Andreza Amaral da Silva

Osteoarthritis (OA) is a degenerative joint disease that affects humans and animals worldwide. Its early diagnosis remains challenging due to subtle clinical signs and late radiographic changes. This study aimed to explore candidate biomarkers associated with spontaneous OA and to investigate their correlation with ultrasonographic scores to support early diagnosis. Clinical, radiographic, and ultrasonographic evaluations were performed on 52 equine metacarpophalangeal joints, with and without OA, allowing joint scoring and classification into osteoarthritis (OAG) and control groups. Synovial fluid samples were analyzed for cartilage degradation (C2C), untargeted ¹H NMR-based metabolomics, and lipid peroxidation (TBARS). Statistical analyses included Student’s t-test, Mann–Whitney U test, univariate and multivariate metabolomic analyses, and Spearman’s correlation (p < 0.05). Ultrasonography revealed higher scores in the synovial fold, membrane, and fluid, indicating synovitis as the predominant finding in the acute phase. C2C and TBARS concentrations were significantly higher in the OAG. Seven metabolites differed between groups, with citrate and TBARS showing the strongest correlations with ultrasonographic scores. These findings suggest increased metabolic activity and lipid peroxidation in early OA and highlight citrate and TBARS as potential auxiliary biomarkers for early diagnosis associated with synovitis.

Goats have been proposed for hyperimmune serum production (HSP) in vaccine quality control, and ozone therapy may enhance immunoglobulin levels. This study aimed to establish an HSP protocol and evaluate whether major ozone autohemotherapy (O3-MAHT) could increase antibody titers in goats. A 2×2 crossover design was used, involving 10 goats inoculated with attenuated mumps (n=5) or measles (n=5) antigens. Twelve hours after booster inoculation, 240mL of whole blood were collected into bags, which either received 240mL of an O2/O3 gas mixture (20μg/mL; Gozone; n=5) or remained untreated (Gcontrol; n=5) before reinfusion. TBARS, MDA, complete blood count, and anti-measles and anti-mumps antibody titers were analyzed. A time effect was observed for TBARS, MDA, HCT, TPP, WBC, lymphocyte, segmented neutrophil, and monocyte indices. Antibody titers were affected by the study phase. In phase 1, there was a trend for time, treatment, and time × treatment interaction effects on viral particle inactivation. Additionally, Gozone tended to show greater viral particle inactivation at T21 compared to Gcontrol. All goats responded to the HSP protocol, and O3-MAHT showed a potential trend toward improving viral particle inactivation, supporting further investigation.

Cryoprotectants are required to reduce damage caused to the cells due to low temperatures during the cryopreservation. Antifreeze proteins (AFP) have a well-known role in cell membrane protection, while resveratrol is a potent antioxidant. This study assessed the effect of the association of resveratrol concentrations and AFP I in a ram semen extender. Pooled semen of four rams was allocated into six treatments in a factorial arrangement: (CONT, only the semen extender); only AFP I (ANT: 0.1 µg/mL of AFP I), only resveratrol, one treatment with two levels (10 µM/mL or 50 µM/mL of resveratrol); and two treatments with the interactions, with one AFP I and one of the two levels of resveratrol (0.1 µg/mL of AFP I with 10 µM/mL resveratrol; 0.1 µg/mL of AFP I with 50 µM/mL resveratrol). No interaction between factors was observed on sperm kinetics, plasma membrane integrity, hypo-osmotic test, and mitochondrial activity parameters. There was a high probability (P = 0.06) of reducing sperm cells with functional membrane percentage in the hypo-osmotic test and increasing the percentage of sperm with high mitochondrial activity (P = 0.07) was observed in AFP presence. An interaction of AFP and resveratrol was observed in non-capacitated sperm (P = 0.009), acrosomal reaction (P = 0.034), and sperm binding (P = 0.04). In conclusion, the association of resveratrol and AFP did not improve the quality of frozen-thawed semen and even promoted deleterious effects compared to their single addition in the semen extender. The supplementation of 50 µM/mL of resveratrol improved the outcomes of frozen-thawed ram sperm, being a potential cryoprotectant.

