Porcine Germ Cells Phenotype during Embryonic and Adult Development.

Primordial germ cells (PGCs) are the precursors of gametes. Due to their importance for the formation and reproduction of an organism, understanding the mechanisms and pathways of PGCs and the differences between males and females is essential. However, there is little research in domestic animals, e.g., swine, regarding the epigenetic and pluripotency profiles of PGCs during development. This study analyzed the expression of epigenetic and various pluripotent and germline markers associated with the development and differentiation of PGCs in porcine (pPGCs), aiming to understand the different gene expression profiles between the genders. The analysis of gonads at different gestational periods (from 24 to 35 days post fertilization (dpf) and in adults) was evaluated by immunofluorescence and RT-qPCR and showed phenotypic differences between the gonads of male and female embryos. In addition, the pPGCs were positive for OCT4 and VASA; some cells were H3k27me3 positive in male embryos and adult testes. In adults, the cells of the testes were positive for germline markers, as confirmed by gene expression analysis. The results may contribute to understanding the pPGC pathways during reproductive development, while also contributing to the knowledge needed to generate mature gametes in vitro.

Impact of cryopreservation protocols (one- and two-step) on boar semen quality at 5 °C and post-thawing.

The two-step protocol (2 S) is currently used for boar semen cryopreservation. In this method, the cryoprotectant penetrant agents (CPAs) are added at 5 °C to reduce the toxicity of CPAs. An alternative is the one-step protocol (1 S), which is easier, cheaper, and reduces the necessity of equipment, but could increase the toxicity of CPAs. Currently, there are no studies that compared both protocols for boar semen cryopreservation. This experiment aimed to study the effect of cryopreservation protocol (1 S vs 2 S) on boar spermatozoa. In the one-step protocol, after centrifugation, the spermatozoa pellet was resuspended at 17 °C in the extender containing CPAs to achieve a concentration of 1 × 109 spermatozoa/mL and then submitted to cryopreservation. For the two-step protocol, the sperm pellet was resuspended in fraction A at 17 °C to achieve a concentration of 1.5 × 109 spermatozoa/ mL, and then allowed to cool to 5º C before fraction B with CPA was added to the sample to achieve a final concentration of 1 × 109 spermatozoa/mL and followed by freezing. The cryopreservation protocol did not impact total motility at 5 °C (1 S: 78.5 % vs 2 S: 79 %, p > 0.05). After thawing, the two-step protocol improved (p < 0.05) total (1 S: 18.2 % vs 2 S: 29.5 %) and progressive motility (1 S: 9 % vs 2 S: 15%). Further, the 2 S protocol increased (p < 0.05) the percentage of rapid spermatozoa (1 S: 8.7 % vs 2 S: 14.6 %) and spermatozoa with intact plasma and acrosomal membrane (IAIP) (1 S: 40.5 % vs 2 S: 61.5 %), and increased (p < 0.05) live sperm cells with high mitochondrial potential (MHIP) (1 S: 42.9 % vs 2 S: 60 %). The boar semen cryopreservation method (TRT) did not (p > 0.05) alter membrane lipid disorder, lipid peroxidation, and superoxide anion. Thus, the best method for boar semen cryopreservation is the two-step protocol.

Effect of cow's milk with different PUFA n-6: n-3 ratios on performance, serum lipid profile, and blood parameters of grower gilts.

The concern with human health has increased the interest in producing foods enriched with polyunsaturated fatty acids (PUFA), directly or naturally, by inclusion in the animals' diet. The positive effects such as antithrombotic, anti-inflammatory, and hypolipidemic have been observed in pigs and rats, used as human models for study. The present study evaluated the effect of cow's milk with different lipid profiles on performance, serum fatty acid profile, biochemical analysis, and a complete blood count of gilts used as a human model. At 34 days, thirty gilts were equally distributed in three treatments. Experimental treatments were milk from cows without the oil supplementation (C), milk from cows fed an enriched diet with linseed oil (n-3), and milk from cows fed an enriched diet with soybean oil (n-6). Milk supplementation was performed until 190 days old, provided once in the morning. The n-3 and n-6 milk reduced the concentration of myristic acid in the blood and increased the leukocytes. Milk enriched with n-3 compared to n-6 reduced the stearic acid. In conclusion, milk with a better PUFA profile can reduce saturated fatty acids in the blood and alter the concentration of cells in the defense system.

