PERSPECTIVE: Potential and reality of cryopreserving somatic cells of wild felids for conservation.

The loss of biodiversity caused by anthropogenic actions is also a reality for the members of the Felidae family. Except for the domestic cat, all felid species have some degree of threat of extinction in their natural habitat. For this reason, felids have been included in conservation-related studies. This scenario has aroused increasing interest in the formation of somatic cell banks, which when efficiently implemented can be used in preservation strategies for the species. Nevertheless, one of the important steps in the formation of these banks is the understanding of the technical principles and variations involved in cryopreservation techniques, especially because cryopreservation increases the possibilities for Assisted Reproduction Technologies (ARTs) by making the use of biological materials independent of time and space. In wild felids, several species already have promising results in the formation of somatic cell banks, and studies aimed at better viability rates have been constantly proposed, as well as new species have been studied. In some species, aspects involved in successful cryopreservation are already well defined, and slow freezing associated with cryoprotectant solutions composed of intra- and extracellular substances is the most useful approach. The aim of this review was to present the main parameters involved in the elaboration of a somatic cell cryopreservation protocol and their effects, as well as to address the main results achieved for different wild felids. Doi.org/10.54680/fr23510110112.

Interactions among sucrose and concentrations of serum fetal bovine on the cryopreservation of somatic cells derived from red-rumped agoutis.

BACKGROUND: The synergistic action among the different extracellular cryoprotectants could improve somatic cell quality after thawing and provide bases for the formation of biobanks for red-rumped agoutis. OBJECTIVE: This study evaluated the interactions among sucrose (SUC) and concentrations of serum fetal bovine (FBS) on the cryopreservation of somatic cells derived from red-rumped agoutis. MATERIALS AND METHODS: Cells were cryopreserved with 10% dimethyl sulfoxide and different concentrations of FBS (10%, 40%, and 90%) with or without 0.2 M SUC, totaling six comparison groups. Non-cryopreserved cells were used as a control. Cells were evaluated for viability, metabolic activity, proliferative activity, reactive oxygen species (ROS), mitochondrial membrane potential and apoptosis levels. RESULTS: No difference was observed among cryopreserved with DMSO containing (10FBS, 10FBS-SUC, 40FBS, 40FBS-SUC, 90FBS, 90FBS-SUC) and non-cryopreserved groups for viability, metabolic activity, proliferative activity, and ROS levels. Interestingly, only cells cryopreserved with 90% FBS and SUC maintained the mitochondrial membrane potential like the control. This indicates that at high concentrations of FBS, SUC contributes to the maintenance of this parameter in cryopreserved cells. Moreover, at concentrations of 10% and 40% of FBS, SUC contributed to the maintenance of viability evaluated by the levels of apoptosis evaluated after thawing. In summary, we verified that 90% FBS and 0.2 M SUC promote greater ability of cells after thawing. Additionally, SUC positively acts in cryopreservation solutions containing 10% and 40% FBS. CONCLUSION: This information is essential to an understanding of the mechanisms involved in the interactions of extracellular cryoprotectants in somatic cell cryopreservation solutions of red-rumped agoutis. DOI: 10.54680/fr23210110212.

Effect of carvacrol antioxidant capacity on oocyte maturation and embryo production in cattle.

Carvacrol (C10H14O), an efficient phenolic antioxidant substance for several cell types, may become a useful antioxidant for female germ cells and embryo culture. This study investigates the effects of carvacrol supplementation on bovine oocytes in in vitro maturation (IVM) and embryo production. In total, 1222 cumulus-oocyte complexes were cultured in TCM-199+ alone (control treatment) or supplemented with carvacrol at the concentrations of 3 µM (Carv-3), 12.5 µM (Carv-12.5), or 25 µM (Carv-25). After IVM, the oocytes were subjected to in vitro fertilization and embryo production, and the spent medium post-IVM was used for evaluating the levels of reactive oxygen species and the antioxidant capacity (2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2'-azinobis-3-ethyl-benzothiozoline-6-sulphonic acid quantification). A greater (P < 0.05) antioxidant potential was observed in the spent medium of all carvacrol-treated groups compared with the control medium. Moreover, the addition of carvacrol to the maturation medium did not affect (P > 0.05) blastocyst production on days 7 and 10 of culture; however, the total number of cells per blastocyst was reduced (P < 0.05) in two carvacrol-treated groups (Carv-3 and Carv-25). In conclusion, carvacrol demonstrated a high antioxidant capacity in the spent medium after oocyte maturation; however, although embryo production was not affected, in general, carvacrol addition to IVM medium reduced the total number of cells per blastocyst. Therefore, due to the high antioxidant capacity of carvacrol, new experiments are warranted to investigate the beneficial effects of lower concentrations of carvacrol on embryo production in cattle and other species.

