CLOSTAT® Direct Fed Microbial

CLOSTAT® Active Microbial

CLOSTAT® contains a proprietary, patented strain of Bacillus subtilis, PB6. PB6 is a unique, naturally occurring, spore-forming microorganism. Kemin has identified and selected the specific strain of PB6 isolated from the gut of a healthy chicken surviving a necrotic enteritis outbreak. PB6 secretes an active substance which helps maintain the balance of microflora in the intestinal tract of livestock, poultry and equine.

PB6 has been tested by Kemin in various species of animals to show its benefits in the inhibition of pathogens. By inhibiting pathogens in the intestinal tract of livestock and poultry, PB6 can help improve gut health and overall efficiency.

In 2002, Kemin Industries isolated a Bacillus bacterium, which was identified and registered as Bacillus subtilis, PB6.1 That same year, a study was conducted that provided the Kemin R&D laboratory 13 isolates obtained from diagnostic field cases. The objective was to evaluate the inhibitory activity of PB6 on clostridial isolates from poultry via well diffusion assays.

As shown in Table 1, PB6 inhibited growth of all poultry isolates provided for analysis. The degree of inhibition is noted in clearing area (mm) and as a percentage compared to the antibiotic chloramphenicol (positive control). The degree of growth inhibition varied with each isolate.

Table 1. Effect of PB6 on growth inhibition of C. perfringens, C. septicum and C. sordellii poultry isolates. Well diameters represent single to an average of three measurements2

SOURCE ORGANISM WELL DIFFUSION
(mm)
PERCENT INHIBITORY
RESPONSE COMPARED
TO POSITIVE CONTROL
AGAR
UGA Clostridium perfringens
TCP3
23 100% TSAYE
UGA Clostridium perfringens
CP6
17 68% TSAYE
UGA Clostridium perfringens
CP2
28 112% TSAYE
UGA Clostridium perfringens
49824
15
16
63%
94%
TSAYE
Blood Agar
UGA Clostridium septicum
40592
27 90% Blood Agar
UGA Clostridium septicum
51181
26 89% Blood Agar
UMN Clostridium septicum
D07-20687
23
25
74%
97%
Blood Agar
RCM
UMN Clostridium septicum
D07-21793
25
36
86%
100%
Blood Agar
RCM
UARK Clostridium perfringens
ARK07AM
17 68% RCM
UARK Clostridium septicum
ARK124
28 90% RCM
UARK Clostridium septicum
ARK126
16 73% RCM
UARK Clostridium septicum
ARK307
16 70% RCM
UARK Clostridium sordelli
ARK0818
16 80% RCM

UGA: University of Georgia, Poultry Diagnostic Research Center
UMN: University of Minnesota, Veterinary Diagnostic Lab
UARK: University of Arkansas

Again in 2002, Kemin R&D conducted a similar study with a total of 7 isolates (C. perfringens and C. difficile) obtained from diagnostic field cases. The objective of the study was to evaluate the inhibitory activity of PB6 against clostridial isolates from swine via counter streak and/or well diffusion assays.

The objective of these assays was to evaluate the ability of PB6 to inhibit the growth of Clostridium spp. isolated from swine diagnostic cases, not to compare the sensitivity of one Clostridium spp. to another. Examples of each assay are shown in Figure 1. Clostridium perfringens inhibition is noted by the zones of inhibition (A) in a well diffusion assay and areas of clearing (B) as noted in a streak assay.

Examples of C. perfringens inhibition by PB6 as observed in both well diffusion (A) and streak (B) assays.

Figure 1. Examples of C. perfringens inhibition by PB6 as observed in both well diffusion (A) and streak (B) assays3,4

In another study, a series of microbial inhibition function assays were conducted to evaluate PB6 as compared to four commercially available, direct-fed microbial (DFM) materials (noted as A, B, C and D).

The inhibition by the bacterial cultures of the DFM Material (D) and PB6 toward the growth of Clostridium perfringens UGA49824 as tested by a double bacterial counter streak assay is presented in Figure 2. The streak assay shows that PB6 had a larger zone of inhibition than DFM Material (D) in this assay.

Zones of inhibition against growth of C. perfringens UGA49824 by a streak line assay (Material D noted as [1] and PB6 noted as [2]).

Figure 2. Zones of inhibition against growth of C. perfringens UGA49824 by a streak line assay (Material D noted as [1] and PB6 noted as [2])

Suggested Feeding Rates

  • Dairy: 13g/head/day
  • Swine and Poultry: 1 lb per ton of finished or complete feed
  • Equine: 2.5-4 lbs per ton of finished or complete feed

CLOSTAT is sold in 25 kg bags. Store in a cool, dry place out of direct sunlight for maximum shelf-life.


References
1PTA-6737ATCC. American Type Culture Collection, Manassas, Virginia.
2Kemin Internal Document, 09-00010.
3Kemin Internal Document, 09-00008.
4IMG-KANA-00101.
5Kemin Internal Document, 09-00004.
6IMG-KANA-00102.

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