Do dairy cows with mastitis show signs of sickness behavior? - and why should we care? Mette S. Herskin

Presentation Ethologist, study animal behaviour PhD: Pain sensitivity and responses toward acute stress in dairy cows (1999-2004) Special interests: pain, injury & misery Recent bovine projects Pain associated with liver biopsy sampling Behavioural changes and pain during Today: mastitis basic biology and the science behind - not recommendations regarding drugs or other therapy

Outline Disease behaviour - what is it? - why are animals changing behaviour? - what happens in the body? - where is this known from? - what about mastitis? - why should we care?

Interesting topic. Google: > 15 mio hits 300 250 Number of papers 200 150 100 50 0 1980-1981 1990-1991 2000-2001 2010-2011 Publication year

Disease behaviour Earlier: behaviour during disease = no function Sleeps Unpleasant, but banal part of being sick Tremble Has not received same focus as other types of behaviour Interest started in the 80ies concept disease behaviour No appetite Common term for behavioural changes during disease Avoid company Hygiene

Seek company Motivated to exercise Seek isolation Immobile Likes to eat at lot Do not experience hunger Disease changes motivation and needs

In nature Selection pressure from disease = large consequences for the evolution of behaviour Δ behaviour and fever = consistent response across animal groups and pathogens evolutionary biological advantage Sleeps, no appetite, trembling, isolation, hiding, immobile evolutionary survival value ( spread of diease, risk of predation) Fighting pathogen, energy to increased metabolism, heat production Well organised favored biological strategy (behaviour, physiology, metabolism), facilitates the fight against pathogens and can be crucial for survival in nature

How is this known? Experiments with mice/rats as part of studies of disease biology and the immune system - Behaviour before/after disease - Spontaneous/induced (LPS) LPS: lipopolysaccharide = parts of cell walls of pathogens symptoms and fever Activity in the immune system (hormone-like substances = cytokines)

What happens in the body? Brain Periphery Invading organisms cascade af immun. responses = acute phase response Initiating metabolic and behavioural changes Secretion of inflamm. mediators CYTOKINES Disease behaviour Fever Pain sensitivity Disease behaviour facilitates survival behaviour is a component of the immune system

What do sick rodents do? Typical behavioural changes during disease activity, exploration, (apathy) grooming/comfort dirty fur feed intake water intake social behaviour memory and learning capacity Both spontaneous and in tests, not specific for animal species or pathogen pain sensitivity sleep confusion trembling depressed mood (anhedonia)

How do they feel? Methods from psychology Aversion Conditioned taste/aversion paradigm Disease is aversive Rats receive LPS in water tasting like sugar (which they like very much) Later they will avoid sugar-taste Animal priorities and needs Operant conditioning work for access to ressources Disease social animals seek isolation Δ motivational priorities Sensitive towards the environment: nestbuilding at 24, but not at 6, where the survival of the pups are threatened Disease facilitates survival by re-organising needs ~ a motivational system, compeeding with others

What about cattle? What happens when they get sick?

Design of sickness pens Seek company Likes to exercise Seeks isolation Immobile Eats a lot No hunger Disease changes motivation and needs

Mastitis

Is mastitis painful? Huxley & Whay (2006) Kielland et al. (2007) Thomsen et al. (2012) Veterinarians Vet. students Veterinarians Farmers Toxic mastitis 7 (1-10) 7 (3-10) 9 (1-10) 9 (1-10) Clinical (clots) 3 (1-10) 5 (1-10) 2 (1-10) 3 (1-10) Yes, but even veterinarians disagree as to how painful

Research in mastitis? Publications: Web of Science (Oct. 2012): - 5399 papers on mastitis and cow - a handful about behaviour during mastitis (<5 years old) - only 23 included pain as a topic - none has integrated the two concepts Analogy: humans Pain & illness during human mastitis, especially during nursing = taken for granted

Behaviour during mastitis? Dairy cows in tie stalls Mastitis induced by injection of live E. coli or LPS in the udder High degree of control Normal behaviour + pain indicators Compare days ~ clinical signs of disease Fogsgaard et al., 2012; Siivonen et al., 2011

Fogsgaard et al. (2012): 20 dairy cows (1. lactation, 3-6 w after calving) Tie stalls, empty neighbouring stalls TMR, maize silage Milked in the home stall twice/d Controlled health status (SCC < 27,000 cells/ml)

Rectal Temperature Feed intake 40.5 35 18 Rectal Body Temperature, C 40.0 39.5 39.0 38.5 Feed intake, Kg/day 30 25 20 15 Milk yield, L/milking 16 14 12 10 8 6 38.0-12-6 036 121824 36 48 60 72 10-36 -12 12 36 60 4-36 -24-12 Time relative to infection, hours Time relative to infection, hours Time relat E. coli Count in Milk Somatic Cell Count in Milk E. coli bacterias in milk, log10cfu/ml 1e+7 1e+6 1e+5 1e+4 1e+3 1e+2 1e+1 1e+0 Milk somatic cell count, x 10 3 SCC/mL 5000 4000 3000 2000 1000 0 1e-1-6 036 1218 24 36 48 60 72-12-6 036 12 18 24 36 60 Systemic Time relative to infection, symptoms hours Time of relative mastitis to infection, hours

