Lymphocyte immunophenotyping and concentration of MMP-9 in transudates and exudates in horses.

This study is the first to provide information on the lymphocyte subpopulations in peritoneal effusions in horses. Peritoneal transudates (n = 12), peritoneal exudates (n = 6) and a pleural exudate (n = 1) were analyzed. The total nucleated cell count (TNCC), total protein (TP) and matrix metalloproteinase-9 (MMP-9) concentration determined by ELISA were measured and routine cytological evaluation was performed. CD3, CD4, CD8 and CD21 positive cells were detected by flow cytometry. A higher percentage of neutrophils (P < 0.05) and higher MMP-9 (P < 0.01) levels were found in exudates. A higher percentage of macrophages (P < 0.05) and lymphocytes (P < 0.01) were found in transudates. CD4 + lymphocytes were the most common lymphocyte subpopulation in all samples. CD21 + lymphocytes were the least common in all samples. A large variability in the percentage of CD21 + lymphocytes was found in exudates. The percentage of CD21 + lymphocytes positively correlated with the level of total protein (r = 0.5704, P < 0.05). The correlation was even stronger in the group of exudates. The percentages of lymphocyte subpopulations did not correlate with the level of MMP-9 or with cytological findings. The level of MMP-9 positively correlated with the percentage of neutrophils (r = 0.4980, P < 0.05), the level of TP (r = 0.7855, P < 0.01) and TNCC (r = 0.6129, P < 0.01). A significantly higher level of MMP-9 was detected in euthanized horses than in horses that survived (P < 0.05). However, it was shown that the level of MMP-9 in the peritoneal fluid can change significantly in a short time. More studies on repeated abdominocentesis could contribute to elucidating the role of MMP-9 as a prognostic indicator.

Regulatory T lymphocytes and selected cytokines in horses with mild to severe equine asthma and different inflammatory cytology profiles.

This study determined Tregs and inflammatory cytokines in BALF and peripheral blood (PB) of adult horses with mild and severe asthma and different BALF cytological inflammation profiles. Horses of diverse breeds with asthma (age range: 2-20 years, n = 24) were divided into groups according to the number of points obtained in a standardized clinical scoring system (mild-moderate equine asthma - MEA, severe equine asthma - SEA) and according to the inflammation type based on cytological finding. Plasma levels of IFN-γ, IL-4, IL-10, IL-17 and MMP-9 in the BALF were determined by ELISA. Tregs in the BALF and PB were detected by flow cytometry. Horses with SEA were older than horses with MEA and had higher concentrations of MMP-9 in the BALF (P < 0.05). The neutrophilic inflammation group was characterised by higher age, grade of clinical score, percentage of Tregs and MMP-9 concentration in the BALF compared to the mastocytic inflammation group (P < 0.05). Age positively correlated with neutrophils, grade of score, Tregs in the BALF and in the PB. The grade of score positively correlated with MMP-9. The percentage of Tregs in the BALF positively correlated with Tregs in the PB. Tregs in the BALF and PB positively correlated with neutrophils and MMP-9 in the BALF and with IFN-γ, IL-4 and IL-10 in the plasma. IFN-γ and IL-10 in the plasma positively correlated with the age. The results indicate that the evaluation of selected parameters in PB, the collection of which is less invasive than the collection of BALF, is of potential importance.

Lymphocyte immunophenotyping in dogs with lymphopenia of common causes.

Lymphocyte immunophenotyping can be useful for evaluating immune competence and predicting the disease prognosis. It is essential to gain knowledge about canine lymphocyte immunophenotypes in various conditions. The study deals with the characteristics of lymphopenia in dogs, with an emphasis on lymphocyte immunophenotyping by flow cytometry. Blood samples from 44 dogs with lymphopenia were included in the study. All lymphopenias sent from veterinary clinics to the diagnostic laboratory were analyzed. The hematological and biochemical abnormalities were investigated, as well as the effect of the age. Lymphopenias were classified according to the level of C-reactive protein (CRP). The percentage of T cells, B cells, Th cells and Tc cells, and T/B and Th/Tc ratios were determined by flow cytometry. Lymphopenias often occurred in dogs over 7 years of age (79.5 %). The most common were postoperative lymphopenia (31.8 %) and inflammatory diseases (29.5 %), most commonly affecting the gastrointestinal tract. Frequent abnormalities were monocytosis (56.8 %), increased CRP (72.7 %) and decreased albumin/globulin ratio (50.0 %). The percentage of Th lymphocytes was significantly lower in the group with elevated CRP than in the group with basal CRP (P = 0.0329). A negative correlation was found between the level of CRP and the percentage of Th lymphocytes (r = -0.3278, P = 0.0390). This study provided new insights into the appearance, incidence and classification of canine lymphopenia.

