Effectiveness of Cikrón V to prevent postpartum metritis in sows

2. Abstract
3. Introduction
4. Hypothesis
5. Objectives
6. Literature
   review
7. Materials and
   methods
8. Results and
   discussion
9. Conclusion
11. References

Abstract.

In Cuba, the
prevalence of postpartum metritis in sows is high. This illness
is associated fundamentally with mastitis and
agalactia, as a well-known syndrome called
Metritis-Mastitis-Agalactia (MMA). This entity causes big
economic losses to swine breeders, since it influences directly
on the reproductive and productive behavior of the animal,
reducing litter size and vigor. In this respect, one management
procedure that helps to compensate for the damage is to regroup
piglets, positioning them in litters living in a better
physiological state. However, this solution is not 100% percent
effective for it does not target PM incidence clinically, nor it
reduces it down to desirable levels.

For this reason, other solutions to this serious health
problem are currently under study. At the moment, research is
being carried out by Cuban institutions to produce a new
generation of drugs with medicinal plants to cure different
animal and human illnesses of this kind. So far, results
compensate for the deficit of generic drugs, unavailable under
our present economic conditions, so that new drugs of natural
origin, as Cikrón V, are being extended. In this respect,
the present research is intended to prove the effectiveness of
this drug for preventing and curing PM in sows.

Key words: Cikrón V, postpartum metritis
(PM) in sows, Metritis-Mastitis-Agalactia (MMA).

Introduction.

In Cuba, the prevalence of postpartum metritis in sows
is high. This illness is associated fundamentally with mastitis
and agalactia, as a well-known syndrome called
Metritis-Mastitis-Agalactia (MMA). This entity causes big
economic losses to swine breeders, since it influences directly
on the reproductive and productive behavior of the animal,
specially when litters cannot be fed properly.

Antiseptic and healing drugs for veterinary use have
been among the least available veterinary drugs in Cuba.
Rhizophora mangrove L constitutes a natural antiseptic, known for
its ethno-botanical uses. It is colloquially known as Red Mangrove, whose bark
yields an extract obtained for veterinary purpose at the National
Center of Plant and Animal Health. The extract is produced as
Cikrón V for veterinary commercial use.

Rhizophora mangrove L. grows on the Cuban coastline, as
well as on the borders of channels and lagoons. It is one of the
traditionally healing plants, to which healing properties are
conferred. In Cuba, it is reported as astringent (Roig, 1974).
Red Mangrove bark liquid extract is known to stop hemoptisis. It
is used against asthma and breathing disorders, while the water
extracted from the shell is used to avoid some animal
plagues.

Derivatives of this plant are also used against angina
and illnesses of the throat (Friedman, 1984). The concoction of
its leaves is described to be effective against intoxication by
polluted fish ulcers. Extracts obtained from the bark are also
used to mitigate breathing and digestive ailments, venereal
illnesses and infections on the skin, as well as the leprosy
(Cáceres and col., 1993).

Omen, 1998, outlines that the treatment of both
endometritis and metritis is very wide and varied. Apart from
antibiotics, sulfonamides, hormones, antiseptic and disinfectant
solutions have also been used, but because of their residual
effect in the food as well as their secondary effects, especially
those associated with bioresistance in animals and man, they all
bring about, in one way or another, a lot of all
inconveniences.

At the moment, research is being carried out by Cuban
institutions to produce a new generation of drugs with medicinal
plants to cure different animal and human illnesses of this kind.
So far, results compensate for the deficit of generic drugs,
unavailable under our present economic conditions, so that new
drug of natural origin, such as Cikrón V, are being
extended. In this respect, the present research is intended to
prove the effectiveness of this drug for preventing and curing PM
in sows.

Hypothesis.

• The application of Cikrón-V favorably
  influences on the prevention of PM in sows.

General
objective.

• To evaluate the effectiveness of a drug of natural
  origin, Cikrón V, based on Rhizophora mangrove L., Red
  mangrove, extracts for preventing PM in sows.

Specific objectives.

• To determine the prevalence of the illness in the
  herd under study.
• To determine the main microorganisms, causing the
  illness.
• To identify the antimicrobial activity of
  Cikrón V.
• To evaluate the effectiveness of Cikrón-V for
  preventing PM.

Literature
review.

Among the pharmaceutical preparations used in the
prevention and treatment of the illness are antibiotics. The
literature outlines that good results have been obtained with
Neometrina vaginally, applying 40ml of the solution, preferably
by washing the infected area with a lukewarm saline
solution.

