RESUMO: O presente estudo objetivou avaliar a eficácia de um protocolo one-step de congelamento do sêmen suíno utilizando dimetilacetamida (DMA) e etilenoglicol como crioprotetores. Durante 10 semanas, a fração rica dos ejaculados de dois machos suínos foram coletados, uma vez por semana. Após a coleta, os ejaculados foram diluídos (1:1; v/v) no diluidor de resfriamento. Após a avaliação da concentração espermática, os ejaculados foram agrupados em um pool com o mesmo número de espermatozoides de cada macho e estabilizados a 20 °C por 120 min. Os criopropetores foram adicionados ao diluidor de congelamento a 20 °C, em diferentes concentrações, compondo seis tratamentos: glicerol (controle), 1,25% e 2,5%; etilenoglicol, 1,5% e 2,5%; e DMA, 2,5% e 5,0%. As amostras foram armazendadas em palhetas de 0,25 mL contendo 35 x 106 espermatozoides. Após 90 min a 20 °C as palhetas foram submetidas a uma curva de resfriamento até 5 °C (0,3 a 0,5 °C/mim) e mantidas a 5 °C por 60 min. O congelamento foi realizado a partir da colocação das palhetas horizontalmente a 5 cm acima do nitrogênio líquido por 10 min, com sua posterior imersão no nitrogênio líquido. Após o descongelamento a 37 °C por 30 segundos a qualidade espermática foi avaliada através de um sistema computadorizado e por citometria de fluxo. A motilidade espermática foi maior (P < 0,05) nos tratamentos com 5,0% e 2,5% DMA (22,2 ± 2,6% e 20,0 ± 2,8%, respectivamente) do que no tratamento com 2,5% etilenoglicol (8,2 ± 1,0%). A integridade da membrana plasmática (P = 0,08) e potencial de membrana mitocondrial (P = 0,27) foi similar entre os tratamentos. O tratamento com 2,5% de etilenoglicol foi menos eficiente em manter membrana acrossomal intacta (P< 0,01). Alguns parâmetros de cinética espermática (DAP, DCL, DSL, VAP, VCL, VSL e ALH) foram afetados positivamente pelo uso de DMA a 5.0%. O protocolo simplificado para congelamento de sêmen suíno resultou em motilidade espermática insatifatória após o descongelamento, independente do crioprotetor utilizado. RESUMO onestep one step (DMA crioprotetores 1 semanas coletados semana coleta 11 (1:1 v/v vv v 2 C 12 concentrações controle, controle , (controle) 125 25 1,25 15 1,5 2,5 50 0 025 0,2 3 9 0,3 03 (0, 05 0, °C/mim Cmim mim 6 fluxo P 0,05 005 5,0 22,2 222 22 (22, 26 2,6 200 20, 28 8 2,8% respectivamente 8,2 82 (8, 1,0%. 1,0% . 1,0%) 0,08 008 08 0,27 027 27 P< 0,01. 001 0,01 01 0,01) DAP, DAP (DAP DCL DSL VAP VCL ALH 5.0% utilizado (1: (controle 1,2 1, 2, 02 (0 0,0 00 5, 22, (22 2,8 8, (8 1,0 5.0 (1 ( (2 5.

