RU2372051C1 - Early diagnostic technique for tracheobronchitis in calves - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: invention refers to veterinary science. The technique implies intravenous introduction of 0.6% hydrogen peroxide in 0.9% saline solution in a dose 0.4 ml per kg of body weight. In case cough reaction is observed 1-7 minutes after introduction, tracheobronchitis is diagnosed.

EFFECT: invention allows accelerating diagnostics of tracheobronchitis in calves.

8 tbl, 3 ex

Description

The invention relates to the field of veterinary medicine and relates to a method for early diagnosis of tracheobronchitis in calves.

Tracheitis is an inflammation of the mucous membrane of the trachea. In animals, in most cases, it is registered not as an independent disease, but as a complication of laryngitis (laryngotracheitis) or bronchitis (tracheobronchitis).

Bronchitis is an inflammation of the mucous and submucous membranes of the bronchi. With the course, acute and chronic bronchitis are distinguished. Depending on the localization of the inflammatory process in the bronchi of one size or another, there are macrobronchitis, when the inflammatory process is localized in large bronchi, microbronchitis, when small bronchi are involved in the process, and diffuse bronchitis, when inflammation spreads throughout the bronchial tree. Depending on the nature of the inflammatory exudate, bronchitis can be catarrhal, fibrinous, purulent, putrefactive and hemorrhagic [1, 2].

Among respiratory diseases of calves of viral etiology, parainfluenza-3 (PG-3) and infectious rhinotracheitis (IRT) are of great importance, which occur in the form of both mono- and mixed infections, manifested by keratoconjunctivitis, rhinitis, tracheitis, bronchitis, catarrhal bronchopneumonia, often turning into interstitial with complications of conditionally pathogenic microflora [3, 4].

A known method for the diagnosis of tracheitis: the diagnosis is made on the basis of examination, palpation, auscultation and x-ray examination.

In case of infectious rhinotracheitis, parainfluenza-3, serological, virological and (or) molecular genetic studies are carried out, and in case of complications by bronchopneumonia, bacteriological studies are also carried out [2, 5].

However, this method includes clinical and laboratory studies and does not allow to identify the subclinical course of tracheobronchitis.

A known method for detecting tracheobronchitis, including palpation of the first tracheal ring, in which, in case of hypersensitivity of the mucous membrane, a cough is recorded [6].

The disadvantage of this method is that the increased sensitivity of the first tracheal ring and the entire trachea is recorded with severe tracheitis and bronchitis (dry or wet cough, hard vesicular breathing, wheezing, expiratory dyspnea, etc.).

It is known that the sensitivity of the mucous membrane of the larynx, trachea and bronchi with inflammation is increased. Palpation in this case causes a cough and a general motor reaction of animals. In healthy cattle, a cough reaction is almost impossible to cause [6].

There is also a method for the diagnosis of pulmonary hypertension. The method is based on the intravenous administration of a 0.3% hydrogen peroxide solution and is used in medical practice [7].

The closest in essence to the claimed method is a method involving palpation of the last tracheal ring, in case of hypersensitivity, early manifestation of tracheobronchitis is diagnosed [8].

The disadvantage of this method is that palpation of the last tracheal ring does not allow to detect an increase in sensitivity of the mucous membrane of the bronchi, which ultimately makes it impossible to diagnose tracheobronchitis at earlier stages of the disease.

The technical result of the invention is the acceleration of the diagnosis of tracheobronchitis in calves.

The method is as follows. Calves are injected intravenously with a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg of body weight. In the event of a cough reaction, 1-7 minutes after administration, tracheobronchitis is diagnosed.

An analysis of the data in the scientific, technical and patent literature did not reveal a method for the early diagnosis of tracheobronchitis in calves using intravenous administration of a 0.6% hydrogen peroxide solution, which indicates compliance with the claimed solution to the novelty criterion.

The invention can be used for early diagnosis of tracheobronchitis in calves both in large livestock complexes and in small farms, which is very important for livestock breeders and veterinarians for the timely adoption of treatment and prophylactic measures for stopping tracheobronchitis, monitoring the effectiveness of treatment of calves with respiratory pathology.

Using the method allows to detect tracheobronchitis in calves 2-6 days earlier than when using the prototype [8]. In addition, intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution promotes the activation of day-to-day metabolism.

