Artigos - Reprodução Animal e Radiologia Veterinária - FMVZ

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Evaluation of biochemical and electrolytic components of semen from ram supplemented with different concentrations of selenium and its correlation with sperm quality
(2022-01-01) Silva, D. C. [UNESP]; Codognoto, V. M. [UNESP]; Piagentini, M. [UNESP]; Dantas, A. [UNESP]; Sousa, G. C. [UNESP]; Silva, L. S. [UNESP]; Souza, E. R. [UNESP]; Filho, R. A. Almeida [UNESP]; Denadai, R. [UNESP]; Oba, E. [UNESP]; Universidade Estadual Paulista (UNESP)
The aim of the study was to evaluate whether supplementation with different levels of selenium (Se) can change the biochemical and electrolytic components of semen, causing an improvement in seminal quality in rams. Thirty animals were kept in an intensive pen system, fed with hay and commercial ration, allocated into five groups (six animals/pen) and supplemented with a mineral mixture containing 0mg (G1), 5mg (G2), 10mg (G3), 15mg (G4) and 20mg (G5) of Se/kg. Each group received a different treatment every 56 days and treatments were rotated between groups following a dynamic sequence. Semen samples were collected by electroejaculation after the end of each treatment to evaluate the levels of fructose, citric acid, potassium (K), sodium (Na), calcium (Ca), Se, zinc (Zn), manganese (Mn), sulfur (S) and lead (Pb). The statistical design was a 5x5 Latin square. The different levels of Se supplementation evaluated maintained the concentrations of electrolytes and minerals in the semen at the required levels and did not change the sperm quality, concluding that higher intakes of Se do not cause antagonistic effects on the absorption and subsequent action of other essential minerals supplied to the animals and still maintains electrolyte balance
Ability of donkey sperm to tolerate cooling: Effect of extender base and removal of seminal plasma on sperm parameters and fertility rates in mares
(2022-09-26) Gobato, Mariana L. M. [UNESP]; Segabinazzi, Lorenzo G. T. M. [UNESP]; Scheeren, Verônica F. C. [UNESP]; Bandeira, Rafael S. [UNESP]; Freitas-Dell'Aqua, Camila P. [UNESP]; Dell'Aqua, José A. [UNESP]; Papa, Frederico O. [UNESP]; Universidade Estadual Paulista (UNESP); Ross University School of Veterinary Medicine
Artificial insemination using cooled-transported semen has marked importance in equine breeding programs around the world, and the high value of mules has generated avid interest in donkey semen biotechnology. However, donkey semen cools poorly in commercially available equine extenders. Therefore, this study aimed to develop approaches to improve the ability of donkey semen to tolerate cooling. Ejaculates of seven donkeys (n = 21) were cooled at 5°C for 48 h in three different extenders (milk-based, SM; sodium caseinate-based, SC; or egg yolk-based, EY) in the presence or absence of seminal plasma (centrifugation, C). Sperm motility, plasma membrane integrity (PMI), plasma membrane stability (PMS), mitochondrial membrane potential (HMMP), intracellular hydrogen peroxide (H2O2), and intracellular superoxide ((Formula presented.)) were assessed before, 24 h, and 48 h post-cooling. In addition, 15 mares (163 estrous cycles) were randomly inseminated with semen from two jacks (Jack 1, n = 90; Jack 2, n = 73) previously cooled for 24 h under one of the treatments (SM, SC, EY, SM-C, SC-C, or EY-C). Groups EY, SC-C, and EY-C (P < 0.05) demonstrated superior sperm analytical parameters to SM at 24 and 48 h. Centrifugation positively affected sperm analytical parameters in cooled donkey semen extended in SM and SC (P < 0.05). Mares bred with semen extended in SC (67%, 18/27), SC-C (89%, 24/27), EY (89%, 25/28), or EY-C (74%, 20/27) had significantly greater conception rates than mares bred with SM (33%, 9/27; P < 0.05). Mares bred with SM-C had intermediate conception rates (59%, 16/27). In conclusion, SC and EY improved the cooling ability and fertility of donkey semen in horse mares, and centrifugation positively affected donkey semen extended in SM.
