ANIMAL nutrition Image: Scott Olson, Getty Images

SWINE DIARRHEA :
A WORLDWIDE PROBLEM
Introduction - Diarrhea is one of the most common occurrences and probably the most important challenge for swine
producers. Common causes of diarrhea are viruses (e.g., rotavirus, coronavirus), bacteria (e.g., Escherichia coli, Clostridium difficile
and Clostridium perfringens), and even protozoans. Diarrhea (i.e., disease outbreak) are often times complex and multi-factorial
in nature1. In many cases, diarrhea may be caused by multiple disease agents (e.g. co- or mixed-infections)2–4, making proper
diagnostic and treatment difficult. Other conditions such as oversupply of nutrients and poor nutrient digestibility can disrupt the
balance of gut microbiota, which in turn encourage the proliferation of pathogenic bacteria. Untreated animals often experience
high morbidity and mortality due to dehydration or complication with other diseases (e.g., secondary infections and necrotizing
lesions). This eventually leads to the spread of diseases in the herd, resulting in drop in animal growth performance and economical
loss.
Clostridial species and swine diarrhea - Diarrhea caused by enteric pathogens, especially clostridial species is common
in commercial farms. Two clostridial species namely Clostridium difficile and Clostridium perfringens are well known causative
agents of swine diarrhea and neonatal enteritis, with high isolation rate of these two bacterium from many pig farms in the Asia
Pacific regions, i.e, Australia5, Thailand6, Taiwan7, Japan8. Some studies reported as many as >50% of fecal and rectal samples
collected from animals were tested positive for either one of the two clostridial species5,8; with up to 90% of these isolates
being toxigenic strains5, indicating that the two clostridial species including their toxigenic subtypes may have high prevalence

worldwide. The most common C. perfringens serotypes isolated
from animals belong to serotype A and C4,6. C. perfringens type
C often causes high morbidity and high mortality in neonates
and suckling piglets, with syndromes ranging from bloody
diarrhea to necrotic enteritis, and sometimes even sudden
deaths. Contrary, animals infected by C. perfringens type A
usually experience moderate morbidity and lower mortality
compared to infections by C. perfringens type C. Outbreaks
of C. perfringens type C are acute and occur in piglets less
than one week old, chronic cases have been recognized with
increasing frequency in pigs up to several weeks after birth. For
C. difficile, the most common isolates in most parts of the world
are ribotype RT014 and RT0788,9. Infections by C. difficile is
often characterized by diarrhea with variable morbidity and
mortality.
The pathogenicity of these two clostridial species are
largely attributed to their capability to produce toxins which
can be detrimental to the intestinal lining. Animals, especially
neonatal and suckling piglets which lack immunity against
infections are susceptible to infections; though, infected
animals can also become asymptotic without showing
symptoms of infection10,11. Risk factors for infection of young
animals include contact through farm environments12,13 and
transmission from infected sow4, but vector of transmission
may vary from farms to farms. Sick and asymptotic but
otherwise healthy animals can shed the disease agents through
their feces, increasing transmission rate in the litter and herd14.
The implication of managing clostridial species in farms and
animals go beyond animal health and welfare as transmission
to human, i.e., through meat and direct contact is possible7.
More worrying is the fact that hypervirulent strains of C. difficile
isolated from animals and human (identical genotypes), as
well as C. perfringens which only infects animals, have become
resistant to many antibiotics6. Effective management of disease
agents such as clostridial species in farm environments and
animals is not only important to maintain healthy herd, but also
crucial to prevent the spreading of this disease agent14,9.
Probiotic for diarrhea management - Current approach for
swine producers to control diarrhea is palliative and antidotal,
i.e. the use of antibiotics at therapeutic dose. Antibiotics are also
often used in sub-therapeutic doses for prophylaxis. The use of
antibiotics at sub-therapeutic doses is not the desired approach
as it creates bacterial resistances. Other means for the prevention
of post-weaning diarrhea is through diet supplementation of
zinc oxide15; though a more recent study has highlighted that
high dosage (i.e., 2.5g/ kg feed) in diet for over two weeks
may promote the development of bacterial resistance to some
antibiotics such as tetracycline and sulfonamide16.
Another effective alternative for diarrhea management relies
on a holistic approach to biosecurity which also necessitates diet
intervention. Diet is an important aspect in maintaining gut
health as excess supply of nutrition may promote the growth of
pathogens. Precision nutrition and good quality raw materials
are important to improve digestibility and reduce wastage.
Precision nutrition requires the accurate use of proper ingredients
and nutrients for optimal animal performance. Using the right
additives such as probiotic, organic acids, essential oils and
prebiotic (Figure 1) may reduce diarrhea incidence and reliance
on antibiotics17.

