Current measures do not provide sustainable control of the disease, although they are beneficial from an economic point of view,efforts to develop a more effective vaccine against swine mycoplasma have been proposed and vaccines developed using recombinant DNA technology represents a viable alternative

1. Vaccine updates for important swine Mycoplasmas
Assignment
on
DIVISION OF BACTERIOLOGY AND MYCOLOGY, IVRI
Mamta singh
Phd Scholar
Roll no. 1773

2. Swine mycoplasma
• M .hyopneomoniae-enzootic pneumonia
• M .hyosynoviae-arthritis
• M .hyorhinis- mainly commensels, rhinitis
Vaccines –M .hyosynoviae-no commercial vaccine
A vaccine specific for M.
hyosynoviae was investigated in
Denmark and induced a high level of
specific serum IgG in all animals one
week post booster (Lauritsen, K. T.
et al. 2010)
• M .hyorhinis-no commercial vaccine
• M .hyopneomoniae
commercially available vaccines
Experimental vaccines

3. Whole-cell bacterins
• major advantages of M. hyopneumoniae vaccination include improvement
of daily weight gain(2–8%) and feed conversion ratio (2–5%), shorter time
to reach slaughter weight, reduced clinical signs, lung lesions and less
treatment costs (Maes et al., 1999)
• Despite the beneficial effects, bacterins provide only partial protection,do
not prevent colonization of M. hyopneumoniae on the epithelial cells
(Thacker et al., 2000)
• vaccination is still considered the most effective practice for controlling this
infection (Mateusen et al., 2002)
• commercially available M. hyopneumoniae vaccines are made from
killed mycoplasma cultures (bacterins) but have different adjuvants
,mild like aluminum salts, whereas the oil-based ones, usually in the
form of emulsions (oil and water mix) are more reactive and may take a
longer time to be taken up, giving a more prolonged release and
stimulation (Yuki and Kiyono, 2003)
• Vaccination with inactivated, adjuvanted whole-cell bacterins (alone or in
combination with antibiotics) is frequently used worldwide to control M.
hyopneumoniae infections (Haesebrouck et al., 2004)

4. Continued….• efficacy of vaccination has been variable, may result from different factors like
infection level, age of infection, complicating factors and variability between
different isolates of M. hyopneumoniae (Calus et al., 2007; Villarreal et al., 2011)
• studies indicate that currently available commercial vaccines can reduce the
number of organisms in the respiratory tract (Meyns et al., 2006; Villarreal et al.,
2011), and consequently, the level of infection in the animals, and by extension
likely also in the herd (Sibila et al., 2007)
• vaccination significantly reduces clinical symptoms and lung lesions, only limited
reduction occurs in transmission of M. hyopneumoniae (Meyns et al., 2006;
Villarreal et al., 2011)
• current commercial vaccines are mostly based on the J strain, which was originally
isolated from a pig herd in the UK, therefore be questioned whether this strain
has similar characteristics as the M. hyopneumoniae strains circulating in pig
herds in other parts of the world (Stakenborg et al., 2005; Vranckx et al., 2011)
• maternal antibodies interference a single dose of M. hyopneumoniae bacterin to
pigs approximately 1 week of age induced long-lasting protective immunity, with
reduction in the extent of lung lesions after challenge with a virulent strain of M.
hyopneumoniae (Reynolds et al., 2009)

5. Circumvent PCV M- merck intervet
Circovirus and Mycoplasma combination swine vaccine(porcine
circovirus vaccine, type 2, mycoplasma hyopneumoniae bacterin,
adjuvanted with Microsol Diluvac Forte® Killed Baculovirus Vector)
• in healthy swine 3 weeks of age or older as an aid in the
prevention of viremia, as an aid in the reduction of virus shedding
caused by Porcine Circovirus Type 2, and as an aid in the control
of pneumonia caused by Mycoplasma hyopneumoniae.
•Dosage: 2 ml IM, repeat in 3 weeks. 21 day slaughter withdrawal
•ready-to-use combination vaccine
* Provides maximum performance and less variation in finishing
Ingelvac MycoFLEX- inglovac
Highly syringeable bacterin recommended for the
vaccination of healthy, susceptible swine 3 wks of age
or older as an aid in the reduction of
enzootic Pneumonia caused by Mycoplasma
hyopneumoniae
Dosage: 1 ml IM. Duration of immunity is at least 26
wks. Semi-annual revaccination is recommended for
breeding swine. 21 day slaughter withdrawal

