FOR RUMINANTS

kemin.com/guthealth

What is CLOSTAT®?CLOSTAT contains a proprietary, patented strain of Bacillus subtilis PB6. PB6 is a unique, naturally occurring,

spore-forming microorganism. Kemin has identified and selected PB6 as an active substance that helps maintain

the balance of microflora in the intestinal tract of livestock (Figure 1).

Figure 1. Bacillus subtilis PB6 in the intestine.

Features

• Contains PB6, a unique, patented strain of Bacillus subtilis

• Stable during processing and packaging

• Stable when blended with other feed ingredients

• Stable under normal commercial pelleting conditions

• Compatible with antibiotics, coccidiostats and organic acids commonly used in the animal feed industry

• Proven efficacy in research

Stability • Mineral premix concentrate (oxides, sulfates) for three weeks

• Long term - temperatures ranging from 5°C (41°F) to 65°C (149°F) for one year

• Short term - 194°F (90°C) for 10 minutes

• Resistant to acid conditions of pH 2.0 for 90 minutes

• Stable during processing and packaging, when blended with other feed ingredients, and under normal

commercial pelleting conditions

Mode of ActionBacillus subtilis PB6 secretes active substances that are inhibitory toward various strains of pathogens including Clostridium,

Salmonella and E.coli spp1 (Figure 2).

Zone of Inhibition and Bacillus subtilis PB6A zone of inhibition test examines bacteria sensitivity and resistance

to certain compounds. The size of the zone surrounding the compound

disk on the plate is an indication of microbial susceptibility to the

compound (Figure 3).2

Enteric Disease PathogensThere are many different pathogens causing intestinal health issues in ruminants. The most notable are E.coli, Salmonella spp,

Clostridial spp, Cryptosporidium, Rotavirus and Coronavirus (Table 1). Of particular interest are E.coli, Salmonella and

Clostridial bacteria. Each of these types of bacteria, while different in many aspects, can similarly target the intestinal tract, cause

death by the systemic effects of toxins and show up clinically as bloody diarrhea.4

Pathogen Epidemiology

E. Coli Attaches to intestinal wall and secretes toxins that disrupt normal gut function.5

Salmonella spp.

Invades the mucosa of the small and large intestine and produces toxins. Invasion of epithelial cells

stimulates the release of pro-inflammatory cytokines, which induce an inflammatory reaction. The acute

inflammatory response causes diarrhea and may lead to ulceration and destruction of the mucosa.6

Clostridia spp.Secretes toxins that destroy intestinal lining and increase permeability of blood vessels. Overgrows in

environments rich in protein, readily available energy.7

Cryptosporidium parvum Causes changes in intestinal mucosa, including partial villous atrophy, crypt lengthening and inflammation.8

Rotavirus and CoronavirusInfection of the small intestinal mucosa leads to loss of surface area and malabsorption of nutrients by

intestinal villi.9

Figure 2. Pathogen cell wall disruption.

Figure 3. Bacillus subtilis PB6 and Clostridium

perfringens inhibition.3

Table 1. Enteric disease pathogens.

Bacillus subtilis PB6 ResearchResearch conducted with Bacillus subtilis PB6 has demonstrated antagonistic activity against a wide range of pathogens

(Table 2) (Figure 4).10

Bacterial Strains Inhibition Zone Diameter (mm)

Clostridium perfringens ATCC13124 17.79 ± 0.44

Escherichia coli CVCC1555 13.40 ± 0.14

Salmonella typhimurium ATCC14028 13.49 ± 0.11

Staphylococcus aureus CVCC1885 13.78 ± 0.26

Listeria monocytogenes CMCC54002 14.34 ± 0.76

Vibrio alginnolyficus 1609 13.46 ± 0.43

Table 2. Characteristics of Bacillus subtilis PB6.10

It is important to note Salmonella is rarely considered part of the normal flora of the intestinal tract.7 Asymptomatic cattle can shed

Salmonella for prolonged periods after infection and, in the case of Salmonella Dublin infection, some cattle may be intermittent

shedders for life.7 Clostridial organisms are normally found in the digestive tract of cattle and only become problematic with dietary

stress, injury, management changes, parasitism or other stress related events.11 Rapid growth of C. perfringens results in the

production of toxins, which can lead to intestinal tract disorders both in young and mature ruminants.

Figure 4. Characteristics of Bacillus subtilis PB6 on A) Clostridium perfringens ATCC13124, B) Escherichia

CVCC1555, C) Salmonella typhimurium ATCC14028.12

References

1. Yeow-Lim Teo, Alex., Tan, Hai-Meng. Applied and Environmental Microbiology, Aug. 2005, p.4185-4190.

2. Todar, K., Bacterial Resistance to Antibiotics. Todar’s Online Textbook of Bacteriology. Accessed 5-6-16.

3. Kemin Internal Image IMG-KANA-00211.

4. McGuirk, Sheila M., Dealing with Salmonella and Clostridium Problems on our Dairies. University of Wisconsin, School of Veterinary Medicine.