Beneficial Impact of Hypercapnic Conditions During Early Incubation on Broiler Hatchability. Embryo Mortality and Postnatal Performance
(2023-01-01) Kroetz Neto, Felipe Lino; Gonzales, E. [UNESP]; Novaes, G. A.; Pereira, R. J.G.; Aviagen America Latina Ltda; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP)
We assessed the extent to which CO2 levels altered different hatching and chick parameters. In Experiments 1 and 2, a total of 16,184 eggs from Cobb 500 breeders were incubated in single stage incubators under three different conditions: (a) standard ventilated incubator (CON, Exp.1 and 2); (b) increasing CO2 levels during the first 10 days of incubation until 0.7% (V7000, Exp. 1) and (c) until 0.8% (V8000, Exp. 2). High levels of CO2 improved hatchability, possibly due to lower embryo mortality from ED18 to ED21. Internal and external pipping in experiment V8000 started later than in CON; nevertheless, the hatch still occurred before in V8000 as a result of the shorter durations of external pipping and hatch. In Experiment 3, a total of 12,138 eggs from Cobb 500 were incubated in single stage incubators under three different conditions: (a) standard ventilated incubator (CON); (b) increasing CO2 levels until 1.0% with ventilation (V10000); and (c) increasing CO2 levels until 1.0% without ventilation (NV10000). Hypercapnic conditions led to better hatchability and lower embryo mortality from ED18 to ED21. Internal pipping started earlier in NV10000, but only V10000 differed from CON in terms of the average time for hatch. Hypercapnic groups also showed shorter durations of external pipping and hatch when compared to CON. Post-hatch analysis revealed no differences among incubation conditions in terms of body weight gain, feed conversion ratio, mortality by sudden death syndrome, and production factor. Nevertheless, V10000 showed a lower mortality by ascites and a better viability when compared to CON, while NV10000 presented a higher mortality by other causes. Altogether, our findings indicate that in addition to not being detrimental to embryo survival, high CO2 levels reduce embryonic mortality at 18-21 days of incubation and increase hatchability.
The Challenge of Coexistence: Changes in Activity Budget and Ranging Behaviour of Brown Howler Monkeys in Response to the Presence of Conspecifics and Heterospecifics
(2023-01-01) Sobral, Gisela; Fuzessy, Lisieux F. [UNESP]; de Oliveira, Claudio Alvarenga; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP)
The presence of other animals, both conspecifics and heterospecifics, is a major driving force for how animals organize themselves in space and time. Although theoretical models are available to explain the role of each in animal movement, fine-scale assessments of daily movement are scarce, particularly for primates. Hence, our goal was to assess whether and how the presence of conspecifics and heterospecifics influence spatiotemporal landscape use in two, wild, howler monkey (Alouatta guariba) groups. We followed the groups for 14 months in a large, continuous forest, during which we recorded their daily path length (DPL), home range, activity budget, feeding, and the presence of other groups (conspecifics) and other species (heterospecifics). The two groups differed in DPL, home range, proportion of fruits ingested, and time devoted to moving and resting. Partial least squares path modelling showed that variation in DPL was explained by the percentage of leaves or fruits ingested and by the presence of conspecifics, but not of heterospecifics. Group differences in several ecological variables emphasise the need to conduct further studies of space use with more groups in the same area to understand the underlying mechanisms of these differences. Moreover, our analysis shows that within-species interactions may be a stronger force in spatiotemporal organisation than interspecies interactions, at least in this folivorous primate. This is relevant from both a theoretical standpoint, and also when considering the consequences of habitat fragmentation and reduction. Deforestation leads to decreased resource availability and increased likelihood of encounters with conspecifics, which ultimately alters the proportion of food items ingested and increases the DPL, disrupting energy balance.
All-triploid offspring in the yellowtail tetra Astyanax altiparanae Garutti & Britski 2000 (Teleostei, Characidae) derived from female tetraploid × male diploid crosses
(2023-04-01) Alves, Andreoli Correia; Yasui, George Shigueki; do Nascimento, Nivaldo Ferreira; Monzani, Paulo Sérgio [UNESP]; Senhorini, José Augusto [UNESP]; Pereira Dos Santos, Matheus; Federal Rural University of Rio de Janeiro (UFRRJ). Animal Science Graduate Programme; Centro Nacional de Pesquisa e Conservação da Biodiversidade Aquática ContinentalTA/ICMbio; Universidade de São Paulo (USP); Unidade Acadêmica de Serra Talhada; Universidade Estadual Paulista (UNESP)
This study aimed to evaluate the ploidy and survival of larvae resulting from crosses between tetraploid females and diploid males of yellowtail tetra Astyanax altiparanae, both females (three diploids and three tetraploids) and males (n = 3 diploids). Breeders were subjected to hormonal induction with pituitary gland extract from common carp fish (Cyprinus carpio). Females received two doses at concentrations of 0.3 and 3.0 mg/kg -1 body weight and at intervals of 6 h. Males were induced with a single dose of 3.0 mg/kg -1 applied simultaneously with the second dose in females. Oocytes from each diploid and tetraploid female were fertilized with semen from the same male, resulting in two crosses: cross 1 (diploid male and diploid female) and cross 2 (diploid male and tetraploid female). The procedures were performed with separate females (diploid and tetraploid) and diploid males for each repetition (n = 3). For ploidy determination, 60 larvae from each treatment were analyzed using flow cytometry and cytogenetic analyses. As expected, flow cytometry analysis showed that progenies from crosses 1 and 2 presented diploid and triploid individuals, respectively, with a 100% success rate. The same results were confirmed in the cytogenetic analysis, in which the larvae resulting from cross 1 had 50 metaphase chromosomes and those from cross 2 had 75 chromosomes. The oocytes have a slightly ovoid shape at the time of extrusion. Diploid oocytes had a size of 559 ± 20.62 μm and tetraploid of 1025.33 ± 30.91 μm. Statistical differences were observed between eggs from crosses 1 and 2 (P = 0.0130). No significant differences between treatments were observed for survival at the 2-cell stage (P = 0.6174), blastula (P = 0.9717), gastrula (P = 0.5301), somite (P = 0.3811), and hatching (P = 0.0984) stages. In conclusion, our results showed that tetraploid females of the yellowtail tetra A. altiparanae are fertile, present viable gametes after stripping and fertilization using the 'dry method', and may be used for mass production of triploids. This is the first report of these procedures within neotropical characins, and which can be applied in other related species of economic importance.
