Challenges & Opportunities with Feed Ingredients to Maintain Gut Microbiome

Martin Clunies, Ph.D., Bruce Schumann, M.Sc. and Natalie Litvak, M.Sc.

Monogastric Nutrition Group

Health Canada – CFIA Initiatives for the Feed Industry

•  In late 2017 2018, Health Canada will be moving all “Medically Important” antimicrobials (Category I, II and III) to the prescription drug list (i.e. will require a veterinary prescription).

–  Includes all routes of administration (In-feed, In-water & Injectable) •  Category I - None •  Category II – Growth 24 – Veterinary Oversight - 52 •  Category III – Growth 37 – Veterinary Oversight - 91

•  Category IV are low importance and will continue to be able to be used without a veterinary prescription

CFIA Modernization of Feed Regulations

•  Current Table IV guidelines – Min 100 ppm & Max 500 ppm zinc •  November 2016 – Proposal for Maximum Nutrient Values for Swine

–  1000 ppm

150 ppm

Zinc-resistance gene czrC identified in methicillin-resistant Staphylococcus hyicus isolated from pigs with exudative epidermitis (Slifierz et al., 2014)

High dietary zinc supplementation increases the occurrence of tetracycline and sulfonamide resistance genes in the intestine of weaned pigs (Vahjen et al., 2015)

Modernization of the Feed Regulations

Veterinary Biologics

•  Reduce compliance burden •  Support innovation •  Ensure feeds are safe •  Contribute to production and maintenance of healthy livestock •  Safe foods of animal origin •  Do not pose a significant risk to the environment

Veterinary Biologics

•  Develop a list of Veterinary Health Products (VHP’s), e.g. probiotics, direct fed microbials

•  Work with Health Canada to differentiate products as drugs or feeds –  Compendium of Non-Feed Products Authorized for Use in

Livestock Feeds

Future Changes to the Feed Regulations

•  Discussions between the CFIA Animal Feed Division & Canadian Center for Veterinary Biologics to open up ingredients for registration that have a physiological benefit (gut modulation, increased immune function, prevents E-coli, etc.) – Current fall under a Drug classification

•  Easier approval process for the use of alternatives to antimicrobials: –  Prebiotics and Probiotics –  Enzymes –  Herbal Extracts –  Acidifiers

10 Years Experience With More Than 2.4 Million Hogs Marketed

Metrics Natures Blend Medicated1

Pigs BA/litter 14.33 11.98

#Wean/Sow/Year 29.91 26.53

Nursery Livability, % 98.5 98.5

Nursery ADG, kg/d 0.450 0.480

Nursery Feed/gain 1.58 1.65

Finisher Livability, % 98.54 97.20

Finishing ADG, kg/d 0.986 0.989

Finishing Feed/gain 2.76 2.67

1 Medication Program included, all nursery feeds medicated, Paylean in the finisher last 21+ days.

Focusing on Gut Health as a Strategy to Replace Antimicrobial Agents

•  Concerns of bacteria developing resistance to commonly used antimicrobial agents.

•  Precautionary measures to eliminate the agricultural use of certain classes of drugs important in human medicine.

•  The scientific basis for such decisions is the subject of heated debates.

Our US Neighbours Are Leading the Charge Toward Producing Livestock Without the

Use In-Feed Antibiotics.

•  The trial that got their attention was a large multi-discipline collaborative study in which they fed a combination of Chlortetracycline, Sulfamethazine and Penicillin for a period of 14 days.

–  Using genetic markers, they found that untreated pigs had a diverse population of bacteria, some of which had genes for antibiotic resistance.

–  In most cases, these bacteria had single marker genes for antibiotic resistance.

–  After only 14 day of feeding the medicated feed, there was a huge shift in the bacterial populations, a 20-100 fold increase in the population of E coli.

–  A larger proportion of the bacterial population now had genes indicating resistance to antibiotics.

–  More bacteria seemed to acquire multiple antibiotic resistant genes. –  Bacteria even acquired resistance genes for antibiotics not fed in this trial.

•  Results like these have set us on a path of what is commonly referred to as the “post-antibiotic age” of animal agriculture.

The Function of the Digestive System and Gut Microbiome.