The sheep experimental model is increasingly being used in laparoscopy, and there is a need to ensure animal welfare during and after the procedures . Therefore, the present study aims to evaluate postprocedural clinical and laboratory variables of sheep submitted to different intrabdominal pressures during laparoscopy. Ten female nonpregnant healthy adult Santa Ines ewes (48.8 ± 5.6 kg) were randomly submitted to a crossover study using different intrabdominal pressures (IAPs): 0 mmHg (G1), 10 mmHg (G2), 12 mmHg (G3), and 15 mmHg (G4). They were anaesthetised and mechanically ventilated (VT=15 ml.kg−1) with positive end-expiratory pressure (3 cmH2O) and FiO2=1.0. At the same time, pneumoperitoneum was induced and maintained by Hasson's trocar cannula attached to an insufflator with no invasive visceral intervention. Physical examination and malondialdehyde levels were gathered immediately before (T0), 24h (T1), 72h (T2), 7 days (T3) and 14 days (T4) after procedure. Haematology and fibrinogens variables were gathered at T0, T2 and T3, while serum biochemistry at T0 and T2. In the group with 15 mmHg of IAP, fibrinogen levels suggest a late inflammatory response persisted for up to 7 days (from 310±179 to 610±335 mg/dL). Malondialdehyde levels in the pneumoperitoneum persisted in ascending levels for up to 14 days (from 257.2±136.5 to 369.7±201.4 nM/g Hb). White blood cell (WBC), neutrophils, lymphocytes, and N:L ratios increased above normal levels in 72h post-procedure and returned to normal within 7 days, except in the group of 15 mmHg of IAP which the highest value was reached 7 days post-procedure. Alterations in clinical parameters were mild and within normal values for the specie and breed in all groups. Biochemical analysis for all parameters was within the normal range in all groups. Still, monitoring aspartate aminotransferase (AST) up to 7 days after laparoscopic procedures is advisable due to its 72h increase. Although no significant clinical and haematological alterations were observed, biochemical alterations and oxidative stress were greater up to 14 days after the procedure in the group with 15 mmHg IAP.

The experiment was conducted at Araí & Zumbi farm on sixty healthy Dorper ewes to compare blood glucose, hormonal profile, and insulin resistance evaluation in sheep from conception until 48 h postpartum in single and twin pregnancies. All experimental ewes raised under semi-intensive management system. Sixty animals were selected from 150 estrous synchronized and pregnant ewes. The animals were divided into two groups based on single (G1, n = 30) and twin pregnancies (G2, n = 30). Blood samples were collected at nine time points: immediately after fixed-time artificial insemination (D0); at 30 days (D30), 90 days (D90), 120 days (D120), 130 days (D130), and 140 days (D140) of pregnancy; on the delivery day (DD); and at 24 h (PD1) and 48 h (PD2) postpartum. The results of blood glucose, insulin, glucagon, cortisol, and thyroid hormones (T3 and T4) levels showed significant differences over the analyzed sample times; however, only cortisol showed differences within groups, with the G1 having higher values than the G2 group. The interaction of the groups × nine sample times showed a significant result (P = 0.001) only for glucagon. The number of fetuses directly interfered with the glucagon profile throughout gestation. The glucose, cortisol, glucagon, and the homeostatic model assessment for insulin resistance (HOMA IR) concentrations increased at DD and decreased at PD1 and PD2. T3 and T4 showed different behaviors among the sample times. T3 values presented a decrease from D0 to D90, followed by an increase from D90 to DD. Otherwise, for T4 values, a decrease from D90 to D130 was observed, followed by an increase from D130 to D140. Despite the changes found in the endocrine system and metabolism in Dorper ewes throughout pregnancy, the nutritional management ensured a healthy status during pregnancy, delivery, and postpartum in single and twin gestation, whose HOMA IR profiles remained identical.

The hematopoietic system changes during the pregnancy to carry fetal development and maternal needs. This study compared the hematological parameters between ewes with single and twin pregnancies during gestation, delivery, and postpartum. The experiment was conducted on 60 healthy pregnant Dorper ewes that were divided into two experimental groups: Group 1 (G1), with single pregnancies (n=30), and Group 2 (G2), with twin pregnancies (n=30). Blood samples were collected from all ewes at different times: immediately before fixed-time artificial insemination (AI); on day 30, 90, 120, 130, and 140 of pregnancy; immediately after delivery; and at 24h and 48h postpartum. Statistical analysis compared the two groups at different times (P<0.05). Mild, normocytic, and hypochromic anemia was detected in all ewes from AI time and throughout pregnancy from both groups, but did not prove to be of clinical relevance. In the peripartum stage (from the 140th day of pregnancy to 48h postpartum), the ewes with twin pregnancies (G2) exhibited higher erythrogram values and neutrophil:lymphocyte ratio than did ewes with single pregnancies (G1). This indicated a greater hematopoietic adaptation in the body during the development of two fetuses. Except for the eosinophil numbers, all leukogram parameters were influenced by pregnancy in a similar way in both groups, and was characterized mainly by leukocytosis with neutrophilia during peripartum due to the high presence of endogenous cortisol at delivery. Thus, these findings showed that pregnancy was a stressful physiological event that increased the leukocyte count with a slight alteration in the erythrogram of Dorper ewes.

