JP2005097225A - Prophylaxis and treatment of livestock uteritis by activated chlorine dioxide - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a washing method having high bactericidal and disinfecting effect by eliminating bacteria in the uterus of livestock affected with endometritis or disinfecting the interior of uterus of the livestock. <P>SOLUTION: The method for efficiently eliminating a phlogogenic microorganism comprises washing the interior of uterus of livestock affected with uteritis by using a chlorine dioxide agent containing activated chlorine dioxide. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

Detailed Description of the Invention

The present invention relates to prevention and treatment of domestic animal uteritis by activated chlorine dioxide.
Livestock as used herein includes ungulates, such as horses, cows, sheep, goats, etc., and chlorine dioxide stabilized and activated in the uterus of the livestock is used directly or in combination with a pharmacologically acceptable carrier. It is characterized by the treatment and prevention of uterine inflammation by sterilization and sterilization of U. pneumoniae bacteria and disinfection of septic exudates associated therewith.
The sterilization and sterilization disclosed in this specification includes concepts such as “sterilization” and “disinfection”.

Uteritis refers to a case where an inflammatory change extends from the mucous membrane to the muscle layer, and is therefore classified into endometritis, uterine myositis, and periuteritis depending on the site of inflammation. This inflammatory change can also reach the serosa and is often referred to as septic uteritis due to the transition to sepsis.

The main cause of this septic endometriosis is born by infection during the period of birth with bacteria belonging to the family Enterobacteriaceae such as staphylococci, streptococci, Escherichia coli, Citrobacter and Klebsiella.

The uterine mucosa infected by the causative fungus becomes necrotic, the mucosa is detached, and a septic exudate containing necrotic products is discharged into the vagina. The muscle layer is also edematous, with granule-presenting sphere infiltration throughout the uterine wall, and changes in inflammatory thrombus in the uterine vein.

This endometritis occurs in horses, cattle, pigs, sheep, goats and other livestock and dogs, and is a major cause of infertility and miscarriages. That is, when suffering from this disease, sperm motility is suppressed, ascending in the uterus is prevented, implantation of fertilized eggs to the endometrium is inhibited, and even after implantation, the embryo is killed early, Inducing miscarriage and premature birth.

This disease is caused by Staphylococcus, which is a causative bacterium such as uteritis, pneumonia, sepsis, etc. Streptococcus aureus, nephritis, pharyngitis, and streptococci frequently detected in septic exudates of horses with uteritis, such as S. aureus equi, S .; agalactias, S .; pyogenes, S .; E. coli, Citrobacter, Klebsiella detected from endometriosis, bovine mastitis, or Pseudomonas spp. which is a major cause of opportunistic infections, sepsis, and sores It is a non-infectious disease caused by. In particular, Klebsiella and Pseudomonas have strong expression of resistance mechanisms against antibiotics, and excessive antibiotic administration causes serious disease accompanied by the onset of bacterial substitution by resistant bacteria.

On the other hand, there is equine infectious uteritis (Contagious Equine Metritis CEM), which is designated as a reported infectious disease. The disease is a reproductive infection that is highly contagious, prevalent in the breeding series, and causes endometriosis, infertility and miscarriage in mares.

Toyolorella equigenitalis, the main causative agent of this disease, is transmitted directly or indirectly by the mating of horses, human fingers, instruments, etc., and after an incubation period of 2 to 14 hours, intrauterine with cervicitis and vaginitis Develops meningitis. In 1980, it became a pandemic in the racehorse production area in Japan and caused serious damage to producers.

Infectious disease and non-infectious uteritis prevention and treatment methods include cleaning and disinfecting the genital organs of the uterus and stallions with an antiseptic, followed by sterilization of the causative bacteria by administration of antibiotics.

Infectious and non-infectious disease-causing bacteria in mares are mainly carried by the clitoral sinus and clitoral fossa, and the bacteria are inhabited in the accumulated plaque. It is common practice to remove waste products accumulated by washing the clitoral fossa and lateral clitoral sinus, injecting disinfectants and antibiotics, and removing the causative bacteria by infiltration.