Changes in rabbit lymphocyte subpopulations and activation following long-term administration of meloxicam.

Meloxicam is a commonly used analgesic in rabbits. However, its possible impact on lymphocyte subpopulations remained unknown. The aim of the study was to investigate a possible effect of long-term administration of meloxicam on rabbit lymphocyte subpopulations. The study included 8 rabbits given meloxicam orally once daily (1 mg/kg BW) for 14 days and 8 rabbits as a control group. Peripheral blood samples were collected on day 0 (before the first dose of meloxicam), day 3, 7 and 14. Samples were evaluated with a haematology analyser and a flow cytometer. A significant decrease in T: B cell ratio was found in all samples taken during meloxicam administration compared to day 0, as well as in comparison with the control group (P < 0.01). A significant increase (P < 0.05) in proportion of CD5 +CD8 + lymphocytes occurred by day 3. Subsequently, although the values slightly decreased, they still remained elevated throughout all the experiment compared to the values from day 0 (P < 0.05). A slight decrease in T and B cell activation (CD5 +CD25 + and IgM+CD25 +) noticed by day 3, declined during the next days of administration and became more and more significant (finally, P = 0.0078). Since a high significant decrease (P < 0.01) in both T and B cell activation as well as a significant increase (P < 0.05) in CD5 +CD8 + T cells proportion were observed after meloxicam administration, a predicted effect of long-term administration of meloxicam on rabbit lymphocytes was confirmed.

Electrolyte, Oestradiol and Progesterone Concentrations in Ovarian Cyst Fluid in Guinea Pigs (Cavia porcellus).

Despite their high prevalence, the aetiopathogenesis of ovarian cysts in guinea pigs (Cavia porcellus) is unknown. We examined tissue samples of cystic ovaries from 20 guinea pigs by histopathology and immunohistochemistry for CD10 antigen, and determined the concentrations of Na+, K+, Cl-, oestradiol and progesterone in the cyst fluid. Cystic rete ovarii were found in all 20 guinea pigs but no follicular cysts. Concentrations of Na+ (142.2 ± 5.04 mmol/l), K+ (5.6 ± 0.76 mmol/l) and Cl- (116.1 ± 5.25 mmol/l) corresponded to extracellular, rather than intracellular, fluid indicating that rete ovarii cysts in guinea pigs are transudative in nature. A positive correlation was found between Na+ and Cl- concentrations (r = 0.7713, P <0.0001). The mean concentrations of oestradiol (61.9 ± 17.44 pg/ml) and progesterone (2.7 ± 5.42 ng/ml) in the cyst fluid were higher than expected because the rete epithelium is not known to have steroidogenic activity. A negative correlation was found between oestradiol and progesterone concentrations in cyst fluid (r = -0.4296, P = 0.0178). Possible pathogenetic mechanisms of cystic rete ovarii in guinea pigs are discussed.

General Caspase Inhibition in Primary Chondrogenic Cultures Impacts Their Transcription Profile Including Osteoarthritis-Related Factors.

OBJECTIVE: The knowledge about functions of caspases, usually associated with cell death and inflammation, keeps expanding also regarding cartilage. Active caspases are present in the growth plate, and caspase inhibition in limb-derived chondroblasts altered the expression of osteogenesis-related genes. Caspase inhibitors were reported to reduce the severity of cartilage lesions in osteoarthritis (OA), and caspase-3 might represent a promising biomarker for OA prognosis. The objective of this investigation was to decipher the transcriptomic regulation of caspase inhibition in chondrogenic cells. DESIGN: Limb-derived chondroblasts were cultured in the presence of 2 different inhibitors: Z-VAD-FMK (FMK) and Q-VD-OPH (OPH). A whole transcriptome RNA sequencing was performed as the key analysis. RESULTS: The analysis revealed a statistically significant increase in the expression of 252 genes in the FMK samples and 163 genes in the OPH samples compared with controls. Conversely, there was a significant decrease in the expression of 290 genes in the FMK group and 188 in the OPH group. Among the top up- and downregulated genes (more than 10 times changed), almost half of them were associated with OA. Both inhibitors displayed the highest upregulation of the inflammatory chemokine Ccl5, the most downregulated gene was the one for mannose receptors Mrc1. CONCLUSIONS: The obtained datasets pointed to a significant impact of caspase inhibition on the expression of several chondro-/osteogenesis-related markers in an in vitro model of endochondral ossification. Notably, the list of these genes included some encoding for factors associated with cartilage/bone pathologies such as OA.

