We Take pride in our Denagard Injection

Subjects covered an overview

Formulation / product characteristics Pharmacokinetic / pharmacodynamic relationships Microbiological activity Efficacy against enteric / respiratory / systemic infections Safety Comparison vs. generic products/formulations

Denagard Injection: range

Range of presentations of tiamulin base in sesame oil are available:

10% base equivalent to 12.3% tiamulin hydrogen fumarate (international approved/supported formulation A 8813C) 16.2% base - equivalent to 20% tiamulin hydrogen fumarate (international approved/supported formulation A 20001B)

Denagard Injection a different class

Tiamulin hydrogen fumarate C32H51NO8S = 610 Soluble in water pH 3.1 to 4.1 Tiamulin base C28H47NO4S = 493.8 Lipid soluble enhanced cell penetration

Denagard Injection formulation characteristics

Most advanced Pleuromutilin antibiotic injectable for pigs Tiamulin levels in lung and other target tissues exceed the mean MICs for major pathogens involved in enteric / respiratory / systemic infections The state-of-the-art medication that delivers more active, effective and reliable control of swine dysentery, colitis, ileitis, enzootic pneumonia, pleuropneumonia and mycoplasmal arthritis

Denagard product description

Active ingredient: Tiamulin - semisynthetic bacteriostatic antibiotic derived from basidiomycete (Clitopilus scyphoides) Antibiotic family: Pleuromutilin Developed specifically for use in animals Pleuromutilin antibiotics not used in human medicine for treatment of respiratory and enteric diseases Tiamulin hydrogen fumarate (all oral formulations); Tiamulin base (oily injectable formulations)

Denagard

Pharmacokinetic (PK) / Pharmacodynamic (PD) relationships

Basic PK/PD relationships

PK of drug - concentrations in plasma/serum usual; extracellular fluids/plasma linked; intestinal contents (ileum, colon); intracellular penetration lipid solubility PD of drug minimum inhibitory concentration (MIC); minimum bactericidal concentration (MBC); also killing rate = concentration and time factors Clinical response; clinical cure; bacterial elimination

Basic PK/PD relationships

Cmax maximum concentration

For bactericidal antimicrobials (aminoglycosides, fluoroquinolones) Look for Cmax/MIC ratio of 10-12: 1 for clinical kill of bacterium

AUC area under the curve (time & concentration)

For bactericidal antimicrobials (penicillins, trimethoprim/sulphas) AUC/MIC ratio of 100-120 over 24 hours for clinical kill of bacterium Equivalent to 4-5 times MIC (=MBC)

Basic PK/PD relationships

Steady state usually AUC/24h (following feed and water medication). Time >MIC also helpful - aim for >18h+post antibiotic effect (PAE)
Useful for bacteriostatic antibiotics (tiamulin, tetracyclines, macrolides) Inhibitory effect above MIC Bactericidal effect above MBC (Eliminatory 4-5 times MBC)

Denagard Injection

Tiamulin parenteral pharmacokinetics in pigs

McKellar et al. 2004 Anderson et al. 1994 Forster et al. 1982

Denagard injection - pharmacokinetics

Plasma concentrations are relatively low Cmax 0.61g/ml; AUC 12.82 g.h/ml Lung concentrations high Cmax 9.6g/g; AUC 232g/g Colon contents also high Cmax 12.75g/g; AUC 314g/g In feed or drinking water Denagard concentrations lower and flatter vs. parenteral injection concentrations.

Denagard injection - pharmacokinetics

Ratios Lung/plasma AUC ratio = 18.1 : 1 Intracellular/extracellular concentration ratio for leucocytes also = 18.2 : 1 (Nielsen and Szancer, 1998) Plasma protein binding low 30-40% (Gadebusch, 1976)

Pharmacokinetics: UK study results (McKellar et al. 2004)

Aim to determine in 25 landrace pigs (bw 20-25kg) the plasma and target tissue distribution of Denagard injectable following a single i/m injection of 15mg tiamulin hydrogen fumarate per kg bodyweight. AUC - area under plasma concentration time curve T max - Time of C max AUMC - area under moment curve MRT - mean residence time

Denagard unique injection PK plasma, lung, colon contents (single i/m injection: 15mg/kg bw)
(McKellar et al, 2004)

Pharmacokinetics: UK study results

(McKellar et al. 2004)

Pharmacokinetics of Denagard in tissues calculated from mean concentrations from each time of kill following single i/m injection of tiamulin (15mg/kg bw)

Denagard - unique injection PK - study results

(McKellar et al. 2004)

Pharmacokinetics of Tiamutin (tiamulin) in tissues calculated from mean concentrations from each time of kill following single i/m injection of tiamulin
AUC(gh/ml) AUMC(gh2/ml) MRT(h) Cmax(g/ml) tmax(h) Plasma 12.82 252.02 19.66 0.61 4.00 Colon wall 64.51 1252.78 19.42 2.27 6.00 Colon contents 314.23 9013.0 28.68 12.75 24.00 Lung 231.52 3868.1 16.71 9.60 4.00

The C max and AUC values were very much higher for colon wall, colon contents and lung, than for plasma.

Denagard Injection

Tiamulin parenteral pharmacokinetics in pigs

Anderson et al. 1994

Pharmacokinetics: US study results

(Anderson et al. 1994)

Groups of five pigs received intramuscular (i/m) doses of tiamulin base 11mg/kg and 22mg/kg body weight (equivalent to 13.6 and 27.2mg thf/kg body weight respectively) once daily for four consecutive days. On the last day of medication the pigs were euthanized and the tissues harvested for microbiological assay. Body weight pigs: 42kg

Pharmacokinetics: USA study results

(Anderson et al.1994)

Parenteral dosage rate (thf) equivalent

Denagard (tiamulin) activity (g/g) Lung Tonsils Colon mucosa 2.58 8.99 Colon contents 3.09 24.9

13.6mg/kg 27.2mg/kg

26.9 71.0

3.23 8.44

Tiamulin base 11mg/kg and 22mg/kg body weight

PK/PD relationship of tiamulin (injection) in lung and intestine

Tiamulin parenteral pharmacokinetics: summary

The concentrations of Denagard in the lung tissues were much higher than in plasma. The ratio of mean lung/plasma concentrations were between 15:1 and 19:1 from 2 to 4 hours post-injection and even larger at 24 hrs and 32 hrs post-injection. Mean lung concentrations were quite constant at approx 8.0 g/ml between 2 and 8 hours post-injection and were still in excess of 3.0 g/ml at 32 hours post-injection.

Tiamulin parenteral pharmacokinetics: summary

The lung/plasma AUC ratio is 18.1:1 which is almost identical to the intracellular/extracellular ratio for leucocytes (18.2:1) Tissue concentrations in target tissues such as lung, colon and colon contents, achieved with a single i/m injectable dose of 15mg thf/kg bodyweight, is high and exceeded the MIC values for key respiratory and enteric pathogens e.g. M. hyopneumoniae, Actinobacillus pleuropneumoniae, Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis.

