Exposure to progesterone before an ovulation synchronization protocol increases the follicular diameter and fertility of multiparous suckled Bos taurus cows.

The objective of this study was to evaluate the effect of administering injectable progesterone (P4i) before a timed artificial insemination (TAI) protocol on the follicular growth, ovulation, and pregnancy rate of Bos taurus suckled cows. The effect of P4i administration before the TAI on the pregnancy rate (P/AI) was evaluated in 576 suckled Bos taurus cows at 30-90 days postpartum. In addition, the effect of P4i administration before TAI on follicular dynamics was evaluated in subgroup of 401 suckled Bos taurus cows. On Day -10 (D-10), cows were divided into two experimental groups (Control and P4i). In this moment, P4i cows received i.m. 150 mg of injectable long-action progesterone. After that, both experimental groups received a synchronization protocol (Day 0; D0) that consisted of administration i.m. of 2 mg of estradiol benzoate and a progesterone intravaginal insert on D0. On Day 8 (D8), the progesterone insert was removed, and the cows received 500 µg of cloprostenol, 400 IU of eCG, and 1 mg of estradiol cypionate. TAI was performed 48 h after the removal of the progesterone insert. The ultrasound exams were performed in a subgroup of cows on Days 0, 8, 10 and 12 to evaluate the diameter of the largest follicle, rate of follicular growth and risks of single and double ovulation. The pregnancy diagnosis was performed 30 days after TAI in all cows to determine the pregnancy rate. The diameter of the largest follicle, on D10 (P = 0.84), rate of follicular growth (P = 0.14), ovulation rate (P = 0.40) and double ovulation rates (P = 0.23) did not differ between experimental groups. The pregnancy rate was greater in the P4i group [Control 46.2 % (133/288) vs. P4i 55.6 % (160/288); P = 0.03]. The diameter of the largest follicles (LF) on D0 (Control 11.6 ± 0.2 vs. P4i 13.3 ± 0.3) was greater (P = 0.01) in the P4i group. In conclusion, injectable progesterone before the ovulation synchronization protocol increased the diameter of the largest follicle on the D0 and the pregnancy rate in multiparous Bos taurus suckled beef cows.

Women's cardiovascular health - the cardio-oncologic jigsaw.

Improvements in cancer care have led to an exponential increase in cancer survival. This is particularly the case for breast cancer, where 5-year survival in Australia exceeds 90%. Cardiovascular disease (CVD) has emerged as one of the competing causes of morbidity and mortality among cancer survivors, both as a complication of cancer therapies and because the risk factors for cancer are shared with those for CVD. In this review we cover the key aspects of cardiovascular care for women throughout their cancer journey: the need for baseline cardiovascular risk assessment and management, a crucial component of the cardiovascular care; the importance of long-term surveillance for ongoing maintenance of cardiovascular health; and strong evidence for the beneficial effects of physical exercise to improve both cancer and cardiovascular outcomes. There is general disparity in cardiovascular outcomes for women, which is further exacerbated when both CVD and cancer co-exist. Collaboration between oncology and cardiac services, with an emergence of the whole field of cardio-oncology, allows for expedited investigation and treatment for these patients. This collaboration as well as a holistic approach to patient care and key role of patients' general practitioners are essential to ensure long-term health of people living with, during and beyond cancer.

The impact of potassium peroxymonosulphate and chlorinated cyanurates on biofilms of Stenotrophomonas maltophilia: effects on biofilm control, regrowth, and mechanical properties.

The activity of two chlorinated isocyanurates (NaDCC and TCCA) and peroxymonosulphate (OXONE) was evaluated against biofilms of Stenotrophomonas maltophilia, an emerging pathogen isolated from drinking water (DW), and for the prevention of biofilm regrowth. After disinfection of pre-formed 48 h-old biofilms, the culturability was reduced up to 7 log, with OXONE, TCCA, and NaDCC showing more efficiency than free chlorine against biofilms formed on stainless steel. The regrowth of biofilms previously exposed to OXONE was reduced by 5 and 4 log CFU cm-2 in comparison to the unexposed biofilms and biofilms exposed to free chlorine, respectively. Rheometry analysis showed that biofilms presented properties of viscoelastic solid materials, regardless of the treatment. OXONE reduced the cohesiveness of the biofilm, given the significant decrease in the complex shear modulus (G*). AFM analysis revealed that biofilms had a fractured appearance and smaller bacterial aggregates dispersed throughout the surface after OXONE exposure than the control sample. In general, OXONE has been demonstrated to be a promising disinfectant to control DW biofilms, with a higher activity than chlorine. The results also show the impact of the biofilm mechanical properties on the efficacy of the disinfectants in biofilm control.