6. Circumvent PCV-M G2 swine vaccine- merck intervet -for use in
healthy swine as an aid in the prevention of viremia, as an aid in the reduction
of virus shedding, as an aid in the reduction of lymphoid infection caused
by Porcine Circovirus Type 2 (PCV2), and as an aid in the reduction of lung
lesions due to Mycoplasma hyopneumoniae. Demonstrated PCV2 efficacy for
at least 20 weeks following completion of vaccination
Dosage:
Option 1 - single 2 ml dose IM for pigs 3 weeks of age or older.
Option 2 - 1 ml IM for pigs as early as 3 days of age, repeat in 3 weeks.
21 day slaughter withdrawal
• only PCV2 vaccine with one- or two-dose options that can be administered
as early as 3 days of age
* Long, strong 5-month duration of immunity lasts 25% longer than the
competition
* Ready-to-use combination vaccine for Circovirus and M.
hyopneumoniae...no mixing, combining or risk of contamination from
vaccination process
M+PAC –Intervet Schering-Plough- Recommended for use as an
aid in the prevention ofPneumonia caused by Mycoplasma
hyopneumoniae infection in swine. Dosage: 1 ml IM or subcut to
pigs 7-10 days of age, repeat in 14 days. For herds that use a single-
dose program, vaccinate pigs at 6 wks of age or older with a single
2 ml IM dose. 21 day slaughter withdrawal

7. RespiSure – zoetis -Highly antigenic whole cell
inactivated Mycoplasma hyopneumoniae bacterin.
Adjuvanted with an oil adjuvent, Amphigen to enhance
and prolong the immune response without causing
detectable tissue damage at injection site. Dosage:
Shake well and inject IM. Pigs - 2 ml at 1 wk, repeat in 2
wks; Sows - 2 ml at 6 wks and 2 wks prior to farrowing.
Revaccinate annually. 21 day slaughter withdrawal
RespiSure One – zoetis
For vaccination of healthy swine 1 day of age or older as an
aid in preventing chronic Pneumonia caused
by Mycoplasma hyopneumoniae. Dosage: 2 ml IM. Semi-
annual revaccination with a single dose of Respi Sure One
is recommended. 21 day slaughter withdrawal
* More convenient, more flexibility
* Vaccinate pigs as early as one day of age
* One-dose Mycoplasma vaccine

8. RespiSure-One ER Bac Plus – zoetis - For vaccination
of healthy swine 3 wks or older as an aid in preventing
disease caused by Erysipelas rhusiopathiae for a period of
20 wks, and Mycoplasma hyopneumoniae for a period of 23
wks
•Dosage: 2 ml IM, revaccinate with erysipelas bacterin in 3
wks. Revaccinate semi-annually. 21 day slaughter withdrawal
Suvaxyn RespiFend MH/HPS- zoetis - vaccination of
healthy swine for prevention of clinical signs, disease and
death due to porcine Polyserositis & Arthritis
disease (Glasser's disease) associated with Haemophilus
parauis serovars 4 & 5, and as an aid in the prevention of
respiratory disease associated with Mycoplasma
hyopneumoniae
•Dosage: Baby pigs - 2 ml IM at 7-10 days of age, repeat in 2-3
wks; Feeder pigs - 2 ml IM at time of arrival, repeat at 2-3
wks; Breeding stock - two 2 ml IM doses 2-3 wks apart prior
to introduction to herd. 21 day slaughter withdrawal
• Anaphylactic reactions may occur

9. New and experimental vaccines
• Whole cell vaccines also have high production costs because of the difficulty of
cultivating M. hyopneumoniae in vitro (Kobisch and Friis, 1996)
• Recombinant DNA technology can be used to overcome problems encountered
with conventional vaccines
• However, Mycoplasma sp. uses an unusual genetic code. The amino acid
tryptophan is not encoded by TGG, as in most organisms, but by TGA, which is a
stop codon (Razin et al., 1998)
• difference has hampered the expression of genes of M. hyopneumoniae
containing TGA codons in Escherichia coli, the most attractive system used for
production of recombinant proteins (Nuc and Nuc, 2006)
• Simionatto et al. (2009) described optimization of a PCR protocol for site directed
mutation of TGA codons to replace TGA codons with TGG in 14 genes from M.
hyopneumoniae, allowing the cloning and expression of these genes in E. coli

