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Coriander (Coriandrum sativum) is a species highly appreciated in Brazilian cuisine and of high nutritional importance due to the presence of vitamins, calcium, and iron. Leafy vegetables are considered nutritionally demanding as their cycles are relatively short and the lack of any essential element affects their growth, yield and quality. This study aimed to evaluate the response of coriander plants (Coriandrum sativum L.) to foliar spraying of boron doses. The experiment was carried out in a greenhouse at The State University of Goiás, Campus Ipameri, in Ipameri-GO. The experimental design was completely randomized, with five treatments and four replications. The treatments consisted of five boron doses applied by foliar spray (0, 1.0, 1.5, 2.0 and 3.0 kg ha-1) using boric acid (17% of Boron) as the source. Fresh and dry matter of shoot and root and leaf boron contents were evaluated. Data were subjected to analysis of variance (F test) and the effect of boron doses were evaluated by regression analysis. The application of 3.0 kg ha-1 of boron increases the boron content of coriander plants, and the fresh and dry matter of roots and shoots was not significantly increased.

Campomanesia adamantium (Cambess.) Berg O. is a native of Savanna, popularly known as gabiroba, guavira, or gabiroba-do-campo, and has many uses, with its fruit being consumed fresh or processed. This study aimed to evaluate different doses of phosphorus and nitrogen in the initial development of gabiroba (Campomanesia adamantium) cultivated in pots. Five doses of phosphorus (0, 100, 200, 300, and 400kg ha-1 of P2O5) and three doses of nitrogen (0, 100, and 200 kg ha-1of N) were tested. The experimental design used was completely randomized in a 5x3 factorial scheme with six replications. The evaluations were carried out 120 days after plant emergence, and the variables analyzed were stem diameter, plant height, and number of leaves. Data were subjected to analysis of variance and F-test. The means were compared by the Tukey test at p0.05. Regression analyzes were adjusted for phosphorus doses. The gabiroba responds significantly to the addition of phosphorus, with the dose of 400 kg ha-1 of P2O5 providing the highest height and the dose of 300 kg ha-1 of P2O5 providing the greatest stem diameter and the number of leaves. Up to 120 days old, gabiroba seedlings do not respond significantly to nitrogen fertilization.

O principal objetivo da educação na atualidade é promover uma formação crítica/cidadã, sendo necessário para isso que ocorram modificações no processo de ensino e aprendizagem. Dessa forma, buscando a promoção dessa formação diferenciada, os subprojetos do Programa Institucional de Bolsas de Iniciação à Docência dos cursos de Química, Matemática, História, Geografia e Biologia da Universidade Federal de Catalão (UFCat), Universidade Federal de Catalão em implantação desenvolveram como atividade de extensão a 2ª Feira de Ciências da UFCat: “Ciência a Flor da Pele”, que contou com a participação 126 alunos e 15 professores, de 10 escolas públicas e privadas da cidade de Catalão e região. Este trabalho objetiva analisar como a participação de alunos da Educação Básica em uma Feira de Ciências de nível regional pode impactar no seu processo de ensino e aprendizagem. Para a coleta dos dados fez-se uso de questionários que foram respondidos pelos discentes da Educação Básica que participaram das atividades; os questionários apresentavam questões abertas e foram analisados mediante Análise Textual Discursiva. A partir da análise dos dados, percebeu-se ganhos significativos no processo de ensino e aprendizagem dos alunos participantes, pois estes foram estimulados a elaborar seus próprios trabalhos de forma crítica, a trabalhar em grupo, a analisar/selecionar informações, a se comunicar em público, dentre outros. Conclui-se assim que a 2ª Feira de Ciências da UFCat proporcionou que os participantes desta atividade fossem atuantes e críticos no meio em que vivem, além da apropriação de conhecimentos acadêmicos, sociais e ambientais.