In vitro induced pluripotency from urine-derived cells in porcine.

BACKGROUND: The generation of induced pluripotent stem cells (iPSC) has been a game-changer in translational and regenerative medicine; however, their large-scale applicability is still hampered by the scarcity of accessible, safe, and reproducible protocols. The porcine model is a large biomedical model that enables translational applications, including gene editing, long term in vivo and offspring analysis; therefore, suitable for both medicine and animal production. AIM: To reprogramme in vitro into pluripotency, and herein urine-derived cells (UDCs) were isolated from porcine urine. METHODS: The UDCs were reprogrammed in vitro using human or murine octamer-binding transcription factor 4 (OCT4), SRY-box2 (SOX2), Kruppel-like factor 4 (KLF4), and C-MYC, and cultured with basic fibroblast growth factor (bFGF) supplementation. To characterize the putative porcine iPSCs three clonal lineages were submitted to immunocytochemistry for alkaline phosphatase (AP), OCT4, SOX2, NANOG, TRA1 81 and SSEA 1 detection. Endogenous transcripts related to the pluripotency (OCT4, SOX2 and NANOG) were analyzed via reverse transcription quantitative real-time polymerase chain reaction in different time points during the culture, and all three lineages formed embryoid bodies (EBs) when cultured in suspension without bFGF supplementation. RESULTS: The UDCs were isolated from swine urine samples and when at passage 2 submitted to in vitro reprogramming. Colonies of putative iPSCs were obtained only from UDCs transduced with the murine factors (mOSKM), but not from human factors (hOSKM). Three clonal lineages were isolated and further cultured for at least 28 passages, all the lineages were positive for AP detection, the OCT4, SOX2, NANOG markers, albeit the immunocytochemical analysis also revealed heterogeneous phenotypic profiles among lineages and passages for NANOG and SSEA1, similar results were observed in the abundance of the endogenous transcripts related to pluripotent state. All the clonal lineages when cultured in suspension without bFGF were able to form EBs expressing ectoderm and mesoderm layers transcripts. CONCLUSION: For the first time UDCs were isolated in the swine model and reprogrammed into a pluripotent-like state, enabling new numerous applications in both human or veterinary regenerative medicine.

Porcine Primordial Germ Cell-Like Cells Generated from Induced Pluripotent Stem Cells Under Different Culture Conditions.

Culture conditions regulate the process of pluripotency acquisition and self-renewal. This study aimed to analyse the influence of the in vitro environment on the induction of porcine induced pluripotent stem cell (piPSCs) differentiation into primordial germ cell-like cells (pPGCLCs). piPSC culture with different supplementation strategies (LIF, bFGF, or LIF plus bFGF) promoted heterogeneous phenotypic profiles. Continuous bFGF supplementation during piPSCs culture was beneficial to support a pluripotent state and the differentiation of piPSCs into pPGCLCs. The pPGCLCs were positive for the gene and protein expression of pluripotent and germinative markers. This study can provide a suitable in vitro model for use in translational studies and to help answer numerous remaining questions about germ cells.

Copper and zinc hydroxychloride cosupplementation improve growth performance and carcass and reduce diarrhea frequency in grower-finisher pigs.