34 LIVESTOCK & FEED Business / JULY 2017

 CLOSTATTM stands out to be one of the most effective
antibiotic alternatives for diarrhea management through early
diet intervention. CLOSTATTM, which is one of Kemin’s
product in the Antibiotic Alternative platform (ABA; Figure
1) contains an active probiotic Bacillus subtilis PB6, isolated
from the gut of healthy animal. Bacillus subtilis PB6 is a Gram
positive bacterium and it is capable of forming endospore - a
dormant and tough structure capable of withstanding extreme
environment. Spore-forming is an important attribute as this
allows the organism to survive the low acidic conditions in the
stomach, as well as pelleting conditions (e.g. 90°C for 5 – 10
mins) used in manufacturing pelleted feed by the feed industry.
Upon ingested by the animals and passage into the intestinal
compartment, the endospore of strain PB6 will germinate into
vegetative cells where the bacterium exerts its antagonistic
effects against potential pathogens through competitive
exclusion and production of antimicrobial peptides.
Figure 1: Kemin’s product portfolio supporting the
Antibiotic Alternative platform (ABA). Kemin provides a
series of solutions to replace Antimicrobial Growth Promoters
(AGPs) in a progressive manner. Products in the ABA
platform can be used individually or in combination to replace
AGP in animal feeds to maintain healthy herd and promote
animal performance. CLOSTATTM belongs to the category
of Probiotics and contains an active probiotic Bacillus subtilis
PB6 that is effective against many Gram positive pathogens
(e.g., Clostridium perfringens and Clostridium difficile) known to
be the causative agent of swine diarrhea. The in vivo study
presented in this article shows that supplementing diet with
CLOSTATTM can be an effective way to manage diarrhea
occurrence for nursey piglets.
Bacillus subtilis PB6 (in CLOSTATTM) is effective at
destroying C. difficile and C. perfringens. Figures 2.I and
2.II illustrate the antagonistic effect of strain PB6 against
C. perfringens isolated from pig farms in Australia. This
antagonistic effect is partly attributed to the actions of
antimicrobial peptides, also known as bacteriocins (cell free
supernatant as in Figure 2.II) produced by strain PB6. Strain
PB6 is not only effective against Clostridial species but also
other Gram positive pathogens including Staphylococcus
aureus and Streptococcus suis, although the latter was mostly
associated with causing respiratory diseases. The mechanisms
of strain PB6 in causing cell death of many Gram positive
pathogens has been deciphered through whole genome
sequencing and peptide identification using MALDI-TOF;
with evidence showing that strain PB6 can produce multiple
bacteriocins including surfactin, difficin, bacilysin, mersacidin
and macrolactin, etc. The ability of strain PB6 in producing
different types of bacteriocins is noteworthy, as each bacteriocin
works differently to cause cell lysis and death. Mersacidin,
for example, targets peptidoglycan biosynthesis pathway
and inhibits cell wall synthesis, which can be detrimental to
many Gram positive pathogens18. On the other hand, other
bacteriocins such as surfactin forms pores on the cell membrane,
leading to cystoplasm leakage and cell death. Together, these
different bacteriocins act synergistically in destabilising the cell
membrane of pathogens, thereby achieving killing, illustrated
in Figure 2. III.B for C. perfringens.
Figure 2. Antagonistic effect of Bacillus subtilis PB6 against
Clostridium perfringens isolated from pig farms in Australia,
determined using (I) streak line assay. Horizontal streak