10. Recombinant subunit and recombinant vector vaccine
• adhesin P97 protein, identified as an important adhesion of M.
hyopneumoniae (Zhang et al., 1995) has been the most studied and
best defined potential protective antigen against M.
hyopneumoniae
• King et al. (1997) reported that a subunit vaccine based on
recombinant adhesin P97 did not induce significant protection in
challenged pigs, However, when the C-terminal region of P97 (R1)
was fused to Pseudomonas toxin A, immunized mice and pigs
produced specific immune responses against R1 (Chen et al., 2001)
• Oral vaccination with recombinant E. rhusiopathiae strains
expressing the P97 protein reduced the severity of pneumonic lung
lesions caused by M. hyopneumoniae infection, showing that E.
rhusiopathiae may be a promising vaccine vector for delivering
foreign antigens to the immune systems of pigs (Ogawa et al., 2009)

11. Recombinant vector vaccine
• Mucosal adjuvants and intranasal vaccine delivery have received special
attention as alternatives that may increase the mucosal immunity
• Conceicao et al. (2006) have reported that a recombinant subunit vaccine
containing the R1 subunit of P97 fused to the B subunit of thermolabile
enterotoxin of E. coli, a parenteral and mucosal adjuvant (rLTBR1) induced
systemic and mucosal antibodies against R1 in mice
• Shimoji et al. (2003) reported that a YS-19 attenuated strain of Erysipelothrix
rhusiopathiae expressing the R1 region of P97 was able to reduce lung lesions
in intranasally (IN) immunized pigs when they were challenged but havig no
humoral or cell-mediated immune response
• Okamba et al. (2007) showed that mice immunized IN with defective
adenovirus expressing the C-terminal region of P97 (rAdP97c) induced a
systemic Th1/Th2 immune response and local immunity ,However bacterin-
based commercial vaccine was more effective in inducing protective
immunity (Okamba et al., 2010)

12. Recombinant vector vaccine
• Mice immunized orally with Salmonella typhimurium
aroA CS332 harboring the R1 region of P97 (Chen et
al., 2006a) and the R2 subunit of ribonucleotide
reductase (NrdF) of M. hyopneumoniae (Chen et al.,
2006b) showed a Th1-based immune response, but
no humoral or mucosal immune responses
• However, when the NrdF R2 subunit was expressed
in S. typhimurium aroASL3261, a mucosal immune
response was observed in mice (Fagan et al., 1997)

13. DNA Vaccines
• Significant immune responses have been observed in response to
DNA vaccines based on heat shock protein P42
• beside induction of both Th1 and Th2 immune responses in mice
, antiserum from the immunized animals was found to inhibit the
growth of M. hyopneumoniae (Chen et al., 2003)
• Similar results were described using a DNA vaccine with the
antigen P46, Both Th1 and Th2 immune responses were obtained
in mice, with an increase in the IFN-g level (Galli et al., 2012)
• DNA vaccines might represent a promising strategy for
developing more effective vaccines against EP. Despite the
immunizing properties of these antigens and the reduced severity
of lung lesions, none of them is currently able to offer total
protection or a similar protection as the commercial vaccines

14. Multivalent vaccines
• Multivalent formulations , another alternative in offering better protection
against M. hyopneumoniae (Chen et al., 2008)
• Five recombinant antigens of M. hyopneumoniae (P97, P97R1, NRDF, P36,
and P46) were evaluated as DNA and protein vaccines
• analysis of mice immunized with the cocktail of antigens administered as
DNA vaccine revealed that only P97 and P36 induced IgG in the serum
• Intramuscular immunization with a cocktail of antigens (P97, P97R1, NRDF,
P36, and P46) as a DNA vaccine induced a Th1 immune response, while
antibody responses appeared to be antigen dependent
• Subcutaneous immunization with the same cocktail administered as a
recombinant subunit vaccine induced humoral and Th1 immune responses
• combination DNA and subunit vaccine was used, both humoral and Th1
responses were obtained

16. Conclusion and future perspectives
• current measures do not provide sustainable control of the
disease, although they are beneficial from an economic point
of view
• Commercial vaccines are used to limit M. hyopneumoniae
infection, however little is known about the exact
mechanisms of protection
• Efforts to develop a more effective vaccine against
mycoplasmas have been proposed and vaccines developed
using recombinant DNA technology represents a viable
alternative
• Furthermore, it is clear that investigations into the host
immune response are required in order to understand the
role it plays, both in protection and in the induction of lesion