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PRODUCTION ANIMALS
doi: 10.1111/avj.12184
ovine respiratory disease complex (BRDC) is the most signicant health problem aecting intensively nished cattle.1 It is
a multifactorial disease, with many factors contributing to its
aetiology and pathogenesis. In feedlot cattle, management and environmental factors such as pathogen exposure, stocking density, cattle
mixing, dust, transport time, nutritional changes and other stressors
*Corresponding author.
a
Animal Science, Queensland Department of Agriculture, Fisheries & Forestry, St
Lucia, Queensland, Australia
b
The University of Queensland, School of Veterinary Science, Gatton, Queensland,
Australia
c
The University of Queensland, Queensland Alliance for Agriculture and Food
Innovation, Centre for Animal Science, St Lucia, Queensland 4072, Australia;
t.mahony@uq.edu.au
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REVIEW
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Mycoplasma bovis
Mycoplasma bovis belongs to the genus Mycoplasma within the Class
Mollicute. All mycoplasma species are pathogens or commensals of
vertebrate hosts.8 They are the smallest prokaryotic cells known to be
capable of self-replication and are characterised by the absence of a
cell wall, dependence on cholesterol for growth, structural simplicity,
small genome (0.61.3 megabases) and limited metabolic
capability.911 The absence of a cell wall results in pleomorphic cell
shapes, prevents Gram-stain analysis and makes treatment with many
of the common antimicrobials, such as penicillin and other -lactams,
ineective.10
Mycoplasma bovis was rst isolated in the USA in 1961, following an
outbreak of severe mastitis in a diary cattle herd.12 Subsequently,
strains have been isolated from cattle worldwide,13 with only Norway
and New Zealand being free of infection.14 It was initially assigned the
name Mycoplasma agalactiae var. bovis because of clinical similarities
to contagious agalactia of small ruminants caused by M. agalactiae.12
It was not until 1976, following total DNADNA hybridisation experiments,15 that M. bovis was elevated to species rank and given its
current name. Today, little is known about the prevalence of this
organism in Australian cattle, despite M. bovis having a worldwide
distribution and being recognised as the most frequently occurring
pathogenic bovine mycoplasma in Europe and the USA.16
In Australia, the rst reported isolation of M. bovis in 1970 was from
a case of mastitis in a dairy cow.17 Since then, research on M. bovis in
Australia has been limited to its association with mastitis in dairy
cattle.18 The involvement of M. bovis in BRDC in Australian beef
cattle, including the feedlot sector, has not been addressed in the
scientic literature.
Epidemiology
In enzootically infected areas, M. bovis has been isolated from the
lungs of a high percentage of calves in the absence of clinical signs,
especially within the rst 34 months of age.19 Mycoplasma bovis can
also be isolated from the URT of cattle in the absence of disease,
allowing it to persist within herds without detection.20 Consequently,
the movement of infected animals between properties and countries is
thought to be the most important method of dissemination of
M. bovis.16
Animals subclinically infected with M. bovis can act as reservoirs,
shedding the organism sporadically for many months to years via
mucosal secretions from the URT or genital tract, or in milk.9,21 Shedding is thought to increase considerably at the beginning of clinical
disease among infected animals21 When exposed to mucosal secretions, the URT mucosa is the primary site of M. bovis colonisation.13
Once established where there are cattle of multiple ages, M. bovis is
dicult to eradicate.16
Historically, environmental persistence by M. bovis was not considered a major source of infection compared with animal to animal
transmission.22 The absence of a protective cell wall can render
mycoplasma species susceptible to inactivation by environmental
conditions such as temperature uctuations, ultraviolet light and desiccation.23,24 However, it has been demonstrated that M. bovis produces a biolm that provides it with a degree of protection from these
186
The gross pathological lung lesions associated with M. bovis pneumonia have been described as a caseonecrotic bronchopneumonia.40,31
Multifocal nodules of caseous necrosis within the lesions of
cranioventral bronchopneumonia have been described as unique to
the lung lesions associated with M. bovis infection.31 The circular,
raised white-yellow nodules containing dry, friable foci of caseous
material that is often seen with M. bovis-associated lung lesions diers
to the non-raised, red or pink, irregularly shaped, non-friable foci of
coagulative necrosis typical of Mann. haemolytica or H. somni.31
Experimental and clinical studies have conrmed an association
between these lesions and M. bovis infection.31,40 Lung lesions associated with M. bovis also lack the liquid purulent material seen in lung
abscesses and within lesions of chronic undierentiated bacterial
bronchopneumonia.31
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success in the control of M. bovis-associated disease has been achieved
using other antimicrobial therapies, because of the incomplete clearance of M. bovis from the respiratory tract of treated animals. The
alleviation of clinical signs without eradication of the organism is a
common result from antimicrobial studies involving commonly used
antimicrobials for the treatment of BRDC, including spectinomycin,45
tilmicosin,38 tulathromycin,53 enrooxacin and valnemulin.44
second Australian study, Fell et al.64 reported the experimental preparation of cattle (n = 209) for feedlot placement with matched industry
cattle. Again, age was not specied, but based on the information
provided it is estimated the age range of these cattle was between 390
and 450 days of age. As M. bovis has historically been considered most
important in younger cattle, this may be one reason why it has not
been considered as a contributor to cases of BRDC in Australia.