Use of protein kinase C and phospholipase A2 inhibitors in bovine endometrial cells treated with estradiol and calcium ionophore
(2021-03-23) Maldonado, Mariângela Bueno Cordeiro [UNESP]; Henry, Francine Messias Ciríaco; Ferreira, Teissiane Fernanda de Vasconcelos [UNESP]; Mello, Barbara Piffero; Binelli, Mario; Membrive, Claudia Maria Bertan [UNESP]; Universidade Estadual Paulista (UNESP); Texas Tech University; Universidade de São Paulo (USP); University of Florida
The release of endometrial prostaglandin-F2α (PGF2α) in bovine females can be induced in vivo by estradiol (E2). However, its role in this mechanism has not been clarified. We hypothesized that E2 stimulates the activity and abundance of protein kinase C (PKC) and phospholipase A2 (PLA2). Our objective in this study was to analyze the effects of PKC and PLA2 inhibitors on PGF2α synthesis induced by E2 and calcium ionophore (CI) in bovine endometrial cells (BEND cells; Experiment 1). Additionally, we evaluated the abundance of PKC and PLA2 in endometrial explants of cows treated or not with E2 17 days after estrus (D17, D0 = estrus; Experiment 2). In Experiment 1, BEND cells were submitted to a PKC inhibitor (10 µM of C25 H24 N4 O2; bisindolylmaleimide I, or BIS I), a PLA2 inhibitor (20 µM of arachydoniltrifluoromethane or AACOCF3), or none. The BEND cells were subsequently treated with E2 and CI, and PGF2α concentrations were measured in the culture medium through radioimmunoassay. For DIF-12 (PGF2α concentration 12 h after treatment subtracted from PGF2α concentration at hour 0), no PKC inhibitor effect was observed (P= 0.2709). However, DIF-12 was lower (P < 0.05) for groups treated with the PLA2 inhibitor and PLA2 inhibitor + CI + E2 groups than the control and CI + E2 groups. Thus, AACOCF3 was an efficient PLA2 inhibitor in the BEND cells culture system, and E2 did not stimulate the synthesis of PKC and PLA2. In Experiment 2, cyclic Nellore heifers received none (n = 5) or 3 mg (n = 6) of 17β-E2 on D17 and were slaughtered 2 h after administration. The abundance of PKC and PLA2 in the endometrial tissue was evaluated using Western blotting analysis. No E2 effect was observed on PKC (P = 0.08) and PLA2 (P = 0.56). We concluded that E2 did not stimulate the activity and abundance of PKC and PLA2.
Eleven-year retrospective analysis of acquired diaphragmatic hernia in 49 dogs and 48 cats
(2023-02-01) Pereira, Geovane J.; Rahal, Sheila C.; Melchert, Alessandra; Abibe, Rebeca B.; Brandão, Cláudia Valéria S; Quitzan, Juliany G.; Mesquita, Luciane R.; Mamprim, Maria J.; Universidade Estadual Paulista (UNESP)
Data on dogs and cats that underwent surgery for a traumatic diaphragmatic hernia were retrospectively collected and analyzed, with an objective to identify factors that influenced the survival rate. Forty-nine dogs and 48 cats were included. The predominant respiratory clinical sign was dyspnea, and the interval from trauma to development of clinical signs did not influence patient survival. Concurrent orthopedic and/or soft tissue injuries were identified in 48.45% of the animals. Intraoperative complications occurred in 14 dogs and 5 cats, and postoperative complications in 7 dogs and 6 cats. Intraoperative death occurred in 6.2% and postoperative death in 8.3% from 1 h to 10 d after surgery. Animals that received surgical treatment within 48 h after diagnosis had a lower risk of death. Conversely, concurrent injuries and intraoperative and postoperative complications were the main factors associated with a higher risk of death.