O presente trabalho teve como objetivo avaliar de forma isolada a aplicação de fontes e doses de nitrogênio, modos de aplicação e doses de potássio e doses e modos de aplicação de nitrocálcio, nas características produtivas da cultura da cenoura. Os experimentos foram realizados em campo na Universidade Estadual de Goiás, Câmpus de Ipameri. O delineamento experimental utilizado em ambos foi o de blocos casualizados em esquema fatorial 2 x 5, com quatro repetições. Os dados foram submetidos à análise de variância (teste F) e as médias comparadas pelo teste de Tukey a 5% de probabilidade e foram realizadas análise de regressão. As fontes de nitrogênio e épocas não influenciaram nas características produtivas da cultura da cenoura em nenhuma das variáveis analisadas. A utilização de doses de nitrogênio até 103 kg ha-1 aumenta a produção comercial de cenoura. As doses de potássio influenciaram nas características do comprimento e diâmetro de raiz, classificação comercial, matéria seca da parte aérea e raiz além da produção comercial até a dose de 87,75 kg ha-1. As doses de nitrocálcio influenciaram positivamente a altura da parte aérea, matéria fresca da parte aérea, produção comercial e produção não comercial da cultura da cenoura até a dose de 311 kg ha-1 do fertilizante.

O ensino de química ainda se baseia em métodos tradicionais de transmissão/recepção/memorização dos conhecimentos. É imprescindível superar este modelo através da utilização de metodologias e recursos didáticos diversificados nas aulas, como os jogos didáticos e as atividades lúdicas. Estes garantem aos alunos a oportunidade de aprender e se divertir ao mesmo tempo e ainda que o professor avalie esse aprendizado. Neste contexto, o presente trabalho tem como objetivo avaliar a elaboração/desenvolvimento de uma atividade lúdica aplicada após o desenvolvimento de aulas expositivo/dialogadas/contextualizadas que abordaram a temática Químico-social: “Catalisadores e o ar que respiramos”. Com o intuito de analisar o que havia sido modificado cognitivamente e socialmente após o desenvolvimento das aulas e atividade lúdica. Para tanto, fez uso de uma pesquisa de caráter qualitativo/quantitativo, onde utilizou-se questionários como instrumento de construção de dados. Este trabalho busca mostra a atividade lúdica como um recurso didático facilitador do processo de ensino e aprendizagem e ainda como instrumento que permite avaliar este processo.

A cultura do repolho é realizada principalmente em pequenas áreas e depende de grande mão de obra, sendo na maior parte conduzida pela agricultura familiar. O objetivo deste trabalho foi identificar a melhor época de aplicação e doses de boro para as características produtivas e qualidade de cabeças de repolho. O experimento foi realizado na fazenda experimental da Universidade Estadual de Goiás, Câmpus de Ipameri (UEG/Ipameri). O delineamento experimental utilizado foi o de blocos casualizados no esquema fatorial 2 x 5, sendo duas épocas de aplicação (15 e 25 dias após o transplante (DAT)) e cinco doses de boro (0; 2,5; 5,0; 7,5 e 10,0 kg ha-1), com quatro repetições. As parcelas foram constituídas por quatro linhas de 1,6 m dispostas no espaçamento de 0,8 x 0,4 m, considerando como área útil as quatro plantas centrais. Foram realizadas as seguintes avaliações: teores de boro foliar, altura e área das plantas, altura da cabeça, diâmetro da cabeça, relação da altura da cabeça e diâmetro da cabeça, número de folhas externas e internas, massa fresca e seca de folhas externas e internas, classificação do peso da cabeça, compacidade, teor de clorofila e produtividade. Nas condições em que este experimento foi conduzido a aplicação de boro aos 25 dias após o transplante proporcionou maior teor de boro foliar na cultura do repolho. A dose indicada para a máxima produção de cabeças de repolho é a de 6,8 kg ha-1 de boro.

Contents A new device for storage and shipping of cell cultures - the Petaka G3 cell management device - was tested for its applicability for cooled-storage of equine semen. Semen from three stallions was processed with EquiPro extender either without antibiotics (three ejaculates per stallion) or with gentamicin (250 mg / l; three ejaculates per stallion). Semen was either stored at five (anaerobic conditions) or 15°C (aerobic conditions) in syringes or cell culture devices. Total and progressive motility, as well as membrane integrity of spermatozoa, were evaluated from days 1 to 7 after collection with computer-assisted semen analysis. In experiment 1 (extender without antibiotics), total motility, progressive motility and viability of spermatozoa significantly decreased over time (p < 0.05). The decrease was significantly faster at 15°C than at 5°C (p < 0.05). In the presence of gentamicin (experiment 2), this difference was no longer present. It can be concluded that cooled-storage of equine semen in sophisticated devices for cell culture is not advantageous to syringes for successful maintenance of semen longevity. [ABSTRACT FROM AUTHOR]