5. Enterohemorrhagic E. coli (EHEC) pathogenesis. Front Cell Infect Microbiol. 2012; 2: 90. Published online 2012 Jul 12. Prepublished online 2012 May 30. doi: 10.3389/fcimb.2012.00090.

Y Nguyen1 and Vanessa Sperandio.

6. Ralph A. Giannella. Chapter 21 Salmonella. Medical Microbiology, 4th edition. 1996.

7. McGuirk, Sheila M., Managing Clostridial Diseases in Cattle. University of Wisconsin, School of Veterinary Medicine.

8. Ernest A. Meyer. Chapter 80 Other Intestinal Protozoa and Trichomonas Vaginalis. Medical Microbiology, 4th edition. 1996.

9. Rotaviral and coronaviral diarrhea. Vet Clin North Am Food Anim Pract. 1985 Nov;1(3):471-93. Torres-Medina A, Schlafer DH, Mebus CA.

10. Kemin Internal Document, 16-00084.

11. Songer, J. Glenn, Clostridial Enteric Diseases of Domestic Animals., Clinical Microbiology Reviews, Apr. 1996, p. 216-234.

12. Kemin Internal Image used with Permission.

13. P. R. Broadway, CLOSTAT® Reduces the Negative Impacts of a Salmonella Challenge in Weaned Holstein Steers. J. Anim. Sci. Volume 95, Supplement 1, 2017. This material is based upon work

supported by the U.S. Department of Agriculture, Bioscience Research Lab, Agricultural Research Service, Lubbock, TX under agreement No. KANA6344-16. Any opinions, findings, conclusion,

or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture.

14. Kluger, M.J. and B.A. Rothenburg. 1979. Fever and reduced iron: their interaction as a host defense response to bacterial infection. Science 203(4378):374–376.

15. Patent U.S.7,247,299.

A B C

Reducing the Negative Impact of Pathogenic Salmonella Typhimurium in Weaned Holstein SteersA research trial conducted with the United States Department of Agriculture at the Livestock Issues Research Unit in Lubbock, TX evaluated

the potential for CLOSTAT, a patented Bacillus subtilis active microbial, to reduce the severity of salmonellosis in weaned Holstein steers

challenged with Salmonella Typhimurium.13 Calves were fed either control diets (no CLOSTAT) or 13 g/h/d CLOSTAT in a starter ration for

35 days. Calves were then assigned to one of four treatments, consisting of CLOSTAT or no CLOSTAT and Salmonella (1.6 x 106 Salmonella

Typhimurium) or no Salmonella. The CLOSTAT calves displayed decreased rectal temperatures (P< 0.001) after the study compared to the

control calves challenged with Salmonella.

Mounting an immune response to a pathogen challenge requires a significant amount of energy. It has been estimated that an increase

in core body temperature by 1oC requires an increase of 10-13 percent in an animal’s metabolic rate.14 Mediating this change in body

temperature would potentially spare glucose, allowing energy to be put towards other productive functions.

Summary• CLOSTAT® is an active microbial containing a proprietary, patented strain of Bacillus subtilis PB6.

• Over 200 internal R&D documents referencing PB6 research trial work.

• North America

• China

• Europe

• India

• SE Asia

• In vitro analysis evaluating PB6 efficacy against multiple strains within bacterial organisms.15

• Commercial animal research trials demonstrating consistent response.

• Gut health is a highly complex system including the structural integrity of the intestine, the balance of microflora and the status of the

immune system. Kemin offers a comprehensive platform of leading solutions impacting key areas of gut health.

Figure 5. Effect of CLOSTAT on rectal temperatures of weaned Holstein steers challenged with

Salmonella Typhimurium.

Time Relative to Salmonella Challege, h

Recta

l Tem

pera

ture

, °C

NoSal Control

Sal Control

NoSal CLOSTAT

Sal CLOSTAT

BR-2017-00001© Kemin Industries, Inc. and its group of companies 2017. All rights reserved. ® ™ Trademarks of Kemin Industries, Inc., U.S.A.

Comprehensive approach to gut health by strengthening intestinal integrity, microbial

balance and immune function.

Integrity: Maintaining the strength of the intestinal barrier

Protection: Supporting immune function

Balance: Diversifying the population of microbes

Gut health is a highly complex system including the structural integrity of the

intestine, the balance of microflora and the status of the immune system. Kemin

offers a comprehensive platform of industry leading solutions for livestock, poultry

and equine.

Kemin offers a range of nutritional solutions for raising healthy animals. We

understand your need to raise healthy livestock that gives consumers the nutritional

and health benefits they are looking for, while also returning a profit. Our products

and services help you with:

• Nutrition

• Feed Quality

• Gut Health

• Pathogen Control

KEMIN.COM/GUTHEALTH1-800-752-2864