ABSTRACT: The present study evaluated the cryoprotectant efficacy of dimethylacetamide (DMA) and ethylene glycol in a one-step protocol to freeze boar sperm. The sperm-rich portion of the ejaculates from two boars were collected once a week, for 10 weeks. After collection, the ejaculates were diluted (1:1; v/v) in the cooling extender. After determining their spermatozoa concentration, the ejaculates were pooled with the same number of spermatozoa from each boar and stabilized at 20°C for 120 min. Distinct cryoprotectants were added to the cooling extender at 20 °C, at different concentrations, composing six treatments: 1.25% and 2.5% glycerol (control); 1.25% and 2.5% ethylene glycol; 2.5% and 5.0% DMA. The samples were stored in 0.25 mL straws, containing 35 × 106 spermatozoa. After 90 min at 20 °C, the straws were submitted to a cooling curve until 5 °C (0.3 to 0.5 °C/min) and kept at 5°C for 60 min. Freezing was conducted by placing the straws horizontally 5 cm above the liquid nitrogen for 10 min, followed by immersion on liquid nitrogen. After thawing at 37 °C for 30 seconds, sperm quality was evaluated through a computer-assisted semen analysis system and flow cytometry. Sperm motility was greater (P< 0.05) in treatments with 5.0% and 2.5% DMA (22.2 ± 2.6% and 20.0 ± 2.8%, respectively) than in treatment with 2.5% ethylene glycol (8.2 ± 1.0%). The integrity of the plasma membrane (P = 0.08) and mitochondrial membrane potential (P = 0.27) was similar among the treatments. The treatment with 2.5% ethylene glycol was the least efficient to maintain intact acrosome membrane (P< 0.01). Some kinetics parameters (DAP, DCL, DSL, VAP, VCL, VSL e ALH) were positively affected by 5.0% DMA. The one-step freezing protocol resulted in unsatisfactory boar sperm motility after thawing, regardless of the cryoprotectant. ABSTRACT (DMA onestep one step spermrich rich week 1 weeks collection 11 (1:1 v/v vv v concentration 20C C 12 2 concentrations 125 25 1.25 2.5 control (control) 50 0 5.0 025 0.2 3 9 0.3 03 (0. 05 0. °C/min Cmin 5C 6 seconds computerassisted computer assisted cytometry P< P 0.05 005 22.2 222 22 (22. 26 2.6 200 20. 28 8 2.8% respectively 8.2 82 (8. 1.0%. 1.0% . 1.0%) 0.08 008 08 0.27 027 27 0.01. 001 0.01 01 0.01) DAP, DAP (DAP DCL DSL VAP VCL ALH (1: 1.2 2. (control 5. 02 (0 0.0 00 22. (22 2.8 8. (8 1.0 (1 1. ( (2

ABSTRACT Artificial insemination success in swine is mainly associated with semen dose quality. Thus, this study compared quality control parameters in 11 Brazilian boar studs after applying audit services for 24 months (1,650 boars). An extensive checklist was applied in each audit, registering ‘compliance’ or ‘noncompliance’ for 75 items. Semen doses produced were analyzed as regards volume and sperm concentration, and microbiological analyses were conducted for semen and water samples collected at distinct production stages. On average, boar studs produced 112.9 semen doses per boar per month, and the odds of raw semen contamination increased when boars were inadequately housed and doses were collected under increased temperatures, with no anti-slip rubber mat or after a poor prepuce cleaning (p < 0.05). Collection from boars with locomotor problems and no regular change of reverse osmosis filters increased the contamination odds in semen doses produced and stored at the stud (p < 0.05). As regards the water submitted to the osmosis reverse process, contamination odds increased as a result of deficient cleaning and disinfection of the purification equipment (p < 0.05). Risk factors for reduced sperm motility (< 70 %) were: no anti-slip rubber mat for semen collection, no cleaning program for automatic feeding system (drops) and bins, and inadequate intervals between semen collections (≤ 2 days or > 7 days; p < 0.05). Two boar studs had the best results for compliance with the checklist items. Constant monitoring, appropriate hygiene of facilities and equipment, and periodical staff training are highlighted as non-negotiable points for boar semen dose quality. Thus 1 1,650 1650 650 (1,65 boars. . boars) ‘compliance ‘noncompliance noncompliance items concentration stages average 1129 112 9 112. month temperatures antislip anti slip 0.05. 005 0.05 0 05 0.05) process ( % collection drops (drops bins ≤ monitoring nonnegotiable non negotiable 1,65 165 65 (1,6 00 0.0 1,6 16 6 (1, 0. 1, (1

Abstract The female reproductive function is coordinated by the endocrine system driven by the hypothalamic-pituitary-gonadal (HPG) axis. While not directly part of the female reproductive system, the gut microbiome plays a crucial role in overall health, including reproductive health. The gut microbiome communicates bidirectionally with the brain via the gut-brain axis, influencing stress levels, mood, and hormonal balance, which can impact reproductive health and fertility. In addition to that, the vaginal and uterine microbiome are directly involved with the reproductive success of farm animals, including female fertility and offspring development. In this paper, we summarize some of the effects of bacterial contamination in the female reproductive tract and their association with reproductive performance in farm animals. hypothalamicpituitarygonadal hypothalamic pituitary gonadal HPG (HPG axis gutbrain levels mood balance that animals development paper