Tiamulin parenteral pharmacokinetics: Summary

Tiamulin is primarily bacteriostatic. To exert bactericidal (mycoplasmacidal) effect it must be present at site of infection during a long time and at a sufficient concentration The area under the curve (AUC 24h) is the most suitable PK parameter to determine the potential antibacterial (antimycoplasmal) effect The tiamulin plasma concentration is the most significant PK parameter for the correlation with the PD of organisms like M.hyopneumoniae

Tiamulin parenteral pharmacokinetics: Summary

Tiamulin gut contents concentration is most applicable for enteric infections with the PD for organisms like B. hyodysenteriae and L. intracellularis Denagard parenteral application gives higher lung / colon levels than oral application

Denagard

Microbiological activity

Denagard The Pride: Tiamulin MIC range for Respiratory pathogens

on day4 Dosage 13.6 mg/kg mg/ml M. hyopneumoniae Respiratory pathogens (tiamulin MIC range g/ml) M. hyorhinis
0.024-0.39 (Thongkamkoon 2005) 0.048 - 0.097 (Thongkamkoon 2010)

M. hyosynoviae
0.0025-0.025 (Windsor 1996) 0.0025-0.01 (Hannan 1997) 0.0156-0.0625 (Aarestrup&Friis 1998) 0.03-0.25 (Stipkovits 2004) 0.048-0.097 (Thongkamkoon 2010)

App
32-64 (Aitken 1999) 2-4 Fodor 2004) 0.25-16.0 (VetPath 2009) 0.5-64.0 (Kucerova 2011)

P.multo
1.0-8.0 (Fodor 2004) 4.0-64.0 (VetPath 2009)

S.suis
1.0-2.0 (Aitken 1999) 0.015-0.5 (Fodor 2004) 0.125-8 (VetPath 2009)

Lung 26.9 0.01-0.05 (Windsor 1996) Tonsils 0.06-1.0 3.23 (Stipkovits 2002) 0.048 (Thongkamkoon2002) <0.015-0.12 (Vicca 2004) 0.015-0.12 (Maes 2007) 0.03-0.125 (Kobayashi 2008) 0.048 - 0.19 (Thongkamkoon 2010)

27.2 mg/kg

Lung 71.0 Tonsils 8.44

H.parasuis 1-8 (Fodor 2004)

Denagard The Pride: Tiamulin MIC range for Enteric pathogens

on day4 Dosage Colon mucosa Colon B.hyo contents Enteric pathogens (tiamulin MIC range g/ml) B.pilo L.intra C.perfring C.difficile Salmo

13.6 mg/kg

2.58

3.09

0.025-0.2 (Kajiwara 2004) 0.031-0.5 (Karlsson 2004) <0.03-2 (Vyt 2006) <0.016-2.0 (2000-04) <0.016-2.0 (2006-07) (Hildago 2009) 0.063-2.0 (Magistrali 2010)

0.041 (Ripley 1998) 0.067 (Ripley1998) 0.125 (Kinyon 2002) 0.125 (Karlsson 2004)

Intra 0.125 (Suphot 2009) Extra 1-4 (Suphot 2009)

0.125-128 (Italy) 4-32 0.25-8 (DK) (Prapasaraku (Agnoletti 2010) l,2007) 0.125-16 (I&DK) (Agnoletti 2010)

27.2 mg/kg

8.99

24.9

Denagard Injection

Tiamulin parenteral pharmacodynamics in pigs Mycoplasma spp.

Kobayashi et al. 2008 Thongkamkoon et al. 2010 Maes et al. 2007 Pridmore et al. 2008 Aarestrup and Friis 1998 Stipkovits et al. 2004 Vicca et al. 2004

AB MIC data Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma synoviae

Mycoplasma isolation / strain specification / MIC testing Mycoplasma AB MIC monitoring investigations ongoing (Europe, Asia) Europe: specialized researchers (Hungary, United Kingdom, Germany, Spain, France) isolation & specification of porcine / avian / bovine Mycoplasma strains. Routine MIC work. Asia: specialized researchers in Thailand & Japan. Isolation & specification of porcine Mycoplasma strains. MIC testing USA: no group currently doing routine Mycoplasma work!

Denagard

Pharmacodynamics
Tiamulin MIC data (M.hyopneumoniae) Kobayashi et al. 2008 Thongkamkoon et al. 2010 Maes et al. 2007 Vicca et al. 2004 Pridmore et al. 2008

MICs (g/ml) M. hyopneumoniae strains from Japan (n=90) (Kobayashi et al, 2008)
Antimicrobial Tiamulin Valnemulin Oxytetracycline Lincomycin Tilmicosin Tylosin Enrofloxacin MIC 50 0.06 0.002 1.0 0.25 0.25 0.06 0.125 MIC 90 0.125 0.004 2.0 0.5 0.5 0.25 0.125 Range 0.03-0.125 0.002-0.004 0.25-4.0 0.125->64 0.25->16 0.06->64 0.06-1.0

Use clinical breakpoints / plasma levels - Resistance

MICs (g/ml) M. hyopneumoniae Thailand (n=10 2008, n=10 2009) (Thongkamkoon et al, 2010)
Antimicrobial Tiamulin Valnemulin Doxy Tylosin Linco Enrofloxacin Florfenicol MIC50 2008 0.048 <0.006 3.12 0.19 0.19 3.12 1.56 MIC50 2009 0.048 <0.006 3.12 0.097 0.19 1.56 1.56 MIC90 2008 0.048 <0.006 6.25 0.19 0.39 6.25 1.56 MIC90 2009 0.097 <0.006 6.25 0.097 0.78 6.25 3.12 MIC range 2008 0.048-0.097 <0.006 3.12-6.25 0.097-0.19 0.19-0.39 0.048-6.25 1.56-3.12 MIC range 2009 0.048-0.19 <0.006 1.56-6.25 0.048-0.19 0.19-0.78 0.024-6.25 1.56-3.12

Use clinical breakpoints / plasma levels - Resistance

MICs (g/ml) M. hyopneumoniae strains from Belgium (n=21) (Maes et al. 2007)
Antimicrobial Tiamulin Lincomycin Tylosin Tilmicosin Enrofloxacin MIC 50 0.03 0.06 0.06 0.5 0.06 MIC 90 0.12 0.12 0.12 0.5 0.5 MIC range 0.015-0.12 0.06->8.0 0.015->1.0 0.25->16.0 0.03->1.0

MICs (g/ml) M. hyopneumoniae strains from Belgium (Vicca et al. 2004)

Antimicrobial Tiamulin Oxytetracycline Lincomycin Tilmicosin Tylosin MIC 50 0.015 0.12 0.06 0.25 0.03 MIC 90 0.12 1.0 0.06 0.5 0.06 Range 0.015-0.12 0.03-2.0 0.06->8.0 0.25->16 0.015->1.0

Minimum inhibitory concentration results for tiamulin against 43 isolates of M.hyopneumoniae from Europe (Pridmore et al. 2008)
Strains M.hyopneumoniae (United Kingdom, 19 strains) M.hyopneumoniae (Spain, 24 strains) MIC 50 0.016 0.016 MIC 90 0.031 0.031 MIC range 0.008-0.031 0.004-0.062

Denagard PK/PD relationship for M. hyopneumoniae - CONCLUSIONS

Several publications report on high sensitivity of M. hyopneumoniae strains against tiamulin. The MIC distribution patters are similar with only one dilution difference. There is no evidence of a resistance pattern developing to tiamulin. Recommended dose rates of tiamulin for parenteral application (treatment interval 3 applications over 3 consecutive days, 15mg/kg bwt) give tiamulin levels in plasma (Cmax 0.61g/ml) far in excess of the MIC 90 for M. hyopneumoniae (range MIC 90 0.031-0.12g/ml).