The coordinated and synchronized movement of animals in groups often referred to as collective motion emerges through the interactions between individual animals within the group. Factors which affect these interactions have the potential to shape collective movement. One such factor is familiarity, or the tendency to bias behaviour towards individuals as a result of social recognition. We examined the effect of familiarity on the expression of collective motion in small shoals of female guppies (Poecilia reticulata). Groups comprising familiar individuals were more strongly polarized than groups of unfamiliar individuals, particularly when in novel surroundings. The ability to form more strongly polarized shoals potentially promotes information transfer and enhances the anti-predator benefits of grouping.

Para promover a motivação dos alunos e a melhoria nos processos de ensino e aprendizagem, duas licenciandas do Curso de Licenciatura em Química da Universidade Federal de Goiás/Regional Catalão, durante a disciplina de Estágio de Licenciatura 2, desenvolveram um Projeto Temático que abordava o tema lixo, por ser uma discussão atual e de suma importância para a sociedade. Neste, as licenciandas desenvolveram aulas expositivo/dialogadas guiadas pelos princípios de Ensino CTS (Ciência, Tecnologia e Sociedade), em que buscaram relacionar a problemática aos seus aspectos sociais, científicos e tecnológicos, com vistas à formação para a cidadania. Também propuseram outras atividades como o uso de vídeos, experimentações, a construção de um painel lúdico e uma dinâmica com o objetivo de promover a construção de conhecimentos cognitivos e outros, fundamentais para a formação de cidadãos críticos. Com as atividades buscaram promover a construção de um conhecimento efetivo com os alunos da Educação Básica. Para avaliar o desenvolvimento do projeto foi desenvolvida uma pesquisa de caráter qualitativo/quantitativo, utilizando-se questionários como instrumento de coleta de dados. Após a análise dos resultados obtidos avalia-se que com o desenvolvimento do projeto as licenciandas conseguiram promover a motivação e a construção de conhecimentos para uma formação cidadã dos alunos.