Effects of vitrification techniques on the formation of skin cryobank of the ocelot (Leopardus pardalis).

BACKGROUND: Skin cryobanks represent important tools for the conservation of the maximum genetic representation of a population, especially those with a certain degree of threat to extinction, such as the ocelot. A relevant step towards the proper establishment of these banks is the definition of adequate cryopreservation techniques for the conservation of the skin. OBJECTIVE: We evaluated the effects of two different techniques [direct vitrification in cryovials (DVC) and solid-surface vitrification (SSV)] for the preservation of ear skin derived from ocelot. MATERIALS & METHODS: For both techniques, we vitrified the ear skin using Dulbeccos modified Eagles medium with 3.0 M dimethyl sulfoxide, 0.25 M sucrose, and 10% fetal bovine serum. Non-cryopreserved tissues were used as control (control group). All tissues were analyzed for their morphometric characteristics by conventional histology and morphological / functional analysis by cell ability during the culture. RESULTS: While tissues cryopreserved by DVC showed similar values for dermis thickness and number of perinuclear halos to the control, tissues cryopreserved by SSV showed similarities to the control regarding the number of melanocytes, percentage of collagen fibers, and numbers of viable cells by apoptosis analysis. Additionally, none of the vitrification techniques affected stratum corneum thickness, number of keratinocytes, tissue proliferative activity, cell viability, or metabolism. CONCLUSION: Both vitrification techniques (DVC and SSV) can be used for the conservation of ocelot skin; however, SSV guarantees a higher cellular quality after in vitro tissue culture in most of the parameters evaluated, such as viability, metabolism, and apoptosis analysis. doi.org/10.54680/fr23110110412.

Oocyte in vitro maturation with eugenol improves the medium antioxidant capacity and total cell number per blastocyst.

This study investigates the impact of eugenol (EU) supplementation on bovine oocyte in vitro maturation (IVM) and antioxidant capacity, as well as in vitro embryo production and quality after conventional in vitro fertilization (IVF). A total of 1077 cumulus oocyte complexes were cultured in TCM-199+ without EU supplementation (control treatment) or supplemented with EU at the concentrations of 10 µM (EU-10), 20 µM (EU-20), or 40 µM (EU-40). After IVM, the oocytes were subjected to IVF and embryo culture. The addition of EU at 40 µM to the IVM medium improved (P < 0.05) the antioxidant capacity and cleavage rate when compared to the control treatment. Moreover, a positive correlation (r = 0.61, P < 0.03) was observed between cleavage rate and EU concentration. The addition of EU at concentrations of 10 and 20 µM decreased (P < 0.05) the calreticulin (CALR) levels in expanded blastocysts when compared to the control treatment and EU-40 treatment. However, the EU-10 and EU-20 treatments had a greater (P < 0.05) mean total cell number (TCN) per expanded blastocyst when compared to the control treatment and EU-40 treatment. In conclusion, the addition of EU to the enriched culture medium during IVM of bovine oocytes improved the antioxidant capacity of the spent medium, as well as the cleavage rate and embryonic quality (i.e., TCN/expanded blastocyst), and reduced the endoplasmic reticulum stress (i.e., CALR levels) in the embryos. Thus, we recommend enriching the IVM medium with 10 µM EU for in vitro bovine embryo production.

Evaluation of different cryoprotectant solutions for the cryopreservation of somatic tissues of Dasyprocta leporina (Linnaeus, 1758).