Early diagnosis of tracheobronchitis using the proposed method was carried out in LLC Voronezhpishcheproduct Novousmansky district of the Voronezh region on 64 calves 1-1.5 months of age.

The essence of the method is illustrated by examples.

Example 1. The rationale for the use of hydrogen peroxide to detect tracheobronchitis in calves.

On a farm that was long-term unsuccessful in PG-3 and IRT, often occurring in the form of tracheobronchitis, 40 calves of 1-1.5 months of age were selected, 14 of them with respiratory pathology (tracheobronchitis, bronchopneumonia) - the first group and 26 clinically healthy calves - the second group.

The calves were injected intravenously with a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg of body weight. In the event of a cough reaction, tracheobronchitis was diagnosed 1-7 minutes after administration.

The animals were monitored continuously for 7 days, body temperature, the number of respiratory movements, the condition of the mucous membranes, the sensitivity of the larynx and trachea, the nature of coughing, wheezing, the presence or absence of shortness of breath, nasal discharge were recorded, the time of the appearance of sensitivity of the last tracheal ring, reaction were recorded calves for the introduction of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. The results of the experiment are presented in table 1.

Table 1 The reaction of calves to the introduction of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. Indicators Calves 1st group (patients) 2nd group (healthy) The number of calves, goals (%) 14 (100) 26 (100) Of these, reacted to the introduction of a 0.6% solution of hydrogen peroxide, heads (%) 14 (100) 12 (46.2) Of these, responded to palpation of the first tracheal ring, goals (%) 14 (100) - palpation of the last tracheal ring, goals (%) 14 (100) - The time of the appearance of sensitivity of the last tracheal ring, day on palpation 3.8 ± 0.46

From table 1 it is seen that clinically healthy calves did not respond to palpation of both the first and last tracheal rings. However, for the introduction of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg of body weight of 26 calves in 12 (46.2%) cough was recorded (from 2 to 7 cough tremors). Whereas the sensitivity of the last tracheal ring in these calves after administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution was manifested on 3.8 ± 0.46 days.

In calves with a clinical form of tracheobronchitis, both a palpation of the first and last tracheal rings and intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution showed a cough (from 3 to 9 coughing tremors) with sputum discharge .

That is, the intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution to calves 3.8 ± 0.46 days earlier reveals a subclinical course of tracheobronchitis in comparison with palpation of the last tracheal ring.

Clinical and biochemical parameters are presented in table 2.

table 2 Clinical and biochemical parameters of healthy and sick calves. Indicators Calves Sick Healthy Responsive to 0.6% hydrogen peroxide Non-reactive 0.6% hydrogen peroxide Body temperature, ° C 39.8 ± 0.22 * 39.0 ± 0.06 38.7 ± 0.06 Respiratory rate, in minutes 54 ± 4.7 * 35 ± 3,2 29 ± 3.1 Heart rate, in min. 118 ± 10.3 * 91 ± 9.3 86 ± 5.2 White blood cells, 10 ⁹ / L 8.3 ± 0.90 10.1 ± 1.01 8.9 ± 0.68 Red blood cells, 10 ¹² / L 5.4 ± 0.11 6.0 ± 0.14 5.9 ± 0.11 Hemoglobin, g / l 81.2 ± 1.97 * 83.7 ± 3.70 88.2 ± 3.72 Proteins, g / l 61.72 ± 2.61 63.14 ± 2.37 62.31 ± 1.83 Lipids, g / l 3.25 ± 0.34 3.43 ± 0.17 3.72 ± 0.29 Cholesterol, mmol / l 3.17 ± 0.34 * 4.00 ± 0.46 4.31 ± 0.42 Creatinine, mM / L 78.3 ± 4.59 71.3 ± 4.29 72.6 ± 3.05 Glucose, mmol / l 2.20 ± 0.43 * 3.31 ± 0.38 * 4.72 ± 0.44 Urea, mmol / l 3.90 ± 0.28 * 3.57 ± 0.71 * 2.19 ± 0.23 Catalase, μM H ₂ O ₂ / (l × min) 31.83 ± 1.40 * 33.66 ± 1.19 35.54 ± 1.69 SOD, cu 0.52 ± 0.06 * 0.75 ± 0.10 0.78 ± 0.14 GPO, mm GSH / (l × min) 25.34 ± 0.75 26.36 ± 1.13 24.92 ± 2.10 MDA, μM / L 1.17 ± 0.07 * 1.18 ± 0.09 * 0.87 ± 0.05 MSM, cu 20.47 ± 0.97 * 16.17 ± 0.75 14.69 ± 0.54 SSE,% 53.83 ± 1.89 50.42 ± 1.94 50.61 ± 1.69 Note: * - P <0.05-0.001 compared with calves that do not respond to intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution.

