Denagard Injection

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

• 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)

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

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


• 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)


• 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

• McKellar et al. 2004

• Anderson et al. 1994

• Forster et al. 1982

Tiamulin parenteral pharmacokinetics in pigs

Denagard injection - pharmacokinetics

• Plasma concentrations are relatively low – Cmax 0.61µg/ml; AUC 12.82 µg.h/ml

• Lung concentrations high – Cmax 9.6µg/g; AUC 232µg/g

• Colon contents also high – Cmax 12.75µg/g; AUC 314µg/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)

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

Pharmacokinetics of Tiamutin® (tiamulin) in tissues calculated from mean concentrations from each time of kill following single i/m injection of tiamulin

PlasmaColonwall

Coloncontents

Lung

AUC (μg h/ml) 12.82 64.51 314.23 231.52

AUMC (μg h2/ml)

252.02 1252.78 9013.0 3868.1

MRT (h) 19.66 19.42 28.68 16.71

C max (μg/ml) 0.61 2.27 12.75 9.60

t max (h) 4.00 6.00 24.00 4.00

Denagard Injection

• Anderson et al. 1994

Tiamulin parenteral pharmacokinetics in pigs

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)

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

Parenteraldosage rate (thf) equivalent

Denagard® (tiamulin) activity (μg/g)

Lung Tonsils Colonmucosa

Coloncontents

13.6mg/kg 26.9 3.23 2.58 3.09

27.2mg/kg 71.0 8.44 8.99 24.9

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 µg/ml between 2 and 8 hours post-injection and were still in excess of 3.0 µg/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 Respiratory pathogens (tiamulin MIC range μg/ml)

Dosage mg/ml M. hyopneumoniae M. hyorhinis M. hyosynoviae App P.multo S.suis

13.6 mg/kg

Lung 26.9

Tonsils

3.23

0.01-0.05 (Windsor 1996)

0.06-1.0

(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 (Thongkamkoon2010)

0.024-0.39 (Thongkamkoon 2005)

0.048 - 0.097 (Thongkamkoon 2010)

0.0025-0.025

(Windsor 1996)

0.0025-0.01 (Hannan1997)

0.0156-0.0625 (Aarestrup&Friis 1998)

0.03-0.25 (Stipkovits 2004)

0.048-0.097 (Thongkamkoon 2010)

32-64

(Aitken 1999)

2-4 Fodor 2004)

0.25-16.0 (VetPath 2009)

0.5-64.0 (Kucerova 2011)

1.0-8.0 (Fodor 2004)

4.0-64.0 (VetPath 2009)

1.0-2.0 (Aitken 1999)

0.015-0.5 (Fodor 2004)

0.125-8 (VetPath 2009)

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 Enteric pathogens (tiamulin MIC range μg/ml)

Dosage

Colon mucosa

Colon contents

B.hyo 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)

0.25-8 (DK)

(Agnoletti 2010)

0.125-16(I&DK) (Agnoletti 2010)

4-32

(Prapasarakul,2007)

27.2 mg/kg

8.99 24.9

Denagard Injection

• 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

Tiamulin parenteral pharmacodynamics in pigs – Mycoplasma spp.

AB MIC data Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma synoviae

• 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!

Mycoplasma isolation / strain specification / MIC testing

Denagard

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

Pharmacodynamics

MICs (µg/ml) M. hyopneumoniae strains from Japan (n=90) (Kobayashi et al, 2008)

Antimicrobial MIC 50 MIC 90 Range

Tiamulin 0.06 0.125 0.03-0.125

Valnemulin 0.002 0.004 0.002-0.004

Oxytetracycline 1.0 2.0 0.25-4.0

Lincomycin 0.25 0.5 0.125->64

Tilmicosin 0.25 0.5 ≤0.25->16

Tylosin 0.06 0.25 0.06->64

Enrofloxacin 0.125 0.125 0.06-1.0

Use clinical breakpoints / plasma levels - Resistance

MICs (µg/ml) M. hyopneumoniae Thailand (n=10 2008, n=10 2009) (Thongkamkoon et al, 2010)