BACKGROUND: Somatic tissue banks represent important tools for the conservation of wild mammals, aiming at the immediate maintenance and safeguarding of biological samples. For agouti, Dasyprocta leporina, studies on the formation of these banks are still scarce, especially regarding protocols of the best cryoprotectant solution employed. OBJECTIVE: To optimize the cryoprotectant solution [ethylene glycol (EG), dimethyl sulfoxide (DMSO), sucrose (SUC)] used for the cryopreservation of agouti somatic tissues. MATERIALS AND METHODS: We treated ear tissues with various cryoprotectant solutions: 3.0 M EG (EG group), 3.0 M EG and 0.25 M SUC (EG-SUC group), 3.0 M DMSO (DMSO group), 3.0 M DMSO and 0.25 M SUC (DMSO-SUC group), 1.5 M EG and 1.5 M DMSO (EG-DMSO group) and 1.5 M EG, 1.5 M DMSO and 0.25 M SUC (EG-DMSO-SUC group). Non-cryopreserved tissues were used as controls. All tissues were analyzed for their ultrastructural and morphometric characteristics by scanning electron microscopy and conventional histology. RESULTS: EG-DMSO-SUC was found to be the optimal cryoprotectant solution in terms of the evaluated parameters, such as thickness of the dermis and skin, number of perinuclear halos, proliferative potential, number of empty lacunas and degenerated chondrocytes. CONCLUSION: Agouti somatic tissue cryopreservation may serve for its conservation and as an experimental model for the development of preservation methods for species of the same genus that are either vulnerable or critically endangered.

Pyroligneous acid from Mimosa tenuiflora and Eucalyptus urograndis as an antimicrobial in dairy goats.

AIMS: To evaluate the applicability of the Mimosa tenuiflora and Eucalyptus urograndis pyroligneous acids (PAs) as alternative antiseptics in dairy goats. METHODS AND RESULTS: Cytotoxicity was evaluated in vitro using bacteria, as well as in vivo using goats, and the influence of PAs on the physicochemical parameters of fresh milk were examined. The cytotoxicity of PAs was evaluated in terms of morphology, cell viability and metabolic activity of goat tegumentary cells. The PA of M. tenuiflora had results similar to those of 2% iodine. For the in vitro tests, strains of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa were used with the well technique, demonstrating inhibition halos greater than 9 mm. In the in vivo test, 15 animals were used per phase of the experiment, and the plate counting technique showed that there was antiseptic action of both extracts, with emphasis on the M. tenuiflora PA. Physicochemical analysis of the milk showed that neither PAs interfered with its physical-chemical parameters. CONCLUSIONS: The PA of M. tenuiflora presented potential as an alternative antiseptic in dairy goats. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates the use of PA as an antimicrobial agent in animals.

Amifostine protects from the peripheral sensory neuropathy induced by oxaliplatin in mice.

Sensory neuropathy is a dose-limiting side effect of oxaliplatin-based cancer treatment. This study investigated the antinociceptive effect of amifostine and its potential neuroprotective mechanisms on the oxaliplatin-related peripheral sensory neuropathy in mice. Oxaliplatin (1 mg/kg) was injected intravenously in Swiss albino male mice twice a week (total of nine injections), while amifostine (1, 5, 25, 50, and 100 mg/kg) was administered subcutaneously 30 min before oxaliplatin. Mechanical and thermal nociceptive tests were performed once a week for 49 days. Additionally, c-Fos, nitrotyrosine, and activating transcription factor 3 (ATF3) immunoexpressions were assessed in the dorsal root ganglia. In all doses, amifostine prevented the development of mechanical hyperalgesia and thermal allodynia induced by oxaliplatin (P<0.05). Amifostine at the dose of 25 mg/kg provided the best protection (P<0.05). Moreover, amifostine protected against neuronal hyperactivation, nitrosative stress, and neuronal damage in the dorsal root ganglia, detected by the reduced expression of c-Fos, nitrotyrosine, and ATF3 (P<0.05 vs the oxaliplatin-treated group). In conclusion, amifostine reduced the nociception induced by oxaliplatin in mice, suggesting the possible use of amifostine for the management of oxaliplatin-induced peripheral sensory neuropathy.