Follicular wave synchronization and follicular superstimulation with FSH are commonly used in OPU-IVP programs to increase oocyte developmental competence. Factors like Growth Differentiation Factor 9 (GDF9) and Bone Morphogenetic Protein 15 (BMP15), from the TGF beta superfamily, are produced by the oocyte and modulate follicular function. The aim of this study was to analyze the FSH-induced effects on (1) embryo production in dual-purpose Simmental cattle, and (2) TGF beta-mediated effects on oocyte-granulosa cell communication. Simmental heifers (n = 12, age 484 ± 62 days) underwent two OPU-IVP cycles in a cross-over design. Follicular waves were synchronized using 0.5 mg cloprostenol on Day 0, followed by 10 μg buserelin on Day 2. Subsequently, half of the heifers were randomly assigned to receive FSH/LH (four injections of 75 IU FSHp and 75IU LHp, 12 h apart on Days 4 and 5) before the first OPU, while the remaining heifers received FSH/LH before the second OPU. At the time of OPU, i.e. 7 days after the start of synchronization, granulosa cells were collected for RT-qPCR analysis. FSH treatment did not affect the number of oocytes collected (17.3 vs. 13.3, P > 0.05), but increased the percentage of quality 1 oocytes compared to controls (45.7 % vs. 22.0 %, P < 0.001). Neither cleavage (86.4 % vs. 85.7 %), nor blastocyst (42.1 % vs. 39.3 %) rate, or the number of transferable embryos produced by IVP (4.1 vs 4.8) was influenced by FSH treatment (P > 0.05 in all cases). FSH treatment increased HIF1A and FSHR levels in granulosa cells, while STAR was decreased (P = 0.008 in all cases). FSH treatment did not affect BMP15 or GDF9 mRNA expression (P > 0.05) but appeared to modulate the expression of genes involved in the BMP signaling pathway. Transcriptional levels of BMP15 receptor (BMPR1A, P = 0.016), and its downstream signaling factor SMAD1 (P = 0.008) were affected by FSH treatment. Our results demonstrated no benefit of this FSH stimulation protocol on IVP results in Simmental heifers. Further, our results suggest that the effects of FSH on bovine oocytes during acquisition of developmental competence may be mediated through BMP, but do not involve the regulation of transcriptional availability of GDF9, providing new insights into possible paracrine effects of the oocyte on granulosa cells., Competing Interests: Declaration of Competing interest The authors declare that they have no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Transition period is a critical time for dairy cows because a large proportion of clinical and subclinical diseases are observed in the first month after parturition. Occurrence of negative energy balance is associated with depressed immunity and these conditions can affect oocyte quality and further embryonic development. The aim of this study was to assess the effects of negative energy balance-associated disorders on in vitro embryo production (IVP) in dairy cattle. We hypothesized that subclinical metabolic and/or inflammatory disorders have a negative effect on oocyte developmental competence and morphokinetic parameters of the resulting embryos. The study was conducted on 30 lactating Holstein-Friesian cows which were assigned into four groups: healthy (HEAL, n  = 6), metabolic disease (META, n  = 8), inflammatory disease (INFL, n  = 8), or combined metabolic and inflammatory disease (COMB, n  = 8). Ovum pick-up (OPU) was performed twice weekly on all cows over a period of four weeks ( n  = 8 OPU sessions/cow) starting on the fifth week postpartum, and the collected oocytes were subjected to routine IVP. Donor's health status did not affect the number of oocytes/OPU or the recovery rate ( p  > 0.05). The number of quality 1 oocytes collected from INFL and COMB cows was lower compared to HEAL cows ( p  < 0.05). Also, the percentage of quality 1 embryos was reduced in META and COMB compared to HEAL cows ( p  < 0.05). Cleavage, blastocyst and hatching rates were similar among groups ( p  > 0.05). Presence of disease did not affect the time required by zygotes to reach specific developmental stages, as recorded by means of time-lapse monitoring. Nevertheless, there was a higher probability of direct cleavage after IVF in oocytes of COMB cows compared to those of HEAL cows ( p  < 0.05). In conclusion, oocytes and embryos derived from dairy cows diagnosed with subclinical metabolic and/or inflammatory diseases during the transition period showed reduced quality but similar developmental potential and morphokinetics when compared to healthy cows. These results shed light on the consequences of subclinical disease on embryonic development in dairy cows which might be important for embryo transfer programs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Serbetci, González-Grajales, Herrera, Ibanescu, Tekin, Melean, Magata, Malama, Bollwein and Scarlet.)