Antimicrobial MIC50 2008

MIC50 2009

MIC90 2008

MIC90 2009

MIC range2008

MIC range2009

Tiamulin 0.048 0.048 0.048 0.097 0.048-0.097 0.048-0.19

Valnemulin <0.006 <0.006 <0.006 <0.006 <0.006 <0.006

Doxy 3.12 3.12 6.25 6.25 3.12-6.25 1.56-6.25

Tylosin 0.19 0.097 0.19 0.097 0.097-0.19 0.048-0.19

Linco 0.19 0.19 0.39 0.78 0.19-0.39 0.19-0.78

Enrofloxacin 3.12 1.56 6.25 6.25 0.048-6.25 0.024-6.25

Florfenicol 1.56 1.56 1.56 3.12 1.56-3.12 1.56-3.12


MICs (µg/ml) M. hyopneumoniae strains from Belgium (n=21) (Maes et al. 2007)

Antimicrobial MIC 50 MIC 90 MIC range

Tiamulin 0.03 0.12 0.015-0.12

Lincomycin 0.06 0.12 0.06->8.0

Tylosin 0.06 0.12 0.015->1.0

Tilmicosin 0.5 0.5 0.25->16.0

Enrofloxacin 0.06 0.5 0.03->1.0


Tiamulin ≤0.015 0.12 ≤0.015-0.12


Lincomycin ≤0.06 ≤0.06 ≤0.06->8.0

Tilmicosin 0.25 0.5 ≤0.25->16

Tylosin 0.03 0.06 ≤0.015->1.0

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

Minimum inhibitory concentration results for tiamulin against 43 isolates of M.hyopneumoniae from Europe (Pridmore et al. 2008)

Strains MIC 50 MIC 90 MIC range

M.hyopneumoniae(United Kingdom, 19 strains)

0.016 0.031 0.008-0.031

M.hyopneumoniae(Spain, 24 strains)

0.016 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 0.61µg/ml) far in excess of the MIC 90 for M. hyopneumoniae (range MIC 90 0.031-0.12µg/ml).

Denagard

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

Pharmacodynamics

MICs (µg/ml) M. hyorhinis strains from Thailand (n=11 2008, n=9 2009) (Thongkamkoon et al, 2010)

Antimicrobial MIC50 2008

MIC50 2009

MIC90 2008

MIC90 2009

MIC range2008

MIC range2009

Tiamulin 0.048 0.097 0.097 0.097 0.048-0.097 0.048-0.097

Valnemulin <0.006 <0.006 <0.006 <0.006 <0.006 <0.006

Doxy 0.78 1.56 3.12 1.56 0.78-3.12 0.39-1.56

Tylosin 3.12 3.12 3.12 6.25 0.078-3.12 1.56-6.25

Linco 0.78 1.56 0.78 3.12 0.39-1.56 0.78-3.12

Enrofloxacin 1.56 3.12 3.12 50 0.78-25 0.78-50

Florfenicol 3.12 3.12 3.12 3.12 1.56-3.12 1.56-3.12


Denagard

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

Pharmacodynamics

MICs (µg/ml) M. hyosynoviae strains from Denmark (n=42) (Aarestrup & Friis, 1998)


Tiamulin 0.0313 0.0625 0.0156-0.0625

Lincomycin 1.0 2.0 0.5-4.0

Tylosin 2.0 16 0.125-16

Tetracycline 1.0 2.0 0.5-2.0

Enrofloxacin 0.5 0.5 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 (µg/ml) M. hyosynoviae strains from Hungary (Stipkovits et al 2004)


Tiamulin 0.125 0.25 0.03-0.25

Valnemulin 0.06 0.125 0.015-0.125

Lincomycin 2.0 4.0 0.5-8.0

Tylosin 4.0 16.0 2.0-32

Doxycycline 1.0 2.0 0.25-4.0

Enrofloxacin 0.5 0.5 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

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

Tiamulin concentrations in joint fluid

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.