At present, it is essential to disinfect light horses before and after mating, and hypoallergenic properties such as chlorohexidine and reverse soap under mild hot water are regularly used. Even this hypoallergenic disinfectant, Round disinfection causes mucosal itch.

Also, chlorohexidine has no effect on rotavirus, herpes virus, adenovirus, and reverse soap does not show efficacy against gram-negative and spore bacteria and viruses.
Currently, stallions are eagerly aspiring for drugs with a broad bactericidal spectrum, including viruses, with no or low irritation to equine genitals.

In the intrauterine cleaning described above, first, a large amount of sterile physiological saline or Ringer's solution heated to 38 ° C. to 40 ° C. is injected, and irrigation is repeated to wash the uterus. Currently, antibiotics are generally used as injecting drugs, and iodine drugs in addition to the above-mentioned drugs are used as disinfectants.

Frequent administration of chloramphenicol, which is commonly used as an antibiotic, has been reported to cause myelosuppression syndrome, suppression of previous immune response, and collapse and hemolysis due to rapid intravenous injection. The prohibition of continuous use for more than 4 days is taken. Aminoglucoside antibiotics (for example, gentamicin), beta-lactam antibiotics, and quinolone antibacterial agents are also regulated for frequent use due to the appearance of side effects or resistant bacteria.

On the other hand, in the local sterilization with a disinfectant, selection of a disinfectant must be considered in view of its effectiveness and toxicity. That is, the cleanliness requirement of the object to be disinfected, biological disinfection and environmental disinfection, basic conditions such as pH, action time, temperature, concentration, range of disinfection spectrum, toxicity of disinfectant to living body, sustainability of action, absorbability It must be selected based on permeability, effectiveness in the presence of organic substances, emergence of resistant bacteria, odor and pigment deposition, ease of use, and ease of exhaust treatment.

As a uterine washing disinfectant that is widely used at present, iodine agents, especially popidone iodine, are used because of the wide disinfection spectrum. In general, iodine is used for disinfection of fingers, but it is highly corrosive and has side effects such as diarrhea, diarrhea, dyspnea, and headache. It is considered unsuitable. Onset of dermatitis to the vagina, oral mucosa, burn site and normal skin of newborns has been reported with this drug.

The minimum lethal bacterial concentration of popidone iodine is 10 ppm or more for Escherichia coli and 1000 ppm for Aspergillus niger, and chlorine dioxide has a bactericidal power 10-100 times stronger than this bacterial power (Japan Food Analysis Center, 1995). In addition, intensive studies were carried out on the sterilization method of uterine lavage using activated chlorine dioxide, which was shown to have extremely low skin toxicity.

There has been no case disclosed to date of uterine lavage of livestock with activated chlorine dioxide. In addition, chlorine dioxide has been conventionally considered to be unsuitable as a disinfectant, but rather deodorant, because the reaction is generally slow-acting in spite of its strong bactericidal power. The inventors have improved the slow-acting properties of chlorine dioxide and devised it to produce immediate action, and have repeatedly studied livestock uterine cleansing agents using activated chlorine dioxide that have never been reported. I came.

Activated chlorine dioxide, unlike sodium hypochlorite, does not produce carcinogenic compounds such as trihalomethanes by reacting with serum proteins or other organic substances, and its action is not affected by pH and has a wide pH range. To demonstrate effectiveness.

Examination of the irritancy test of activated chlorine dioxide on human skin revealed no irritation at high concentration for one week, and no effect on the uterine mucosa even at the actual administration concentration. .

Problems to be solved by the invention

Considering this point, we have made extensive studies on the prevention and treatment of livestock, especially light stallion, with activated chlorine dioxide.
The activated chlorine dioxide of the present invention has almost no side effects on human skin or light horse endometrium due to concentration control, has no acquired resistance to chlorophyll of U. pneumoniae, and has a high protein content. Below, it shows bactericidal and disinfecting effects against bacteria, molds and viruses. Based on these characteristics, a trial was conducted in which activated chlorine dioxide was injected into the bred horse uterus.