Table 2 shows that in clinically healthy calves responding to the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution (positive cough reaction), compared with calves that do not respond to the administration of a 0.6% peroxide solution hydrogen in a 0.9% sodium chloride solution, body temperature is higher by 0.3 ° C, respiratory rate by 6 respiratory movements per minute, heart rate by 5 contractions per minute, white blood cell count is 1.2 · 10 ⁹ / l (13.5%), hemoglobin concentration at 4.5 g / l (5.1%), cholesterol at 0.31 mmol / l (7.2%), although these differences with not statistically significant. At the same time, blood glucose was significantly 1.43 times lower (P <0.05), and urea was 1.63 times higher (P <0.05), respectively.

- супероксид аниона,

- гидроксильного радикала и H₂O₂ - пероксида водорода) у телят, реагирующих на внутривенное введение 0,6% раствора перекиси водорода на 0,9% растворе хлорида натрия. В физиологических концентрациях

не вызывает изменения диаметра трахеи и бронхов, а H₂O₂ и

вызывают контрактацию и релаксацию соответственно. В патологических концентрациях

вызывает релаксацию трахеи и бронхов, а Н₂О₂ и

вызывают преобладающую контрактацию, что и проявляется в виде кашлевой реакции [9, 10]. У клинически здоровых телят, не реагирующих на введение 0,6% раствора перекиси водорода на 0,9% растворе хлорида натрия, его введение не приводит к превышению физиологической концентрации Н₂О₂ и, следовательно, не вызывает контрактации трахеи и кашлевой реакции. У клинически здоровых телят, реагирующих на введение 0,6% раствора перекиси водорода на 0,9% растворе хлорида натрия, дополнительное введение пероксида водорода в кровяное русло приводит к созданию патологической концентрации Н₂О₂ и, следовательно, вызывает преобладающую контрактацию трахеи и бронхов, что на фоне субклинического повышения чувствительности слизистой оболочки трахеи и бронхов провоцирует кашлевую реакцию.There were no significant differences between the activity of antioxidant enzymes (catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPO)) in clinically healthy calves responding to a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution and in non-reacting calves. While the concentration of malondialdehyde (MDA), a secondary product of lipid peroxidation (LPO), in healthy calves, responding to the introduction of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, is 1.36 times (P <0.05) is higher compared with calves that did not respond to the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. Indirectly, this may indicate an increased formation of reactive oxygen species (

- superoxide anion,

- hydroxyl radical and H ₂ O ₂ - hydrogen peroxide) in calves responding to intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. In physiological concentrations

does not cause a change in the diameter of the trachea and bronchi, but H ₂ O ₂ and

cause contracting and relaxation, respectively. In pathological concentrations

causes relaxation of the trachea and bronchi, and H ₂ O ₂ and

cause predominant contracting, which is manifested in the form of a cough reaction [9, 10]. In clinically healthy calves that do not respond to the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, its administration does not exceed the physiological concentration of H ₂ O ₂ and, therefore, does not cause tracheal contraction and a cough reaction. In clinically healthy calves that respond to the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, additional administration of hydrogen peroxide into the bloodstream leads to a pathological concentration of H ₂ O ₂ and, therefore, causes a predominant contraction of the trachea and bronchi , which against the background of a subclinical increase in the sensitivity of the mucous membrane of the trachea and bronchi provokes a cough reaction.

Indicators characterizing the state of the POL-AOD system 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution are presented in table 3.

Table 3. The activity of the main antioxidant enzymes and the concentration of secondary lipid peroxidation products in the blood of calves 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. Indicators Clinically Healthy Calves Responsive to 0.6% hydrogen peroxide Non-reactive 0.6% hydrogen peroxide Catalase, μM H ₂ O ₂ / (l × min) 42.79 ± 2.19 * 37.55 ± 2.74 SOD, cu 0.81 ± 0.21 0.63 ± 0.16 GPO, mm GSH / (l × min) 26.07 ± 0.73 28.96 ± 0.37 MDA, μM / L 0.98 ± 0.08 * 0.88 ± 0.05 Note: * - P <0.05-0.001 compared with the initial values.