There are no studies that have been conducted to estimate the prevalence or genotypic diversity of M. bovis in Australian feedlot cattle, or
beef cattle in general. Ghadersohi et al. estimated M. bovis exposure
and subsequent mastitis among dairy cattle populations in Victoria
and northern Queensland.18,65,66 Using a conventional PCR assay it
was determined that M. bovis was present in 43% and 62% of Victorian and north Queensland dairy herds, respectively. Although that
conrms M. bovis is present and circulating in Australian dairy cattle
herds, it does not provide any insight into the situation for the Australian beef cattle population.
Recent studies using recombinant M. bovis glyceraldehyde-3phosphate dehydrogenase protein61 and M. bovis extracts, including
membrane fractions,62 as experimental vaccines reported that both
were able to elicit strong humoral and weak cell-mediated responses.
However, neither study reported protection from M. bovis infection.
The immunogen composed of the membrane fractions alone was not
as eective as the total cell extract in inducing antibody titres.62 Future
research is required to identify the type of immune response required
to protect cattle against M. bovis infection and which antigens are
required to stimulate it.
Mycoplasma bovis in Australia
There are no studies to date that have investigated the role of M. bovis
in the development of BRDC in Australian feedlot cattle, which is
surprising, given the importance placed on this pathogen in other
countries. One contributing factor could be perceived dierences in
the cattle populations arriving at feedlots in Australia and in the US.
One anecdotal example of this is the generally held view that cattle
arriving at Australian feedlots are older than those entering feedlots in
the USA. This industry view is supported by a 15-year study of 18,112
calves placed in US feedlots, where the average age of the study population was 176 days and the average placement weight was 205 kg.2
Although no comparable study has been completed in Australia to
date, Cusack et al.63 reported a study of 2,468 cattle with an average
placement weight of 340 kg. Although age was not reported in their
study, the higher placement weight is indicative of older cattle. In a
188
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Date
April 2008
July 2008
February 2009
June 2009
June 2009
September 2009
July 2010
November 2010
Feedlot
1
1
2
3
4
5
3
3
49
64
7
46
47
2
67
33
Pathogen
BVDV-1
BoHV-1
BPI3
BRSV
Mycoplasma bovis
Co-infectiona
1 (2%)
0
1 (14%)
2 (4%)
0
1 (50%)
6 (9%)
10 (10%)
5 (10%)
10 (15%)
1 (14%)
11 (24%)
24 (51%)
0
11 (16%)
8 (24%)
0
0
0
0
3 (6%)
0
0
0
0
0
0
4 (9%)
6 (13%)
1 (50%)
13 (19%)
0
42 (86%)
51 (78%)
3 (43%)
45 (98%)
40 (85%)
2 (100%)
7 (10%)
19 (58%)
6 (12%)
9 (14%)
1 (14%)
16 (35%)
29 (62%)
2 (100%)
8 (8%)
7 (21%)
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Table 1. Summary of results from the nasal swab samples collected from cattle diagnosed with bovine respiratory disease complex in Australian
feedlots
Extracts from nasal swabs were tested for the presence of bovine herpesvirus 1 (BoHV-1), bovine viral diarrhoea virus (BVDV-1), bovine respiratory
syncytial virus (BRSV), bovine parainuenza virus 3 (BPIV-3) and Mycoplasma bovis using real-time PCR. The number of positive samples is shown
for the respective pathogen with the percentage of positives shown in parentheses.
a
The number of samples testing positive for M. bovis in combination with one or more viruses, with the percentage in parentheses.