Cryopreservation of Collared Peccary (Pecari tajacu Linnaeus, 1758) Somatic Cells is Improved by Sucrose and High Concentrations of Fetal Bovine Serum.

BACKGROUND: The formation of somatic cell banks is affected by, amongst other factors, the cryoprotectant solution used. The selection of an effective solution, therefore, is a primary parameter. OBJECTIVE: We optimized the cryoprotectant used for collared peccary somatic cell cryopreservation. MATERIALS AND METHODS: We categorized cells into different groups based on their cryopreservation and evaluated the morphology, viability, proliferative activity, metabolism, and oxidative stress. One group was cryopreserved in 10% DMSO with 10% fetal bovine serum (DMSO-10FBS), and another with 50% FBS (DMSO-50FBS). The cryopreservation of both groups included the presence of 0.2 M sucrose (DMSO-SUC-10FBS and DMSO-SUC-50FBS). Non-cryopreserved cells and cells cryopreserved with 10% DMSO (DMSO) supplemented with 0.2 M sucrose (DMSO-SUC) were used as controls. RESULTS: There was no difference observed in morphology or viability among the groups. Proliferative activity was reduced in DMSO-10FBS when compared to controls. Although cryopreservation reduced metabolism, no difference was observed among solutions. A lower level of reactive oxygen species was observed in cells of DMSO-SUC-50FBS when compared to other cryoprotectants. Only cells of DMSO-SUC-50FBS had mitochondrial potential similar to non-cryopreserved cells. CONCLUSION: 10% DMSO supplemented with 50% FBS and 0.2 M SUC was observed to be the most efficient cryoprotectant for preserving collared peccary somatic cells.

Amifostine protects from the peripheral sensory neuropathy induced by oxaliplatin in mice

Sensory neuropathy is a dose-limiting side effect of oxaliplatin-based cancer treatment. This study investigated the antinociceptive effect of amifostine and its potential neuroprotective mechanisms on the oxaliplatin-related peripheral sensory neuropathy in mice. Oxaliplatin (1 mg/kg) was injected intravenously in Swiss albino male mice twice a week (total of nine injections), while amifostine (1, 5, 25, 50, and 100 mg/kg) was administered subcutaneously 30 min before oxaliplatin. Mechanical and thermal nociceptive tests were performed once a week for 49 days. Additionally, c-Fos, nitrotyrosine, and activating transcription factor 3 (ATF3) immunoexpressions were assessed in the dorsal root ganglia. In all doses, amifostine prevented the development of mechanical hyperalgesia and thermal allodynia induced by oxaliplatin (P<0.05). Amifostine at the dose of 25 mg/kg provided the best protection (P<0.05). Moreover, amifostine protected against neuronal hyperactivation, nitrosative stress, and neuronal damage in the dorsal root ganglia, detected by the reduced expression of c-Fos, nitrotyrosine, and ATF3 (P<0.05 vs the oxaliplatin-treated group). In conclusion, amifostine reduced the nociception induced by oxaliplatin in mice, suggesting the possible use of amifostine for the management of oxaliplatin-induced peripheral sensory neuropathy.

The Comet assay for detection of DNA damage in canine sperm.

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno-, terato- or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A-median: 31.55%, interquartile range (IQR): 30.18-38.01; group B-median: 0.90%, IQR: 0.65-1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = -.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non-normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.

Caracterização histomorfológica do sistema tegumentar auricular de cateto - Pecari tajacu Linnaeus, 1758) / Histomorphological characterization of collared peccary (Pecari tajacu Linnaeus, 1758) ear integumentary system