Optimizing gastro-intestinal health and function is essential for sustained animal production systems. •  The digestion of ingested feeds and absorption of nutrients is essential

to the animals survival, growth and development. •  The digestive system contains approximately 70% of the animals

immune cells and plays a crucial role in the early development of a healthy animal.

•  The digestive system is home to a widely diverse population of bacteria in which there are over 1000 species within the gut environment.

•  Referred to as the “gut microbiome” these bacteria are crucial to the overall health of the animal as they provide signals to the animal of its environment.

Intestinal villi, are the Powerhouse of the Digestive Tract

•  The intestinal villi are the work-horse of the gut.

•  A collection of many specialized cells.

•  Protecting the function and health of the villi is the ultimate objective of good nutrition.

Healthy Villi Damaged Villi

Potential Feed Additives for Reducing Anti-Microbial Use in Pork Production

The Most Promising Technologies include: •  Acidifiers •  Probiotics •  Herbal Extracts •  Enzymes •  Omega-3 Fatty acids

In young pigs to increase stomach acidity Reduce pathogenic bacteria in the gut

The Use of Acidifiers in Swine Production Systems

Effect of Acidification of Swine Diets, Organic Acid Effect.

1.  Reducing the pH has a bacteriostatic effect – i.e. reduces further growth of bacteria.

2.  The presence of undissociated organic acids at a low pH has a bactericidal effect – i.e. killing susceptible bacteria.

Clostridium perfringens --- 6.0-7.6 8.5

Escherichia coli 4.3-4.4 6.0-8.0 9.0-10.0

Pseudomonas aeruginosa 4.4-5.6 6.6-7.0 8.9-9.0

Salmonella sp. 4.5-5.0 6.0-7.5 8.0-9.6

Staphylococcus sp. 4.2 6.8-7.5 9.3

Organism Minimum pH Optimum pH Maximum pH

Banmart, 1981.

pH Ranges For Optimum Growth of Pathogenic Bacteria

Citric Acid (%) Fumaric Acid (%) Malic Acid (%)

- - -

1.0 - -

- 0.7 -

- -

0.9

Coliforms (x 10 /gm fresh weight)

Caecum 21 13 9.6 23.0

Colon 8 6.0 1.8 7.0

Rectum 6.0 2.1 2.2 4.2

Scipioni, R. et al., (1978)

Effect of Dietary Fumaric Acid, Malic Acid & Citric Acid on the Level of Coliforms in the

Intestinal Tract of Starter Pigs

To seed the gut with beneficial bacteria Control pathogenic bacteria Stimulate immune system Optimize feed intake

The Use of Probiotics in Swine Production Systems

Effect of Probiotics in Swine

•  Reduces the amount of pathogen bacteria in the gut such as, E.coli, Salmonella sp.

•  Reduce post-weaning scours. •  Improve feed intake and daily gain of pigs. •  The probiotics allow better transition between different feeds. •  Each time you switch feeds, change formulations the microbial

population and the villi adapt.

The Function of the Digestive System and Gut Microbiome.

•  The presence of lactic acid producing bacteria are linked to the health of the gut microbiome.

•  Lactic acid bacteria attach themselves to the wall of the digestive tract.

•  Provide critical roles in communicating with the immune system of the animal.

Effect of Feeding a Bacillus sp. on Bacterial Pathogens in the Pig’s Gut.

Item Bacillus + Control - Enterobacter sp. 5.4 6.2 Clostridia sp. ND 2.3 (3/12) Bifidobacter.sp. 7.9 (5/12) ND (0/12)

Marubashi et al., 1999.

Enterobacteria include species of Escherichia coli, Salmonella, Shigella, Klebsiella, Enterobacter, Serratia, Proteus, and Yersinia

Effect of Probiotic on Piglet Performance.

Control Probiotic

# of Piglets 1,438 1,399

Initial Wt, kg 7.8 7.4

Average days 40.81 39.29

Mortality, % 1.1 0.7

ADGn, kg 0.412 0.452

Natural substitutes for antibiotics Improved enzyme function Natural palatants

The Use of Herbal Extracts® in Swine Production Systems

What Do Herbal Extracts® Do?

•  Better feed utilisation. •  Increased performance.

