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.

References.

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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).