This study investigated copper (Cu) and zinc (Zn) hydroxychloride cosupplementation on the growth performance, diarrhea frequency, carcass, meat quality, and antioxidant activity in grower-finisher pigs. A total of 256 pigs were used from 70 to 154 days (d) of age, distributed in four treatments, with eight pigs in each pen and eight replications per treatment. Diets were provided to grower pigs from 70 to 112 days old and in the finisher, 112 to 154 days old. Copper was considered the low level at 100 mg Cu/kg and 90 mg Cu/kg, respectively, and 150 mg Cu/kg in both periods as high in the grower and finisher periods. In the grower and finisher period, zinc was cosupplemented in the diet at 80 mg Zn/kg and 70 mg Zn/kg, respectively. In the diets, T1 and T2 groups are the traditional inorganic sources for minerals (copper sulfate, CuSO4; zinc oxide, ZnO) and T3 and T4 hydroxychloride sources (copper hydroxychloride, CHC, and zinc hydroxychloride, ZHC). The flavomycin was associated with treatments with low Cu content in the inclusion of 50 g/ton. The experimental design was in randomized blocks, the data were submitted to analysis of PROC MIXED in SAS, the PDIFF test analyzed the treatment effect. At the finisher period, pigs fed both minerals from hydroxychloride source had a higher BW 154 d, average daily gain (ADG) 70 to 154 d, the hot and cold carcass weight and frequency of normal feces than those fed 150 mg Cu/kg and Zn from a traditional inorganic source (P < 0.05). The animals fed low Cu levels of the sulfate source had a higher ADG 70 to 154 d than those fed high Cu levels of the same source (P < 0.05). Pigs fed 150 mg Cu/kg cosupplemented with Zn from a hydroxychloride source had the highest carcass length (P < 0.05). There was no difference among the treatments for meat quality (P > 0.05). Pigs fed 150 mg Cu/kg and Zn from a traditional inorganic source had a higher superoxide dismutase (SOD) activity than the other treatments (P < 0.05). Animals fed low Cu levels from hydroxychloride had a higher malondialdehyde (MDA) formation than those fed sulfate source, regardless of the Cu levels and those fed high Cu levels of hydroxychloride (P < 0.05). In conclusion, 150 mg Cu/kg as copper sulfate cosupplemented to zinc oxide in the diet of growing and finishing pigs impairs the growth performance, carcass and increases diarrhea frequency, and copper and zinc hydroxychloride cosupplementation improves these characteristics.

Maternal Supplementation with Cow's Milk Naturally Enriched with PUFA Alters the Metabolism of Sows and the Fatty Acid Profile of the Offspring.

The study aimed to evaluate the supplementation of gilts with cow's milk naturally enriched with n-3 and n-6 polyunsaturated fatty acids (PUFA) on reproductive outcomes, and the serum biochemical and FA profile of swine females and their offspring. During 316 days, 30 gilts were distributed into three groups: (1) Control, fed a basal diet + milk from cows without oil; (2) n-3, fed a basal diet + milk from cows fed a diet enriched with linseed oil; (3) n-6, fed a basal diet + milk from cows fed a diet enriched with soybean oil. The gilts receiving the diets containing PUFA had higher serum urea and very-low-density lipoprotein levels and lower serum total protein and low-density lipoprotein levels compared to the Control group. Females supplemented with n-3 presented higher serum palmitic acid and γ-linolenic acid levels than those fed n-6. Piglets from the Control group were heavier at birth than those from females supplemented with enriched milk. The piglets from females receiving enriched milk had 140 g higher body weight from 1 to 21 days old compared to the Control group, and greater average daily weight gain from 7 to 14 days old. The serum eicosapentaenoic acid level of piglets fed n-3 was 69% higher than those fed n-6, which reduced the AA/EPA ratio. Gilts supplemented with PUFA-enriched cow's milk showed changes in their serum palmitic and γ-linolenic acid levels, in addition to improved performance, EPA concentration and consequently reduced AA/EPA ratio in their piglets, demonstrating beneficial results for their progeny.

Altrenogest during early pregnancy modulates uterine glandular epithelium and endometrial growth factor expression at the time implantation in pigs.