line represents culture of strain PB6, whereas vertical line
represents culture of Clostridium perfringens. A clear zone of
inhibition was observed for C. perfringens,
indicating inhibition for growth by strain
PB6. (II) well diffusion assay using culture
supernatant of an overnight culture of
strain PB6. Clear zone of inhibition was
visible around a lawn of C. perfringens,
indicating that the secretions of PB6 contains
antimicrobial factors that are capable of
inhibiting the growth of C. perfringens.
(III) Transmission electron micrograph
(x29500): action of B. subtilis PB6 on C.
perfringens cell lysis. A: 1 hour after adding
PB6 supernatant. B: 8 hours after adding
PB6 supernatant.
Animal trial - An animal trial was
conducted at the University of the Philippines Los Baños to
investigate the interactive effects of Bacillus subtilis PB6 (in
CLOSTATTM) with and without in-feed antibiotics or zinc
oxide on the diarrhea occurrence in nursery pigs. A total of
144 weanling pigs were divided into 6 treatment groups (4
pigs/pen for 6 pens per treatment) receiving different diets in
a randomized complete block trial (Table 1). Diets with high
protein content to induce diarrheic conditions were fed from day
42 to 70 post-weaning, and diarrhea score and frequency were
collected on day 0, 7, 14, 21 and 28 post-weaning. Overall, pigs
fed with diet supplemented with zinc oxide experienced fewer
diarrhea days and frequency, and had the lowest diarrhea score
compared to piglets fed with only basal diet (Figure 3). Zinc
oxide is routinely used to prevent diarrhea (e.g., post-weaning
diarrhea) in piglets, with evidence showing improved animal
performance15. However, this benefit of zinc oxide is even
more apparent when used in combination with CLOSTATTM.
Overall, supplementation of diets with CLOSTATTM resulted
in litter having significantly lower diarrhea frequency and
diarrhea days (p-value<0.05) compared to animals fed with diet
not supplemented with CLOSTATTM.
Table 1: Details of experimental group
Experimental Diet
Group
T1 Basal feed
T2 Basal feed + 3000 ppm zinc oxide
T3 Basal feed + 500 g/MT CLOSTAT™ 11 Dry
T4 Basal feed + 3000 ppm zinc oxide + 500 g/MT CLOSTAT™ 11 Dry
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 Figure 3: Effects of treatments on diarrhea frequency
in weaning pigs. All treatments supplemented with (w)
CLOSTATTM showed significant reduction (p-value <0.05) in
diarrhea frequency compared to treatments without (w/o)

Figure 4: Effects of treatments on diarrhea day in weaning

pigs. All treatments supplemented with (w) CLOSTATTM
showed significant reduction (p-value <0.05) in diarrhea frequency
compared to treatments without (w/o) CLOSTATTM.
Conclusion - Managing diarrhea occurrence in swine
production can be challenging due to complexity of infections.
Young animals can be exposed to pathogens through contacts
with the environment or transmission from infected animals.
Antimicrobial growth promoters, i.e. antibiotic or other means
(e.g., zinc oxide) are used to control the disease in farms, but tighter
regulations requires effective alternatives to manage diarrhea
occurrence. CLOSTATTM is an obvious alternative that can be
used to prevent and reduce diarrhea occurrence in pigs, especially
for weaning and nursery piglets where growth rate at this stage is
critical. CLOSTATTM, when used at the recommended dosage,
effectively and significantly reduces diarrhea occurrence, helping
farmers to maintain healthy herd and reduce treatment cost.

- TAN BOON FEI, Ph. D and

CHAN POH SOON, DVM.,
Kemin Animal Nutrition and Health, Asia Pacific.

- THE WRITERS -

TAN BOON FEI CHAN POH SOON

36 LIVESTOCK & FEED Business / JULY 2017