The disinfection and bactericidal efficacy of chlorine dioxide is pH 3.5-7 depending on the compound selected from organic acids, mineral acids, hydrogen peroxide, alcohols and activators using these in combination (for example, ME series manufactured by Sukegawa Chemical Co., Ltd.). It is possible to activate chlorine dioxide by adjusting to pH 4.0, more preferably pH 4.0 to 6.5.

Means for solving the problem

Concentration-controlled activated chlorine dioxide is 1000 ppm for human skin, has little effect on cumulative pasting for one week, is very irritating to the mucous membrane of the eye, is resistant to microorganisms It exhibits a wide range of bactericidal poisons, exhibits activity over a wide pH range, exhibits activity even under high proteins, does not allow the formation of carcinogenic trihalomethanes, generates chlorine gas, and binds to organic substances. Focusing on the excellent properties of activated chlorine dioxide, such as the fact that it does not generate off-flavors, and as a result of intensive studies, activated chlorine dioxide for the treatment and prevention of uteritis that significantly impairs productivity for livestock producers The first successful use of uterine irrigation drugs.

Embodiment of the Invention

The activated chlorine dioxide in the present specification refers to a substance containing chlorine dioxide, which can be generated by adding an inorganic acid such as hydrochloric acid or sulfuric acid to chlorous acid such as sodium chlorite and potassium chlorite. it can.

The chlorine dioxide may be generated from a stabilized chlorine dioxide agent obtained by stabilizing chlorite with a stabilizer. Here, examples of the stabilizer include NaCO ₃ .3H ₂ O, NaHCO ₃ , and NaBO ₃ .

Furthermore, the activated chlorine dioxide may generate chlorine dioxide by an activating agent selected from organic acids, inorganic acids, hydrogen peroxide and alcohols. Typical examples of organic acids here include citric acid and acetic acid. Examples of the alcohols include ethanol and methanol.

In order to enhance the disinfection and bactericidal effect of the activated chlorine dioxide on general bacteria, coliform bacteria, Staphylococcus aureus, and group A streptococcus, the pH is preferably 7.0 or less. Is adjusted to pH 5.5-6.5.

In the present invention, the activated chlorine dioxide concentration is 0.1 to 100 ppm, more preferably 10 to 75 ppm, and further preferably 25 to 50 ppm.

The sterilization and sterilization mechanism of activated chlorine dioxide is an important metabolic-related enzyme involved in the growth of microorganisms and tyrosine, methionine and cystine residues in the cell membrane composition are specifically oxidized to inactivate their functions. Is to collapse or kill. This oxidizing ability is more specific than ozone, and therefore provides excellent sterilization and sterilization. Therefore, the side effects caused thereby are extremely weak. Moreover, the appearance of resistant bacteria is hardly seen. Chlorine dioxide, unlike sodium hypochlorite, has a relatively slow binding with organic substances, and therefore retains its efficacy for a long time even under high protein.

In the method for sterilizing livestock in the uterus using activated chlorine dioxide according to the present invention, a rubber, plastic or metal tubular instrument (uterine lavage tube) is carefully inserted into the uterus. This uterine lavage tube can be used in accordance with the structure of each accessory genital organ such as the uterus and cervix so as not to be damaged.
Further, a connecting portion may be attached so that the medicine can be injected after the cleaning is completed.
Thus, the injection method is not limited to self-made works or commercial products.
The injection method may be drop injection or electric injection, and is not limited to the injection means.

The cleaning period for treatment is injection, cleaning and disinfection in the part affected by microorganisms at the time of puerperal infection, several hours after seeding, septic exudation, and diagnosing inflammatory thrombus abnormality of uterine vein.

Embodiment of the Invention

The present invention will be described more specifically by the following specific experimental examples and examples, but the present invention is not limited thereto.