•  Improving appetite and activating enzymes

•  Fighting a wide range of pathogenic micro-organisms

Through

100

110

120

130

140

150

160

Control +

Control 0.50 kg/T

1.00 kg/T

1.25 kg/T

153

122 128

152

134

Effect of Herbal Extract on Nursery Pig Performance

1.6

1.65

1.7

1.75

1.8

1.85

1.9

1.95

Control +

Control 0.50 kg/T

1.00 kg/T

1.25 kg/T

1.72

1.93

1.88

1.8

1.85

ADG Feed/Gain

gm/d

ay

Herbal Extract Radford, et al.

Herbal Extract

gm/d

ay

0

10

20

30

40

50

60

Control + Control 0.50 kg/T 1.00 kg/T 1.25 kg/T

34

55

35

27 23

Effect of Herbal Extract on Nursery Pig Fecal Parameters

1.15

1.2

1.25

1.3

1.35

1.4

1.45

1.5

Control +

Control 0.50 kg/T

1.00 kg/T

1.25 kg/T

1.33

1.54

1.36 1.33

1.24

% Diarrhea Fecal score

%

Herbal Extract Radford, et al.

%

Herbal Extract

0

1

2

3

4

5

6

7

Negative control HERB EXTRACT 200 g/t

5,27a

6,34b 6.5 6.02

Lactobacillus

E. Coli

Trial # 1589

Source: Manzanilla et al. Department of Animal Nutrition and Production, Autonomous University of Barcelona, Spain 2000

Confirmation of the Effect of Herbal Extract on Lactobacillus in Piglet

Variation in number of colony forming units (CFU), expressed in Log10 CFU/g of fresh material (jeujunum), in piglet in age of 40 days, in a post weaning experiment of 18 days.

Log10 CFU/g

Supplement to endogenous enzyme secretion Improved nutrient utilization Improved gut health

The Use of Enzymes in Swine Production Systems

Effect of Galactosidase/Invertase Cocktail on % Improved Nutrient Utilization

•  Source of Galactosidase and invertase.

•  Targets Soybean meal oligosaccharides.

•  Increases ME and dig of CP of Soybean meal.

-4

-2

0

2

4

6

8

10

ME

Dig CP

FCR

% Utilization improvement

Effect of Enzyme on Nursery Pig Performance

Control Enzyme

Initial wt, kg 6.7 6.7

Final wt, kg 19.6 20.5

ADGn, kg/d 0.368 0.394

Feed:gain 1.43 1.49

Feed costs,$/kg 0.394 0.350

Supplement high energy to young piglets Omega-3 FAs improve health status Omega-3 increased nutrient use Improved Biosecurity

The Use of Vegetable Oils in Swine Production Systems

The Effect of Omega-3 FA on Protein Deposition of Piglets

Measurement 10:1 O-6:O-3 FA

5:1 O-6:O-3 FA

Protein deposition, g/d 61.3 87.8

Protein deposition, g/kg gain 155.9 222.2

Eastwood and Beaulieu, 2014

Effect of Omega-3 Level on Performance of Market Hogs

Level 1 Level 2 Level 3 Level 4

Start wt 27.2 27.2 27.3 27.2

ADGn 0.908 0.943 0.933 .945

ADFInt 2.50 2.57 2.53 2.55

Fd/Gain 2.76 2.73 2.73 2.72

Carc.Index 105.7 107.8 107.4 107.2

All diets contained; Corn 740 kg, SBl 200 kg Bionic Hog Px 40 kg and 20 kg Bionic Oil to Market – Ridgetown Agriculture College 1995.

Combines proven feed technologies Manipulates the gut microbiome Improves nutrient utilization

Natures Blend Feeds & Premixes

Natures Blend Starters & Premixes

Producing Pork & Poultry Without Anti-microbial Agents

•  The production of pork without the use of antimicrobial agents has been a challenge in most intense production systems.

•  These use of single technology approaches have met with very limited success.

•  Multi-factorial strategies have been much more successful in most swine & poultry operations.

•  Natures Blend Program has been successfully producing pork, chicken and turkey products for the better part of 10 years, with results comparable to more traditional programs.

The Strategy of the Natures Blend Feed Program is a Healthy Gut Produces a Healthy

Pig or Chicken.