Denagard

Pharmacodynamics
Tiamulin MIC data (M.hyorhinis) Thongkamkoon et al. 2010

MICs (g/ml) M. hyorhinis strains from Thailand (n=11 2008, n=9 2009) (Thongkamkoon et al, 2010)
Antimicrobial Tiamulin Valnemulin Doxy Tylosin Linco Enrofloxacin Florfenicol MIC50 2008 0.048 <0.006 0.78 3.12 0.78 1.56 3.12 MIC50 2009 0.097 <0.006 1.56 3.12 1.56 3.12 3.12 MIC90 2008 0.097 <0.006 3.12 3.12 0.78 3.12 3.12 MIC90 2009 0.097 <0.006 1.56 6.25 3.12 50 3.12 MIC range 2008 0.048-0.097 <0.006 0.78-3.12 0.078-3.12 0.39-1.56 0.78-25 1.56-3.12 MIC range 2009 0.048-0.097 <0.006 0.39-1.56 1.56-6.25 0.78-3.12 0.78-50 1.56-3.12

Use clinical breakpoints / plasma levels - Resistance

Denagard

Pharmacodynamics
Tiamulin MIC data (M.hyosynoviae) Aarestrup and Friis 1998 Stipkovits et al. 2004

MICs (g/ml) M. hyosynoviae strains from Denmark (n=42) (Aarestrup & Friis, 1998)
Antimicrobial Tiamulin Lincomycin Tylosin Tetracycline Enrofloxacin MIC 50 0.0313 1.0 2.0 1.0 0.5 MIC 90 0.0625 2.0 16 2.0 0.5 MIC range 0.0156-0.0625 0.5-4.0 0.125-16 0.5-2.0 0.25-1.0

MIC data confirmation based on Hungarian M.hyosynoviae strains (Stipkovits et al 2004)

Susceptibility pattern of Danish M. hyosynoviae isolates to tiamulin and tylosin (Aarestrup & Friis, 1998)

MICs (g/ml) M. hyosynoviae strains from Hungary (Stipkovits et al 2004)

Antimicrobial Tiamulin Valnemulin Lincomycin Tylosin Doxycycline Enrofloxacin MIC 50 0.125 0.06 2.0 4.0 1.0 0.5 MIC 90 0.25 0.125 4.0 16.0 2.0 0.5 MIC range 0.03-0.25 0.015-0.125 0.5-8.0 2.0-32 0.25-4.0 0.25-1.0

Tiamulin parenteral pharmacodynamics in pigs (Mycoplasma spp.) - conclusions

Mycoplasma hyopneumoniae strains isolated in Europe and Asia show high and consistent sensitivity to tiamulin. No trend of sensitivity reduction or resistance development is found. Some Mycoplasma hyopneumoniae strains isolated in Europe and Asia show reduced sensitivity to Macrolids and Lincosamids. Limited number of strains show resistance.

Tiamulin parenteral pharmacodynamics in pigs (Mycoplasma spp.) - conclusions

High tiamulin sensitivity was found for Mycoplasma hyorhinis strains. Higher and broader range of MICs for Macrolides vs. Pleuromutilins. High and consistent sensitivity of Mycoplasma hyosynoviae to tiamulin. MIC data from Europe confirm the resistance existence against Macrolides and Lincosamides.

Denagard

Tiamulin concentrations in joint fluid

Skov and Nielsen 1988 McKellar et al. 2004 Adams and Klein 2010

Denagard for systemic infections

Tiamulin concentration in synovial fluid (Skov and Nielsen 1988, i.m. injection 15mg thf / kg bw; one 43kg pig; 3.23 ml Denagard 20% Injection) 2 hours p.i.
Tiamulin conc. Knee 0.13 g/ml Stifle 0.17 g/ml Hock 0.10 g/ml Serum 0.22 g/ml Lung 17.1 g/ml

Conclusions: Takes time to penetrate to joint fluid from serum Range of tiamulin concentrations found in joint fluid (0.10-0.17 g/ml) Remarkably high lung tissue concentrations found

Denagard for systemic infections

Tiamulin concentration in joint fluid (McKellar 2004, i.m. injection 15mg thf kg/bw)
Tiamulin conc. 4 hours p.i. 0.19 g/ml 8 hours p.i. 0.38 g/ml

other data (Skov and Nielsen 1988) confirm results Conclusions: Takes time to penetrate to joint fluid from serum Sufficient tiamulin concentrations found for M. hyosynoviae MIC90 0.025 g/ml & M. hyorhinis MIC90 0.25 g/ml in joint fluid Denagard Injection study (Adams & Klein 2010): conc range 0.41-0.70 g/ml (2-12 h p.i.) joint fluid at dosage 18mg thf kg/bw

PK/PD relationship of Tiamulin (parenteral application) in plasma and joint fluid

(Makhanon, Burch and Klein, 2011)

Denagard PK/PD relationship for M. hyorhinis / M. hyosynoviae - CONCLUSIONS

Recommended dose rates of tiamulin for parenteral application (15 mg/kg bwt) give tiamulin levels in plasma (Cmax 0.61g/ml) far in excess of the MIC 90 for M. hyorhinis and M. hyosynoviae Tiamulin distributes well into joint fluid after intramuscular administration

Denagard PK/PD relationship for M. hyorhinis / M. hyosynoviae - CONCLUSIONS

Tiamulin concentrations in the joint fluid are approximately 40% (range 29-59%) of the tiamulin plasma concentration. They appear likely to cover 21 hours of the 24 hour dosing period at the MIC 90 concentration. Denagard Injection at 15 mg/kg bw correlates well with its indications for the treatment of mycoplasmal pneumonia & arthritis

Denagard Injection arthritis studies

Skov and Nielsen 1998 McKellar 1993 Burch and Goodwin 1984 Blowey and Pott 1992

Determination of tiamulin in various organs and body secretions in pigs following i.m. administration of single dose
B.Skov and B.H.Nielsen Leo Research Report, 1988.

Materials and methods

One pig (body weight 43kg, 10-12 weeks old) was treated (15 mg/ kg bw) with a single dose of Denagard Injection. Two hours after administration the animal was killed and tissue samples taken. Microbiologic assays were used to determine the tiamulin hydrogen fumarate concentrations (detection limit 0.1 g/ml in serum and 0.3 g/ml in tissues).

Results

Serum

Synovial fluid (knee)

Synovial fluid (knee)

Synovial fluid (knee)

Muscle

Lung

Tiamulin hydrogen fumarate (g/ml)

0.22

0.13

0.17

0.10

0.74

17.1

Discussion/Conclusions

The tiamulin hydrogen fumarate concentration in serum was 0.22 g/ml and varied in synovial fluids from 0.10-0.17 g/ml. Remarkable high tissue concentrations were found in the lungs (17.1 g/ml). The tiamulin hydrogen fumarate concentrations in the kidneys (3.3 g/ml) and in the liver (1.8g/ml) were higher than in the muscle (0.74 g/ml).