The antral follicle count (AFC) and Anti-Müllerian hormone (AMH) concentration are validated markers for ovarian reserve in cattle, but their use as fertility markers is controverse. Here we assessed the effects of postpartum diseases on AFC and AMH concentration, as well as the influence of parity and breed on these parameters. Cows (n = 513, mostly Holstein Friesian and Brown Swiss, parity 3.0 ± 1.8) underwent a single ultrasonography examination 28-56 days after parturition and categorized as having low (n ≤ 15 follicles), intermediate (n = 16-24 follicles), or high (n ≥ 25 follicles) AFC based on objective video analysis of recorded sequences. Blood samples for AMH determination were collected at the time of examination and animals divided into low (< 0.05 ng/ml) and high AMH (≥ 0.05 ng/ml) group, respectively. No effects of postpartum diseases or breed on either AFC or AMH groups could be observed. There was a strong interaction between parity and AFC, primiparous cows having less follicles (13.6 ± 6.2 vs. 17.1 ± 7.0, P < 0.001) than pluriparous cows. The AFC did not affect reproductive parameters or productivity of the cows. In comparison, pluriparous cows with high AMH concentration had shorter calving to first service (86.0 ± 37.6 vs. 97.1 ± 46.7 days, P < 0.05) and calving to conception (123.8 ± 51.9 vs. 135.8 ± 54.4 days, P < 0.05) intervals, but lower milk yield (8440.3 ± 2292.9 vs. 8927.9 ± 2192.5 kg, P < 0.05) compared to cows with low AMH. In conclusion, no effect of postpartum diseases on AFC or AMH concentration of dairy cows could be observed. However, an interaction between parity and AFC, as well as associations of AMH with fertility and productivity in pluriparous cows, were demonstrated., Competing Interests: Declaration of Competing Interest The authors declare that they have no financial or personal relationship with other people or organizations that could inappropriately influence or bias the publication of this manuscript., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Scrotal swelling is a clinical situation which can be caused by different aetiologies. In this case report, we describe a multi-week episode of unilateral and bilateral scrotal swelling in boars at an Austrian boar stud and its diagnostic work-up., Case Presentation: In the summer of 2020, the herd veterinarian of an Austrian boar stud reported that over a period of six weeks, five out of 70 boars presented with unilateral severe swelling of the left scrotum and three out of 70 boars with bilateral severe swelling of the left and moderate swelling of the right scrotum, respectively. A complete history was obtained and an on-site evaluation of the facility was done. Five boars were necropsied, and a variety of samples harvested for further diagnostic investigations. Infectious differential diagnoses associated with unilateral swelling of the scrotum or the testis were excluded through serological and tissue testing. In three of the five boars, histopathology revealed complete acute haemorrhagic necrosis of the left testis concurrent with strongly congested blood vessels. Review of the collected information with a group of experts in the field of boar stud management resulted with consensus that, most likely, trauma was the etiologic event causing the clinical signs and pathology. Coincident with discussion of implementing video recording cameras in the boar housing area, no further clinical cases followed. As this case occurred during the first lockdown of the COVID-19 pandemic, we propose that the distress and travelling restrictions may have contributed to frustration among boar stud workers, which was consequently expressed as misbehaviour against boars., Conclusions: Once all known infectious causes of unilateral swelling of the scrotum were excluded, a critical diagnostic work-up focused on non-infectious causes. Non-infectious causes, such as trauma, need to be carefully evaluated, as it may also include human misbehaviour against boars. Summarizing all findings of this case report, the authors hypothesize that a blunt trauma was the reason for the series of mainly unilateral swelling of the scrota of boars., (© 2023. The Author(s).)

Persistent breeding-induced endometritis (PBIE) is a major cause of subfertility in horses and the susceptibility is increased by several factors. The aim of this study was to determine the effects of clinical uterine findings and PBIE therapies, respectively, on pregnancy rate in mares. The analysis included records from 220 mares (390 cycles) inseminated at an artificial insemination (AI) center in Switzerland. Gynecological examinations were performed repeatedly before and after AI to determine cervical tone, uterine edema, and intrauterine fluid accumulation. Pregnancy rate was lower ( p < 0.001) in barren mares compared to mares of other reproductive status. A more flaccid cervix ( p = 0.009) was observed at the time of ovulation in pregnant cycles, but there was no difference ( p > 0.05) regarding uterine edema. Intrauterine fluid accumulation reduced pregnancy rate ( p = 0.002). Oxytocin administration had beneficial effects on pregnancy rate ( p = 0.015), especially for barren mares, while uterine lavage did not have any effect ( p > 0.05). The results show that cervical tone and intrauterine fluid accumulation, but not its degree, are useful parameters for assessment of fertility in mares. Oxytocin treatment improved pregnancy rates in mares with PBIE while uterine lavage had a limited effect.

It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy were detected during this time. Tubular structures of the fetal testes were present from day 50 onwards, whereas cell clusters dominated in the fetal ovary. The genital ducts were well-differentiated and presented a lumen in all samples. No sign of mesonephric or paramesonephric duct degeneration was detected. Expression of AMH was strong in the fetal testes but absent in ovaries. Irrespective of sex, primordial germ cells selectively expressed LIN28. Migration of primordial germ cells from the mesonephros to the gonad was detected at 45 days, but not at 60 days of development. Their number and distribution within the gonad were influenced ( p < 0.05) by fetal sex. Most primordial germ cells (86.8 ± 3.2% in females and 84.6 ± 4.7% in males) were characterized as pluripotent according to co-localization with CD117. However, only a very small percentage of primordial germ cells were proliferating (7.5 ± 1.7% in females and 3.2 ± 1.2% in males) based on co-localization with Ki67. It can be concluded that gonadal sexual differentiation in the horse occurs asynchronously with regard to sex but already before 45 days of gestation.