This study evaluated the effects of supplying altrenogest from day 6-12 of pregnancy on the endometrial glandular epithelium, corpora lutea (CL) morphology, and endometrial and CL gene expression. A total of 12 crossbred females (Landrace × Large White) were used. The females were assigned to 4 treatments according to a random design with a 2 × 2 factorial arrangement, with two categories (sow or gilt) and two treatments (non-treated and treated with altrenogest). On day 6 of pregnancy, animals were allocated to one of the following groups: non-treated (NT, n = 6; 3 sows and 3 gilts), and (T, n = 6; 3 sows and 3 gilts) treated daily with 20 mg of altrenogest, from day 6-12 of pregnancy. All animals were euthanized on day 13 of pregnancy. All CLs were individually weighed, and their volume were determined. The endometrial glandular density (GD), mean glandular area (MGA), and vascular density (VD) were determined by histomorphometric and immunohistochemical analyses. Endometrium samples were collected and analyzed by qRT-PCR to evaluate the abundance of transcripts for VEGF and IGF-I. Females in the T group had higher MGA (P < 0.05) compared to the NT group. There was no effect of treatment on GD or VD for both experimental groups. Sows in the T group had augmented expression of IGF-I (P < 0.05). Progestagen had no detrimental effect on CL morphology. In conclusion, altrenogest improves the uterine environment during the peri-implantation period in pigs without compromising corpora lutea development.

The use of resveratrol decreases liquid-extend boar semen fertility, even in concentrations that do not alter semen quality.

In vitro and in vivo assays were conducted to investigate the effects of trans-resveratrol (RVT) on liquid-extended boar semen during 72 h of storage at 17 °C. Thirty-six ejaculates were collected from six boars, evaluated, and extended. RVT was then added at the indicated treatment concentration (0, 0.01, 0.1 or 1 mM), and the ejaculates were cooled to 17 °C and evaluated at 0, 24, 48, and 72 h. Samples were evaluated for sperm motility, kinetics, plasma and acrosome integrity, mitochondrial membrane potential, anion superoxide levels, lipoperoxidation, and antioxidant enzyme activity. In the follow-up experiment, twenty-eight gilts were fixed-time inseminated with 0 or 0.01 mM RVT liquid-extended boar semen. After five days, they were slaughtered, and their reproductive tracts were recovered. The embryos were collected, and the pregnancy, fertility, and viable embryo rates were calculated. In the in vitro assays, total motility, plasma and acrosome membrane integrity, mitochondrial membrane potential, anion superoxide levels, and lipoperoxidation did not change at any of the evaluation times with the use of RVT up to 0.01 mM. RVT decreased SOD activity without changes in GPx. RVT used at 1 mM showed harmful effects for almost all evaluated parameters. For the in vivo assay, the same pregnancy and fertility rates were observed for both groups, while the viable embryo rate was three-fold lower in the 0.01 mM group than in the 0 mM group. The results showed a dichotomous effect of RVT; a low concentration was not harmful in vitro but was catastrophic for embryo viability.

Generation of neural progenitor cells from porcine-induced pluripotent stem cells.

In this study, porcine embryonic fibroblasts (pEFs) were reprogrammed into porcine-induced pluripotent stem cells (piPSCs) using either human or mouse specific sequences for the OCT4, SOX2, c-Myc, and KLF4 transcription factors. In total, three pEFs lines were reprogrammed, cultured for at least 15 passages, and characterized regarding their pluripotency status (alkaline phosphatase expression, embryoid body formation, expression of exogenous and endogenous genes, and immunofluorescence). Two piPSC lines were further differentiated, using chemical inhibitors, into putative neural progenitor-like (NPC-like) cells with subsequent analyses of their morphology and expression of neural markers such as NESTIN and GFAP as well as immunofluorescent labeling of NESTIN, ß-TUBULIN III, and VIMENTIN. NPC-like cells were positive for all the neural markers tested. These results evidence of the generation of porcine NPC-like cells after in vitro induction with chemical inhibitors, representing an adequate model for future regenerative and translational medicine research.

Removal of seminal plasma prior to liquid storage of boar spermatozoa: A practice that can improve their fertilizing ability.