A criopreservação de tecido somático derivado da pele de catetos consiste numa alternativa para a conservação da biodiversidade por meio da associação com a transferência nuclear. Nesse contexto, a manipulação de tecidos da pele é uma etapa crucial para o sucesso dessa biotécnica. Portanto, o objetivo do presente estudo, foi caracterizar o sistema tegumentar auricular periférico de catetos, visando aprimorar a conservação tecidual. Para tanto, fragmentos auriculares de oito animais foram avaliados quanto às camadas teciduais, aos componentes, à atividade proliferativa e à viabilidade metabólica, usando-se as colorações hematoxilina-eosina e tricrômico de Gomori, quantificação de AgNORs e microscopia eletrônica de transmissão. Assim, tamanhos de 104,2µm e 222,6µm foram observados para epiderme e derme, com uma proporção volumétrica de 36,6% e 58,7%, respectivamente. Além disso, na epiderme, foram evidenciadas as camadas basal (22,5µm), intermediárias (53,5µm) e córnea (28,2µm), com valores médios de 65,3 células epidermais, 43,4 melanócitos e 14,8 halos perinucleares. Já a derme apresentou 127 fibroblastos, com 2,5 AgNORs/nucléolo. Adicionalmente, a atividade metabólica foi de 0,243. Em conclusão, o sistema tegumentar auricular periférico de catetos possui algumas marcantes variações em relação a outros mamíferos, quanto ao número de camadas e espessura da epiderme, quantidade de células epidermais, melanócitos e parâmetros proliferativos.(AU)

The cryopreservation of somatic tissue derived from skin of collared peccaries is an alternative for biodiversity conservation through association with nuclear transfer. In this context, tissue manipulation of skin is a critical step for the success of this biotechnique. Therefore, the aim was to characterize the peripheral ear integumentary system derived from collared peccaries, directing to improve tissue conservation. Thus, ear fragments of eight animals were evaluated for tissue layers, components, proliferative activity and metabolic viability, using hematoxylin-eosin and Gomori Trichrome, AgNORs quantification and transmission electronic microscopy. Hence, sizes of 104.2 µm and 222.6 µm were observed in the epidermis and dermis, with a volumetric ratio of 36.6% and 58.7%, respectively. Moreover, basal layer (22.5 µm), intermediate (53.5 µm) and cornea (28.2 µm), with mean values of 65.3 epidermal cells, 43.4 melanocytes and 14.8 perinuclear halos were evidenced in the epidermal. Already the dermis has 127 fibroblasts with 2.5 AgNORs/nucleolus. Additionally, the metabolic activity was 0.243. In conclusion, the peripheral ear integumentary system derived from collared peccaries possessed some important variations compared to other mammals, as the number of layers and thickness of the epidermis, number of epidermal cells, melanocytes and proliferative parameters.(AU)

PLGA Implants containing vancomycin and dexamethasone: development, characterization and bactericidal effects.

Post-operative endophthalmitis is an infection and an inflammation of the eye following a surgical procedure. Its treatment is based on drug injections into the eye. However, this treatment can lead to ocular complications. Intraocular implants could substitute the conventional therapy. Poly(lactic-co-glycolic acid) (PLGA) implants comprising on vancomycin and dexamethasone were evaluated as drug delivery system to treat endophthalmitis after cataract surgery. Implants were characterized by drug content uniformity, Fourier Transform Infrared Spectroscopy (FTIR), Differential Scanning Calorimetry (DSC), Wide Angle X-ray Scattering (WAXS), Scanning Electron Microscopy (SEM) and in vitro drug release. The bactericidal effect of vancomycin, eluted from the implants, was demonstrated against Staphylococcus aureus and Staphylococcus epidermidis. The drugs were uniformly distributed in the polymer. The analytical techniques revealed the chemical integrity of the drugs incorporated into the polymer and the modification of dexamethasone semi-crystalline nature. Drugs were controlled released from implants; and the eluted vancomycin showed bactericidal effects. In conclusion, PLGA implants containing vancomycin and dexamethasone may represent a therapeutic alternative to treat post-operative endophthalmitis.

Predicting cortical bone adaptation to axial loading in the mouse tibia.

