The goal of this study was to develop a method for the diagnosis of latent respiratory failure in calves with bovine respiratory disease (BRD) in farm conditions. The study involved 48 calves of the red-motley Holstein breed aged 14-28 days, with no previously experienced BRD, no dyspnea at rest, and a lack of anemia. Group I consisted of 24 calves with spontaneous or induced cough and WI clinical score of ≥5. Group II included 24 calves without spontaneous and induced cough and WI clinical score of ≤3. The determination of respiratory rate (RR), respiratory minute volume, and tidal volume in all calves was performed before and after functional exertion with a 15-min run and 30-s apnea. Blood samples were obtained from the jugular vein before 30-s apnea, then immediately after, and finally 1 min after apnea; blood gases and acid-base status were analyzed on an ABL-330, while total hemoglobin content was analyzed on a Micros-60. Tachypnea and dyspnea appeared after the 15-min run in group I calves, lasting 7.2 ± 0.4 and 3.7 ± 0.2 min. In group II calves, tachypnea lasted 3.8 ± 0.2 min, and no dyspnea occurred. Within 1 min, hypoxemia and hypercapnia caused by 30-s apnea in group II calves were compensated for due to an increase in pulmonary ventilation; in group I calves it was compensated for within more than 1 min, with presence of shallow breathing and dyspnea. The relation of RR immediately after 30-s apnea to RR before apnea showed excellent diagnostic value for detecting latent respiratory failure in the calves: area under the curve was 0.981, with cutoff point of ≥1.47, sensitivity of 95.8%, and specificity of 87.5%.

The paper demonstrates a new approach to identify healthy calves (“healthy”) and naturally occurring infectious bronchopneumonia (“sick”) calves by analysis of the gaseous phase over nasal secretions using 16 piezoelectric sensors in two portable devices. Samples of nasal secretions were obtained from 50 red-motley Holstein calves aged 14–42 days. Calves were subjected to rectal temperature measurements, clinical score according to the Wisconsin respiratory scoring chart, thoracic auscultation, and radiography (Carestream DR, New York, USA). Of the 50 calves, we included samples from 40 (20 “healthy” and 20 “sick”) in the training sample. The remaining ten calves (five “healthy” and five “sick”) were included in the test sample. It was possible to divide calves into “healthy” and “sick” groups according to the output data of the sensor arrays (maximum sensor signals and calculated parameters Ai/j) using the principal component linear discriminant analysis (PCA–LDA) with an accuracy of 100%. The adequacy of the PCA–LDA model was verified on a test sample. It was found that data of sensors with films of carbon nanotubes, zirconium nitrate, hydroxyapatite, methyl orange, bromocresol green, and Triton X-100 had the most significance for dividing samples into groups. The differences in the composition of the gaseous phase over the samples of nasal secretions for such a classification could be explained by the appearance or change in the concentrations of ketones, alcohols, organic carboxylic acids, aldehydes, amines, including cyclic amines or those with a branched hydrocarbon chain.

Background and Aim:Calfhood disease is an important problem in dairy farming that could cause significant effects on heifer survival and productivity and has economic and welfare effects. Total protein concentration in the blood serum could be one of the predictors of bovine respiratory disease (BRD) in newborn calves. The number of active nucleolus organizers could be used to assess the viability of the protein synthesis system in cells and tissues. We aimed for a comparative assessment of the dynamics of the main indicators of protein metabolism and nucleolus organizer regions (NORs) activity in the lymphocytes of healthy calves (Group I) and calves with BRD (Group II) during the 1st month after birthMaterials and Methods:This study included 30 calves of the red-motley Holstein breed. Venous blood samples were taken from all calves on the 1st, 7th, 14th, and 28th days after birth. Quantitative analysis of total protein (Serum total protein [STP]), immune globulin (Serum immune globulin [SIg]), urea, and creatinine in serum and transcriptionally active chromosome NORs in the interphase nuclei of lymphocytes was conducted using receiver operating characteristic analysis and factor analysis.Results:In Group I, the STP levels decreased during the 1st month of life, and in Group II, the STP levels were variable. The STP levels in both groups remained within the reference intervals. During the first 2 weeks after birth, the calves’ SIg fluctuated within the statistical error limits and did not significantly differ between the groups. On the 28th day, SIg increased in both the groups (by 42.8% for Group I and 33.7% for Group II). The creatinine concentration showed a decrease but did not go beyond the range of reference values. Urea concentration in Group I markedly decreased and remained below the reference values; it did not change in Group II over the entire observation period. The number of NORs in 1-day-old calves did not significantly differ between the groups and amounted to 2.43 in Group I and 2.59 in Group II. A significant increase in the number of active NORs was found in calves in both groups at the ages of 14 and 28 days. Early BRD predictors (at 1-14 days) could not be identified among the studied indicators. The urea and creatinine concentrations and the NOR activity on day 28 after birth could be late BRD predictors. Protein metabolism in the newborn calves’ organisms is regulated by three types of factors: Maintenance of a constant protein concentration in the plasma, protein decomposition, and de novo synthesis.Conclusion:There were no observed significant differences in the protein metabolism values and dynamics of indicators between healthy calves and calves with developed BRD. Alterations in the studied characteristics are the result, but not the cause of BRD. The increase in active NORs under BRD could be a favorable forecasting indicator. Protection against foreign protein and genetic material is a more important task for the organism than ensuring growth processes during the neonatal period.