Conclusions:• Takes time to penetrate to joint fluid from serum• Range of tiamulin concentrations found in joint fluid

(0.10-0.17 µg/ml)• Remarkably high lung tissue

concentrations found

Tiamulin conc. Knee Stifle Hock Serum Lung

0.13 µg/ml 0.17 µg/ml 0.10 µg/ml 0.22 µg/ml 17.1 µg/ml

Denagard for systemic infections

• Tiamulin concentration in joint fluid (McKellar 2004, i.m. injection 15mg thf kg/bw)

• other data (Skov and Nielsen 1988) confirm resultsConclusions:• Takes time to penetrate to joint fluid from serum• Sufficient tiamulin concentrations found for

M. hyosynoviae MIC90 0.025 µg/ml & M. hyorhinis MIC90 0.25 µg/ml in joint fluid

Denagard Injection study (Adams & Klein 2010): conc range 0.41-0.70 µg/ml (2-12 h p.i.) joint fluid at dosage 18mg thf kg/bw

Tiamulin conc. 4 hours p.i. 8 hours p.i.

0.19 µg/ml 0.38 µg/ml

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 0.61µg/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 μg/ml in serum and 0.3 μg/ml in tissues).

Results

Serum Synovial fluid (knee)

Synovial fluid (knee)

Synovial fluid (knee)

Muscle Lung

Tiamulin hydrogen fumarate(μg/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 μg/ml and varied in synovial fluids from 0.10-0.17 μg/ml.

• Remarkable high tissue concentrations were found in the lungs (17.1 μg/ml).

• The tiamulin hydrogen fumarate concentrations in the kidneys (3.3 μg/ml) and in the liver (1.8μg/ml) were higher than in the muscle (0.74 μg/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.


• 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 (μg/ml)

Joint fluid (μg/ml) Ratio – Joint fluid/plasma (μg/ml)

4 0.56 0.19 0.339

8 0.65 0.38 0.585


• 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.


• 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

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

Avg. lameness score day 0


Diff. day 0 to 3


Diff. day 0 to 7

Control 2.10 1.50 0.60 1.20 0.90

Tiamulin 10 mg/kg bw(9 pigs†)

2.35 1.00 1.35** 0.30** 2.05**

Tiamulin 15 mg/kg bw(10 pigs)

2.30 1.10 1.20* 0.30** 2.00**

Results

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

Lameness score reduction by 87%

Results

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

Avg. weight day 0

Avg. weight day 3

Diff. day 0 to 3 (kg)

Avg. weight day 7

Diff. day 0 to 7 (kg vs.control)

Control 62.00 62.18 0.18 63.59 1.59

Tiamulin 10mg/kg (9 pigs†)

72.52 74.41 1.89** 77.45 4.93** (+3.34)

Tiamulin 15 mg/kg (10 pigs)

63.41 64.59 1.18* 67.28 3.87** (+2.28)

Results


• 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.


• 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

* p<0.001

Number of total pigs

Pigs with clinical arthritis

Untreated control 210 77 (37%)

Tiamulin 15 mg/kg bw

204 27* (13%)


• 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.


• 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.


• Farm history: A farrow to fattening farm was affected by serious polyarthritis problems, 20-30% morbidity rate and 5-10% 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 Observed day Mean Joint Diameter (inch)

Denagard 20% Injection Tiamulin generic

Right foreleg Day 1 2.73 + 0.33 2.83 + 0.23

Day 3 2.53 + 0.21a 2.82 + 0.14b

Day 5 2.56 + 0.25 2.78 + 0.20

Left foreleg Day 1 2.76 + 0.39 2.91 + 0.20

Day 3 2.70 + 0.34 2.86 + 0.14

Day 5 2.50 + 0.25a 2.86 + 0.17b

Right hind leg Day 1 3.21 + 0.56 3.40 + 0.33

Day 3 2.96 + 0.41 3.24 + 0.18

Day 5 2.90 + 0.38a 3.30 + 0.17b

Left hind leg Day 1 3.41 + 0.70 3.33 + 0.25

Day 3 3.12 + 0.59 3.20 + 0.13

Day 5 3.10 + 0.69 3.26 + 0.20

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

Average reduction in joint diameter by 9.3% after 5 days


• 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

• Actinobacillus pleuropneumoniae

• Streptococcus suis

Tiamulin parenteral pharmacodynamics in pigs – other respiratory/systemic pathogens