Seminal plasma (SP) plays a vital role in the maintenance of sperm function and integrity along with being involved in their communication with the female reproductive tract. Under in vitro conditions, although it is generally accepted that boar semen is better preserved when SP is diluted (extended) or removed (cryopreserved), its role during storage is not completely elucidated. In this context, the current study sought to determine the role of SP during storage of boar spermatozoa at 17 °C for 72 h. Thus, two treatments were prepared with semen from the sperm-rich fraction (SRF) of boar ejaculate previous to storage in liquid state: 1) PSP: semen directly extended in Beltsville Thawing Solution (BTS), and 2) ASP: semen first centrifuged with subsequent removal of supernatant (containing SP) followed by suspension of sperm in BTS. From this, two experiments were conducted separately in this work: 1) in vitro and 2) in vivo assays. The former aimed to evaluate how sperm capacity responds to in vitro capacitation (IVC) and whether their quality is affected by previous exposure to SP. In the latter, the objective was to understand how important these previous conditions can be for reproductive performance after artificial insemination (AI). According to our results, the previous removal of SP does not affect sperm quality and the response of these cells to IVC (P > 0.05) along with resulting in a lower percentage of acrosome damage in them [12.87 ±â€¯0.76 (ASP) vs. 16.38 ±â€¯0.73 (PSP)] (P < 0.05). This improved preservation of acrosome integrity in the absence of SP can explain the higher fertility rate (%) [63.27 ±â€¯23.47 (ASP) vs. 38.57 ±â€¯16.30 (PSP)] and number of implanted embryos at 28 days after AI (13.71 ±â€¯4.88 (ASP) vs. 7.16 ±â€¯4.02 (PSP)] (P < 0.05) presented by gilts inseminated with seminal doses of ASP. In conclusion, removal of SP prior to liquid storage of boar sperm for 72 h can be beneficial for their fertilizing ability.

Novel Flow Cytometry Analyses of Boar Sperm Viability: Can the Addition of Whole Sperm-Rich Fraction Seminal Plasma to Frozen-Thawed Boar Sperm Affect It?

Boar semen cryopreservation remains a challenge due to the extension of cold shock damage. Thus, many alternatives have emerged to improve the quality of frozen-thawed boar sperm. Although the use of seminal plasma arising from boar sperm-rich fraction (SP-SRF) has shown good efficacy; however, the majority of actual sperm evaluation techniques include a single or dual sperm parameter analysis, which overrates the real sperm viability. Within this context, this work was performed to introduce a sperm flow cytometry fourfold stain technique for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential. We then used the sperm flow cytometry fourfold stain technique to study the effect of SP-SRF on frozen-thawed boar sperm and further evaluated the effect of this treatment on sperm movement, tyrosine phosphorylation and fertility rate (FR). The sperm fourfold stain technique is accurate (R2 = 0.9356, p > 0.01) for simultaneous evaluation of plasma and acrosomal membrane integrity and mitochondrial membrane potential (IPIAH cells). Centrifugation pre-cryopreservation was not deleterious (p > 0.05) for any analyzed variables. Addition of SP-SRF after cryopreservation was able to improve total and progressive motility (p < 0.05) when boar semen was cryopreserved without SP-SRF; however, it was not able to decrease tyrosine phosphorylation (p > 0.05) or improve IPIAH cells (p > 0.05). FR was not (p > 0.05) statistically increased by the addition of seminal plasma, though females inseminated with frozen-thawed boar semen plus SP-SRF did perform better than those inseminated with sperm lacking seminal plasma. Thus, we conclude that sperm fourfold stain can be used to simultaneously evaluate plasma and acrosomal membrane integrity and mitochondrial membrane potential, and the addition of SP-SRF at thawed boar semen cryopreserved in absence of SP-SRF improve its total and progressive motility.

Probiotics association in the suckling and nursery in piglets challenged with Salmonella typhimurium

This work aimed to study the probiotics association in 144 piglets from birth to 62 days old. In lactation, the design was completely randomized with two treatments, CTL, 1 mL of distilled water and ProbA, 5g in 15 ml of distilled water, both orally, and in the nursery in randomized block design, with 2x3 factorial arrangement of treatments, ProbA ProbB: 30g/ton of ProbB in the ration; CTL ProbB: 30g/ton of ProbB in the ration; ProbA ProbA, CTL CTL; ProbA CHA (challenged); CTL CHA. At 35 days of age the animal of the nursery were inoculated with Salmonella typhimurium orally. There was no effect of the parameters evaluated during the maternity. In nursery, the feed conversion was favorable to the ProbA. In the evaluation of fecal score, the challenged group had more diarrhea and increased elimination of S. typhimurium. Results showed the positive action of probiotics when applied at birth by the direct influence on the formation of the intestinal microbiota.