In embryos subjected to assisted reproductive techniques, epigenetic modifications may occur that can influence embryonic development and the establishment of pregnancy. In horses, the storage temperature during transport of fresh embryos before transfer is a major concern. The aim of this study was, therefore, to determine the effects of two storage temperatures (5 °C and 20 °C) on equine embryos, collected at day seven after ovulation and stored for 24 h, on: (i) morphological development; (ii) expression of candidate genes associated with embryo growth and development, maternal recognition of pregnancy, methylation and apoptosis, and (iii) gene-specific and global DNA methylation. Embryos ( n = 80) were collected on day seven or day eight after ovulation and assigned to four groups: day seven control (E7F, fresh); day seven, stored for 24 h at 5 °C (E5C); day seven, stored for 24 h at 20 °C (E20C) and day eight control (E8F, fresh 24h time control). The embryos and the storage medium (EquiHold, holding medium, Minitube, Tiefenbach, Germany) from all treatment groups were analyzed for (i) medium temperature, pH, and lipid peroxidation (malondialdehyde; MDA) and (ii) embryo morphology, mRNA expression and DNA methylation (immunohistochemistry and gene-specific DNA methylation). The size of embryos stored at 5 °C was larger ( p < 0.01), whereas embryos stored at 20 °C were smaller ( p < 0.05) after 24 h. There were no changes in pH and MDA accumulation irrespective of the group. The mRNA expression of specific genes related to growth and development ( POU5F1, SOX2, NANOG ), maternal recognition of pregnancy ( CYP19A1, PTGES2 ), DNA methylation ( DNMT1, DNMT3A , DNMT3B ) and apoptosis ( BAX ) in the E5C and E20C were either up or downregulated ( p < 0.05) when compared to controls (E7F and E8F). The immune expression of 5mC and 5hmC was similar among treatment groups. Percentage of methylation in the CpG islands was lower in the specific genes ESR1 , NANOG and DNMT1 ( p < 0.001) in E20C embryos when compared to E8F (advanced embryo stage). Therefore, our study demonstrates for the first time the gene-specific and global DNA methylation status of fresh equine embryos collected on days seven and eight after ovulation. Although our results suggest some beneficial effects of storage at 20 °C in comparison to 5 °C, the short-term storage, regardless of temperature, modified gene expression and methylation of genes involved in embryo development and may compromise embryo viability and development after transfer.

The aim of the present study was to characterise key enzymes involved in polyunsaturated fatty acid (PUFA) synthesis in the testis and epididymis collected from 2-year-old healthy warmblood stallions (n=10). The mRNA expression of fatty acid synthase, the Δ9-, Δ6-, Δ5- and Δ4-desaturases and elongases 6, 5 and 2 (encoded by the fatty acid synthase (FASN), the stearoyl-CoA desaturase (SCD), the fatty acid desaturase 2 (FADS2), the fatty acid desaturase 1 (FADS1), the delta 4-desaturase, sphingolipid 1 (DEGS1), ELOVL fatty acid elongase 6(ELOVL6), ELOVL fatty acid elongase 5 (ELOVL5), ELOVL fatty acid elongase 2 (ELOVL2) genes respectively) was determined in equine testis and epididymis. All enzymes were present in testicular tissue and along the epididymis, but mRNA expression differed among localisations. The protein localisation of FADS1, FADS2 and ELOVL5 was determined by immunohistochemistry. In the testes, FADS1 was expressed in the germinal cells and ELOVL5 was expressed in germinal and Leydig cells; FADS2 was not detected. In the epididymis, FADS1 and FADS2 were expressed in the principal and basal cells, whereas ELOVL5 was found only in the principal cells of the caput. All three enzymes were present in epididymal vesicles secreted by an apocrine mechanism. These results suggest active PUFA metabolism during spermatogenesis and epididymal sperm maturation in stallions.