Experimental example 1

Table 1 shows the results of the bactericidal efficacy of activated chlorine dioxide for each microorganism. The reaction conditions in this case are a reaction time of 5 minutes and 25 ° C.
Citrobacter and Klebsiella included in Staphylococcus aureus, Streptococcus, Escherichia coli and Escherichia coli, which are major infectious agents of equine uteritis and are frequently detected in uterine exudates, are all reacted in 5 minutes. Completely killed.
Table 1 shows the bactericidal efficacy of the activated chlorine dioxide for each microorganism.

血清蛋白共存下での軽種馬子宮炎主要起炎菌に対する活性化二酸化塩素殺菌力について

Activated chlorine dioxide bactericidal activity against major causative bacteria of light horse equine uteritis in the presence of serum proteins

It describes.

Experimental example 2

Bactericidal efficacy of activated chlorine dioxide in the presence of bovine serum albumin (BSA 0.5% Wako Pharmaceutical Co., Ltd.) against S. aureus, Group A Streptococcus and E. coli The test results are shown in Table 2.

この表が示すように、ＢＳＡの共存下での活性化二酸化塩素はＢＳＡフリーの場合とほぼ同じ活性を示す。 即ち、活性化二酸化塩素は、次亜塩素酸ナトリウムと異なり共存する蛋白と結合することなく、子宮内でも試験管レベル（ｉｎ ｖｉｖｏ）と同じ除菌・殺菌活性が生体内（ｉｎ ｖｉｖｏ）でも発現する事を示唆している。

As this table shows, activated chlorine dioxide in the presence of BSA shows almost the same activity as in the case of BSA free. That is, activated chlorine dioxide does not bind to coexisting protein unlike sodium hypochlorite, and the same sterilization and bactericidal activity as in vitro is expressed in vivo as well as in vivo. Suggests to do.

Experimental example 3

A safety test of activated chlorine dioxide on human skin was carried out in accordance with the method of Kawamura (Taro Kawamura standardization test for standardization of the patch test 80 301 1970).
The application condition was single and continuous application for one round (cumulative application test), and changes in the skin surface were visually observed and evaluated according to the criteria described in the above literature. As an experimental method, in the single and cumulative patch test, patch adhesive bandage (1.5 x 1.5 cm) is impregnated inside the human upper arm with water as a negative control, and activated chlorine dioxide 200 ppm, 500 ppm and 1000 ppm each as 100 μl as a subject. In the single test, the contact time with the skin was occluded for 24 hours, and in the cumulative application, the occlusion for 24 hours was repeated continuously for 1 week. The evaluation was determined by standing for 30 minutes after removing the adhesive bandage and visually observing.
Table 3 shows the results of single-ply stimulation.

活性化二酸化塩素累積貼布によるヒト皮膚の刺激試験結果については

About result of irritation test of human skin by activated chlorine dioxide cumulative patch

Describe in.

この結果、活性化二酸化塩素の実使用濃度の１０〜２０倍の濃度で１〜７日間の単回および累積貼付で全くヒトの皮膚に対して刺激性は認められなかった。Table 5 shows the evaluation criteria at this time.

As a result, no irritation was observed on human skin at single to cumulative application for 1 to 7 days at a concentration of 10 to 20 times the actual use concentration of activated chlorine dioxide.

Uterine lavage ducts were inserted into 13 mares that developed endometriosis where exudate in the uterus leaked intermittently from the perineum and repeated early estrus, and warmed while observing with a uteroscope 100 to 200 ml of sterilized physiological saline was injected into the uterus, and the uterine horn and uterine body were lightly pressed and collected from the rectum, and the collected solution was macroscopically and observed with a microscope as a pre-treatment test solution. Next, activated chlorine dioxide (Biosukegawa design # 1000) was adjusted to 50 to 1000 ppm and 1 to 18 L with sterilized physiological saline, and then filled into an irrigator, followed by using a uterine endoscope and a uterine lavage tube. The sample was injected into the uterus while being kept at 40 ° C. After being left for 1-2 days, it was washed once or twice with the same amount of physiological saline, and microorganisms were detected in the washing solution. The control detection microorganism is E. coli which is the main causative bacterium of endometritis. Detection of Coli, Streptococcus, Klebsiella, Citrobacter Pseudomonas and Staphylococcus was performed. The classification and identification of the detected bacteria followed a conventional method. The results are shown in Table 6. In addition, the effect judgment is obtained from the disappearance result of the target bacilli and the exudate appearance.