Combined effects feeding restriction and flushing on fertility of gilts inseminated at different estrous cycles / Efeitos combinados da restrição alimentar e flushing sobre a fertilidade de marrãs inseminadas artificialmente em diferentes ciclos estrais

The study evaluated the effect of dietary restriction in prepubertal gilts associated with the use of flushing in the preceding cycle of artificial insemination, 2nd or 3rd estrus on productive performance, number of corpora lutea and fetus, besides the fetus survival rate. A total of 96 gilts were distributed to a completely randomized design with 2x2x2 factorial arrangement of treatments, consisting of: nutritional strategy, ad libitum or restricted feeding, use or not of flushing in the preceding cycle AI, and time of AI, held on the 2nd or 3rd estrus. The age at which gilts expressed the 1st estrus was not influenced by nutritional strategy, but the body weight and weight gain were higher (P 0.05) in gilts submitted to ad libitum feeding. No interactions were observed as treatment effect on number of corpora lutea and fetus survival rate. The number of fetuses in females on ad libitum feeding was not influenced by time of AI, but the females feed restriction showed an increase (P 0.05) in the number of fetuses when inseminated on 3rd estrus. Thus, it is concluded that the use of ad libitum nutritional strategy allows for insemination forward 2nd estrus without reducing the number of fetuses, reducing the non-productive days gilt. However, this strategy reduces the rate of fetal survival until day 35 of gestation...

Foi avaliado o efeito da restrição alimentar na fase pré-púbere associada ao emprego do flushing no ciclo precedente ao da inseminação artificial, sendo realizada no 2 ou 3 estro, sobre o desempenho produtivo, numero de corpos lúteos e de fetos, além da taxa de sobrevivência fetal. Um total de 96 fêmeas foi distribuído em delineamento inteiramente casualizado com arranjo fatorial 2x2x2, sendo considerados os fatores: estratégia nutricional, alimentação ad libitum ou restrita; utilização ou não de flushing no ciclo precedente a IA, e momento da IA, realizada no 2 ou 3 estro, constituindo oito tratamentos. A idade em que as marras manifestaram o 1 estro não foi influenciada pela estratégia nutricional, mas o peso e o ganho de peso foram maiores (P 0,05) na alimentação ad libitum. Não foram observados interação e tão pouco efeito de tratamento no numero de corpos lúteos e a taxa de sobrevivência dos fetos. O numero de fetos em fêmeas mantidas em alimentação ad libitum não foi influenciado pelo momento da IA, contudo as fêmeas em restrição alimentar apresentaram aumento (P 0,05) no numero de fetos quando inseminadas no 3 estro. Desse modo, conclui-se que o emprego da estratégia nutricional ad libitum possibilita adiantar a inseminação para o 2 estro sem redução no numero de fetos, diminuindo os dias não produtivos da marra. Porém, essa estratégia reduz a taxa de sobrevivência fetal até 35º dia de gestação...

Exploring the effects of second estrus synchronization and dietary flushing on the incidence of ovarian cysts in gilts by using exogenous gonadotropins / Explorando os efeitos da sincronização do segundo estro e flushing alimentar sobre a incidência de cistos ovarianos em marrãs utilizando gonadotrofinas exógenas

Estrus stimulation by exogenous gonadotropins (EG) in association with dietary flushing is an important tool for theimprovement of gilt reproductive performance. However, there is evidence associating both flushing and EG with a disturbance in the endocrine balance that could lead to increased ovarian cysts. The aim of this study was to evalu- ate whether flushing or EG might affect the ovulation rate and the incidence of ovarian cysts. Seventy-one gilts were randomly distributed into 2x2 factorial design with four treatments: flushing and hormone (wFwH); no flushing and hormone (nFwH); flushing without hormone (wFnH); and neither flushing nor hormone (nFnH). Gilts were slaughtered for macroscopic and histopathological ovary examination approximately five days after AI. The characterization of these cysts was performed by optical microscopy in the following: follicular cysts (FC), luteinizedcysts (LC) or cystic corpora lutea (CCL). The number of ovulations did not differ between treatments. There was no interaction between the factors in any analyzed variable. The frequency of gilts with CCL and LC was not affected by flushing and EG. No difference was found in the incidence of FC, with 12.5% and 5.88% in gilts from wFwH and nFwH treatments, respectively. There were no differences in the proportion of CCL between FC and LC (9.85 vs. 4.22 and 4.22%, respectively). In conclusion, the use of exogenous gonadotropins for second estrus synchronization in gilts, either alone or in association with dietary flushing, does not increase the incidence of ovarian cysts, nor does it decrease the ovulation rate.