Unilateral orchitis-epididymitis and diffuse periorchitis in a dog due to β-haemolytic streptococcal infection
(2023-01-01) Fuchs, Kárita da Mata [UNESP]; de Sousa, Gabriela Carneiro [UNESP]; Denadai, Renan [UNESP]; de Souza, Fabiana Ferreira [UNESP]; de Meira, Cezinande [UNESP]; Universidade Estadual Paulista (UNESP)
Epididymitis can be associated with orchitis and periorchitis, and can be classified as acute or chronic, and of an infectious or non-infectious aetiology. This study aims to describe a case of orchitis-epididymitis and diffuse periorchitis due to β-haemolytic streptococcal infection. The dog presented at the veterinary service with an enlarged, hyperaemic scrotum with regional acute pain, lethargy and hyporexia. In the ultrasound exam, an abnormal amount of peritesticular anechoic fluid and enlargement of the left epididymis was observed. Treatment consisted of a bilateral orchiectomy and scrotal ablation. The histopathological exam indicated left orchitis-epididymitis and diffuse periorchitis, with neutrophilic inflammatory cell infiltrate and the presence of coccoid bacteria. The microbiological exam isolated β-haemolytic streptococci. Bilateral orchiectomy and the use of a specific antimicrobial was effective in treating the condition.
Effects of anethole supplementation on bovine embryo production and quality
(2023-08-01) Janini, Ludimila Cardoso Zoccal [UNESP]; Dellaqua, Thaisy Tino [UNESP]; Membrive, Claudia Maria Bertan [UNESP]; Oba, Eunice [UNESP]; Nichi, Marcilio; Rizzoto, Guilherme [UNESP]; da Cruz Landim-Alvarenga, Fernanda [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP)
Improvements are necessary to further expand the potential of in vitro embryo production (IVP). Anethole, a natural antioxidant, was shown to be a low-cost alternative with an important role in the gene modulation of oxidative stress, and it can be used to produce higher quality embryos. The aim of this study was to evaluate the anethole effect during in vitro maturation (IVM) on the oocyte nuclear maturation, and then, during IVM and/or in vitro culture (IVC) on the production and quality of bovine embryos. In Experiment 1, cumulus-oocyte complexes (COCs) were matured in IVM medium in the following groups: control group (MC); supplemented with 300 µg/ml anethole (M300); and supplemented with 3,000 µg/ml anethole (M3000). The three groups were matured for 24 h to assess nuclear maturation (metaphase II) and gene expression of cumulus cells (CCs). The group that obtained the best response was M300, which was chosen for Experiment 2, together with MC. In this experiment, maturation was carried out either in IVM medium with 300 µg/ml anethole (M300) or not (MC). Subsequently, the COCs were submitted to in vitro fertilization and then divided into four groups for IVC: MC–CC (IVM and IVC controls); MC–C300 (control IVM and IVC with 300 µg/ml); M300-CC (IVM with 300 µg/ml and control IVC); and M300-C300 (IVM and IVC with 300 µg/ml anethole each). On the third day (D3) of the culture, cleavage was evaluated, and on D8, the production and classification of blastocysts were assessed. It was observed that 3,000 µg/ml anethole reduced the percentage of COCs that had reached nuclear maturation. The genes GREM1, GSTA1, SOD1, and CAT, related to oxidative stress resistance, and COX2 and HAS, involved in cell metabolism, showed increased abundance in CCs in the anethole presence. Regarding embryo production, the use of 300 µg/ml anethole during IVM and IVC improved the percentage of cleaved embryos. In addition, anethole treatment during IVM and IVC increased the expression of GPX1 gene while decreasing IFNT2α gene in the embryos produced. In conclusion, the addition of 300 µg/ml anethole during IVM modulated the expression of genes related to oxidative stress and oocyte development, and its use during IVM and IVC modulated the gene expression related to embryo quality. However, further studies need to be performed to better understand anethole's impact on embryo development.
Renal resistive index in obese and non-obese cats
(2022-01-01) de Souza, Fúlvia Bueno [UNESP]; Gonçalves, Natália Volpi [UNESP]; Bonatelli, Shayra Peruch [UNESP]; Belotta, Alexandra Frey [UNESP]; Geraldes, Silvano Salgueiro [UNESP]; Mamprim, Maria Jaqueline [UNESP]; Guimaraes-Okamoto, Priscylla Tatiana Chalfun [UNESP]; Lourenço, Maria Lúcia Gomes [UNESP]; Ramos, Paulo Roberto Rodrigues [UNESP]; Rahal, Sheila Canevese [UNESP]; Melchert, Alessandra [UNESP]; Universidade Estadual Paulista (UNESP)
This study aimed to compare renal function between obese and normal-weight healthy cats, using intrarenal resistive index (RI), serum symmetric dimethylarginine (SDMA), and serum creatinine, and to identify the variables that might influence intrarenal RI. Thirty crossbred client-owned cats met the inclusion criteria and were allocated into two groups: Control and Obese. Body weight, body mass index (BMI), body condition score (BCS), SAP, serum SDMA, urea, and creatinine were evaluated. B-mode and Doppler ultrasound of the kidneys were done. RI evaluation was in the interlobar artery. SDMA and intrarenal RI were compared between groups, also considering the gender of the cats. A correlation analysis between intrarenal RI with the other parameters was performed. SDMA was higher in the Obese group. Intrarenal RI was higher in females than males in the Obese group. Obese females presented higher RI and SDMA than Control females. A positive correlation was observed between RI, age, body weight, and BMI. Six obese cats (40%) showed increased RI. The increase in body weight, BCS, and BMI resulted in a simultaneous increase in RI and SDMA. The RI may assist in monitoring renal function, and may be associated with preclinical kidney changes in obese cats.
