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Mycoplasmas, Ureaplasmas & Haemoplasmas

Illustration by David S. Goodsell, the Scripps Research Institute

Week 4, Lecture 20: PJL

Mycoplasma Group: Overview Facultative anaerobes, stain poorly

Tiny cell (0.3 – 0.8 um)

Trilaminar plasma membrane Lipoprotein & phospholipid

No peptidoglycan layer β-lactams – Penicillins & Cephalosporins

highly pleomorphic

Closely related to Streptococcus Cannot synthesize cholesterol

Infections are varied: Respiratory (Upper & Lower Resp. Tract)

Mastitis

Arthritis, Chondropathy, Otitis media

Urogenital tract infections

Week 4, Lecture 20: PJL

Transmission Electron Microscope

Scanning Electron Microscope

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Mycoplasma Group: Virulence

Highly host-adapted: prolonged carrier states Epi/Intracellular association

Insidious infections – chronicity

No LPS-endotoxin: no local or systemic signs

Strain virulence variation

Virulence Factors Antigenic variation

Biofilms (in-vivo & in-vitro)

H2O2 – ciliotoxic and hemolytic

Adhesins – varied non-fimbrial types

Capsules

Urease - ureaplasmas , IgA protease

Superantigens – T-cell and antigen presenting cell activation Week 4, Lecture 20: PJL

M. gallisepticum

- tip organelle

- trachea

Mycoplasmas: Culture-based Dx Culture followed by specific Ab-based and/or PCR based identification Gold-Standard but ….. Time ……. Weeks for some

Overgrowth or pre-sample Abx affect culture outcome

Dx samples: resp. tract (TTA, lung tissue), joint fluid, milk, conjunctiva

Week 4, Lecture 20: PJL

Pure

Culture

Mycoplasma

“fried-egg”

microcolonies

Digitonin Sensitivity Assay

DSA

Broth

Amplification

Digitonin binds membrane cholesterol Mycoplasma’s cannot synthesize – cell death

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Mycoplasma Group: Rapid Dx

Week 4, Lecture 20: PJL

PCR Serology - cheaper

Multiplex PCR

Microarray coming

Pathogenic Mycoplasmas Bovine

M. bovis

M. mycoides subsp. mycoides (SCV)– Contagious Bovine Pleuropneumonia (CBPP) Reportable

Porcine

M. hyopneumoniae

M. hyorhinis – polyserositis & arthritis in 3-10 wk old pigs (sporadic)

M. hyosynovitis – polyarthritis in 12-24 wk pigs (sporadic)

Avian

M. gallisepticum

M. synoviae – Infectious synovitis (chickens & turkeys); air-sacculitis, anemia

M. meleagridis – air-sacculitis & osteodystrophy (turkeys)

M. iowae – reduced hatchability, embryo mortality (turkeys)

Laboratory Animal Infections

M. pulmonis – mouse/rat pneumonia

M. neurolyticum – conjuctivitis & “Rolling Disease” in mice (neurotoxin)

Miscellaneous & Haemoplasmas

Global

Most relevant but: - sub-Saharan Africa, Asia

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Evolution of Mycoplasma bovis

ACVIM Consensus Statement: M. bovis Infections in Cattle. J Vet Intern Med 2011: 25-772

1961 USA

1964 Israel

1970 Australia

1974 France, UK

1989 Brazil

2005 S. Africa

2008 China

Mb-free N.Z & Norway

Pathogenesis

Dairy Beef

Otitis

Media?

Mastitis Herd

Waste

Milk

Calves/Heifers

Colonization

Mastitis

Pneumonia

BRDC Pneumonia

Feedlot

2%-to-100%

Proximity &

repeated contact

30-60%

UK

30-40%

Italy

60-80%

CPPS – tenosynovitis/arthritis

Dairy: 7-20%

Bacteremia

Dissemination

3-4 weeks

after arrival

Otitis, arthritis

Arthritis

Nicholas, R. (2011) Bovine mycoplasmosis: silent and deadly. Vet. Record 168:459.

Mycoplasma bovis ~ $150 million/yr (US) – dairy & beef

Caseonecrotic bronchopneumonia

Cranioventral lobes

Subclinical–to-clinical insidious

Acute: multifactorial (BVDV, Mh/Pm/Hs or A. pyogenes

Chronic Pneumonia & Polyarthritis Syndrome

Dx – serology, PCR (TTA, conjunctiva), culture

Intervention: Management – Mb-free stockers, stress (shipping, cold)

Vaccine (Pulmo-GuardJ MpB)

Not in Canada

Metaphylaxis - Draxxin

Quarantine

Label use Abx: Tulathromycin (Draxxin)

Gamithromycin (Zactran)

Week 4, Lecture 20: PJL

Nicholas, R. (2011) Vet. Record, 168:459.

Otitis media

Eustachian tube

Insidious

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Mycoplasma bovis - Mastitis

Variable presentations: Hx of Abx failure Severe mastitis outbreaks “contagious agalactia”

Endemic subclinical (+/- changes in SCC)

Mastitis Signs: Hx of Abx failure Multiple quarters, not painful, mild systemic signs

Milk changes

Agalactia can resolve (often permanent)

Transmission Milk, resp. secretions, frozen semen

Infusion cannula misuse, damp sponges

Calves - pneumonia, arthritis, otitis media

Week 4, Lecture 20: PJL

Mycoplasma bovis - Mastitis Dx - Abx unrewarding, agalactia, no fever Milk culture (Hayflick’s) – PCR rapid, available, technical challenges

Intervention: Herd-size is risk factor

Eliminating from animal &/or herd challenging Clinical signs can resolve but : chronic shedders

Dairy Biosecurity – Closed-herd concept Bulk tank hx of farm of origin pathogen free stock-sources Quarantine and check

ACVIM- Treatment … not recommended. Confirm i.d., quarantine, cull(?)