Plasma and tissue kinetic study of Tiamulin in pigs (determination of tiamulin in plasma and joint fluid)
Q.A.McKellar Leo Research Report, 1993.

Materials and methods

Twenty-five male pigs. Three pigs killed at following time points: 2, 4, 6, 8, 24, 32, 48, 72h. Denagard Injection dosage: 15 mg/kg bw once. Microbiologic assay was used to determine the tiamulin hydrogen fumarate concentrations Sufficient synovial fluid sampled from two pigs

PK relationships between plasma and joint fluid

Time (h)

Plasma concentration (g/ml) 0.56

Joint fluid (g/ml)

Ratio Joint fluid/plasma (g/ml) 0.339

0.19

0.65

0.38

0.585

Discussion/Conclusions

The tiamulin joint fluid concentrations are approx. 46% of the plasma concentration (range 34-59%) (McKellar et al 2004). Additional data from Skov and Nielsen (1988a, 1988b) gave simuilar results from three more pigs at a 2 hour timepoint (average 35%, range 28-59%).

Use of tiamulin in a herd of pigs seriously affected with Mycoplasma hyosynoviae arthritis
D.G.S. Burch and R.F.W. Goodwin Veterinary Record 115, 594-595, 1984.

Materials and methods

Forty young gilts from a sow herd (220 sows) with lameness problems were allocated to one untreated control and three treatment groups and treated three consecutive days (Denagard Injection dosage: 10 & 15 mg/kg bw). Pigs were weighed at start of trial (day 0) and on days 3 and 7. Pigs were scored on severity of lameness on days 0, 3 and 7 (0-normal, 1-slightly lame, moderately lame, 3-severely lame) M. hyosynoviae was isolated (synovial fluid) from joints of autopsied pigs with lameness (mild to moderate signs)

Results
Avg. lameness score day 0 Avg. lameness score day 3 Diff. day 0 to 3 Avg. lameness score day 7 Diff. day 0 to 7

Control

2.10

1.50

0.60

1.20

0.90

Tiamulin 10 mg/kg bw (9 pigs) Tiamulin 15 mg/kg bw (10 pigs)

2.35

1.00

1.35**

0.30**

2.05**

2.30

1.10

1.20*

0.30**

2.00**

* p<0.02; ** p<0.01; One pig removed from trial (autopsy: separation right femoral head)

Results

Mycoplasma arthritis trial (Burch & Goodwin, 1984) Lameness scores tiamulin injection

Lameness score reduction by 87%

Results
Avg. Avg. Diff. day weight day 0 weight day 0 to 3 (kg) 3 Avg. Diff. day weight day 7 0 to 7 (kg vs.control)

Control

62.00

62.18

0.18

63.59

1.59

Tiamulin 10mg/kg (9 pigs) Tiamulin 15 mg/kg (10 pigs)

72.52

74.41

1.89**

77.45

4.93** (+3.34) 3.87** (+2.28)

63.41

64.59

1.18*

67.28

* p<0.05; ** p<0.01; one pig removed from trial (autopsy: separation right femoral head)

Results

Discussion/Conclusions

All the treated groups showed significantly better weight gain during the trial period in comparison with the untreated control group. The group with tiamulin 10mg/kg bw performed best. All treated groups showed a significantly greater reduction in lameness score than the negative control. Results indicate that tiamulin at 10 & 15mg/kg bw, when injected for 3 days, was an effective treatment for pigs affected with arthritis caused by M.hyosynoviae. Tiamulin worked at both tested concentrations well and can be recommended for the treatment of M.hyosynoviae arthritis.

The prevention of Mycoplasmal arthritis in gilts using Tiamulin 200 Injection

R.W. Blowey and J.M. Pott 12th IPVS Congress The Netherlands, Proceedings 625, 1992.

Materials and methods

Sixty gilts (weight range 70-100kg) from a sow gilt multiplication unit (320 sows) selected for future breeding stock were given a single injection (Denagard dosage: 15 mg/kg bw) on day of transfer to selection area. Mycoplasmal arthritis was typically seen 1-3 weeks after selection and transfer (40-50 cases per week). Diagnosis confirmed by isolation of M. hyosynoviae from joint fluid. Diagnosis confirmation: post-mortem of live affected animals; isolation of M. hyosynoviae from joint fluid The incidence of clinical arthritis was recorded

Results

Number of total pigs Untreated control 210

Pigs with clinical arthritis 77 (37%)

Tiamulin 15 mg/kg bw

204

27* (13%)

* p<0.001

Discussion/Conclusions

Arthritis occurred in 77 of 210 control gilts (37%), whereas in the tiamulin-treated goup only 27 of 204 gilts were affected (13%). The single dose of 15 mg thf/kg bw was highly effective in reducing the incidence of arthritis disease. For effective decrease of clinical severity and total elimination of M. hyosynoviae arthritis over a longer time period, daily treatment (15 mg/kg bw) over 3 days is recommended.

Efficacy of Denagard 20% Injection for Treatment of Polyarthritis in Nursery

Dr.S.Talummuk & others 21st IPVS Congress Canada, Proceedings 1012, 2010.

Materials and methods

Twenty pigs with severe swollen joints and respiratory symptoms were divided into two groups and treated three consecutive days (Denagard Injection dosage: 15 mg / kg bw). Joint diameters were determined before onset of treatment, during treatment (day 3) and after withdrawal of treatment (day 5). The pH-value of the injection formulations was determined. Irritation and pain reaction at the injection site was evaluated.

Materials and methods

Farm history: A farrow to fattening farm was affected by serious polyarthritis problems, 20-30% morbidity rate and 510% mortality rate from suckling piglets to nursery Preventive program was water treatment with amoxicillin after weaning for 5-7 days.

Materials and method

20 pigs with severe swollen joints and respiratory signs were selected and divided into two groups. Group 1 was treated by Denagard 20% injection Group 2 was treated by generic tiamulin injection for three days. Nasal swabs were collected from all pigs before onset of treatment to confirm Mycoplasma infection.

Materials and method

Joint diameter of all four legs was measured by Vernier caliper before the first tiamulin injection (day 1), the third injection (day 3) and two days after withdrawal of treatment (day 5). Signs of pain after injection were observed in the nursery. Statistical analysis of the joint diameter was performed by ANOVA at p<0.05.

Bacterial swab for Mycoplasma hyorhinis

Results showed 100% of pigs positive for Mycoplasma hyorhinis

Clinical signs: sample pigs

Swollen joint

Clinical signs: sample pigs

Polyarthritis

Measuring with Vernier caliper

Measuring with Vernier caliper

Injecting Denagard 20%

Reduction of joint swelling

Mean Diameter of joint (inch) before, during and after the treatment (Ab p<0.05)
Legs
Right foreleg

Observed day
Day 1 Day 3 Day 5

Mean Joint Diameter (inch) Denagard 20% Injection

2.73 + 0.33 2.53 + 0.21a 2.56 + 0.25 2.76 + 0.39 2.70 + 0.34 2.50 + 0.25a 3.21 + 0.56 2.96 + 0.41 2.90 + 0.38a 3.41 + 0.70 3.12 + 0.59 3.10 + 0.69

Tiamulin generic
2.83 + 0.23 2.82 + 0.14b 2.78 + 0.20 2.91 + 0.20 2.86 + 0.14 2.86 + 0.17b 3.40 + 0.33 3.24 + 0.18 3.30 + 0.17b 3.33 + 0.25 3.20 + 0.13 3.26 + 0.20

Left foreleg

Day 1 Day 3 Day 5

Right hind leg

Day 1 Day 3 Day 5

Left hind leg

Day 1 Day 3 Day 5

M. hyorhinis arthritis joint swelling reduction after Denagard injection (Talummuk et al, 2010)

Average reduction in joint diameter by 9.3% after 5 days

Discussion/Conclusions

The mean joint diameter of Denagard treated group showed a gradual decrease from the first to the last day of evaluation. Significant differences of joint diameter were found in three legs on day 3 and day 5. The acidic pH and different active ingredient used in the generic products can reduce the performance and efficacy and cause pain at the site of injection. In conclusion Denagard 20% Injectable performed better than generics, significantly reducing swollen joints in nursery pigs suffering from polyarthritis. Its significant efficacy was demonstrated 3 to 5 days after treatment.