Estimation of long-term costs of postacute care in survivors of the methanol poisoning outbreak.

OBJECTIVES: To fill the existing research gap related to long-term costs of postacute care in methanol poisoning survivors, healthcare cost for 6 years after the outbreak has been modelled and estimated. DESIGN: In a prospective longitudinal cohort study, data collected from 55 survivors of the Czech methanol mass poisoning outbreak in 2012 were collected in four rounds (5 months, then 2, 4 and 6 years after the discharge) in the General University Hospital in Prague according to the same predefined study protocol. The collected data were used to inform the cost model. SETTING AND PARTICIPANTS: All 83 patients discharged from a hospital poisoning treatment after the 2012 methanol outbreak were informed about the study and invited to participate. Fifty-five patients (66%) gave their written informed consent and were followed until their death or the last follow-up 6 years later. The costs were modelled from the Czech healthcare service (general health insurance) perspective. MAIN OUTCOME MEASURES: Long-term national budget impact of the methanol poisoning outbreak, frequencies of sequelae and their average costs. RESULTS: The postacute cost analysis concentrated on visual and neurological sequelae that were shown to be dominant. Collected data were used to create process maps portraying gradual changes in long-term sequelae over time. Individual process maps were created for the central nervous system, peripheral nervous system, sequelae detected during eye examinations and sequelae concerning the visual evoked potentials. Based on the process maps the costs of the postacute outpatient care were estimated. CONCLUSIONS: In 2013-2019 the highest costs per patient related to postacute care were found in the first year; the average costs decreased afterwards, and remained almost constant for the rest of the studied period of time. These costs per patient ranged from CZK4142 in 2013 to CZK1845 in 2018, when they raised to CZK2519 in 2019 again.

Onset of calciotropic receptors during the initiation of mandibular/alveolar bone formation.

Mandibular/alveolar (m/a) bone, as a component of the periodontal apparatus, allows for the proper tooth anchorage and function of dentition. Bone formation around the tooth germs starts prenatally and, in the mouse model, the mesenchymal condensation turns into a complex vascularized bone (containing osteo-blasts, -cytes, -clasts) within only two days. This very short but critical period is characterized by synchronized cellular and molecular events. The m/a bone, as others, is subjected to endocrine regulations. This not only requires vasculature to allow the circulation of active molecules (ligands), but also the expression of corresponding cell receptors to define target tissues. This contribution aimed at following the dynamics of calciotropic receptors´ expression during morphological transformation of a mesenchymal condensation into the initial m/a bone structure. Receptors for all three calciotropic systemic regulators: parathormone, calcitonin and activated vitamin D (calcitriol), were localized on serial histological sections using immunochemistry and their relative expression was quantified by q-PCR. The onset of calciotropic receptors was followed along with bone cell differentiation (as checked using osteocalcin, sclerostin, RANK and TRAP) and vascularization (CD31) during mouse prenatal/embryonic (E) days 13-15 and 18. Additionally, the timing of calciotropic receptor appearance was compared with that of estrogen receptors (ESR1, ESR2). PTH receptor (PTH1r) appeared in the bone already at E13, when the first osteocalcin-positive cells were detected within the mesenchymal condensation forming the bone anlage. At this stage, blood vessels were only lining the condensation. At E14, the osteoblasts started to express the receptor for activated vitamin D (VDR). At this stage, the vasculature just penetrated the forming bone. On the same day, the first TRAP-positive (but not yet multinucleated) osteoclastic cells were identified. However, calcitonin receptor was detected only one day later. The first Sost-positive osteocytes, present at E15, were PTH1r and VDR positive. ESR1 almost copied the expression pattern of PTH1r, and ESR2 appearance was similar with VDR with a significant increase between E15 and E18. This report focuses on the in vivo situation and links morphological transformation of the mesenchymal cell condensation into a bone structure with dynamics of cell differentiation/maturation, vascularization and onset of receptors for calciotropic endocrine signalling in developing m/a bone.

Determination of reference intervals of acid-base parameters in clinically healthy dogs.