The development of predictive mathematical models can contribute to a deeper understanding of the specific stages of bone mechanobiology and the process by which bone adapts to mechanical forces. The objective of this work was to predict, with spatial accuracy, cortical bone adaptation to mechanical load, in order to better understand the mechanical cues that might be driving adaptation. The axial tibial loading model was used to trigger cortical bone adaptation in C57BL/6 mice and provide relevant biological and biomechanical information. A method for mapping cortical thickness in the mouse tibia diaphysis was developed, allowing for a thorough spatial description of where bone adaptation occurs. Poroelastic finite-element (FE) models were used to determine the structural response of the tibia upon axial loading and interstitial fluid velocity as the mechanical stimulus. FE models were coupled with mechanobiological governing equations, which accounted for non-static loads and assumed that bone responds instantly to local mechanical cues in an on-off manner. The presented formulation was able to simulate the areas of adaptation and accurately reproduce the distributions of cortical thickening observed in the experimental data with a statistically significant positive correlation (Kendall's τ rank coefficient τ = 0.51, p < 0.001). This work demonstrates that computational models can spatially predict cortical bone mechanoadaptation to a time variant stimulus. Such models could be used in the design of more efficient loading protocols and drug therapies that target the relevant physiological mechanisms.

Phosphorylated H2AX in parthenogenetically activated, in vitro fertilized and cloned bovine embryos.

In vitro embryo production methods induce DNA damage in the embryos. In response to these injuries, histone H2AX is phosphorylated (γH2AX) and forms foci at the sites of DNA breaks to recruit repair proteins. In this work, we quantified the DNA damage in bovine embryos undergoing parthenogenetic activation (PA), in vitro fertilization (IVF) or somatic cell nuclear transfer (SCNT) by measuring γH2AX accumulation at different developmental stages: 1-cell, 2-cell and blastocyst. At the 1-cell stage, IVF embryos exhibited a greater number of γH2AX foci (606.1 ± 103.2) and greater area of γH2AX staining (12923.6 ± 3214.1) than did PA and SCNT embryos. No differences at the 2-cell stage were observed among embryo types. Although PA, IVF and SCNT were associated with different blastocyst formation rates (31.1%, 19.7% and 8.3%, P < 0.05), no differences in the number of γH2AX foci or area were detected among the treatments. γH2AX is detected in bovine preimplantation embryos produced by PA, IVF and SCNT; the amount of DNA damage was comparable among those embryos developing to the blastocyst stage among different methods for in vitro embryo production. While IVF resulted in increased damage at the 1-cell embryo stage, no difference was observed between PA and SCNT embryos at any developmental stage. The decrease in the number of double-stranded breaks at the blastocyst stage seems to indicate that DNA repair mechanisms are functional during embryo development.

Diverse outcomes of Photodynamic Antimicrobial Chemotherapy on five Enterococcus faecalis strains.

OBJECTIVES: In the present study, the effectiveness of Photodynamic Antimicrobial Chemotherapy (PACT) was evaluated on planktonic cells and biofilms of five Enterococcus faecalis clinical isolates. METHODS: Planktonic cells and biofilms of E. faecalis E2, E3, ER3/2s, OS16 and AA-OR34 were grown in SDMY medium plus 0.4% glucose. Approximately 5.0×10(7)CFU planktonic cells and 24h biofilms were subjected to PACT using the combination of Light Emitting Diodes (LEDs, Biotable(®)) and Photogem(®). The metabolic activity of bacterial cells was evaluated by a resazurin assay. Biomass values of the biofilms were determined by a crystal violet assay. RESULTS: Compared to the water-treated control group, gradual increases of light energy led to greater reduction of metabolic activity of planktonic cells and biofilms of E. faecalis when the combination of LEDs and Photogem(®) was applied. Photogem(®) alone significantly reduced the metabolic activity of planktonic cells, whereas LEDs or Photogem(®) alone did not result in biofilm viability changes. PACT yielded similar antimicrobial outcomes on planktonic cells of all tested E. faecalis strains, whereas biofilms of E. faecalis E3, ER3/2s and OS16 were more resistant to PACT than biofilms of E. faecalis E2 and AA-OR34. CONCLUSIONS: The efficacy of PACT on E. faecalis biofilms was strain dependent. PACT demonstrated its potential as an adjuvant antimicrobial treatment by killing of E. faecalis planktonic and biofilm cells.