Denagard

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

Pharmacodynamics

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:

• 8µg/ml (sensitive), 9 - 16 µg/ml (moderately sensitive), >16 µg/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 (µg/ml) A.pleuropneumoniae strains from United Kingdom (n=15) (Aitken et al. 1999, Veterinary Record, 144, p.128)


Tiamulin 32.0 64.0 8.0-64.0

Valnemulin 16.0 32.0 4.0-32.0


Penicillin V 3.6 3.6 1.8-28.8

MICs (µg/ml) A. pleuropneumoniae strains from Hungary (n=10) (Fodor et al. 2004)


Tiamulin 2.0 4.0 2.0-4.0

Doxycycline 0.25 1.0 0.25-8.0

Lincomycin 8.0 16.0 1.0-16.0

Tylosin 16.0 32.0 24.0-32.0

Use clinical breakpoints - ResistanceTiamulin 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 (µg/ml) A.pleuropneumoniae strains from B, DK, F, Ger, NL, Pol, E, UK (n=129) (Felmingham 2009, VetPath II project)

Used CLSI method

Antimicrobial MIC 50 MIC 90 RangeResistance

(%)

Tiamulin 8.0 16.0 0.25-16 0

Tilmicosin 8.0 16.0 4-16 0

Lincomycin >32 >32 2.0-32.0 - - -

Tylosin 32.0 32.0 0.5-32.0 - - -

Tetracycline 1.0 16.0 0.25-32 15%

Amoxycillin 0.5 0.5 0.25-32 - - -

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

Tiamulin effective against APP < 8 µg/ml

0.125 0.25 0.5 1.0 2.0 4.0 8.0 16.0 32.0

B (13 isol.)

- - - - - - - - - - - - - - - - - - 13 - - -

DK (28) - - - 1 - - - - - - - - - - - - 16 11 - - -

F (18) - - - - - - - - - - - - - - - 2 15 1

Ger (21) - - - - - - 1 - - - - - - 4 13 3

NL(7) - - - - - - - - - - - - - - - - - - 7 - - -

Pol (19) - - - - - - - - - - - - 1 4 13 1 - - -

E (8) - - - - - - - - - 1 - - - 1 5 1 - - -

UK (8) - - - - - - - - - - - - - - - 1 14 - - - - - -

(VethPath collection in EU , 2006/2007)

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

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

No resistance pattern observed

Comparative susceptibility patterns of A. pleuropneumoniae to Tiamulin and Tetracycline

(VetPath II project)

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

Resistant tetracycline isolates

Susceptible tiamulin isolates

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

Use clinical breakpoints - ResistanceTiamulin breakpoints: sensitive ≤ 8.0; resistant >16.0

Antimicrobial MIC 50 MIC 90 RangeResistance

(%)

Tiamulin 8.0 16.0 0.5-64 1.7

Tilmicosin 4.0 8.0 1.0-256 1.2

Tulathromycin 8.0 16.0 1.0-32.0 - - -

Tetracycline ≤0.5 16.0 0.5-128.0 24%

Florfenicol 0.5 16.0 0.25-8 0.8%

Amoxycillin/Clavulanic acid

≤0.5 ≤2.0 0.5-0.5 0.8%

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.