OBJECTIVE: To establish reference intervals for traditionally- and Stewart's approach-determined acid-base parameters in a population of clinically healthy dogs. DESIGN: Prospective study (June 2011-September 2012). SETTING: Veterinary teaching hospital. ANIMALS: Two hundred twenty-four client-owned, clinically healthy dogs. INTERVENTIONS: Blood was collected from the jugular vein and the dorsal pedal artery. MEASUREMENTS AND MAIN RESULTS: In the whole blood samples, pH, PCO2 , and PO2 were measured and HCO3- , standard and total bicarbonate, base excess, oxygen content, and alveolar-arterial oxygen differences were calculated. Plasma sodium, potassium, chloride, calcium, albumin, and lactate concentrations were measured and the following parameters were calculated separately for venous and arterial samples: anion gap, anion gap corrected for albumin and phosphate, sodium chloride difference and ratio, chloride gap, strong ion difference, strong ion gap, and unmeasured anions. CONCLUSION: Reference intervals for traditionally- and Stewart's approach-determined acid-base parameters were established for venous and arterial blood in dogs.

C-reactive protein and high mobility group box 1 in dogs with gastric dilatation and volvulus.

OBJECTIVE: To (1) measure C-reactive protein (CRP) and high mobility group box 1 (HMGB1) and (2) evaluate their prognostic value and relationship to severity of systemic inflammatory response syndrome, routine hematological and acid-base parameters in dogs with gastric dilatation volvulus (GDV). DESIGN: Prospective observational study from September 2010 to June 2012. SETTING: Veterinary teaching hospital. ANIMALS: Forty-one client-owned dogs with GDV. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Blood was collected before surgery (baseline), postsurgery, 6-10 hours postsurgery, and 18-22 hours postsurgery. CRP and HMGB1 were measured in all samples, and routine hematological, biochemical, and acid-base analyses were performed. Only baseline and postsurgery samples were used from nonsurvivors (n = 10). CRP increased significantly from postsurgery sampling to 18-22 hours postsurgery, while HMGB1 did not change over time. There was a significant difference in HMGB1 between survivors and nonsurvivors over time. Both proteins correlated with systemic inflammatory response syndrome severity, total leukocyte, segmented neutrophils, and band counts. HMGB1 correlated also with acid-base parameters (pH, bicarbonate, base excess). CONCLUSION: HMGB1 and CRP behaved differently in regards to their kinetic patterns, with HMGB1 appearing to better reflect the severity of tissue injury in dogs with GDV than CRP.

Terlipressin in the treatment of late phase catecholamine-resistant septic shock.

BACKGROUND/AIMS: Norepinephrine, but also dopamine and epinephrine are recommended as first line vasopressors in the treatment of septic shock. In some patients, septic shock deteriorates and becomes to be resistant to catecholamines. In this situation, addition of vasopressin or terlipressin can be advantageous. The aim of our pilot study was to evaluate the impact of terlipressin on open label norepinephrine requirements and mortality. METHODOLOGY: In a randomized, controlled, single centre study we assigned patients with late advanced septic shock refractory to catecholamines (norepinephrine >0.6µg/kg/min for more than 24h) to receive either continuously terlipressin 4mg/24h for 72 hours in addition to open label norepinephrine (TERLI group) or to continue therapy only with catecholamines (CON group). All vasopressor infusions were titrated to maintain a target blood pressure. RESULTS: We enrolled 30 patients, of whom 13 were assigned to terlipressin and in 17 we continued in catecholamine therapy. There was no significant difference in norepinephrine consumption between the groups. Open label norepinephrine infusion rates decreased significantly in the TERLI group as compared with initial consumption, but the decrease we observed only in 7 (54%) patients. There was no significant difference between groups in the rate of death at day 28 (77% in TERLI group and 94% in CON group; p=0.18) or at day 90 (91% vs. 94%; p=0.85). CONCLUSIONS: Continuous terlipressin infusion was not effective in reducing norepinephrine consumption or in the mortality of patients, if administered in late phase of catecholamine refractory septic shock.

Gamma/delta T-cell lymphoma in a dog.

A 15-month-old dachshund was presented for examination because of a cough. Thoracic radiographs revealed the presence of a mass in the upper mediastinum. A diagnosis of gammadelta T-cell lymphoma was made by biopsy and flow cytometry analysis. The dog was treated with chemotherapy and remains asymptomatic after 24 months.

Stress responses in adult cattle due to surgical dehorning using three different types of anaesthesia.