Eating and ruminating behaviour 20 38 % time eating 18 16 14 12 P<0.05 % time ruminating 36 34 32 30 28 26 P<0.05 10-2 -1 0 1 2 24-2 -1 0 1 2 Day after injection with E. coli in one quarter Day after injection with E. coli in one quarter Eating and ruminating reduced during the first 24h, where the clinical symptoms were clearest

Comfort behaviour and standing idle 8 45 Frequence of self-grooming 7 6 5 4 3 P<0.05 % time standing idle 40 35 30 25 P<0.05 2-2 -1 0 1 2 Day after injection with E. coli in one quarter 20-2 -1 0 1 2 Day afterinjection with E. coli in one quarter Reduced comfort behaviour and standing idle in the initial 24h

Behaviour during mastitis? Yes, dairy cows do show typical behavioural changes during disease Mastitis sickness behaviour But was there something that did not fit? Fogsgaard et al., 2012; Siivonen et al., 2011

Siivonen et al. (2011) Fogsgaard et al. (2012) N=6 N=20 Induced by LPS 20-40 cfu live E. coli Observations 2 x 24h 5 x 24h (-2 to +2) Standing idle Self-grooming Rumination Lying time (+ on inflamed side) Location of inflammation on body probably influences the behaviour Knocking out sensory input from infected quarters would help us answer Dairy cows with induced mastitis show signs of disease and pain

Behaviour during mastitis? Classic examples of sickness behaviour eating, feed intake ruminating comfort behaviour Behavioural Δ corr with local and systemic clinical signs - SCC, milk yield and rectal temperature Very low dosis adfærd og klinik normaliseret indenfor 48 h standing idle probably related to pain/soreness of the infected quarter location of infection and flooring might affect the expression of sickness behaviour Yes, cattle do show sickness behaviour

Why should we care? Basic knowledge Immunesystem Motivation Disease models Allow sickness behaviour Promote healing? Meet needs less discomfort and frustration, better welfare Optimize management Identify sick individuals Improve diagnostics Housing of the sick animals? A big potential and lots of work to do Recovery back to duty? (Danish study just started)

Challenge In order to get exploit the potentials within sickness and pain in dairy cow production we need to be able to separate the concepts at least scientifically

Effects of sickness and disease Might mix up results ~ pain / disease the lack of lying in the studies just presented Recent results about changes in pain sensitivity?

Pain sensitivity during mastitis Earlier slide: inflammatory mediators increased pain sensitivity Older studies: hyperalgesia during and after spontaneous mastitis (Fitzpatrick et al., 1998) Mild to moderate spontaneous mastitis What happened during acute clinical E. coli mastitis?

Hyperalgesi and allodyni

Thermal nociceptive stimulation Radiant heat specific for heat sensitive receptors No/very limited handling of the animals Can be applied all over the body

Description of laser Computer controlled, invisible CO 2 -laser Wave length: 10.6 m Diameter: 0.6 cm Laser pointer attached to aim Stimulation stops at preset time or by keypress Placed on trolley

Validation of laser test Latency to move leg, sec. Kicking, % 20 16 40 30 12 8 20 *** *** 10 4 0 0.0 0.6 1.2 1.8 2.4 0.0 0.6 1.2 1.8 2.4 chosen power Laser power output, Watts Valid measure of nociception with large variation Herskin et al., 2003

Pain sensitivity during mastitis A subsample of our cows were tested for thermal pain sensitivity using laser stimulation Showed reduced responses toward stimulation directed at hind legs and udder during days with clinically signs Pos correlations between rectal temperature and latency to respond Here, the disease might have blunted the responses to pain Complex interactions exist between pain and disease behaviour

Effects of sickness and pain Need to integrate studies of disease and pain and be able to separate them Sick animals + nerve block / pain relief Healthy animals + model of udder pain without disease Important for understanding of the concepts and for improvements of practice in the future

Take-home messages Behavioural changes during disease are due to a complex interaction between immunesystem and brain Disease behaviour has a function = promote survival Disease ~ motivation animal needs and priorities change

Curious? Gregory, NG. 1998. Physiological mechanisms causing sickness behaviour and suffering in diseased animals. Anim. Welf. 7: 293-305. Broom DM. 2006. Behaviour and welfare in relation to pathology. Appl. Anim. Behav. Sci. 97: 73-83. Dantzer, R. 2009. Cytokine, sickness behaviour and depression. Immunol Allergy Clin. N. Am. 29: 247-264. Thank you for the attention