Denagard Injection

Tiamulin parenteral pharmacodynamics in pigs other respiratory/systemic pathogens

Actinobacillus pleuropneumoniae Streptococcus suis

Denagard

Pharmacodynamics
Tiamulin MIC data (A.pleuropneumoniae) Aitken et al. 1999 Fodor et al. 2004 VetPath 2004-2006 Variable MIC results reported depending on method CLSI have established a method and breakpoints

Denagard and Porcine Pleuropneumonia (introduction)

Denagard is active against A. pleuropneumoniae in vitro and in vivo A. pleuropneumoniae isolates from many countries have been shown to be sensitive in vitro to tiamulin Breakpoints for tiamulin: 8g/ml (sensitive), 9 - 16 g/ml (moderately sensitive), >16 g/ml resistant - calculated by Casals, 1990

Tiamulin MICs A. pleuropneumoniae - breakpoints

Suggested breakpoints for APP: sensitive (<8.0); intermediate (<9-16); resistant (>16.0) (Casals et al, 1990)

MICs (g/ml) A.pleuropneumoniae strains from United Kingdom (n=15)

(Aitken et al. 1999, Veterinary Record, 144, p.128)

Antimicrobial Tiamulin Valnemulin Oxytetracycline Penicillin V

MIC 50 32.0 16.0 2.0 3.6

MIC 90 64.0 32.0 16.0 3.6

Range 8.0-64.0 4.0-32.0 1.0-16.0 1.8-28.8

MICs (g/ml) A. pleuropneumoniae strains from Hungary (n=10) (Fodor et al. 2004)
Antimicrobial Tiamulin Doxycycline Lincomycin Tylosin MIC 50 2.0 0.25 8.0 16.0 MIC 90 4.0 1.0 16.0 32.0 Range 2.0-4.0 0.25-8.0 1.0-16.0 24.0-32.0

Use clinical breakpoints - Resistance Tiamulin breakpoints: sensitive 8.0; resistant >16.0

EU VETPATH II Project EU

Objectives: To produce a trans-Europe collection of strains from animals not recently exposed to antibiotics. Uses: Use the collection for regulatory purposes CVMP guideline EMEA/CVMP/627/01-Final Guideline for the demonstration of Efficacy for Veterinary Medicinal Products containing AI`s

EU VETPATH II Project EU

Participating companies: Bayer, Ceva, Elanco, Fort Dodge, Intervet/ Schering-Plough, Novartis, Pfizer, Vetoquinol, Virbac Bacteria strain collection (DK, B, NL, GB, D, F, E, POL, CZ): Actinobacillus pleuropneumoniae respiratory (goal:140 samples) Streptococcus suis respiratory/meningitis (goal:140 samples)

MICs (g/ml) A.pleuropneumoniae strains from B, DK, F, Ger, NL, Pol, E, UK (n=129)
(Felmingham 2009, VetPath II project)

Antimicrobial Tiamulin Tilmicosin Lincomycin Tylosin Tetracycline Amoxycillin

MIC 50 8.0 8.0 >32 32.0 1.0 0.5

MIC 90 16.0 16.0 >32 32.0 16.0 0.5

Range 0.25-16 4-16 2.0-32.0 0.5-32.0 0.25-32 0.25-32

Resistance (%) 0 0 ----15% ---

Used CLSI method

Tiamulin MICs for Actinobacillus pleuropneumoniae (VetPath II, individual countries)

0.125 B (13 isol.) DK (28) F (18) Ger (21) NL(7) Pol (19) E (8) UK (8) ----------------0.25 --1 ------------0.5 ------1 --------1.0 ------------1 --2.0 ----------1 ----4.0 ----2 4 --4 1 1 8.0 13 16 15 13 7 13 5 14 16.0 --11 1 3 --1 1 ----------32.0

Tiamulin effective against APP < 8 g/ml

(VethPath collection in EU , 2006/2007)

Comparison of Tiamulin MICs against App vs. Tilmicosin MICs (VetPath II project)

No resistance pattern observed

(EU VethPath collection, 2009, 129 strains B/DK/F/GER/NL/POL/E/UK)

Comparative susceptibility patterns of A. pleuropneumoniae to Tiamulin and Tetracycline

(VetPath II project)

Susceptible tiamulin isolates

Resistant tetracycline isolates

(EU VethPath collection, 2009, 129 strains B/DK/F/GER/NL/POL/E/UK)

MICs (g/ml) A. pleuropneumoniae strains from Czech Republic (n=242, isolated in 2007-2009)
(Kucerova et al. 2011, Veterinary Microbiology 150, p.203-206)

Antimicrobial Tiamulin Tilmicosin Tulathromycin Tetracycline Florfenicol Amoxycillin/Clavul anic acid

MIC 50 8.0 4.0 8.0 0.5 0.5 0.5

MIC 90 16.0 8.0 16.0 16.0 16.0 2.0

Range 0.5-64 1.0-256 1.0-32.0 0.5-128.0 0.25-8 0.5-0.5

Resistance (%) 1.7 1.2 --24% 0.8% 0.8%

Use clinical breakpoints - Resistance Tiamulin breakpoints: sensitive 8.0; resistant >16.0

Evaluation of tiamulin administered i.m. to pigs for treatment of Actinobacillus Pleuropneumonia

G.L. Schultz, R.A. Schultz, M.D.Anderson and T.L.Cue Proceedings IPVS Rio de Janeiro,92, 1988.

Materials and methods

Forty-eight healthy crossbred pigs were infected with APP (serotype 5) culture. Medication was administered when clinical signs of pneumonia were present. Medication was continued once daily until rectal temperatures were normal for 2 consecutive days or until pigs have received injections for four consecutive days (Denagard Injection dosage tiamulin base: 3mg/lb; 6mg/lb; 9mg/lb, non-medicated control = 6.6 mg/kg; 9.9mg/kg; 13.2 mg/kg tiamulin base or 8.1; 12.2; 16.3mg/kg tiamulin hydrogen fumarate)

Materials and methods

Individual pig weights were determined on day 0, 7, 16. Feed consumption was monitored for the same period of time. Post mortem examination were performed on all pigs. Extent & severity of gross pneumonic lesions were diagrammed and scored. Each lung was scored 1, 2, 3 or 4 to indicate no lesions, less than 25%, 25 to 75%, greater than 75% lesion involvement.