Abstract Polyethylenimine (PEI) has been explored as an efficient non-viral system for delivering genes to cells; however, there were no protocols for its use in porcine fetal fibroblasts (PFF). Therefore, we compared different concentrations of FITC-PEI (0.625, 1.25, 2.5, 5, 10, 20, 40, or 80 µg/mL) and incubation times (30 min, 1 h, or 2 h). It was observed that the incubation time did not affect the internalization of the PEI-FITC and that 30 min was sufficient to capture the complex. The concentrations higher than 10 µg/mL could reach many marked PFF (>90%). Then, two PEI concentrations were tested, 10 or 40 µg/mL, combined with an N/P of 2 with the pmhyGENIE-5 for 30 min. The percentage of PFF-GFP positive was similar between the PEI concentrations in the evaluation time points (24 h, 48 h, and 72 h). However, 40 µg/mL caused higher membrane damage rates. Thus, it can be concluded that concentrations between 10 – 80 µg/ml of PEI promote high incorporation rates, even in periods as short as 30 minutes. Furthermore, it can be stated that the transfection condition used in Polyplexes 1 (10 µg/mL of PEI and 37.5 µg/mL of pmhyGENIE-5 for 30 min) efficiently produces genetically edited porcine fetal fibroblasts with low cell damage. (PEI nonviral non viral cells however PFF. . (PFF) Therefore FITCPEI FITC 0.625, 0625 0 625 (0.625 125 25 1.25 5 2.5 20 8 µgmL µg mL (3 h h. h) PEIFITC 3 complex >90%. 90 >90% (>90%) Then tested 4 NP N P pmhyGENIE5 pmhyGENIE pmhyGENIE- PFFGFP GFP 24 (2 7 However rates Thus µgml ml minutes Furthermore (1 375 37 37. (PFF 0.625 062 62 (0.62 12 1.2 2. ( 9 >90 (>90% 0.62 06 6 (0.6 1. >9 (>90 0.6 (0. > (>9 0. (0 (>

Abstract Among the different methods used for semen collection from domestic cats, the pharmacological collection by urethral catheterization becomes disruptive. Medetomidine is the elected α2-adrenoceptor agonist for that, but in several countries, it is not commercially available. This study aimed to evaluate the efficacy of detomidine compared to medetomidine in collecting semen by urethral catheterization in domestic cats. Urethral catheterization was performed on 13 mongrel cats using a disposable semi-rigid tomcat urinary catheter. Of the 19 semen collections performed with medetomidine induction, 94.7% were successful, while with detomidine induction, only 56.3% of 16 were successful. The values semen samples variables were as follows for volume - 10.56 ± 0.4 vs 8.88 ± 0.5 mL, motility - 171.67 ± 0.79 vs 49.77 ± 3.45%, vigor – 4.1 ± 0.03 vs 3.10 ± 0.1 and concentration - 3.24 ± 0.19 vs 2.15 ± 0.13 ×109 sperm/mL respectively for medetomidine and detomidine group. The failure in semen collections with detomidine was mainly due to azoospermic samples, poor urethral relaxation, insufficient volume, or contamination of urine. The sperm concentration was also lower in the detomidine group (P <0.05) when compared to medetomidine. However, when the volume of semen collected was compared, we found no statistical differences. Despite its low performance in collecting semen from cats, detomidine may be an alternative when medetomidine is not accessible.

Abstract Twin birth is a complex condition observed in most livestock animals, when the female gives birth to two or more offspring, generally out of the same mating. In cattle, it is a rare condition (3 to 5%) and depends on the genetic background and environmental factors. Twin birth is a result of multiple ovulations, being more common in dairy rather than in beef cattle. Calves could be monozygous or dizygous, with the same or of different sexes. When twins are born with different sexes, a sexual condition called Freemartinism occurs in between 90 to 97% of pregnancies, causing infertility in the female calf. Knowing that the twin rate is rare in commercial beef cattle, here we present an even rarer case of twin birth from two different sires after natural mating, also called heteropaternal superfecundation.