• 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)


• 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

* p<0.01

Non-medicated control

Dena 3mg/lb(= 8.1 mg THF/kg bw)



Mortality (%) 16 0 * 0 * 0 *

Lung lesion score

17.2 10.2 * 10.0 * 8.4 *

Re-isolation APP (%)

75 33 * 25 * 16 *

ADF lbs d0-16 1.15 2.17 * 2.23 * 1.95 *

ADG lbs d0-16 0.23 0.77 * 0.93 * 0.99 *

F/G lbs d0-16 5.0 2.82 2.39 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 µg/ml, resistant ≥32 µg/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

• Brachyspira hyodysenteriae / Brachyspira pilosicoli

• Lawsonia intracellularis

• Clostridium perfringens / Clostridium difficile

Tiamulin parenteral pharmacodynamics in pigs – enteric pathogens

Denagard – microbiological activity

• Brachyspira hyodysenteriae / Brachyspira pilosicoli

• Lawsonia intracellularis

MICs Brachyspira spp


B. hyodysenteriae Karlsson et al, 2002

Tiamulin 0.125 1.0 ≤0.016-2.0

Lincomycin 16 64 ≤1.0-64

Tylosin >256 >256 ≤2.0->512

B. pilosicoli Kinyon et al, 2002

Tiamulin 0.125 1.0 0.06-8.0

Lincomycin 32 64 4.0->128

Tylosin >512 >512 <16->512

MICs Brachyspira spp


B. hyodysenteriae Hildago et al, 2009

Tiamulin 0.25 2.0 0.016-2.0

Lincomycin 16 128 2.0-128

Tylosin >256 >256 4.0->256

B. hyodysenteriae Magistrali et al, 2010

Tiamulin 0.063-2.0

Lincomycin 4.0-64.0

Tylosin 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.


• 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 of pigs

InfectedTiamulin

Medicated

Number with clinical signs of swine dysentery

Day 0 Day 1 Day 5 Day 12

5 - - 0 0 0 0

5 + - 5 5 1 0

5 + + 5 0 0 0

Re-isolation of B. hyodysenteriae from faeces of experimentally infected pigs (Taylor et al. 1980)

Number of pigs

InfectedTiamulin

Medicated

Number with clinical signs of swine dysentery


5 - - 0 0 0 0

5 + - 5 5 3 0

5 + + 5 1 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.


• 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


Negative control

0.66 0.66 1.83 1.45

Tiamulin 10mg/kg bw

1.50 0.12 0.16 0.08

Tiamulin 15mg/kg bw

1.18 0.40 0.09 0.00

Weight gain (in kg)

Day 1 Day 4 Day 8

Negative control 32.72 31.79 30.57

Tiamulin 10mg/kg bw

32.95 34.03 (+1.08kg) 36.07 (+3.12kg)

Tiamulin 15mg/kg bw

36.85 38.06 (+1.21kg) 40.66 (+3.81kg)

Percentage of pigs positive for B. hyodysenteriae

Day 1 Day 4 Day 8

Negative control 17 42 67

Tiamulin 10mg/kg bw

25 8 0

Tiamulin 15mg/kg bw

36 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 Chlortetracycline

• Lincomycin Tiamulin

• Tylosin 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

• Intracellular MICs• Measure effect of antimicrobial on L.i. after

it is established within the enteroctye – lipid solubility increases cell penetration

Extracellular Lumen

IntracellularEnterocytes

General methods

• Tissue culture system (McCoy cell monolayer)

• Antimicrobials tested in a range of different concentrations

(0.125 – 128 μg/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 (n=6)

EU L.intracellularis isolates (n=4)

Intracellular MIC (μg/ml)

Extracellular MIC (μg/ml)

Intracellular MIC (μg/ml)

Extracellular MIC (μg/ml)

Chlortetracycline 4-64 32-64 0.25-16 16-64

Lincomycin 16->128 >128 8-64 32->128

Tylosin 0.25-32 1->128 0.5-2 2-16

Tiamulin 0.125-0.5 1-32 0.125 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.