Results
Non-medicated Dena 3mg/lb Dena 6mg/lb control (= 8.1 mg THF/kg bw) (= 12.2 mg THF/kg bw) Mortality (%) 16 0* 10.2 * 0* 10.0 * Dena 9mg/lb (= 16.3 mg THF/kg bw) 0* 8.4 *

Lung lesion score 17.2

Re-isolation APP 75 (%) ADF lbs d0-16 ADG lbs d0-16 F/G lbs d0-16
* p<0.01

33 *

25 *

16 *

1.15 0.23 5.0

2.17 * 0.77 * 2.82

2.23 * 0.93 * 2.39

1.95 * 0.99 * 1.97

Efficacy of tiamulin injection against artificial infection with APP ST5 (estimated MIC of 3.0)
(Schultz et al. 1988)

Conclusions Actinobacillus pleuropneumonia studies

Recent MIC data prove consistent sensitivity of tested Actinobacillus pleuropneumoniae strains. No resistant strains were found (CLSI breakpoints for tiamulin sensitive 16 g/ml, resistant 32 g/ml) and it does not seem to develop a resistance pattern to tiamulin unlike for example to tetracycline. In the APP challenge study (serotype 5) a significant linear reduction of lung lesion severity and number of APP positive pigs were found as drug level increased.

Conclusions Actinobacillus pleuropneumonia studies

All the Denagard-treated groups showed significantly reduced mortality vs. non-medicated controls. The medication group with the highest Denagard dose (9mg/lbs = 16.3mg THF/kg bw) performed best. Denagard Injection is an effective treatment against induced Actinobacillus Pleuropneumonia infection in swine.

Denagard Injection

Tiamulin parenteral pharmacodynamics in pigs enteric pathogens

Brachyspira hyodysenteriae / Brachyspira pilosicoli Lawsonia intracellularis Clostridium perfringens / Clostridium difficile

Denagard microbiological activity

Brachyspira hyodysenteriae / Brachyspira pilosicoli Lawsonia intracellularis

MICs Brachyspira spp

Antimicrobial B. hyodysenteriae Tiamulin Lincomycin Tylosin B. pilosicoli Tiamulin Lincomycin Tylosin

MIC 50

MIC 90

Range

Karlsson et al, 2002 0.125 16 >256 1.0 64 >256 Kinyon et al, 2002 0.125 32 >512 1.0 64 >512 0.06-8.0 4.0->128 <16->512 0.016-2.0 1.0-64 2.0->512

MICs Brachyspira spp

Antimicrobial B. hyodysenteriae Tiamulin Lincomycin Tylosin B. hyodysenteriae Tiamulin Lincomycin Tylosin

MIC 50

MIC 90

Range

Hildago et al, 2009 0.25 16 >256 2.0 128 >256 0.016-2.0 2.0-128 4.0->256

Magistrali et al, 2010 0.063-2.0 4.0-64.0 64-128

Denagard Injection swine dysentery studies

Taylor 1980 Burch 1983

Efficacy of parenteral tiamulin medication in the therapy of experimental swine dysentery

Prof.D.J.Taylor 6th IPVS Congress Copenhagen, Denmark, Proceedings p.255, 1980.

Materials and methods

Fifteen crossbred pigs from a herd known to B. hyodysenteriae-free were chosen for the challenge study. The pigs were infected with B. hyodysenteriae (isolate 36/4). Clinical signs of swine dysentery produced were mild and lasted for a relatively short time period. Tiamulin was administered as a single intramuscular injection (10mg thf/kg bw Disappearance of clinical signs of swine dysentery and re-isolation of B. hyodysenteriae from faeces were determined.

Effect of single i/m injection of tiamulin (10mg thf/kg bw) in experimental swine dysentery (Taylor et al.
1980)

Number Infected Tiamulin of pigs Medicated

Number with clinical signs of swine dysentery Day 0 Day 1 Day 5 Day 12

5 5 5

+ +

0 5 5

0 5 0

0 1 0

0 0 0

Re-isolation of B. hyodysenteriae from faeces of experimentally infected pigs (Taylor et al. 1980)
Number Infected Tiamulin of pigs Medicated Number with clinical signs of swine dysentery Day 0 Day 1 Day 5 Day 10

5 5 5

+ +

0 5 5

0 5 1

0 3 0

0 0 0

Conclusions

Faeces returned to normal consistency and appearance within 24 hours of treatment. Appetites and body condition markedly improved and no further signs of clinical swine dysentery were seen after medication. Single i/m injection of tiamulin at a dose of 10mg thf/kg bw could be sufficient for the treatment of naturally occurring swine dysentery with the elimination of the causative organism, Brachyspira hyodysenteriae.

Tiamulin injection for the treatment of swine dysentery

D.G.S.Burch Veterinary Record, 113, 236-237 1983.

Materials and methods

Thirty-six crossbred pigs were purchased from a known swine dysentery farm. When over 50% of the pigs had diarrhoea the trial was started. Scoring for severity of diarrhoea (3 blood in faeces, 2 watery or mucoid, 1 soft faeces, 0 normal). Weighing of the pigs (day 4 & day 8). Post-mortem examination were performed in case of dead pigs.

Diarrhoea Score

Day 1 Negative control 0.66 Tiamulin 10mg/kg bw Tiamulin 15mg/kg bw 1.50 1.18

Day 2 0.66 0.12 0.40

Day 4 1.83 0.16 0.09

Day 8 1.45 0.08 0.00

Weight gain (in kg)

Day 1 Negative control Tiamulin 10mg/kg bw Tiamulin 15mg/kg bw 32.72 32.95 36.85

Day 4 31.79 34.03 (+1.08kg) 38.06 (+1.21kg)

Day 8 30.57 36.07 (+3.12kg) 40.66 (+3.81kg)

Percentage of pigs positive for B. hyodysenteriae

Day 1 Negative control Tiamulin 10mg/kg bw Tiamulin 15mg/kg bw 17 25 36

Day 4 42 8 0

Day 8 67 0 0

Conclusions

Pigs affected with acute swine dysentery were effectively treated with a single dose at a rate of either 10 or 15mg/kg bw The response to treatment appeared very rapid (i.e. in 24 hours) Due to cessation of diarrhoea, the treated pigs gained weight, whereas the untreated pigs did not. Both levels of tiamulin were equally effective treatment for pigs affected with swine dysentery.

Antimicrobial Minimum Inhibitory Concentrations for Lawsonia intracellularis Isolates from the U.S. and Europe
Dr. Suphot Wattanaphansak, Prof. Connie Gebhart, Prof. Randall Singer Veterinary Microbiology 134, 305-310, 2009.