From table 3 it is seen that 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution in calves that respond to its administration by a cough reaction, the catalase activity significantly increases by 27.1% (P <0 , 05), and the concentration of MDA in the blood decreases by 20.4% (P <0.05) compared to the initial values, which indicates an increase in the activity of the enzymatic link of the antioxidant system and a decrease in the content of secondary lipid peroxidation products in the blood.

Indicators of sorption ability of erythrocytes (SSE) and the content of medium molecular weight proteolysis products in the blood of the so-called medium-weight molecules (MSM), reflecting the effect of oxidative stress on biological membranes and the degree of endogenous intoxication, are presented in table 4.

Table 4. Indicators of the sorption ability of erythrocytes (SSE) and the content of medium molecular weight proteolysis products in the blood of calves 48 hours after the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution Indicators Clinically Healthy Calves Responsive to 0.6% hydrogen peroxide Non-reactive 0.6% hydrogen peroxide MSM, cu 14.88 ± 0.91 14.86 ± 0.70 SSE,% 48.16 ± 1.56 47.58 ± 2.45

Table 4 shows that in all clinically healthy calves 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, the sorption capacity of red blood cells tends to decrease, and in calves responding to 0.6 % hydrogen peroxide solution in a 0.9% sodium chloride solution, and the concentration in the blood of MSM, which indicates a decrease in the degree of endogenous intoxication and an improvement in the state of biomembranes.

The content of urea and glucose in the blood 48 hours after the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution is presented in table 5.

Table 5. The level of glucose and urea in the blood 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution. Indicators Clinically Healthy Calves Responsive to 0.6% hydrogen peroxide Non-reactive 0.6% hydrogen peroxide Glucose, mmol / l 5.13 ± 1.20 * 5.37 ± 0.54 Urea, mmol / l 2.35 ± 0.54 * 1.63 ± 0.27 Note: * - P <0.05-0.001 compared with the initial values

From table 5 it can be seen that 48 hours after the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution in calves that respond to it with a cough reaction, the blood glucose level increases by 55.0% (P <0 , 05), and the concentration of urea in the blood serum is reduced by 65.8% (P <0.05). Calves that do not respond to the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution tend to increase glucose levels and lower blood urea. The data presented indicate an improvement in metabolic processes and oxidative-antioxidant status in calves 48 hours after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg of body weight.

Example 2. The rationale for the early diagnosis of tracheobronchitis in calves with antiviral antibody titers.

In a farm that was permanently unfavorable for PG-3 and IRT, 10 calves of 1-1.5 months of age were selected, not responding to palpation of the last tracheal ring, but responding to an intravenous 0.6% hydrogen peroxide solution in a 0.9% chloride solution sodium cough reaction (subclinical tracheobronchitis). The animals were monitored for a month, body temperature, heart rate and respiratory movements were determined, appetite, the condition of the mucous membranes, sensitivity of the larynx, trachea and intercostal spaces, the nature of coughing, wheezing, shortness of breath, nasal discharge, timing of occurrence and duration of the disease were taken into account. tracheobronchitis, took into account complications in the form of bronchopneumonia. Calves aged 1-1.5 and 1.5-2 months were bled to determine antiviral antibodies. The titers of antiviral antibodies in calves are presented in table 6.

Table 6. Antiviral titers in calves with tracheobronchitis. No. p / p Pathogen Antibody Titers PG-3 IRT Subclinical Clinically pronounced Subclinical Clinically pronounced one 1:20 1:20 1: 2 1:32 2 1:40 1:80 1: 4 1: 4 3 1:80 1: 160 1: 8 1:16 four 1:80 1: 160 neg. 1: 8 5 1: 160 1: 320 neg. neg. 6 1: 160 1: 160 1:16 1:16 7 1:80 1: 320 1: 8 1:16 8 1: 160 1: 320 neg. 1: 8 9 1:40 1:80 1: 4 1: 8 10 1: 160 1: 480 1: 4 1:16 Wed 1:98 1: 210 1: 4,6 1: 10.8

From table 6 it is seen that the average titer to the pathogen PG-3 in calves that do not respond to palpation of the last tracheal ring, but respond to intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution in a dose of 0.4 ml per kg of body weight of the animal (subclinical course of tracheobronchitis), is 1:98, in the clinical course - 1: 210, and to the causative agent of the RTI is 1: 4.6 and 1: 10.8, respectively.