In the horse, it is still unclear if and to what extent low progestin concentration contributes to early conceptus loss. In the present study, we have investigated if reduced or elevated progestin concentration in the early luteal phase influences endometrial function and conceptus development. We hypothesized that reduced progestin concentration via delayed downregulation of endometrial progesterone receptors (PR) influences endometrial function in healthy fertile mares while progestin substitution does not. Genitally healthy estrous mares (n = 8; age 4-14 years) were inseminated and treated with either altrenogest (0.044 mg/kg once daily orally) on days 5-10 after ovulation (ALT), cloprostenol (125 μg once daily intramuscularly) on days 0-3 after ovulation (CLO) or left untreated (CON). ALT and CLO treatment were chosen to increase and decrease total peripheral progestin concentration, respectively. Each treatment was given to every mare in consecutive cycles. On day 14 after ovulation, endometrial fluid was collected with a cotton roll inserted into the uterus and an endometrial biopsy for immunohistological demonstration of progesterone (PR) receptor distribution was collected. In endometrial fluid, free amino acid concentrations were analyzed by ion exchange liquid chromatography with an amino acid analyzer. Cell nuclei staining positive for the PR were determined in the luminal and glandular epithelium as well as in the stroma. Pregnancy rate tended to differ among treatments. The percentage of luminal epithelial cells staining positive for PR differed among treatments (p < 0.05) and was higher in CLO (84.1 ± 1.9%) than in ALT (70.7 ± 4.7%) and CON cycles (72.8 ± 4.1%). Concentrations of the amino acids isoleucine (CON 0.17 ± 0.03, CLO 0.14 ± 0.02, ALT 0.23 ± 0.04 μmol) and lysine (CON 0.27 ± 0.08, CLO 0.18 ± 0.05, ALT 0.44 ± 0.13 μmol) were influenced by treatment (p < 0.05) and lower in CLO than in ALT and CON cycles. In conclusion, impaired luteal function due to CLO treatment during the early luteal phase of pregnant mares delayed downregulation of progesterone receptors in the endometrial epithelium on day 14. This influenced endometrial function as reflected in lower concentrations of the amino acids lysine and isoleucine in endometrial secretions. Enhanced progestin concentration had less clear effects in healthy fertile mares., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Anti-Müllerian hormone (AMH) has gained increasing interest as a biomarker for assessment of gonadal activity. The ability to predict the ovarian follicular reserve of prepubertal female horses (fillies) or to identify stallions with testicular pathologies already during their prepubertal life has not been analyzed so far. Both would help to select fertile horses and reduce costs associated with keeping animals. The objectives of the present study were to (1) assess AMH, LH, FSH, progesterone (females) and testosterone (males) dynamics in prepubertal horses from birth onwards and (2) determine whether AMH concentrations detected in plasma of prepubertal female and male horses are correlated with postpubertal gonadal development. Warmblood foals (n = 30, 14 females, 10 normal males and 6 males with abnormal testicular development) born between February and May of two consecutive years (n = 28 in the first year and n = 2 the next year), were included in the study. Information on gestational length, parity of the dam and placental weight was collected for all foals. Blood samples for hormone analysis were collected from birth onwards every four weeks up to the age of one year. At two years, blood samples were collected on the day when antral follicle count (AFC) and total testicular volume (TTV) were assessed. AMH was detectable in the plasma of all animals from birth onwards and its concentration was significantly higher (P < .001) in males than in females, regardless of testicular development. In males, AMH and testosterone concentration were similar for all animals during the first year of life, regardless of testicular development. At two years, AMH concentration was higher (P < .05) in males with abnormal testicular development than in those with normal testes. In females, AMH concentration at two years was correlated with AMH concentration at birth (P < .05) and with AFC (P < .001). At birth, LH concentration was lower (P < .05) in stallions with abnormal testes (0.3 ± 0.2 ng/ml) than in controls (0.6 ± 0.2 ng/ml). A high negative correlation between AMH concentration and gestation length was observed in males during the first eight weeks of life (P < .01, r = -0.64 to -0.71). Elevated progesterone concentrations over 1 ng/ml were observed in several females starting with 20 weeks of age. This was paralleled by an increase in AMH concentration and was preceded by FSH and LH increases. In conclusion, AMH determination can be reliably used from two years onwards to identify stallions with abnormal testicular development, but it is inconclusive before puberty. In female horses, determination of AMH concentration at a prepubertal age allows for prediction of AMH and AFC after puberty. We suggest that premature luteinisation occurs before the onset of puberty in female horses and that LH secretion in the perinatal period is involved in testicular development and descent in the horse., (Copyright © 2018 Elsevier Inc. All rights reserved.)