Testicular alterations and semen quality in a selected group of breeding buffaloes
(2023-01-01) Albuquerque, Rodrigo Dos Santos; Monteiro, Francisco Décio de Oliveira; Lima, Moises Moreira; da SILVA, Aluizio Otavio Almeida; E Cunha, Michel Santos; da CONCEIÇÃO, Maria Eduarda Bastos Andrade Moutinho; Scheeren, Verônica Flores da Cunha [UNESP]; Freitas, Camila de Paula [UNESP]; Papa, Frederico Ozanam [UNESP]; Monteiro, Bruno Moura; Viana, Rinaldo Batista; Coutinho, Leandro Nassar; Miranda, Moysés Dos Santos; Teixeira, Pedro Paulo Maia; Universidade Federal do Pará (UFPA); Federal Institute of Tocantins (IFTO); Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Federal Rural University of the Amazon
Testicular ultrasound enables the evaluation of changes in the testicular parenchyma. This study aimed to report the occurrence of hypoechogenic testicular alterations and their relationship with semen quality in five breeding buffaloes. Two buffaloes presented with hyperechoic points characteristic of fibrosis and anechoic density content between the parietal and visceral tunica. The two bulls without ultrasonographic changes showed higher average trajectory speed, linear velocity, curvilinear velocity, amplitude of lateral displacement of the spermatic head, total motility, progressive motility, fast speed, and acrosomal membrane values within the normal range. The number of spermatozoa with major and total defects was higher in the group of animals without alterations. The three buffaloes that presented with testicular alterations produced semen within established freezing standards.
Ligamentum teres reconstruction using autogenous semitendinosus tendon with toggle technique in rabbits
(2023-03-23) Abibe, Rebeca Bastos [UNESP]; Rahal, Sheila Canevese [UNESP]; dos Reis Mesquita, Luciane [UNESP]; Doiche, Danuta [UNESP]; da Silva, Jeana Pereira [UNESP]; Mamprim, Maria Jaqueline [UNESP]; Pinho, Renata Haddad [UNESP]; Battazza, Alexandre [UNESP]; Alves, Carlos Eduardo Fonseca [UNESP]; Saunders, W. Brian; Universidade Estadual Paulista (UNESP); Texas A&M University
Background: Ligamentum teres (LT) has traditionally been considered a vestigial or redundant structure in humans; however, based on new studies and the evolution of hip arthroscopy, the LT injury has been viewed as a source of hip pain. Therefore, LT reconstruction can be beneficial in some cases. Rabbits have been frequently used as a model for cranial cruciate ligament reconstruction but few studies are available for ligamentum teres reconstruction. Objective: To evaluate the semitendinosus tendon to replace ligamentum teres with the toggle technique, using rabbits as an experimental model. Methods: Twenty-six female Norfolk rabbits with approximately 3 months of age were divided into two equal groups after excision of ligamentum teres (LT) from the right hip joint: G1—no reconstruction of LT and capsulorrhaphy; G2—double—bundle reconstruction of the LT using semitendinosus tendon autograft. In both groups, the LT was removed from the right hip joint. In G2 the autograft was harvested from the left hind limb of the same rabbit. The rabbits were evaluated clinically at different time intervals; before surgery (M1), 48 h (M2), 15 days (M3), 30 days (M4) and 90 days (M5) after surgery. Results: The rabbits supported their limbs on the ground in both the groups. As complications of the procedure, four hip joints showed subluxations in the radiographic evaluation of G1; three at M4 and one at M5. In G2; two luxations of hip joints at M3 and one subluxation at M4 were seen. On ultrasound, irregular articular surface was seen in 30.8% of the rabbits that had subluxation of hip joints. Gross evaluation identified tendon graft integrity in 76.92% of the rabbits. Histological analysis revealed graft adhesion to the bone in the early phase comprised of sharpey-like collagen fibers. Conclusion: The double-bundle reconstruction of the LT using autologous semitendinosus tendon associated with the toggle rod shows an early phase of tendon graft ligamentization at 90 days post-operatively in young rabbits, but biomechanical bias suffered by the tendon during gait must be considered.