However, a detailed analysis found that calves No. 2, 3, 4, 5, 7, 8, 9, 10 have an increase in the titer of antibodies to the antigen of the pathogen PG-3 (on average from 1: 100 to 1: 240). In calves No. 1 and 6, he remained unchanged. In addition, calves Nos. 3, 4, 7, 8, 9, 10 showed an increase in the titer of antihemagglutinins to the antigen of the IRT pathogen, and in calves Nos. 2 and 5, it remained unchanged.

That is, in all calves with the clinical manifestation of tracheobronchitis, either an increase in antibody titers to one or more causative agents of acute respiratory diseases (PG-3 and IRT) is recorded or their value remains unchanged.

The timing and duration of the course of respiratory diseases in the above calves are presented in table 7.

From table 7 it is seen that, on average, in calves that do not respond to palpation, but respond to intravenous administration of a 0.6% hydrogen peroxide solution with 0.9% sodium chloride at a dose of 0.4 ml per kg of body weight by a cough reaction, indicating subclinical tracheobronchitis, the latter was detected on 36.4 ± 1.68 days, and its clinical manifestation (mucous membrane of the nose, conjunctiva, nasolabial mirror hyperemic, with erosion, serous discharge from the nose, increased sensitivity of both the first and last tracheal rings and larynx, with auscultation dry and wet rales are heard, expiratory dyspnea is noted) on 44.4 ± 1.66 days. That is, the time from subclinical tracheobronchitis to its clinical manifestation is on average 8.0 ± 0.33 days. In addition, in 6 (60%) of 10 calves with tracheobronchitis, a complication of bronchopneumonia was recorded.

Example 3. Justification of effectiveness.

On a farm that is permanently unfavorable for tracheobronchitis of viral etiology (PG-3 and IRT), 14 calves of 1-1.5 months of age were selected, not responding to palpation of the last tracheal ring, but responding to intravenous administration of a 0.6% hydrogen peroxide solution to 0.9% sodium chloride solution at a dose of 0.4 ml per kg of body weight by cough reaction. The calves recorded the time of occurrence of a cough reaction after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, as well as the time of appearance of the sensitivity of the last tracheal ring during palpation. The results are presented in table 8.

From table 8 it is seen that in 50% of calves, a cough reaction was recorded 1-2.5 minutes after the administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg body weight, in 28 , 6% after 3-4.5 minutes and in 21.4% of the animals after 6.5-7 minutes. That is, the average time of occurrence of a cough reaction after intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution was 3.5 ± 0.56 minutes.

In 7.1% of calves, the sensitivity of the last tracheal ring (subclinical tracheobronchitis detected by palpation) was recorded on the 2nd day after detecting a cough reaction by the claimed method, in 50% of animals on the 3-4th day and in 42.9% of calves on 5-6th day. That is, the use of the proposed method in comparison with the known one allows earlier to diagnose subclinical tracheobronchitis by 4.1 ± 0.36 days.

Thus, the inflammatory process in the mucous membrane of the trachea and bronchi with the active participation of PG-3 and IRT viruses increased the sensitivity of the first of the bronchial mucosa and the last tracheal rings, then the first tracheal rings, which ultimately manifested tracheobronchitis symptom complex. Against the background of increased sensitivity of the mucous membrane of the trachea and bronchi, the intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution at a dose of 0.4 ml per kg of animal body weight led to contraction of the trachea and bronchi and, as a result, to a cough reactions.

The experiments show the feasibility of using a method that takes into account the reaction of calves to the intravenous administration of a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution, which allows early diagnosis of tracheobronchitis and timely treatment and preventive measures.

Information sources

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Claims (1)

A method for the early diagnosis of tracheobronchitis in calves, characterized in that a 0.6% hydrogen peroxide solution in a 0.9% sodium chloride solution is administered intravenously at a dose of 0.4 ml per kg body weight and, in case of a cough, after 1-7 minutes after administration, tracheobronchitis is diagnosed.