening pigs.

The antibacterial effects of a combination of tiamulin and chlortetracycline in vitro against a number of field isolates of Pasteurella multocida, Haemophilus pleuropneumoniae and Bordetella bronchiseptica were examined. There was a marked synergism between the two antibiotics against all eight isolates of P multocida, against seven of nine isolates of H pleuropneumoniae and against the single strain of B bronchiseptica tested. Two field trials were carried out on a herd with a history of complicated enzootic pneumonia where the presence of Mycoplasma hyopneumoniae and P multocida had been established and subsequently the presence of H pleuropneumoniae was discovered. Feed containing tiamulin at 100 ppm combined with chlortetracycline at 300 ppm was given for seven days to pigs affected with pneumonia, and the results were compared with untreated controls and pigs receiving chlortetracycline at 300 ppm. There was a follow-up observation period of three weeks when all groups received unmedicated feed. During the medication period the combination treated groups showed a statistically significant increase in average daily weight gain of 156 g (20.4 per cent) and in feed conversion efficiency of 0.576 (20.8 per cent) and a numerical improvement in average disease score in comparison with the untreated controls. These improvements were approximately double those observed in the groups treated with 300 ppm chlortetracycline which showed improvements of 93 g (12.2 per cent) in average daily gain and 0.301 (10.9 per cent) in feed conversion efficiency. During the following three weeks most of the initial gains were lost, probably owing to the reinfection of the treated groups by the untreated controls.

Tylosin tartrate and tiamutilin effects on experimental piglet pneumonia induced with pneumonic pig lung homogenate containing mycoplasmas, bacteria and viruses.

The effects of tylosin tartrate and tiamutilin were examined in pneumonias induced experimentally in neonatal piglets with a homogenate of pneumonic pig lung, obtained from pigs with naturally acquired enzootic pneumonia. The homogenate contained mycoplasmas, including Mycoplasma hyopneumoniae (M suipneumoniae) and M hyorhinis, and certain bacteria and viruses. The experimental pneumonias generally resembled mycoplasmal pneumonia histologically but were complicated by aspiration pneumonia in some animals. both tylosin tartrate (50 mg/kg) and tiamutilin (10 mg/kg) administered orally twice daily for 10 days, beginning 14 days after intranasal infection, significantly reduced the incidence and severity of macroscopical pneumonic lung lesions. M hyopneumoniae could be isolated from the lungs of the unmedicated piglets, but not from drug-treated piglets. The numbers of M hyorhinis, Acholeplasma granularum, Haemophilus parasuis, Pasteurella multocida and P haemolytica in the lung tissue of the infected piglets were significantly reduced by drug therapy. The role of bacterial in the experimental infection appeared to be that of secondary invaders.