A estimulação do estro por gonadotrofinas exógenas (GE) associada ao flushing alimentar é uma ferramenta importante na melhoria do desempenho reprodutivo de marrãs. Contudo, há evidência da associação do flushing com GE levando ao desequilíbrio no sistema endócrino que poderia levar ao aumento de cistos ovarianos. O objetivo deste estudo foi avaliar se o flushing ou GE pode afetar a taxa de ovulação e a incidência de cistos ovarianos. Setenta e uma marrãs foramdistribuídas aleatoriamente em arranjo fatorial 2x2 com quatro tratamentos: flushing e hormônio (cFcH); sem flushing e com hormônio (sFcH); com flushing e sem hormônio (cFsH) e sem flushing e hormônio (sFsH). Marrãs foram abatidas para exame macroscópico e histopatológico dos ovários, aproximadamente cinco dias após IA. A caracterização desses cistos foi realizada por microscopia óptica: cistos foliculares (CF), cistos luteinizados (CL) ou corpos lúteos císticos(CCL). O número de ovulações não diferiu entre os tratamentos. Não houve interação entre os fatores em qualquer variável analisada. A frequência de leitoas com CCL e CL não foi afetada pelo flushing e GE. Não houve diferença na incidência de CF, com 12,5% e 5,88 % em leitoas dos tratamentos cFcH e sFcH, respectivamente. Não foram obtidas diferenças na proporção de CCL entre CF e CL (9,85 vs. 4,22 e 4,22%, respectivamente). Em conclusão, a utilização de gonadotrofinas exógenas para sincronização do segundo estro de marrãs, isoladamente ou em associação com o flushing, não aumenta a incidência de cistos ovarianos e não diminui a taxa de ovulação.

Post-thaw addition of seminal plasma reduces tyrosine phosphorylation on the surface of cryopreserved equine sperm, but does not reduce lipid peroxidation.

The objective was to verify the relationship between equine semen cryopreservation and changes related to increased lipid peroxidation. Also, addition of autologous or homologous seminal plasma from a stallion with a good freezing response to post-thawed sperm was tested to determine whether it would confer protection. Frozen-thawed sperm were evaluated and allocated into three groups: without plasma addition, and supplemented with either homologous or autologous seminal plasma. All groups were evaluated at 0, 60 and 120 min after incubation at 37 °C. Cryopreservation did not increase plasma membrane disorders (mean ± SEM 9.48 ± 0.65 and 1.62 ± 0.23% in raw and frozen-thawed sperm, respectively). However, both membrane peroxidation and protein phosphorylation were increased (P < 0.05) compared to raw semen (1.74 and 5.20-fold, respectively). There was a correlation (r = 0.73; P < 0.05) between the increase in lipid peroxidation and tyrosine phosphorylation. Seminal plasma, regardless of origin, reduced (P > 0.05) tyrosine phosphorylation present on the surface of cryopreserved sperm; however, lipid peroxidation was not significantly reduced. In conclusion, we inferred that emergence of phosphorylated proteins on the surface of cryopreserved sperm was due to increased lipid peroxidation that occurred during the freezing/thawing process; however, reduced tyrosine phosphorylation that occurred after addition of seminal plasma was triggered by other mechanisms, apparently independent from the reduction in membrane peroxidation.