Physical characteristics of the eggs of red-legged partridge (Alectoris rufa) reared in captivity / Características físicas dos ovos da perdiz vermelha (Alectoris rufa) criada em cativeiro

O objetivo do presente estudo foi avaliar os efeitos da idade das fêmeas (um, dois e três anos) e do mês de postura (março, abril e maio) sobre as características físicas dos ovos da perdiz vermelha (Alectoris rufa) criada em cativeiro. O peso (W), o comprimento máximo (L) e a largura máxima (B) de 2878 ovos foram determinados diretamente, enquanto o índice de forma (B/L), o volume (V) e a superfície (S) foram calculados com base nos parâmetros determinados diretamente. A análise mostrou diferenças significativas (P<0,01) no peso dos ovos entre as diferentes idades e entre meses de postura, com menor peso nas fêmeas mais jovens. Observaram-se diferenças significativas (P<0,01) no comprimento do ovo entre as classes de idade, mas não entre os meses de postura (P>0,05). Observaram-se diferenças significativas (P<0,01) na largura máxima e no índice de forma do ovo entre as diferentes classes de idades, com valores mais elevados nas fêmeas mais velhas e no período de postura mais tardio. O volume dos ovos estimados por meio de V1= 0,51LB2e V2=0,913W foi afetado significativamente (P<0,01) pela idade e pelo mês de postura, bem como as áreas, S1=4.835W0,662, S2=4,951V10,666e S3=4,951V20,666, as quais apresentaram os mesmos efeitos.

Mensurações ultrassonográficas da cisterna da glândula mamária de caprino transgênico / Ultrasound measurements of the mammary gland of transgenic hormone-induced lactating goat

Milk production of transgenic does was evaluated by ultrasound measurements of the mammary gland. Two Canindé goats, which were nine months of age were used in the trial, one non-transgenic or other transgenic for hG-CSF. For hormone-induced lactation, animals were given estradiol (0.25mg/kg, IM), progesterone (0.75mg/kg, IM), and prednisolone (0.4mg/kg, IM). Ultrasonographic exams were carried out during milking, using a Falcon 100 ultrasound equipment with a 5MHz convex probe and were performed by the same operator. The results were expressed as mean±standard error. The maximum greater length and shorter length of the cistern were respectively 5.14cm and 1.36cm for the transgenic animal and 7.28cm and 2.25cm for non-transgenic, which is consistent with the maximum milk volume produced. The relationship between the average area of cisterns and milk yield was expressed as a linear correlation curve, with a correlation coefficient significantly positive for both transgenic (Y=-1.1314+10.8538*x; r=0.97) and non-transgenic (Y=-21.7551+18.3634*x; r=0.97) animals. In conclusion, the ultrasound is a practice and appropriate technique to evaluate the cisterns in ruminant udders in transgenic animal.

Goat oocyte production by standard or one-shot FSH treatments and quantitative analysis of transcripts for EGF ligands and its receptor after in vitro maturation.

Hormonal ovarian stimulation may affect the success of embryo production by regulating transcripts in recovered cumulus-oocyte complexes (COCs). Here, in parallel to morphological classification and in vitro maturation (IVM) rate analysis, we investigated the expression of epidermal growth factor (EGF) and its receptor (EGFR) in oocytes and cumulus cells from goat COCs recovered by laparoscopy after standard [multi-dose follicle-stimulating hormone (FSH)] or one-shot (single dose FSH plus eCG) treatments. No differences were observed among the number of recovered and morphologically graded COCs or the IVM rates for both gonadotropic treatments. However, the standard protocol produced COCs with higher EGFR expression in the cumulus cells than the one-shot treatment. Additionally, EGF mRNA levels were less than EGFR mRNA levels, and they did not differ among COCs from both treatments. However, during maturation, the EGF transcripts increased in oocytes derived only from the standard protocol. Interestingly, IVM strikingly increased EGFR expression in oocytes and cumulus cells but not in oocytes that fail in first polar body extrusion, irrespective of hormonal treatment. These results appear to be related to the resumption of meiosis and suggest that EGF may act through the cumulus cells or directly on the oocyte receptor.