Validation of the Unesp-Botucatu pig composite acute pain scale (UPAPS) in piglets undergoing castration
(2023-04-01) Robles, I.; Luna, S. P.L. [UNESP]; Trindade, P. H.E. [UNESP]; Lopez-Soriano, M.; Merenda, V. R.; Viscardi, A. V.; Tamminga, E.; Lou, M. E.; Pairis-Garcia, M. D.; North Carolina State University; Universidade Estadual Paulista (UNESP); College of Veterinary Medicine Kansas State University; Prairie Swine Centre
To accurately assess pain and support broadly-based analgesic protocols to mitigate swine pain, it is imperative to develop and validate a species-specific pain scale. The objective of this study was to investigate the clinical validity and reliability of an acute pain scale (UPAPS) adapted for newborn piglets undergoing castration. Thirty-nine male piglets (five days of age, 1.62 ± 0.23 kg BW) served as their own control, were enrolled in the study and underwent castration in conjunction with an injectable analgesic administered one-hour post-castration (flunixin meglumine 2.2 mg/kg IM). An additional 10, non-painful female piglets were included to account for the effect of natural behavioral variation by day on pain scale results. Behavior of each piglet was video recorded continuously at four recording periods (24 h pre-castration, 15 min post-castration, 3 and 24 h post-castration). Pre- and postoperative pain was assessed by using a 4-point scale (score 0-3) including the following six behavioral items: posture, interaction and interest in surroundings, activity, attention to the affected area, nursing, and miscellaneous behavior. Behavior was assessed by two trained blinded observers and statistical analysis was performed using R software. Inter-observer agreement was very good (ICC = 0.81). The scale was unidimensional based on the principal component analysis, all items except for nursing were representative (rs ≥ 0.74) and had excellent internal consistency (Cronbach's alpha ≥ 0.85). The sum of scores were higher in castrated piglets post-procedure compared to pre-procedure, and higher than in non-painful female piglets confirming responsiveness and construct validity, respectively. Scale sensitivity was good when piglets were awake (92.9%) and specificity was moderate (78.6%). The scale had excellent discriminatory ability (area under the curve > 0.92) and the optimal cut-off sum for analgesia was 4 out of 15. The UPAPS scale is a valid and reliable clinical tool to assess acute pain in castrated pre-weaned piglets.
Writing 3D In Vitro Models of Human Tendon within a Biomimetic Fibrillar Support Platform
(2022-01-01) Monteiro, Rosa F.; Bakht, Syeda M.; Gomez-Florit, Manuel; Stievani, Fernanda C. [UNESP]; Alves, Ana L. G. [UNESP]; Reis, Rui L.; Gomes, Manuela E.; Domingues, Rui M. A.; Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine; ICVS/3B’s─PT Government Associate Laboratory; Universidade Estadual Paulista (UNESP)
Tendinopathies are poorly understood diseases for which treatment remains challenging. Relevant in vitro models to study human tendon physiology and pathophysiology are therefore highly needed. Here we propose the automated 3D writing of tendon microphysiological systems (MPSs) embedded in a biomimetic fibrillar support platform based on cellulose nanocrystals (CNCs) self-assembly. Tendon decellularized extracellular matrix (dECM) was used to formulate bioinks that closely recapitulate the biochemical signature of tendon niche. A monoculture system recreating the cellular patterns and phenotype of the tendon core was first developed and characterized. This system was then incorporated with a vascular compartment to study the crosstalk between the two cell populations. The combined biophysical and biochemical cues of the printed pattern and dECM hydrogel were revealed to be effective in inducing human-adipose-derived stem cells (hASCs) differentiation toward the tenogenic lineage. In the multicellular system, chemotactic effects promoted endothelial cells migration toward the direction of the tendon core compartment, while the established cellular crosstalk boosted hASCs tenogenesis, emulating the tendon development stages. Overall, the proposed concept is a promising strategy for the automated fabrication of humanized organotypic tendon-on-chip models that will be a valuable new tool for the study of tendon physiology and pathogenesis mechanisms and for testing new tendinopathy treatments.
Induced membrane technique using bone cement with or without cefazolin in chicken segmental radius defect
(2023-01-01) Campeiro Junior, Luiz D. [UNESP]; Rahal, Sheila Canevese [UNESP]; Souza, Marcos A.; Osowski, Alini [UNESP]; Silva Júnior, José I. S. [UNESP]; Universidade Estadual Paulista (UNESP); Mato Grosso School of Veterinary Medicine
The utilization of antibiotic-loaded cement spacer in the induced membrane development has been a debate topic in human medicine. To the best of the author's knowledge, these combinations have not yet been evaluated in birds. Therefore, this study assessed induced membrane formation using radiography and histology, in a segmental defect of a chicken radius, with or without the addition of cefazolin. Thirty 18-month-old healthy chickens were divided into two equal groups: G1—bone defect filled with bone cement; G2—bone defect filled with cefazolin powder-loaded bone cement. Radiographic examinations of the left forearm were taken immediately after surgery and at 7, 15, and 21 postoperative days. For the collection of the induced membranes, five chickens in each group were euthanized at 7, 15, and 21 days after surgery. Radiographically, the bone cement was identified as a radiopaque structure occupying the bone defect in both groups. Mild new bone formation in at least one of the fractured extremities of the bone defect was seen only 21 days after surgery in most chickens. Histologically, there was no difference in the mean thickness of the induced membrane between groups at all time points. Multifocal multinucleated cells differed between groups at 7 (G1 > G2) and 21 (G2 > G1) days after surgery. Mononuclear inflammatory infiltrate differed between groups only on day 21 (G1 > G2). Fibrous tissue proliferation did not differ between groups at all evaluation times. Blood vessel density differed only at 21 days postoperatively (G2 < G1). Multifocal areas of cartilage differed between groups at all time points (G1 > G2). In conclusion, cefazolin mixed with bone cement did not affect thickness of the induced membrane, but did result in a negative effect on some histological aspects, such as fewer vessels, less multifocal areas of cartilage, and persistence of inflammation.