Efeito de probiótico no desenvolvimento de leitões desafiados com Escherichia coli / Effect of probiotic on the development of piglets challenged with Escherichia coli

Foram utilizados 72 animais aos 21 dias de idade para averiguar o efeito do probiótico no desempenho de leitões desafiados com E. coli. O delineamento foi em blocos casualizados, com arranjo fatorial 2x2. Os quatro tratamentos foram: probiótico com desafio; probiótico sem desafio; sem probiótico com desafio; sem probiótico e sem desafio. O probiótico Protexin™ foi administrado via ração, dos 21 aos 63 dias de idade e aos 35 dias, os animais dos tratamentos 1 e 3 foram desafiados. As variáveis analisadas foram: peso médio aos 35, 49 e 63 dias de idade; ganho diário de peso dos 35 aos 49 dias; dos 49 aos 63 dias; dos 35 aos 63 dias e escore fecal. As variáveis ganho diário de peso, peso médio e escore fecal foram submetidos à análise de variância pelo PROC GLM. Em relação ao peso e ganho de peso verificou-se ação benéfica do probiótico quando se associaram estas características com a ocorrência de diarréia pós-desafio e a redução da E. coli. Concluiu-se que o probiótico no modelo experimental aplicado apresentou efeito notório nas reduções de E. coli e da diarréia, podendo exercer influência no desempenho dos leitões.

We used 72 piglets at 21 days old and the aim was to determine the effect of probiotics on performance of piglets challenged with E. coli. The design was a randomized block design with 2x2 factorial arrangement. The four treatments were: with probiotic and challenge, with probiotic without challenge, without probiotic with challenge, without probiotic and challenge. The probiotic was administered via Protexin ™ diet, from 21 to 63 days old and 35 days, the animals in treatments 1 and 3 were challenged. The variables were: average weight at 35, 49 and 63 days old, daily weight gain from 35 to 49 days, 49 to 63 days, from 35 to 63 days and fecal score. The variables average weight, daily weight gain and diarrhea score were subjected to analysis of variance by PROC GLM. In terms of weight and weight gain was found beneficial effect of probiotic when these characteristics were associated with the occurrence of diarrhea post-challenge and the reduction of E. coli. It was concluded that the probiotic used in the experimental model showed clear effect on the reductions of E. coli and diarrhea, are relatively influenced in the performance of piglets.

Glândula mamária do mocó (Kerodon rupestris -Wied Neuwied, 1820): aspectos morfológicos / Mammary gland of the rock cavy (Kerodon rupestris - Wied Neuwied, 1820): morphologic aspects

Procurando colaborar com a exploração zootécnica racional dos mocós(Kerodon rupestris) foi descrita a morfologia da glândula mamária destes roedores, que por sua vez podem tornar-se uma fonte de alimento para população nordestina. Foram utilizados cinco animais cedidos pelo Centro de Multiplicação de Animais Silvestres (CEMAS) da pela Escola Superior de Agricultura de Mossoró (ESAM) registrado junto ao IBAMA como criadouro científico sob o número 12.492- 0004. As glândulas mamárias foram retiradas através de dissecação e foram fotografadas. Cada animal apresentou duas glândulas mamárias localizadas na região inguinal. Histologicamente a glândula mamária era composta por uma grande quantidade de lóbulos, repletos de glândulas túbulos-alveolares com epitélio simples cúbico. Na papila mamária fibras musculares apresentaram-se dispostas de modo circular, o que possivelmente auxilia na contração para excreção do leite.

In attempt to collaborate with the rational zootecnic exploration of the Rock Cavy (Kerodon rupestris), the morphology of the mammary gland from these rodends was described, so that it can become a food source for north eastern population. Were used five animals yieldedby the Center of Multiplication of Animals Savage (CEMAS), createdby the Superior School of Agriculture of Mossoró (ESAM) registered in IBAMA as scientific breeding with number 12,492- 0004. The mammary glands were remove through dissecation and were photographed. Each animal presented two mammary glands in the base of the insertion of the pelvic members in the abdominal region. Histologicaly the mammary gland is composed of a great amount of lobes, replete of alveolar tubs glands with cubical simple epithelium.Its mammary papilla showed an amount of muscular staple fibres incircular way that assists it in the contraction for excrement of milk.