According to modern phylogenetic theory of general pathology (V.N. Titov, 2003, 2013) inflammation is a nonspecific biological reaction, providing the removal of excessive amount of highmolecular macromolecules and maintenance of metabolic homeostasis (endoecology). The increase of any metabolite concentration above the physiological range is a violation of the «purity» of intercellular medium. This triggers two nonspecific biological reactions, namely the excretion used to remove «biological waste» of a molecular weight less than 70 kD and the inflammation in case the larger molecules and their complexes should be removed via neutrophils, resident macrophages and endothelial cells. Blood concentration of leukocytes has been used for evaluation of inflammation and intoxication activity for many decades. However, at the molecular level there is a requirement in precise differentiation of metabolites, activating leukocytes, and metabolites, excessively forming as a result of activation, because the last disturb molecular homeostasis and may damage cells and tissues. Validity of leucocytal intoxication index (LII) of Ya.Ya. Kalf-Kalif, concentration ratio of low and medium molecular weight substances, average molecular peptides, total and effective albumin as markers of endogenous intoxication (M.Ya. Malakhova, 2005) are considered. It is shown that primary agent, activating neutrophils and some factors of humoral immunity, is a lipopolysaccharide (endotoxin) of gram-negative microflora (M.Yu. Yakovlev, 2003; O.W. McIntyre, 2011). An excess of endotoxin inflow is possible under intestine and liver pathologies, and due to sympathoadrenal system activation, and also with feed and air. The overload of systems and organs of elimination of endotoxin causes secondary immunodeficiency, which becomes the cause of acute and chronic inflammatory processes of various localizations. It is offered to consider lipopolysaccharide of gramnegative microflora as a primary agent of endogenous intoxication and all the metabolites, produces at increased concentrations as a result of polymorphonuclear leukocytes activation such as reactive oxygen species, oxidated proteins, products of lipid peroxidation and proteolysis, as a secondary one. Such an approach allows to single out various points of application for therapy of endogenous intoxication: i) elimination of excessive concentration of lipopolysaccharide in blood by limiting its production and income from internal and external sources, as well as by a variety of methods of active binding and excretion of endotoxin (S.V. Smirnov et al., 2003; K. Battenschoen et al., 2010); ii) reducing the concentration of «secondary» metabolites in blood and tissues.

This research was conducted to study hormonal and metabolic statuses and to determine ultrasound criteria for early diagnosis of fetal growth retardation syndrome (FGRS) in dairy cows of Black-motley (n = 41) and Red-motley (n = 56) Holstein breeds. Concentrations of progesterone, testosterone, 17β-estradiol, dehydroepiandrosterone sulfate, cortisol, and triiodothyronine in blood serum were assessed by immune-enzyme analysis. Concentrations of plasma L-ascorbic acid and nitrogen oxides, serum immune globulin, middle molecular peptides, and bactericidal and lysozyme activity were determined by spectrophotometry. On days 38-40 and 60-65 of gestation, FGRS in cows was associated with hypoprogesteronemia, decrease of adrenal and thyroid gland functions, decreased nitric oxide synthesis, low immune responsiveness, and high endogenous intoxication. In conclusion, body length less than 16 mm on days 38-40 and less than 45 mm on days 60-65 are the criteria for designating underdevelopment of embryos and fetuses in dairy cows. FGRS genesis in dairy cows is determined by the embryo's and fetus's malnutrition at the early implantation and placentation stages, caused by the imbalance of endocrine regulation, nitric oxide systems, and endogenous intoxication.