• 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 β2 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

C.perfringens (n=38) Denmark

C.difficile (n=15) Italy/Denmark

MIC 50 4 2 8

MIC 90 64 4 16

Geometric mean 4.6 2 8.4

MIC range 0,125-128 0.25-8 0.125-16

0

10

20

30

40

50

60

MIC (mcg/ml)

% s

train

s

Danish C. perfringens Italian C. perfringens

MIC50: 2

MIC90: 4

Geom M: 2.0

MIC50: 4

MIC90: 64

Geom M: 4.6

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

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 µg/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

Tiamulin MIC data (Streptococcus suis)

• Aitken et al. 1999• Fodor et al. 2004• VetPath 2004-2006

Pharmacodynamics

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 (µg/ml) Streptococcus suis strains from UK / Hungary

MIC 50 MIC 90 Range

Aitken et al. 1999 2.0 2.0 1.0 – 2.0

Fodor et al. 2004 0.125 0.25 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 0.25 0.5 1.0 2.0 4.0 8.0 16.0

F (19 isol.) 1 2 3 11 2 - - - - - - - - -

D (15 isol.) - - - 1 1 4 7 1 - - - - - -

NL (13 isol.) - - - - - - - - - 8 5 - - - - - - - - -

E (10 isol.) - - - 1 - - - 6 1 - - - 2 - - -

UK(9 isol.) - - - - - - - - - 5 3 1 - - - - - -

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






• Safety


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) Dose volume (ml) 10mg thf/kg bw swine dysentery , ileitis (single dose)

Dose volume (ml) 15mg thf/kg bw multiple dose for PRDC

7 0.35 0.52

10 0.5 0.75

20 1 1.5

40 2 3

50 2.5 3.75

100 5 7.5

200 10 15

250 12.5 18.75

Denagard Injection

Medication programmes used in Thailand

Recommended program: gilts

Gilt management program for PRDC and Mycoplasma spp. control

35 34 333231302928272625AGE

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

Wks

Recommended program: breeders

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

Denagard sensitive pathogens

Pregnancy wks

1 shot ofDenagard20% injection: 10-15mg/kg bw, sow at 10 wk of pregnancy

10

Farrowing

1211 151413

Pregnant

9 16

Recommended program: piglets

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

10 9876543210AGE

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

Sucking piglets Nursery piglets

Wks

Recommended program: mycoplasmal arthritis

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

10 9876543210AGE

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

Sucking piglets Nursery piglets

Wks

Recommended program: PRDC

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

2120181716151413121110AGE

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

Starter Grower

Wks

Recommended program: enteric diseases

2019181716151413121110AGE

Starter

Treatment program for swine dysentery, ileitis and colitis

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

Grower

Wks

Recommended program: APP

2019181716151413121110AGE

Starter

Mass treatment for Acute APP

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

Grower

Wks

Denagard Injection

Medication programmes used in Denmark

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

Animal Disease Dosage Treatment duration

Gilts Lameness (enter of new facilities)

1ml/15kg bw (13.3mg/kg bw) or 1ml/10kg bw (20mg/kg bw)

1-3 days or every 2nd day

Weaners /nursery pig/growers

Ileitis, Colitis, Swine Dysentery

10mg/kg bw 1-3 consecutive days

Nursery pig/growers Pleuropneumonia 15mg/kg bw 3 consecutive days

Suckling piglets Eradication Swine Dysentery, Enzootic Pneumonia

10 mg/kg bw 3-4 times during the treatment period of breeding animals

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

Animal Disease Dosage Treatment duration

Breeding animals, that do not eat and drink during eradications

Eradication Swine Dysentery, Enzootic Pneumonia

1 ml/20kg bw (10mg/kg) Day 1 and day 3

Newborn pigs Mycoplasma suis andClostridium perfringens type A diarrhoea

10 mg/kg bw Once within the first 4 days






• Safety


Denagard Injection

• Study

A study was conducted to quantify any difference between Denagard®‘s 10% Injectable formulation and currently available generic tiamulin injectable formulations

Comparison vs. generic products / formulations


• 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 oily-based 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

GenericGeneric

pH test: customers can test, themselves

Generic Tiamulin

Denagard® 8.5

2

A

B

neutral

neutral

We Take pride in our Denagard® Injection

Documents

Denagard Injection