A decrease in fertility of equine semen during cooled-storage so far has mainly been attributed to changes in sperm membrane function. In the present study we hypothesized that cooled-storage also changes the sperm DNA methylation level. For this purpose, semen was collected from 10 fertile stallions and processed for cooled-storage at 5 °C. Two final concentrations, 50 × 10 6 and 100 × 10 6  cells/mL, were used. Semen was analyzed for total motility, progressive motility, membrane integrity, phosphatidylserine translocation (PST), mitochondrial membrane potential and chromatin condensation, immediately after processing and at 24 h-intervals until 72 h of storage. DNA cytosine methylation was assessed by ELISA after DNA extraction and denaturation. DNA methylation did neither change over time nor was affected by semen concentration. Total motility, progressive motility, membrane integrity, PST, mitochondrial membrane potential and chromatin condensation changed over storage time, but no differences between semen concentrations could be demonstrated. The results demonstrate that cooled-storage of equine semen does not induce changes in sperm DNA cytosine methylation. In cooled-semen of good quality, a concentration of 100 × 10 6 sperm/mL does not affect semen longevity. It can be concluded that a better fertility of cooled-stored than cryopreserved stallion semen is at least in part a result of only minor influences of cooled-storage on DNA integrity and methylation., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Eight-week-old calves were either castrated by partial scrotal resection (SR) without removing the testes (n = 10), Burdizzo (BZ) clamp (n = 10), orchidectomy (OR; n = 10), or were left gonad intact as controls (CO; n = 10). Concentrations of anti-Muellerian hormone (AMH), inhibin A, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) in plasma were determined from 16 to 48 weeks of age. At 18 months, testes of SR, BZ, and CO bulls were obtained and the immunolocalization of LH and FSH receptors and AMH analyzed. Concentration of AMH in plasma of CO and SR bulls decreased with increasing age (P < 0.001). A similar AMH profile in CO and SR indicates that SR did not induce a true cryptorchid state. In groups OR and BZ, AMH was undetectable. Plasma inhibin concentration was higher in groups CO and SR than BZ and OR (P < 0.001). Plasma LH and FSH concentrations decreased over time (P < 0.001) and were higher in groups BZ and OR than SR and CO (P < 0.001). In the testes, immunolabeling for AMH existed in Sertoli cells of CO and SR but not BZ bulls. FSH receptors were localized in Sertoli cells, Leydig cells, spermatocytes, and the epididymis of CO and SR animals, whereas LH receptors were restricted to Leydig cells. In BZ animals, FSH and LH receptors and AMH were absent, indicating complete testicular degeneration. In conclusion, AMH is a more reliable marker for the presence of testicular tissue in bulls than inhibin. Scrotal resection did not induce a true inguinal cryptorchid state but affected testicular responsiveness to gonadotropic stimulation., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The aim of the present study was to characterise receptors for LH and FSH (LHR and FSHR, respectively) and aromatase in epididymal and testicular tissue from stallions of different ages (prepubertal, young, mature and old). Gene and protein expression were assessed by real-time quantitative polymerase chain reaction (real-time qPCR), immunohistochemistry and multiple immunofluorescence labelling. There were no differences in LHR mRNA expression in epididymal and testicular parenchyma in stallions of different age. In contrast, expression of FSHR and CYP19A1 in caput, corpus and cauda epididymis and in testicular parenchyma increased with age (P<0.001). Immunolabelling for LHR, FSHR and aromatase was influenced by puberty. In postpubertal stallions, positive staining for LHR and aromatase was detected in Leydig cells, whereas protein expression of FSHR was present in Sertoli cells and primary spermatocytes. In prepubertal colts, staining for LHR, FSHR and aromatase was detected in seminiferous tubules. In epididymal tissue, aromatase was present in the cauda epididymis only, regardless of age. In conclusion, the results highlight the significance of gonadotropin action and oestrogen production for the maturation of male reproductive tissue in the horse. The presence of FSHR in the seminiferous tubules suggests effects of FSH on spermatogenesis in this species. The importance of oestrogen production for maintenance of testicular function in stallions was confirmed.