100 Holstein calves aged 7-14 days investigated the effectiveness of various cough provocation methods for bovine respiratory disease early detection. The animals were evaluated according to the clinical scoring system WI, to provoke cough in calves, they performed: 15-minute run, 30-second apnea on exhalation, palpated the trachea in its lower third region and intravenously injected 0.6% hydrogen peroxide solution at 0.9 % sodium chloride solution at a dose of 0.4 ml per kg of body weight to induce H2O2-induced bronchoconstriction. Out of 100 calves, 60 were selected without spontaneous cough and with a WI clinical score of 3 points or less, in which the leukocyte count, differential leukocyte count, neutrophil-to-lymphocyte ratio, nuclear shift index, and pH of exhaled breath condensate and the hydrogen peroxide concentration in exhaled breath. For 2 weeks, these animals were monitored daily for the bovine respiratory disease signs. It was established that 30-second apnea and H2O2-induced bronchoconstriction allow diagnosing bronchitis in calves 6-12 days before its symptom complex (peak) development when other symptoms are still absent and WI clinical score is 3 points or less. However, already at this disease stage, animals exhibit leukocytosis, increased serum content of haptoglobin, decreased pH exhaled breath condensate, and increased the hydrogen peroxide concentration in exhaled breath. The trachea lower third cough on palpation appearance indicates the tracheobronchitis early manifestations presence in calves. Trachea lower third Sequential palpation and 30-second apnoea to provoke coughing in calves allows not only the bovine respiratory disease early detection, but also to differentiate early bronchitis and tracheobronchitis manifestations.

A b s t r a c tInflammatory diseases of the respiratory and gastrointestinal tracts are the actual problem of veterinary medicine. Innate or natural immunity plays the leading role in the initial reactions to microbial agents contaminating sterile mucous membranes of newborn animals. It is implemented through the activation of cellular and humoral factors of nonspecific resistance. Leukocyte concentration, leukogram, content of cationic proteins in neutrophils, blood serum lysozyme activity (BSLA) were studied in 20 red-motley calves in 1 hour after their birth on the days 2, 5-7 and 14-15 of life, together with an impact of these indices on the terms of appearance and clinical course of inflammatory diseases of the respiratory and gastrointestinal tracts, under conditions of a large dairy complex environment (Voronezhpishcheprodukt Co Ltd, Novousmansky district, Voronezh region) in 2014. Blood sampling was implemented in the morning before feeding from the jugular vein. Body temperature, pulse and respiratory rate, state of the visible mucous membranes, time of appearance and intensity of sucking reflex, the presence and nature of cough, dyspnea, nasal expiration, reaction to palpation of the larynx, trachea, intercostal spaces, the abdominal wall, percussion and auscultation of the chest were determined in calves. The markers of intestinal inflammation are soluble protein, erythrocytes (hemoglobin), leukocytes (leukocyte elastase), pH. They were identified in feces by dry-chemistry method during the same period and also selectively at the age of 1-1.5 months. According to clinical and hematological indices the animals were retrospectively divided into 2 groups: group A with leukocyte concentration of (11.0-18.3)½10 9 /l and segmentonuclear neutrophils (SN) more than 4½10 9 /L at birth; group B with leukocyte concentration of 6.5-11.3½10 9 /l and SN lesser than 3.9½10 9 /l at birth. Granulocyte blood concentration dynamics in calves of these groups significantly differed both among themselves and from physiological dynamics (adaptive norm) during the first 15 days of their life. This was determined by various phases of adaptation syndrome (mobilization, resistance, exhaustion) and various possibilities of calves' granulocytic system reaction to irritants impact according to J. Wilder's law of initial values (1957). The increased blood level of SN and rod neutrophils (RN) in animals of group A at birth indicates existing activation of bone marrow and may not significantly increase in response to a rather intensive stimulus. Initial level of SN and RN in calves of group B was close to the physiological one, it significantly increased on the day 2 and decreased by the days 5-7 of life. Cationic protein concentration and BSLA in all the calves during the research period were lower than the indices typical of the animals of this age. Diarrhea developed in all the calves on the day 2 of life. Its duration in animals of groups A and B was 8.6±1.1 and 4.2±0.6 days, respectively. When diarrhea symptoms disappeared, inte...

The article discusses general principles of obtaining diagnostic information using eight chemical piezoelectric gas sensors with nanostructural coverings from exhaled breath condensate for health assessment of the upper respiratory tract of pre-month-old calves. Multidimensional information of an e-nose can be presented in various numeric and visualized matrices, characteristics of which are an integral analytical signal of the sensor array. The research devoted to the search for the techniques of extracting analytical information from multidimensional e-nose data to assess upper respiratory tract state from the appearance of the first sign of respiratory disease to tracheobronchitis and bronchopneumonia. The piezoelectric sensor array is characterized by high sorption activity with priority biomolecules (which are the markers of the abnormal metabolic processes), low cost along with reliable repeatability of sorption properties from batch to batch, the simplicity of application, and fast response and recovery time. Here we present the results of simple algorithms used for assessment of upper respiratory tract state by a 2-minute analysis of odour over 1-ml biosample without sample preparation. It was shown that traditional quantitative parameters of e-nose are not adequate for simultaneous sample grouping and volatile compounds qualitative composition establishing. The additionally calculated sorption parameters Aij are more informative in the analysis of biosamples volatile: using for identification of volatile biomarkers, describe the health state correlating with clinical diagnosis. The sequence of information processing: signals of each sensor, integral characteristic, “visual prints,” additional sorption parameters–allows assessing the calf health virtually in situ without transporting samples to specialized laboratories.