Table 2. Summary of results from the tissue samples collected from cattle diagnosed with bovine respiratory disease complex in Australian feedlots
Date
February 2009
February 2009
March 2009
September 2009
Feedlot
2
6
7
5
2
2
7
3
Tissue
Lung
Trachea
Trachea
Trachea
Pathogen
BVDV-1
BoHV-1
BPI3
BRSV
Mycoplasma bovis
Co-infection
0
0
1
0
1
2
2
3
0
0
0
0
0
0
0
0
2
2
5
3
1
2
3
3
Extracts from tissue samples were tested for the presence of bovine herpesvirus 1 (BoHV-1), bovine viral diarrhoea virus (BVDV-1), bovine
respiratory syncytial virus (BRSV), bovine parainuenza virus 3 (BPIV-3) and Mycoplasma bovis using real-time PCR. The number of positive samples
is shown for the respective pathogen with the percentage of positives shown in parentheses.
189
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support the addition of M. bovis to the list of pathogens to be considered in BRDC investigations in Australia.
Acknowledgments
The authors acknowledge the provision of clinical samples and tissues
from Drs Tony Batterham, David Frith, Kev Sullivan and Matt George.
This study was supported by grant B.FLT.0224 from Meat and
Livestock Australia, with matching funds provided by the Australian
Government.
References
1. Taylor JD, Fulton RW, Lehenbauer TW et al. The epidemiology of bovine respiratory disease: what is the evidence for predisposing factors? Can Vet J
2010;51:10951102.
2. Snowder GD, Van Vleck LD, Cundi LV et al. Bovine respiratory disease in
feedlot cattle: phenotypic, environmental, and genetic correlations with growth,
carcass, and longissimus muscle palatability traits. J Anim Sci 2007;85:18851892.
3. Edwards A. Respiratory diseases of feedlot cattle in central USA. Bovine Pract
1996:57.
4. Edwards TA. Control methods for bovine respiratory disease for feedlot cattle.
Vet Clin North Am Food Anim Pract 2010;26:273284.
5. Callan RJ, Garry FB. Biosecurity and bovine respiratory disease. Vet Clin North
Am Food Anim Pract 2002;18:5777.
6. Smith RA. Impact of disease on feedlot performance: a review. J Anim Sci
1998;76:272274.
7. Sackett D, Holmes P, Abbott K et al. Assessing the economic cost of endemic
disease on the protability of Australian beef cattle and sheep producers. Final
Report: Project AHW.087. Meat & Livestock Australia, Sydney, 2006
8. Calderon-Copete SP, Wigger G, Wunderlin C et al. The Mycoplasma conjunctivae genome sequencing, annotation and analysis. BMC Bioinformatics
2009;10:S7.
9. Pitcher DG, Nicholas RA. Mycoplasma host specicity: fact or ction? Vet J
2005;170:300306.
10. Miles RJ, Nicholas RA. Mycoplasma protocols. Humana Press, Totowa, USA, 1998.
11. Bove JM. Molecular features of mollicutes. Clin Infect Dis 1993;17(Suppl
1):S1031.
12. Hale HH, Helmboldt CF, Plastridge WN et al. Bovine mastitis caused by a
Mycoplasma species. Cornell Vet 1962;52:582591.
13. Nicholas RAJ, Ayling RD, Stipkovits LP. An experimental vaccine for calf pneumonia caused by Mycoplasma bovis: clinical, cultural, serological and pathological ndings. Vaccine 2002;20:35693575.
14. Nicholas RA. Bovine mycoplasmosis: silent and deadly. Vet Rec 2011;168:459
462.
15. Askaa G, Erno H. Elevation of Mycoplasma agalactiae subsp. bovis to species
rank: Mycoplasma bovis (Hale et al.) comb. nov. Int J Syst Bacteriol 1976;26:323
325.
16. Nicholas R, Ayling R, McAulie L. Mycoplasma diseases of ruminants. CAB
International, Oxfordshire, UK, 2008.
17. Cottew GS. Mycoplasmas isolated from cattle in Australia. Aust Vet J
1970;46:378-&.
18. Ghadersohi A, Hirst RG, Forbes-Faulkener J et al. Preliminary studies on the
prevalence of Mycoplasma bovis mastitis in dairy in cattle in Australia. Vet
Microbiol 1999;65:185194.
19. Boothby JT, Jasper DE, Zinkl JG et al. Prevalence of mycoplasmas and immune
responses to Mycoplasma bovis in feedlot calves. Am J Vet Res 1983;44:831838.