Dehorning adult cattle is a surgical procedure causing distress of varying intensities that can be reflected in behavioural changes and alterations in plasma cortisol levels. Stress responses during the dehorning process were evaluated in 18 Red Pied cows. The cows were divided into 3 groups of six and kept in tie-stall housing. Those in the first group were dehorned under general anaesthesia (GA) induced by intravenous administration of xylazine and ketamine. The second group was dehorned under sedation and local anaesthesia (SLA) induced by intramuscular administration of xylazine and local anaesthesia with lidocaine. The third group was dehorned under local anaesthesia (LA) with lidocaine. Dehorning was performed with a foetotomy wire. Blood samples were taken 0.5 h before dehorning to determine cortisol levels, and, by means of a central venous catheter inserted into the jugular vein, during surgery at 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 5, 6, 7 and 8 h post-surgery. Concurrently, occurrence of stress behaviours was assessed. Cortisol levels were measured by enzyme immunoassay (EIA). The lowest mean peak levels of plasma cortisol (82.53 +/- 6.04 nmol l(-1)), the most rapid return of plasma cortisol levels to baseline values (1.92 +/- 1.11 h), and the lowest occurrence of stress behaviours (2.38 +/- 5.83%) were noted in the SLA group. The highest mean peak levels plasma cortisol (113.86 +/- 25.65 nmol l(-1)), the slowest return of plasma cortisol levels to baseline values (3.83 +/- 2.18 h) and the most frequent occurrence of stress behaviours (65.48 +/- 28.72%) were observed in the LA group. There were significant differences between the SLA and LA groups in peak plasma cortisol levels (p = 0.011) and in occurrence of stress behaviours (p = 0.003). Sedation induced by intramuscular administration of xylazine in conjunction with local anaesthesia with lidocaine is considered the most suitable method of anaesthesia when dehorning adult cattle. Local anaesthesia with lidocaine alone was least suitable.

Comparison of the effect of amphotericin B desoxycholate and amphotericin B colloidal dispersion on renal functions and renal morphology in rats.

BACKGROUND AND AIM: Amphotericin B (AmB) desoxycholate remains as one of the most efficacious agents currently available for the treatment of systemic fungal infections; however, amphotericin B colloidal dispersion (ABCD) has been developed because of AmB desoxycholate nephrotoxicity. The goal of our study was to compare the effect of administration of AmB desoxycholate and ABCD on renal functions and renal morphology in rats. RESULTS: Amophotericin B desoxycholate as well as ABCD causes damage to renal tubuli and polyuria. Amophotericin desoxycholate causes considerably more severe damage to tubuli than ABCD, but the morphological damage to renal glomeruli is minimal in both formulas. In tubular cells, AmB desoxycholate causes severe damage to mitochondria, vacuolation of cytoplasm, and increased values of volume density of peroxisomes. CONCLUSION: None of these formulas causes a decrease in glomerular filtration in rats when animals are properly hydrated.

Stress ulcer prophylaxis in critically ill patients: a randomized controlled trial.

BACKGROUND/AIMS: Critically ill patients especially who require mechanical ventilation or have coagulopathy are at increased risk for stress-related gastrointestinal hemorrhage. There are conflicting data on the efficacy and complication rates of various prophylactic regimens. METHODOLOGY: Our single-center randomized, placebo-controlled study included 287 patients with high risk for stress-related upper gastrointestinal hemorrhage (>48 h mechanical ventilation, coagulopathy). We compared 3 prophylactic regimens (proton pump inhibitor--omeprazole 40 mg i.v. once daily, n=72; H2 antagonists--famotidine 40 mg twice a day, n=71; and sucralfate 1 g every 6 hours, n=69) with placebo (n=75) in patients with trauma or after major surgery. RESULTS: Of 287 assessable patients, clinically significant stress-related upper gastrointestinal bleeding was observed in 1%, 3%, 4%, and 1% of patients assigned to receive omeprazole, famotidine, sucralfate, and placebo, respectively (p>0.28). Bleeding developed significantly more often in patients with coagulopathy compared with the others (10% vs. 2%; p=0.006). The gastric pH (p>0.001) and gastric colonization (p<0.05) was significantly higher in the patients who received pH increasing substances when compared with the other 2 groups. Nosocomial pneumonia occurred in 11% of patients receiving omeprazole, in 10% of famotidine patients, in 9% of sucralfate patients and in 7% of controls (p>0.34). No statistically significant differences were found for days on ventilator, length of ICU stay, or mortality among all the 4 groups. CONCLUSIONS: We could not show that omeprazole, famotidine, or sucralfate prophylaxis can affect already very low incidence of clinically important stress-related bleeding in high-risk surgical intensive care unit patients. Furthermore, our data suggested that especially gastric pH increasing medication could increase the risk for nosocomial pneumonia. Routine prophylaxis for stress-related bleeding even in high-risk patients seems not to be justified.