Studies carried out with Rhizophora mangrove L watery
and alcoholic roots and shafts extracts demonstrate their
inhibitory activity against both Gram positive and Gram negative
bacteria of human interest. (Red and Limit, 1978). Rhizophora
mangrove L. is reported among the American plants killing fungi.
Sánchez, 1998, outlines that the healing properties
conferred to this plant are due to their composition in tannins
and some compounds.

The essential volatile and compound oils, or
semi-volatiles oils that are also present in Rhizophora mangrove
L. watery bark extract are used for antimicrobial purposes either
against gram-positive and gram-negative bacteria or mushrooms
(Hussein, 1990) ( Mount and col., 1991) (Alade and Irobi, 1993)
(Chinou and col., 1996).

The National Center of Plant and Animal Health has tried
different extracts from the bark of this plant. Among its
aromatic components, appearing in high quantities, there are some
that can be linked to having anti-rust effects (Sánchez
and col., 1998). It was demonstrated that Cikrón V is able
to stimulate healing wounds in rabbits, Figueroa and col. (1995),
and to prevent the umbilical infections in calves (Figueredo and
cabbage., 1995).

According to a study by Figueroa and col. (1995), the
solution was not found to cause irritability, but to act as a
protecting film that accelerates healing in a 100% of the cases
studied, causing no inflammation either and diminishing the
number of ulcers.

Other significant evaluations were carried out by
(García and col., 1998) (Bulnes and col., 2000). By means
of the test of
irritability, carried out after having used Rhizophora mangrove
extract, no hystopathological alterations were
observed.

Materials and
methods.

The present research was carried out in two MINAZ' swine
husbandry companies, located in the municipality of Martí,
north of Matanzas. To determine the prevalence of the illness,
sows were studied in two groups. Group one, 140 sows, was located
in the TO breeding center, while group two, 92 sows, was located
in the center identified as B. Prevalence was calculated by means
of the following equation:

Prevalence = Number of sick animals x
100

Total of potential animals.

Diagnosing the main microorganisms associated with the
illness included a collection of vaginal fluids from ten sows
(n=10), performed endocervically for bacteriology tests.
Collection was made by ahering samples to a thick cotton
collector.

Samples were examined under the microscope to observe
microorganism forms, grouping them accordingly. Cultivation in an
Agar medium was carried out and incubation was performed at
37ºC during 24 hours, to identify the present
microorganisms. Cikrón V antimicrobial activity was
evaluated.

Microorganisms reported as ethiological agents causing
the illness were Escherichia coli (ATCC 25922), indicating the
presence of Gram negative microorganisms, and Staphylococcus
aureus (ATCC 6538), indicating the presence of Gram positive
microorganisms. For this purpose, "in vitro" methods of diffusion
in Agar (D.A.), as suggested by Pons and col. (1992) Majtan and
Majtanova (1997) were used. Antiseptic and disinfectant
solutions, described by Everything and col. (1994), consisted on
diluting the microorganisms in a 0.85% saline solution to obtain
a 1×108 UFC/ml innocuous solution for each microorganism,
respectively.

The innocuous quantity of 0.1 ml was applied on the
surface of a means of cultivation (Nutritious Agar), extending it
with a bar of sterile glass. Later on, perforation of the means
of cultivation with a speaker of 6 ± 1 mm of diameter was
carried out, and 100 ml of Cikrón-V, approximately, was
applied. The means of cultivation was kept under a temperature of
4ºC, during 1 hour, so that the prediffusion of the solution
toward the means of cultivation could be carried out. When this
time period elapsed, we proceeded to incubation at 37ºC
during 24 hours.

Three trials, or replicates, for each microorganism were
carried out and the inhibition halo was measured with a rule
scaled in millimeters.. The value average of the inhibition halo
was calculated both for each replica and totally. To define the
type of antimicrobial activity, the definition approaches used
were those presented by (Everything and col., 1994), see Chart
1.

Chart 1. Criteria about
antimicrobial activity, considering the inhibition
halum

To evaluate the prevention effectiveness of
Cikrón-V against PPM, a total of 35 sows, housed in the
maternity section of the TO and B units, were treated
experimentally. Random distribution was made in four
groups.

Group I: with n=10 Cikrón V, applying
concentration A.

Group II: with n=10 Cikrón V, applying
concentration B.

Group III: with n=10 Cikrón V, applying
concentration C.

Group IV: with n=5, acting as control under
placebo conditions.

The treatment indicated to the groups was applied via
intra-uterus. It was applied with an indicated dose of 80ml
during three days after parturition (24, 48 and 72h postpartum).
The application of these treatments was carried out with the help
of an artificial insemination catheter, coupled to a
syringe.