20. Maunsell FP, Woolums AR, Francoz D et al. Mycoplasma bovis infections in
cattle. J Vet Intern Med 2011;25:772783.
21. Pfutzer H. Epizootiology of the Mycoplasma bovis infection of cattle. Zentrabl
Bakteriol, 1988;20(Suppl):394399.
22. Nicholas RAJ, Ayling RD. Mycoplasma bovis: disease, diagnosis, and control.
Res Vet Sci 2003;74:105112.
23. Pfutzner H, Sachse K. Mycoplasma bovis as an agent of mastitis, pneumonia,
arthritis and genital disorders in cattle. Rev Sci Tech 1996;15:14771494.
24. Caswell JL, Archambault M. Mycoplasma bovis pneumonia in cattle. Anim
Health Res Rev 2007;8:161186.
190
25. McAulie L, Ellis RJ, Miles K et al. Biolm formation by mycoplasma species
and its role in environmental persistence and survival. Microbiology
2006;152:913922.
26. Maunsell FP, Donovan GA, Risco C et al. Field evaluation of a Mycoplasma
bovis bacterin in young dairy calves. Vaccine 2009;27:27812788.
27. Radaelli E, Luini M, Loria GR et al. Bacteriological, serological, pathological and
immunohistochemical studies of Mycoplasma bovis respiratory infection in veal
calves and adult cattle at slaughter. Res Vet Sci 2008;85:282290.
28. Robino P, Alberti A, Pittau M et al. Genetic and antigenic characterization of
the surface lipoprotein P48 of Mycoplasma bovis. Vet Microbiol 2005;109:201209.
29. Perez-Casal J, Prysliak T. Detection of antibodies against the Mycoplasma bovis
glyceraldehyde-3-phosphate dehydrogenase protein in beef cattle. Microb
Pathog 2007;43:189197.
30. Windsor D, Windsor H. Quality-control testing of Mycoplasma medium. In:
Miles R, Nicholas R, editors. Methods in molecular biology. Vol. 104: mycoplasma
protocols. Humana Press, New Jersey, 1998:6167.
31. Gagea MI, Bateman KG, Shanahan RA et al. Naturally occurring Mycoplasma
bovis-associated pneumonia and polyarthritis in feedlot beef calves. J Vet Diagn
Invest 2006;18:2940.
32. Thomas A, Ball H, Dizier I et al. Isolation of mycoplasma species from the lower
respiratory tract of healthy cattle and cattle with respiratory disease in Belgium.
Vet Rec 2002;151:472476.
33. Allen JW, Viel L, Bateman KG et al. The microbial ora of the respiratory tract
in feedlot calves: associations between nasopharyngeal and bronchoalveolar
lavage cultures. Can J Vet Res 1991;55:341346.
34. Stipkovits L, Ripley P, Varga J et al. Clinical study of the disease of calves
associated with Mycoplasma bovis infection. Acta Vet Hung 2000;48:387395.
35. Arcangioli M-A, Duet A, Meyer G et al. The role of Mycoplasma bovis in bovine
respiratory disease outbreaks in veal calf feedlots. Vet J 2008;177:8993.
36. Booker CW, Abutarbush SM, Morley PS et al. Microbiological and
histopathological ndings in cases of fatal bovine respiratory disease of feedlot
cattle in Western Canada. Can Vet J 2008;49:473481.
37. Fulton RW. Bovine respiratory disease research (19832009). Anim Health Res
Rev 2009;10:131139.
38. Gourlay RN, Thomas LH, Wyld SG. Increased severity of calf pneumonia associated with the appearance of Mycoplasma bovis in a rearing herd. Vet Rec
1989;124:420422.
39. Martin SW, Bateman KG, Shewen PE et al. A group level analysis of the associations between antibodies to seven putative pathogens and respiratory disease
and weight gain in Ontario feedlot calves. Can J Vet Res 1990;54:337342.
40. Shahriar FM, Clark EG, Janzen E et al. Coinfection with bovine viral diarrhea
virus and Mycoplasma bovis in feedlot cattle with chronic pneumonia. Can Vet J
2002;43:863868.
41. Prysliak T, van der Merwe J, Lawman Z et al. Respiratory disease caused by
Mycoplasma bovis is enhanced by exposure to bovine herpes virus 1 (BHV-1) but
not to bovine viral diarrhea virus (BVDV) type 2. Can Vet J 2011;52:11951202.