No vaccines in Canada

Management:

Bulk tank monitoring (culture, ELISA, PCR)

Week 4, Lecture 20: PJL

VIDO - GAPDH vaccine research - Glycolytic pathway - Also adhesin

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M. hyopneumoniae – “Enzootic Pneumonia” of Swine

Growing/finishing pigs (25-100 kg) Chronic catarrhal/suppurative bronchopneumonia High morbidity (direct & aerosol) Low mortality

Risk Factors Stress: stocking density, ammonia levels Co-infections – PRRSV, Pm, H. parasuis, APP, Bb

Signs – Subclinical-to-Clinical

Chronic nonproductive cough “cough-score” reduced ADG and FCE anorexia, pyrexia, dyspnea (co-pathogens)

Post mortem: cranial lung lobes Histo - bronchiolar lymphoid infiltration/hyperplasia

Global

M. hyopneumoniae: Dx & Intervention

Antemortem - Herd ELISA, PCR (nasal swab)

Post mortem Culture – (Friis), most definitive but 4-8 weeks

Faster - Immunohistochemistry on lung tissue, PCR

Difficult to eliminate Parenteral - Tulathromycin (Draxxin®)- label-use

Feed – Prescription mix - Lincomycin

Management Strategies: Biosecurity

Herd surveillance program, restock with SPF

Stocking density, ammonia levels,

Vaccines (sows/piglets): RespiSure-One® Co-pathogens vaccine status as well

CFIA (FYI) - Compendium of Medicating Ingredient Brochures - Summary of Feed Drug Clearances

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Severe fibrinous pericarditis in a pig also affected with enzootic pneumonia.

et al. Veterinary Record 2008;162:537-540

Copyright © British Veterinary Association. All rights reserved.

M. gallisepticum

Chronic Respiratory Disease (CRD) in chickens

Infectious Sinusitis (IS) in turkeys

Habitat Colonization of resp./repro. mucosa

Lateral and vertical (Eggs/semen) transmission

Signs (good hygiene equals milder signs) Chickens - nasal discharge, tracheal

rales, coughing sneezing

Turkeys - swelling of infraorbital sinuses, mild conjunctivitis

’d ADG/FCE and egg production

Mortality low but can approach 30%

If co-infections (viruses, or E. coli )

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M. gallisepticum : Dx and Txt

Culture – respiratory tract (Frey’s Media)

Rapid tests – Serology (flock surveillance) Serum Plate Agglutination (SPA) test (Intervet) cheap

mix commercial Antigen (MG, MS, MM) with 0.2 mL serum agglutination = positive

PCR- readily available (PDS) Swabs or broth culture i.d.

Intervention- goal is prevention

MG-free breeder flocks – Egg dipping/heating Biosecurity, (all-in, all-out) Live vaccine available – MycoVAC-L (intervet) ocular instillation Abx in Feed: Tetracycline or Erythromycin

Misc. Non-haemotrophic Mycoplasmas (FYI - except one)

Bovine: many species (mastitis, conjunctivitis, resp./repro. tract infections)

M. bovoculi - keratoconjunctivitis M. dispar – calf pneumonia Ureaplasma diversum – role in vulvitis, infertility

Caprine & Ovine: many species (first two are Notifable)

M. capricolum subsp. capripneumoniae – Contagious Caprine Pleuropneumonia M. agalactiae – contagious agalactia M. capricolum subsp. capricolum – septicemia, mastitis, arthritis & pneumonia M. mycoides subsp. capri - septicemia, pleuropneumonia, arthritis & mastitis M. ovipneumoniae – small ruminant pneumonia

Canine

M. cynos – part of kennel cough

Feline

M. felis – conjunctivitis, foals (pleuritis) & humans (immuncomp) M. gateae - arthritis

Z

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Haemotrophic Mycoplasmas

Obligate RBC parasites (prev. Order Rickettsiales ; Genera Haemobartonella & Eperythrozoon)

Insect vectors (mosquito, tick, flea), direct transmission, transfusion

Cannot culture on artificial media

Subclinical to clinical hemolytic anemias, ill-thrift, infertility

~ 9 species described to date

Haemobartonella felis → M. haemofelis (Feline Infectious Anemia)

Haemobartonella canis → M. haemocanis (canine haemobartonellosis)

Eperythrozoon suis → M. suis (porcine icteroanemia)

Eperythrozoon ovis → M. ovis (Ovine/Caprine eperythrozoonosis)

Eperythrozoon wenyonii → M. wenyonii (Bovine eperythrozoonosis)

Several others that remain poorly characterized

Haemoplasmas

Feline Infectious Anemia

Mycoplasma haemofelis

Prevalence (0-50%) significance ?

Signs: pale m.m., lethargy, anorexia, depression, dehydration, pyrexia – can relapse/recur

Infection route – fleas, fights blood transfusion donors

Diagnosis: anemia Blood smear

PCR-based (commercial & routine Dx labs)

Doxycycline, Enrofloxacin Blood smear: Wright-Giemsa

Global