I summarized it.

Table 6

Table 7 shows the therapeutic effect of light horse endometritis by sterilization and sterilization washing of activated chlorine dioxide shown in Table 7.

有効率７６．９％と高い示し、活性化二酸化塩素の馬子宮洗浄・除菌の有効性が認められた。

The effectiveness rate was as high as 76.9%, and the effectiveness of cleaning and sanitizing equine uterus with activated chlorine dioxide was confirmed.

Reference example 1

Table 8 shows the results of diagnosis of irritation to the horse endometrium by activated chlorine dioxide when equine uteritis was washed and sterilized with activated chlorine dioxide.

この結果、活性化二酸化塩素１〜１０ｐｐｍ濃度、１８Ｌで子宮内を洗浄しても子宮内膜に何ら影響を与えないことを認めた。
この結果、活性化二酸化塩素１〜１０ｐｐｍ濃度、１８Ｌで子宮を洗浄しても子宮内膜に何ら影響を与えず、安全性が高い薬剤であることを認めた。

As a result, it was confirmed that even if the uterus was washed with 18 L of activated chlorine dioxide at a concentration of 1 to 10 ppm, the endometrium was not affected.
As a result, it was confirmed that even if the uterus was washed with activated chlorine dioxide at a concentration of 1 to 10 ppm and 18 L, the endometrium was not affected and the drug was highly safe.

The invention's effect

As described above, by cleaning and sterilizing horse endometrium infected with pathogenic microorganisms with activated chlorine dioxide, an effective rate of 76.9% was shown using disease improvement as an index. In other words, it was suggested that washing and activating dioxygen can be used to treat and prevent equine endometritis. In addition, upon uterine lavage, no side effects such as erythema, papules, edema and blisters of the endometrium caused by activated chlorine dioxide were observed, and it was confirmed that this was a very safe and effective washing and treating agent.
Furthermore, it also means fluorescence as an effective treatment for preventing and treating equine infectious uteritis (CEM), which is included in surveillance infectious diseases that cause fatal damage to livestock producers.
As a result, activated chlorine dioxide enables prevention and treatment of all types of cattle, swine and other livestock, regardless of prevention and treatment of light horse equine uteritis. As a result, sterilization and disinfection during artificial insemination of rare animals It can be used as an agent.

Claims (9)

A preventive / treating agent for livestock uteritis, mainly composed of activated chlorine dioxide and chlorine dioxide. An aqueous solution containing activated chlorine dioxide and chlorine dioxide, which is used at pH 7.0 or lower, preferably pH 6.0, for preventing and treating livestock uteritis. A method for the treatment and prevention of veterinary uteritis characterized by direct injection into the uterus of livestock using a combination of activated chlorine dioxide and a pharmacologically acceptable carrier. The said chlorine dioxide and chlorine dioxide agent contain the dissolved chlorine dioxide gas aqueous solution, chlorous acid, and its salt, The treatment / prevention agent of livestock uteritis of Claims 1-3 characterized by the above-mentioned. 4. The livestock uteritis treatment / prevention agent according to claim 1, wherein the chlorine dioxide and chlorine dioxide are generated from stabilized chlorine dioxide stabilized by a stabilizer. The activated chlorine dioxide is generated from a chlorine dioxide agent which has activated the generation of chlorine dioxide by an activator selected from organic acids, mineral acids, hydrogen peroxide and alcohols. A therapeutic / preventive agent for livestock uterine inflammation. The use / concentration of the chlorine dioxide in the chlorine dioxide agent is 0.1 to 500 ppm. A method for treating and preventing uterine inflammation characterized by injecting the activated chlorine dioxide into a uterus through a uterine lavage fistula and treating the diseased part by disinfection of the diseased part of the uterine microorganism. The said domestic animal is a light stallion, The therapeutic / preventive agent of the uteritis in any one of Claims 1-7.