42. Houghton SB, Gourlay RN. Synergism between Mycoplasma bovis and
Pasteurella haemolytica in calf pneumonia. Vet Rec 1983;113:4142.
43. Thomas LH, Howard CJ, Stott EJ et al. Mycoplasma bovis infection in
gnotobiotic calves and combined infection with respiratory syncytial virus. Vet
Pathol 1986;23:571578.
44. Stipkovits L, Ripley PH, Tenk M et al. The ecacy of valnemulin (Econor) in the
control of disease caused by experimental infection of calves with Mycoplasma
bovis. Res Vet Sci 2005;78:207215.
45. Poumarat F, Le Grand D, Philippe S et al. Ecacy of spectinomycin against
Mycoplasma bovis induced pneumonia in conventionally reared calves. Vet
Microbiol 2001;80:2335.
46. Wawegama NK, Kanci A, Marenda MS et al. Histochemical and morphometric
characterization of broncho-pneumonia in calves caused by infection with
Mycoplasma bovis. Vet Microbiol 2012;158:220224.
47. Haines DM, Martin KM, Clark EG et al. The immunohistochemical detection of
Mycoplasma bovis and bovine viral diarrhea virus in tissues of feedlot cattle with
chronic, unresponsive respiratory disease and/or arthritis. Can Vet J 2001;42:857
860.
48. Caswell JL, Bateman KG, Cai HY et al. Mycoplasma bovis in respiratory disease
of feedlot cattle. Vet Clin North Am Food Anim Pract 2010;26:365379.
49. Bashiruddin JB, Frey J, Knigsson MH et al. Evaluation of PCR systems for the
identication and dierentiation of Mycoplasma agalactiae and Mycoplasma
bovis: a collaborative trial. Vet J 2005;169:268275.
50. Subramaniam S, Bergonier D, Poumarat F et al. Species identication of
Mycoplasma bovis and Mycoplasma agalactiae based on the uvrC genes by PCR.
Mol Cell Probes 1998;12:161169.
51. Cai HY, Bell-Rogers P, Parker L et al. Development of a real-time PCR for
detection of Mycoplasma bovis in bovine milk and lung samples. J Vet Diagn Invest
2005;17:537545.
52. Sachse K, Salam HSH, Diller R et al. Use of a novel real-time PCR technique to
monitor and quantitate Mycoplasma bovis infection in cattle herds with mastitis
and respiratory disease. Vet J 2010;186:299303.
53. Godinho KS, Rae A, Windsor GD et al. Ecacy of tulathromycin in the treatment of bovine respiratory disease associated with induced Mycoplasma bovis
infections in young dairy calves. Vet Ther 2005;6:96112.
54. Behrens A, Heller M, Kirchho H et al. A family of phase- and size-variant
membrane surface lipoprotein antigens (Vsps) of Mycoplasma bovis. Infect Immun
1994;62:50755084.
55. Brank M, Le Grand D, Poumarat F et al. Development of a recombinant
antigen for antibody-based diagnosis of Mycoplasma bovis infection in cattle. Clin
Diagn Lab Immunol 1999;6:861867.
56. Lysnyansky I, Ron Y, Yogev D. Juxtaposition of an active promoter to vsp
genes via site-specic DNA inversions generates antigenic variation in
Mycoplasma bovis. J Bacteriol 2001;183:56985708.
57. Poumarat F, Solsona M, Boldini M. Genomic, protein and antigenic variability
of Mycoplasma bovis. Vet Microbiol 1994;40:305321.
58. Le Grand D, Solsona M, Rosengarten R et al. Adaptive surface antigen variation in Mycoplasma bovis to the host immune response. FEMS Microbiol Lett
1996;144:267275.
59. Howard CJ, Gourlay RN, Taylor G. Immunity to Mycoplasma bovis infections of
the respiratory tract of calves. Res Vet Sci 1980;28:242249.
60. Chima JC, Wilkie BN, Ruhnke HL et al. Immunoprophylaxis of experimental
Mycoplasma bovis arthritis in calves: protective ecacy of live organisms and
formalinized vaccines. Vet Microbiol 1980;5:113122.
61. Prysliak T, van der Merwe J, Perez-Casal J. Vaccination with recombinant
Mycoplasma bovis GAPDH results in a strong humoral immune response but does
not protect feedlot cattle from an experimental challenge with M. bovis. Microb
Pathog 2013;55:18.
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