The diagnosis of recovery of the illness was carried out
on the fifth day postpartum, while the clinical observation of
the animals continued until the 15th day. The rectal temperature
was also controlled with a clinical thermometer, having its scale
graded in Celsius degrees. Temperature values were taken between
8:00 and 9:00 am from birth up to the 5th day
postpartum

Statistical analysis included the use of a 3.1 version
of Statgraphics, Microsoft
Excel and 1987 SAS. A (2 x 2) chart of contingency was used,
while Chi-squared was calculated.

Results and
discussion.

Resulting percentages of prevalence were 22.85% in unit
TO and 21.73 in unit B, see Chart 2. This indicates that the
prevalence of metritis has been high in the units under study,
while no significant differences were observed between these
units.

Chart 2. Prevalence of metritis
in swine production units.

According to Cama, 2000, the illness is a common
dysfunction in the sow, whose prevalence has increased in the
last years. As a consequence, a great negative economic has
affected the sow’s health and caused serious dysfunctions
in the litter.

As a result of the bacteriological sample collection, it
was detected that the main microorganisms present in the metritis
were Escherichia coli, Staphylococcus aureus and Actynomices
pyogenes, coinciding with reports by (Falceto, 2002).

In the diffusion test in agar for the "in vitro"
evaluation of the antimicrobial activity of the medication, it is
observed that results are in the 16 mm range (charts 1 and 3).
According to the approach by Everything and col. (1994), the
inhibitory response is classified as "marked" in this case, so
Escherichia coli and Staphylococcus aureus were identified with
inhibition halos averages of 16±0.36 mm and
17.4±0.36mm, respectively.

Chart 3. Antimicrobial activity
of Cikrón V.

These belong in the same class as those described by
Armenteros (1998), who found a marked antimicrobial activity of
the product on the presence of Staphylococcus aureus, with an
inhibitory halo average of 19.6 Red ± 0.76 mm.
Antimicrobial activity of solutions based on Red Mangrove
(Rhizophora mangrove L.) on the presence of microorganisms is
reported. Authors also point out this drug can inhibit the growth
of pathogen human bacteria in more than 70% of the
cases.

In the case of Escherichia coli, it spreads to be more
resistant, given its Gram negative condition, where a layer in
its cellular wall increases its resistance, in a general way, on
the presence of antimicrobial products (Omen, 1998).

When evaluating the effectiveness of Cikrón-V in
the prevention of puerperal metritis postpartum, it was observed
that, apart from the concentration, the percentage of prevention
was high (90%) in each group. One can also observe that
significant differences exist among the placebo group with the
three groups under treatment.

Chart 4. Efficacy of the Cikrón
V in the prophylaxis of puerperal metritis.

Each column presents different
letters, Significant difference, p<0.01

In chart 5, figures resulted after the temperature had
been taken during the development of the experiment. As it can be
appreciated, in none of the groups, except for the placebo, fever
was present at the beginning of the treatment. Later on,
progressive fever was presented in all the sick animals. In the
healthy ones the behavior was normal, remaining in the normal
range for the species that is from 37.5 to 39.5 º C
(Vizcaíno and col., 2001).

Chart 5. Sows' rectal temperature (in
ºC) during treatment.

We can support that Cikrón V is effective for the
prevention of the metritis in sows right after delivery, being a
highly effective treatment that does not bring about alterations
of the animals’ body temperature.

Conclusion.

• Prevalence of the illness was 22.85% and 21.73 % in
  the TO and B units, respectively.
• The main microorganisms associated with the illness
  were Escherichia coli, Staphylococcus aureus and Actynomices
  pyogenes.
• Cikrón V presents a marked antimicrobial
  activity.
• Cikrón V resulted 90% effective in the
  treatment of postpartum metritis for concentrations A, B and C,
  respectively.

Recommendation.

• Carrying out a pilot clinical trial (Phase III) by
  the National Swine Husbandry Company, to evaluate the
  effectiveness of Cikrón V in production units and extend
  it as a standard drug for preventing and curing postpartum
  metritis islandwide, is recommendable.

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Yuván Contino Esquijeroza *

Félix Aguero Díaz * *

* Veterinary Medicine Major.

School of Veterinary Medicine and Animal
Husbandry.

"Fructuoso Rodríguez Pérez" La Habana
Agrarian University (UNAH).

Researcher in EEPF" Indio Hatuey", Matanzas,
Cuba.

**
Researcher.

Group on Clinical
Reproduction.

National Center of
Plant and Animal Health. (CENSA).