Effect of acupuncture on hematologic, muscular biomarkers, fibrinogen and serum lactate parameters in training rodeo bulls
(2023-05-01) Rosa, Vitor Bruno Bianconi; dos Santos, Ivan Felismino Charas [UNESP]; Souto, Letícia Gondim [UNESP]; de Paiva Porto, Emília; Pizzigatti, Dietrich; Cholfe, Bruno Fornitano; de Almeida, Breno Fernando Martins; Patelli, Thais Helena Constantino; Takahira, Regina Kiomi [UNESP]; Northern Parana State University (UENP); Universidade Estadual Paulista (UNESP); University of the Republic Uruguay; Unilago University; Integrated Colleges of Ourinhos; Rondônia Federal University (UNIR)
The study aimed to evaluated the effects of acupuncture in rodeo bulls in training, by determining hematological variables, creatine kinase (CK), aspartate aminotransferase (AST), fibrinogen, and plasma lactate. Thirty adult healthy bulls, crossbred, were included in the study and randomly allocated into two groups of 15 animals, according to the use of acupuncture treatment for six months (GA) or not (GB). The variables were measured 30 min before (TP0) and 10 min (TP10min), 12 (TP12h), 24 (TP24h), 48 (TP48h), and 72 h (TP72h) after a single episode of jumping emulating rodeo exercise. The GB group showed variations in hemoglobin between TP0 and TP10min (p = 0.002) and TP0 and TP12h (p = 0.004), and the GA presented an increase in eosinophil values between TP0 and TP12h (p = 0.013) and TP0 and TP24h (p = 0.034). Leukopenia was observed in GB between TP10min and TP72h ((p = 0.008). The CK values were high (↑ 300 UI/l) after exercise until the TP24h, and decreased in TP48h, in both groups. The plasma lactate elevation was lower in the GA at TP10min (p = 0.011), TP12h (p = 0.008), TP72h (p < 0.001). The rodeo bulls submitted to acupuncture treatment showed smaller variations in hemogram, elevated eosinophils levels, and lower plasma lactate levels after exercise.
Influence of culture conditions on the secretome of mesenchymal stem cells derived from feline adipose tissue: Proteomics approach
(2023-08-01) Lara, Maria Laura [UNESP]; Carvalho, Marcos Gomides [UNESP]; de Souza, Fabiana Ferreira [UNESP]; Schmith, Rubia Alves [UNESP]; Codognoto, Viviane Maria [UNESP]; De Vita, Bruna [UNESP]; Freitas Dell'Aqua, Camila de Paula [UNESP]; Landim, Fernada da Cruz [UNESP]; Alvarenga, Marina Landim e [UNESP]; Universidade Estadual Paulista (UNESP); Universitat de Barcelona; Omics Animal Biotechnology
This study aimed to describe the secretome of mesenchymal stem cells derived from feline adipose tissue (AD-MSCs) and compare the effects of different culture conditions on AD-MSC proteomics using a shotgun approach. Adipose tissue was collected from 5 female cats and prepared to culture. Conditioned media was collected at third passage, in which the cells were cultured under 4 conditions, normoxia with fetal bovine serum (N + FBS), hypoxia with FBS (H + FBS), normoxia without FBS (N – FBS), and hypoxia without FBS (H – FBS). Then, the secretome was concentrated and prepared for proteomic approaches. Secretomes cultured with FBS-free medium had more than twice identified proteins in comparison with the secretomes cultured with FBS. In contrast, hypoxic conditions did not increase protein amount and affected only a small proteome fraction. Relevant proteins were related to the extracellular matrix promoting environmental modulation, influencing cell signaling pathways, and providing a suitable environment for cell proliferation and maintenance. Moreover, other proteins were also related to cell adhesion, migration and morphogenesis. Culture conditions can influence protein abundance in AD-MSC secretome, and can give also more specificity to cell and cell-free treatments for different diseases.