MIC testing of Lawsonia intracellularis

Limited information on Lawsonia intracellularis (L.i.) MICs available Obligate intracellular bacterium
Difficult to isolate and propagate No standardized MIC protocols applicable Must use a modified cell culture system

Few strains available because of difficulty in initial purification and propagation from intestine

Objective

Determine antimicrobial MICs of 10 L.i. field isolates from pigs in the United States and Europe against 6 antimicrobials commonly used for ileitis control and treatment
Carbadox Lincomycin Tylosin Chlortetracycline Tiamulin Valnemulin

General methods

Extracellular MICs
Measure effect of antimicrobial on L.i. while in the gut lumen prior to enterocyte infection exposure time short only 2 hours MICs tend to be higher

Extracellular Lumen

Intracellular MICs
Measure effect of antimicrobial on L.i. after it is established within the enteroctye lipid solubility increases cell penetration
Intracellular Enterocytes

General methods

Tissue culture system (McCoy cell monolayer) Antimicrobials tested in a range of different concentrations (0.125 128 g/ml) titration of low and high MICs possible Intracellular & extracellullar MIC assays for L.i. performed (mimic real infection situation organism exposed to ABs before/after invasion into intestinal cells) MIC identified as the lowest conc. that inhibited 99% of L.i. growth (compared to AB-free control)

Intracellular and extracellular MIC ranges for L. intracellularis strains from the United States and Europe1
Anti-microbial agent US L.intracellularis isolates EU L.intracellularis isolates (n=6) (n=4) Intracellular MIC (g/ml)
Chlortetracycline Lincomycin Tylosin Tiamulin 4-64 16->128 0.25-32 0.125-0.5

Extracellular Intracellular MIC Extracellular MIC (g/ml) (g/ml) MIC (g/ml)

32-64 >128 1->128 1-32 0.25-16 8-64 0.5-2 0.125 16-64 32->128 2-16 1-4

1. S.Wattanaphansak, R.S.Singer, C.J.Gebhart (2009): In vitro antimicrobial activity against 10 North American and European L.intracellularis isolates. Veterinary Microbiology, 134, 305-310.

Intracellular and extracellular MIC ranges for L. intracellularis strains from the United States and Europe1

1. S.Wattanaphansak, R.S.Singer, C.J.Gebhart (2009): In vitro antimicrobial activity against 10 North American and European L.intracellularis isolates. Veterinary Microbiology, 134, 305-310.

Discussion/Conclusions

Intracellular MICs against L.i. were lower than extracellular MICs for all antimicrobials tested Both intracellular and extracellular MICs for European isolates tended to be lower than those for U.S. isolates. Geographic differences? L.i. strains demonstrate the highest degree of intracellular sensitivity to tiamulin & valnemulin Low susceptibility and high variation in the sensitivity to drugs like tylosin, lincomycin and chlortetracycline situation incidence, future trends Tiamulin and valnemulin are highly effective antimicrobials vs Lawsonia intracellularis

The other pathogens

Gram positive Staphylococcus, Streptococcus, Clostridium perfringens, Listeria monocytogenes, Erysipelothrix spp., Arcanobacterium (Corynebacterium) pyogenes Gram negative Pasteurella spp., Klebsiella pneumoniae, Haemophilus spp., Fusobacterium necrophorum, Bacteroides vulgatus Campylobacter (Vibrio) coli

Pleuromutilin susceptibility of Clostridium perfringens and Clostridium difficile isolates from pigs in Italy and Denmark
Dr. Fabrizio Agnoletti et al. 12th IPVS Congress Vancouver, Canada, Proceedings p.625, 2010.

Clostridium perfringens / Clostridium difficile

Susceptibility testing of 68 C.perfringens (Italy 30 strains recovered 2007-2008, Denmark 38 strains from 2006) & 15 C.difficile (Italy 10 strains, Denmark 5 strains, recovered 2007-2008) strains Strains originated from neonatal healthy pigs or pigs affected by enteritis Genetic characterization of isolates: Clostridium perfringens strains were toxin-typed by a multiplex PCR for the detection of major toxin encoding genes (cpa, cpi, cpetx and cpb) (5) and 2 gene presence was evaluated according to Baums et al. (2004)

Tiamulin MIC ranges for Clostridium perfringens / Clostridium difficile (Agnoletti et al 2010)
C.perfringens (n=30) Italy MIC 50 MIC 90 Geometric mean MIC range 4 64 4.6 0,125-128 2 4 2 0.25-8 C.perfringens (n=38) Denmark C.difficile (n=15) Italy/Denmark 8 16 8.4 0.125-16

C. perfringens susceptibililty to TIAMULIN and origin of isolates MICs differ between countries!

0 0 0 0 % strains 0 0 0 0 0 0 0 0 0
0 0 0 0 0 0 0 ,0 0 0 0, 00 0 0 ,0 0 0 0, 00 0 0 ,0 0 0 0 0 0 ,0 0 0 0 0 0 0 0 0 0 0

MIC50: 2 MIC90: 4 Geom M: 2.0

MIC50: 4 MIC90: 64 Geom M: 4.6

MIC (mcg/ml)

Danish C. perfringens

Italian C. perfringens

PK/PD relationships of Tiamulin (water/parenteral application) in colon contents

Conclusions

C.perfringens strains from DK showed higher susceptibility to tiamulin compared to strains from Italy. Pharmacokinetic studies at tiamulin doses of 15-20mg/kg bw indicate that after oral or parenteral application tiamulin colon concentrations between 13-18 g/ml (Anderson et al.1994, McKellar 2004) can be achieved. All Danish C.perfringens isolates can based on the PK data be considered susceptible to tiamulin. 20% of the Italian C.perfringens isolates (MIC values between 32 and 128) can be considered resistant to tiamulin. All C.difficile isolates showed susceptibility to tiamulin.

Denagard

Pharmacodynamics
Tiamulin MIC data (Streptococcus suis) Aitken et al. 1999 Fodor et al. 2004 VetPath 2004-2006

Denagard and Streptococcus suis

Denagard is active against Streptococcus suis in vitro and in vivo S. suis isolates from all over the world have been shown to be highly sensitive in vitro to tiamulin

MICs (g/ml) Streptococcus suis strains from UK / Hungary

MIC 50 Aitken et al. 1999 Fodor et al. 2004 2.0 0.125

MIC 90 2.0 0.25

Range 1.0 2.0 0.015 0.5

Tiamulin efficacy and breakpoints to be determined many field reports positive

Tiamulin MICs for Streptococcus suis (VetPath II collection 2004-2006)

0.125 F (19 isol.) D (15 isol.) NL (13 isol.) E (10 isol.) UK(9 isol.) 1 --------0.25 2 1 --1 --0.5 3 1 ------1.0 11 4 8 6 5 2.0 2 7 5 1 3 4.0 --1 ----1 8.0 ------2 --16.0 -----------

Comparison of Tiamulin MICs against S. suis vs. Tilmicosin/Tylosin

(EU VethPath collection, 2004/2006

Denagard microbiological activity

Final Conclusions Highly active antimicrobial against Brachyspira spp. Lawsonia intracellularis: Consistently low MICs & narrow MIC ranges Pronounced efficacy in vitro against Mycoplasma spp. High susceptibility of Actinobacillus pleuropneumoniae strains in Europe Pronounced activity against Streptococcus suis

Subjects covered an overview

Formulation / product characteristics Pharmacokinetic / pharmacodynamic relationships Microbiological activity Efficacy against enteric / respiratory / systemic infections Safety Comparison vs. generic products/formulations

Denagard Injection

Safety

Denagard - safety

At single doses of 100mg/kg orally in the pig, hyperpnoea and abdominal discomfort were noted. No CNs effects were noted. Safety studies in the pregnant sow at daily doses of 16mg thf / kg body weight have revealed no effect (Pott et al 1984, Riley et al 1982, Edwards et al 1987). Similarly, this daily dose has no effect on fertility in either the sow or the boar (Edwards et al 1985, Glawischnig 1975, Laber et al. 1985)