RESUMO Nas ovelhas, a demanda de nutrientes aumenta durante a gestação, especialmente nas últimas seis semanas, quando ocorre maior desenvolvimento do feto. Os objetivos deste trabalho foram estudar o perfil bioquímico de ovelhas durante a gestação e no periparto imediato, comparando-se gestação única com gemelar. Foram utilizadas 60 ovelhas, Dorper, divididas em dois grupos experimentais com base no diagnóstico de gestação ultrassonográfico: grupo 1: 30 ovelhas com feto único; grupo 2: 30 ovelhas com gestação gemelar. Em nove momentos de coleta, foram mensurados no soro sanguíneo: ureia, creatinina, proteína total (PT), albumina, globulina, aspartato aminotransferase (AST), gamaglutamiltransferase, fosfatase alcalina, colesterol e triglicérides. O peso vivo das ovelhas do G2 foi maior que do G1 em todos os momentos, inclusive quando estavam vazias. Houve diferença entre as ovelhas do G1 e do G2 nos valores de PT, albumina, AST e triglicérides. A partir de 120 dias de gestação, o perfil proteico se modificou, com menores níveis de ureia, PT, albumina e globulina, mostrando que a demanda metabólica se intensificou no último mês da gestação. Os níveis de colesterol e triglicérides diminuíram de 140 dias de gestação para o parto. Este trabalho mostrou que a gestação proporcionou mudanças significativas no metabolismo da ovelha, principalmente no terço final da gestação, quando há maior necessidade de acompanhamento para evitar a incidência de doenças metabólicas.

This study aimed to assess the effect of percutaneous transthoracic lung biopsy on the oxidative metabolism of sheep by measuring the oxidative stress markers of superoxide dismutase (SOD), total glutathione (GSH-t), peroxidase (GSH-Px) and thiobarbituric acid reactive substances (TBARS) in the red cells of these animals. Blood samples were collected from 20 clinically healthy sheep prior to, and 30 min after, percutaneous transthoracic lung biopsy. After biopsy, there was a significant decrease (p < 0.05) in SOD and GSH-Px activity, with no significant change (p ≥ 0.05) in GSH-t and TBARS concentrations. These results showed that percutaneous transthoracic lung biopsy did not significantly affect the oxidative metabolism of sheep 30 min after the procedure, which may be used widely in this species without causing serious tissue damage.

Estudou-se a broncopneumonia de ocorrência natural em bezerros e a influência da administração de acetato de DL-α-tocoferol na profilaxia e tratamento desta enfermidade. Foram avaliados 60 bezerros machos, com idade máxima de 10 dias, divididos em dois grupos experimentais: GSV (grupo sem vitamina) e GCV (grupo com vitamina). A suplementação com dose única de 4.500 UI de acetato de DL-α-tocoferol por via intramuscular (IM) foi precedida por exame físico, perfil hematológico, determinação de proteínas séricas, globulinas, gamaglutamiltransferase e citologia do lavado traqueobrônquico (D0) para verificar a sanidade e homogeinização dos grupos. Os bezerros foram mantidos em bezerreiro e distribuídos aleatoriamente, 2 a 2 em baias individuais de 2,40m², onde permaneceram até o 21º dia (D0 ao D21). Foram avaliados por exame físico diário e, na presença de sinais clínicos indicativos de broncopneumonia (DX), retirados do bezerreiro, avaliados por hemograma e citologia do lavado traqueobrônquico, e tratados com enrofloxacina (5mg/kg, IM). Durante o tratamento, o exame físico foi realizado diariamente, e o hemograma e a citologia do lavado traqueobrônquico repetidos uma semana após seu término (DY). Nos bezerros sadios foram repetidos hemograma e citologia do lavado traqueobrônquico, no último dia (D21) do experimento. Não houve diferença significativa quanto às variáveis gamaglutamiltrasfrase (p=0,09), proteínas séricas (p=0,27) e globulinas (p=0,10) e a idade dos bezerros (p=0,15) entre os grupos. Os bezerros do grupo GSV e GCV levaram em média 11 e 12 dias para adoecerem, respectivamente. Contudo, não houve diferença estatística significativa entre os grupos (p=0,68). Em 34 bezerros (56,66%) foi diagnosticado broncopneumonia, sendo que 17 destes bezerros (50%) pertenciam ao grupo GCV e 17 (50%) ao grupo GSV. Com relação aos sinais clínicos, não houve diferença significativa entre os grupos em nenhum dos momentos estudados. Dos 34 bezerros que adoeceram 73,52% morreram pela doença, sendo 64,7% do GCV e 82,35% do GSV. A citologia do lavado traqueobrônquico não apresentou diferença significativa entre os grupos em nenhum dos momentos estudados (D0, DX, DY e D21). A suplementação com vitamina E não teve influência na profilaxia e no tratamento de broncopneumonia de bezerros.