In vivo embryo development in bitches inseminated laparoscopically after ovulation time estimated based on a single progesterone determination
(2023-01-01) Alves, Aracelle Elisane; Motheo, Tathiana Ferguson; Apparicio, Maricy Ferreira [UNESP]; Mostachio, Giuliano Queiroz; dos Santos, Ricarda Maria; Vicente, Wilter Ricardo Russiano [UNESP]; Luvoni, Gaia Cecilia; Universidade Federal de Uberlândia (UFU); Universidade de Cuiabá; Universidade Estadual Paulista (UNESP); Centro Universitário de Rio Preto; Università Degli Studi di Milano
Logistic and economical limitations are often the causes of dog owners not accurately monitoring the estrous cycle and the optimal insemination time. The aim of this study was to evaluate in vivo early embryonic development in bitches, after the analysis of sequential vaginal cytologies associated to single progesterone measurement and single laparoscopic insemination with high quality semen (fresh and with high spermatozoa concentration) or low-quality semen (frozen/thawed and with low spermatozoa concentration) at 48 h post-ovulation time predicted on a single progesterone measurement. Ten bitches were inseminated with 250 x 106 fresh spermatozoa (80% motility), and ten with 80 x 106 frozen/thawed spermatozoa (60% motility) in the cranial part of each uterine horn. Seven days later, ovariohysterectomy was performed and the oviducts and uterine horns and body were flushed to recover embryos and unfertilized oocytes. In 80% of the bitches inseminated with fresh and 50% of bitches inseminated with frozen/thawed semen, embryos at 2 to 8 cells stage were recovered mostly from the, oviducts. This study indicates that pregnancies can be obtained with a single laparoscopic intrauterine insemination after single serum progesterone measurement, although with a low number of embryos. This result should be taken into account in case economic or logistic restrictions that affect the possibility of owners to plan an accurate monitoring of the optimal breeding time using fresh and frozen semen.
Two Types of Management for the Noninvasive Treatment of Pectus Excavatum in Neonatal Puppies—Case Reports
(2023-03-01) Pereira, Keylla Helena Nobre Pacífico [UNESP]; Fuchs, Kárita da Mata [UNESP]; Terçariol, Lara Ataídes Arantes; Silva, Renata Cesar [UNESP]; Camargo, Gabriel de Azevedo [UNESP]; Mendonça, Júlia Cosenza [UNESP]; Paulino, Netelin Tainara; Zone, Marcelo Alejandro; Oba, Eunice [UNESP]; Lourenço, Maria Lucia Gomes [UNESP]; Universidade Estadual Paulista (UNESP); Self-Employed Veterinarian
Pectus excavatum is a deformity of the thorax characterized by ventrodorsal narrowing of the sternum bone and costal cartilages, which can lead to compression and cardiopulmonary alterations in dogs, presenting a high prevalence in brachycephalic breeds. The aim of this report was to describe two types of management for the noninvasive treatment of pectus excavatum in newborn puppies of the breeds French Bulldog and American Bully. The puppies presented dyspnea, cyanosis and substernal retraction during inspiration. The diagnosis was performed by physical examination and confirmed by chest X-ray. Two types of splints were performed (a circular splint with plastic pipe and a paper box splint on the chest), aiming at thoracic lateral compression and frontal chest remodeling. The management was effective for the conservative treatment of mild-grade pectus excavatum, resulting in the repositioning of the thorax and improvement of the respiratory pattern.
The Temporal Associations of B-Mode and Power-Doppler Ultrasonography, and Ovarian Steroid Changes of the Periovulatory Follicle and Corpus Luteum During Luteogenesis and Luteolysis in Jennies
(2023-03-01) Magalhaes, Humberto B. [UNESP]; Canisso, Igor F. [UNESP]; Dell-Aqua, Jose A. [UNESP]; Universidade Estadual Paulista (UNESP); University of Illinois Urbana- Champaign
This study aimed to determine the associations between B-mode and Power-doppler ultrasonography and ovarian steroids of the periovulatory follicle and respective corpus luteum (CL) during luteogenesis and luteolysis in jennies. Twenty-four periovulatory follicles/estrus of correspondent one inter-ovulatory interval (n = 12 jennies) were assessed in the study. B-mode ultrasonography and teasing were carried out once day until the detection of a periovulatory follicle (≥28 mm, uterine edema, and signs of estrus). Thereafter, jennies were monitored at 4-hour-intervals by B-mode and Power-doppler ultrasonography. Closer to ovulation, jennies were hourly checked. Each CL was checked daily from luteogenesis to luteolysis. Plasma concentrations of estradiol and progesterone were assessed daily with chemiluminescence immunoassay. Granulosa echogenicity and thickness increased from -36 hour to -1 hour before ovulation in 70% of follicles (P < .05) and were strongly associated with impending ovulation (r = 0.80 and r = 0.70, respectively). The follicular-wall blood flow increased from -72 to -24 hour pre-ovulation, while the estradiol concentration declined from 42 pg/mL by -72 hour to 31.6 pg/mL by 24 hour before ovulation (P < .05). The vascularization of the periovulatory follicle decreased from 62% (-36 hour) to 37% (-1 hour) before ovulation (P < .05). The CL vascularization and progesterone concentration gradually increased, reaching the peak at 11- and 10-day after the ovulation, respectively (P < .05). The CL vascularization started to decline 3 day before luteolysis, while progesterone concentrations started to drop 4 day before luteolysis (P < .05). In conclusion, the structural changes of the periovulatory follicle detected on B-mode and Power-doppler can be used to detect impending ovulation in donkeys; however, Power-doppler, but not B-mode ultrasonography, can be used to assess CL function in jennies.

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