Helicobacter pylori is not a risk factor for stress ulcer bleeding in polytraumatized patients.

BACKGROUND/AIMS: The pathogenesis of acute stress-related hemorrhage in critically ill polytraumatized patients is uncertain and any role of Helicobacter pylori infection is unknown. The aim of our study was to evaluate the relationship between Helicobacter pylori serological status of patients developing stress-related bleeding and those with no appearance of upper gastrointestinal bleeding. METHODOLOGY: In our single-center prospective cohort study we investigated over a 3-year period all consecutive patients with upper gastrointestinal hemorrhage for Helicobacter pylori infection by serology. Control group comprised 101 such patients with no evidence of hemorrhage. RESULTS: Of 396 assessable patients, stress-related upper gastrointestinal bleeding was observed in 11 (3.1%) patients. Six (55%) of the 11 bleeding patients and 45 (45%) of the 101 control group patients had serological evidence of Helicobacter pylori infection (p=0.5). Bleeding developed significantly more often in patients with more serious injury (for ISS, p=0.04, for TRISS p=0.03). Bleeding patients showed insignificantly higher mortality (36% vs. 15%; p=0.09). CONCLUSIONS: Helicobacter pylori infection was not significantly more prevalent in polytraumatized patients with hemorrhage when compared with control group. Our data suggest that the infection with Helicobacter pylori does not play an important role in bleeding, indicating no causative role for it in upper gastrointestinal hemorrhage of polytraumatized patients. The incidence of bleeding is low and bleeding develops usually later, in the meantime is the Helicobacter pylori infection eradicated with the antibiotics used for another purpose.

Componentes químicos selectos de biopsia ósea y sangre durante osteopatías metabólicas subclínicas en bovinos / Selected chemical constituents in bone biopsies and blood during subclinical metabolic osteopathies in bovines

Se determinaron componentes químicos en biopsias de huesos y sangre en toros de engorda y vacas lecheras sanos, y durante osteopatías metabólicas subclínicas. Al muestreo los toros (n = 143 Holstein X Czech Spotted Breed) de 10-15 meses de edad tenían un peso de 300 a 450 kg. Las vacas (n = 118, Czech Spotted Breed), 3-8 años de edad, se encontraban entre dos y 20 semanas posparto. El grupo I de toros (testigo, n = 47) y el grupo de vacas (testigo, n = 57), procedentes de cuatro hatos cada uno. Ninguno de los animales de los hatos presentaban signos clínicos de osteopatías. El grupo II de toros (n = 96) sin signos clínicos de osteopatías provenía de ocho hatos con frecuencias de raquitismo en forma clínica del 21 por ciento al 50 por ciento en otros animales; el grupo II de vacas (n = 61) sin signos clínicos de osteopatías se seleccionó de cinco ranchos con osteopatías generalizadas en 10 por ciento a 18 por ciento de otros animales. La ración alimentaria de los animales de los grupos I estaba balanceada. Para los animales de los grupos II era deficiente en fósforo; adicionalmente, las vacas del grupo II tenían exceso de calcio en la ración. Se tomaron biopsias de hueso en cresta iliaca y muestras de sangre una vez en cada animal. Los valores de Ca2+, P inorgánico y del equilibrio ácido-base en sangre no tuvieron significancia para el diagnóstico de osteopatías subclínicas. En las biopsias de cresta iliaca se determinaron disminuciones significativas (P < 0.05) en g de ceniza/g de materia seca libre de grasa (MSLG) y g de ceniza/cm3 de tejido esponjoso en animales de los grupos II en comparación con los grupos I (testigo). En los toros del grupo II se diagnosticó raquitismo debido a la disminución de cenizas/g de MSLG. Se determinaron osteomalacia y osteoporosis subclínicas en vacas del grupo II por valores disminuidos de cenizas/g de MSLG y por cm3 de tejido esponjoso. El análisis químico de biopsias óseas demostró efectividad para diagnosticar osteopatías metabólicas subclínicas en bovinos.