Denagard Injection 10% (t base) - Dosage and Administration

Single intramuscular injection at 1.0 ml per 12.5 kg bw for Swine dysentery (1-2 days) Multiple intramuscular injection (3 days) at 1.5 ml per 12.5kg bw for treatment of enzootic pneumonia or PRDC Multiple intramuscular injection (3 days) at 1.5 ml per 12.5kg for Mycoplasmal arthritis Multiple intramuscular injection (3 days) at 1.5 ml per 12.5kg for APP Pleuropneumonia

Denagard Injection 20% (thf) - Dosage and Administration

Single intramuscular injection at 1.0 ml per 20kg bw for Swine dysentery (1-2 days) Multiple intramuscular injection (3 days) at 1.5 ml per 20kg bw for treatment of enzootic pneumonia or PRDC Multiple intramuscular injection (3 days) at 1.5 ml per 20kg bw for Mycoplasmal arthritis Multiple intramuscular injection (3 days) at 1.5 ml per 20kg bw for APP Pleuropneumonia

Table 1 Denagard 20% (thf) Schedule for swine

Weight (kg) 7 10 20 40 50 100 200 250 Dose volume (ml) 10mg thf/kg bw swine Dose volume (ml) 15mg thf/kg bw dysentery , ileitis (single dose) multiple dose for PRDC 0.35 0.5 1 2 2.5 5 10 12.5 0.52 0.75 1.5 3 3.75 7.5 15 18.75

Denagard Injection

Medication programmes used in Thailand

Recommended program: gilts

Gilt management program for PRDC and Mycoplasma spp. control

AGE
Wks

25

26

27

28

29

30

31

32

33 34

35

On 2 consecutive days of Denagard 20% injection 10mg/kg bw before mating, at 31 wk old

Recommended program: breeders

Breeder management program for PRRSV positive herds to control secondary infection from Mycoplasma spp. and Denagard sensitive pathogens

Pregnant
Pregnancy wks

1 0

1 1

12

13

Farrowing 1 15 16 4

1 shot of Denagard20% injection: 10-15mg/kg bw, sow at 10 wk of pregnancy

Recommended program: piglets

Weaning program to decrease pathogens in nursery unit : Mycoplasma spp., Streptococcus spp., H. parasuis , etc.

Sucking piglets
AGE
Wks

Nursery piglets 3 4 5 6 7 8 9 10

At weaning day 1 shot of Denagard 20% injection: 15 mg/kg bw

Recommended program: mycoplasmal arthritis

Treatment program for Mycoplasmal arthritis in nursery pigs caused by M. hyorhinis

Sucking piglets
AGE
Wks

Nursery piglets 3 4 5 6 7 8 9 10

3 consecutive days of Denagard 20% injection: 15mg/kg bw

Recommended program: PRDC

Treatment program for PRDC :Mycoplasma spp., H. parasuis, Streptococcus spp., Pasteurella multocida, etc.

Starter
AGE
Wks

Grower 13 14 15 16 17 18 20 21

10

11

12

3 consecutive days of Denagard 20% injection 10 mg/kg bw

Recommended program: enteric diseases

Treatment program for swine dysentery, ileitis and colitis

Starter
AGE
Wks

Grower 13 14 15 16 17 18 19 20

10

11

12

3 consecutive days of Denagard 20% injection 10mg/kg bw

Recommended program: APP

Mass treatment for Acute APP

Starter
AGE
Wks

Grower 13 14 15 16 17 18 19 20

10

11

12

3 consecutive days of Denagard 20% injection 15mg/kg bw

Denagard Injection

Medication programmes used in Denmark

Denagard Injection medication strategies Denmark (Denagard 20% Injection product)

Animal
Gilts

Disease
Lameness (enter of new facilities)

Dosage

Treatment duration
every

1ml/15kg bw (13.3mg/kg 1-3 days or bw) or 1ml/10kg bw 2nd day (20mg/kg bw) 10mg/kg bw 15mg/kg bw 10 mg/kg bw

Weaners /nursery pig/growers Nursery pig/growers Suckling piglets

Ileitis, Colitis, Swine Dysentery Pleuropneumonia Eradication Swine Dysentery, Enzootic Pneumonia

1-3 consecutive days 3 consecutive days 3-4 times during the treatment period of breeding animals

Denagard Injection medication strategies Denmark (Denagard 20% Injection product)

Animal Disease Dosage
1 ml/20kg bw (10mg/kg)

Treatment duration
Day 1 and day 3

Breeding animals, that do Eradication Swine not eat and drink during Dysentery, Enzootic eradications Pneumonia Newborn pigs Mycoplasma suis and Clostridium perfringens type A diarrhoea

10 mg/kg bw

Once within the first 4 days

Subjects covered an overview

Formulation / product characteristics Pharmacokinetic / pharmacodynamic relationships Microbiological activity Efficacy against enteric / respiratory / systemic infections Safety Comparison vs. generic products/formulations

Denagard Injection

Comparison vs. generic products / formulations

Study A study was conducted to quantify any difference between Denagards 10% Injectable formulation and currently available generic tiamulin injectable formulations

Materials and methods

Chemical and physical properties of five generic tiamulin 10% Injectable products (registered in CEE countries) vs. Denagard 10% Injectable were compared Parameters evaluated: appearance, pH-value, water miscibility, water content, tiamulin concentration, efficacy of the preservation system, concentration of related substances and degradation products

Results

Majority of generic products contained tiamulin hydrogen fumarate (THF) as active ingredient. The pH-value of the three water-based formulations was much lower (pH 4.15-4.6) in comparison to Denagard 10% Injection (pH 8.74) which is an oily formulation.

Results

Two of the generic products were oily-based these products were shown to contain higher concentrations of known impurities at levels that exceed the limits specified by EU Pharmacopoeia. Antimicrobial Preservative Effectiveness (APE) results established that the preservative system employed in the Denagard formulation is superior to that used in one oilybased generic products tested.

Conclusions

Generic products which contain tiamulin hydrogen fumarate (THF) are less tolerated. THF formulations are more acidic and cause more pain & irritation at the site of injection. Generic formulations with low pH-value are less stable during storage than oily-based formulations like Denagard. Trials have show that those formulations are likely to cause irritation, pain at the injection site, higher tissue reaction scores and in some cases, post injection lameness.

Conclusions

Denagard 10% Injection has a high tissue tolerance & low pain reaction after injection because it is less acidic and uses tiamulin base as its active ingredient. Denagard was the only oily-based injectable to justify the regulatory requirements for purity. Using generic injectables could expose animals to impurities which do not provide pharmaceutical activity.

Conclusions

Shelf-life and stability of generic injectable products are unlikely to be as good as that of Denagard because the preservation system used in Denagard Injectable products is more effective than that found in generic product tested.

Appearance

A: Denagard 20% with yellowish color B: Tiamulin generic with no color

Sandoz Tiamulin base

pH test

Generic

Generic

pH test: customers can test, themselves

8.5

A
Denagard

neutral

Generic Tiamulin

neutral

We Take pride in our Denagard Injection