Phytochemicals Against Drug-Resistant Bacterial Biofilms and Use of Green Extraction Solvents to Increase Their Bioactivity.

Antimicrobial resistance has become one of the major global public health issues of the twenty-first century. One of the main factors in the limited action of antimicrobials is related to the ability of microorganisms, particularly bacteria, to form biofilms. These complex and well-organized communities allow the colonizing cells to acquire survival advantages over the same cells in suspension, including antibiotic resistance. A huge percentage of bacterial infections in humans are associated with biofilms, and many of them are chronic. Therefore, there is an urgent need to develop new products effective in controlling or eradicating biofilms. Plant secondary metabolites (phytochemicals) have demonstrated their potential as antibacterials against planktonic cells and sessile communities when used alone or in synergy with other molecules. This chapter covers recent advances in the activity of phytochemicals against biofilms, particularly those formed by drug-resistant bacteria. In addition, taking into account that the extraction step is crucial for the successful development of new bioactive compounds, the use of novel solvents that increase the phytochemical effect, such as natural deep eutectic solvents (NADES), as well as the recent applications of these solvents as antimicrobials are discussed.

Use of injectable progesterone at the beginning of the TAI protocol is not necessary in super-early resynchronization started 14 days after artificial insemination in Bos indicus beef heifers.

In a protocol for the resynchronization of ovulation starting 14 days (D14) after timed artificial insemination, the effects of short-action injectable progesterone (P4i) administration and the length of treatment with an intravaginal progesterone (P4) device on follicular dynamics, the conception rate (P/AI) and the percentage of false positives were evaluated in 1065 Nelore heifers previously timed-inseminated. On D14, P4 devices were inserted, and heifers allocated, based on a 2 × 2 factorial design, to receive (P4i) or not 50 mg of P4i (NoP4i), and to remove the P4 device after 6 (6Day) or 8 days (8Day). At the time of P4 device removal (D20 and D22), non-pregnancy diagnosis was performed using vascularization of the corpus luteum (VCL) evaluated. At this time, non-pregnant heifers received 150 µg of D-cloprostenol, 0.6 mg of estradiol cypionate and 300IU of eCG. TAI was performed 48 h after P4 device removal. For these heifers, ultrasound examinations were performed at P4 device removal and at TAI to evaluate follicular dynamics and at 30 days after TAI to evaluate the P/AI. Pregnant heifers based on VCL were evaluated using B-mode ultrasonography 10 days after Doppler ultrasound to evaluate the percentage of false positives. Statistical analyses were performed using the GLIMMIX procedure of SAS. There were no interaction effects between P4i and duration of treatment with a P4 device. The P/AI was diagnosed by Doppler, 1st TAI, 2nd TAI, total and percentage of false positives did not differ between heifers receiving or not P4i. Similarly, duration of treatment with a P4 device did not influence the P/AI by Doppler, 1st TAI, 2nd TAI and total. However, the percentage of false positives diagnoses was higher in 6Day heifers (P = 0.01). The diameter of largest follicle at P4 device removal was greater in the 8Day heifers (P = 0.001), and at TAI was higher in the P4i-treated heifers (P = 0.03). Additionally, the percentage of false positives diagnoses was lower in heifers that ovulated to the 1st TAI protocol (P = 0.001). In conclusion, for resynchronization 14 days after TAI, it is not necessary to inject P4i at the beginning of the protocol. In addition, P4 device removal after 6 instead of 8 days increases the percentage of false positives because of the earlier diagnosis (20 days after TAI), but does not interfere in P/AI of resynchronization protocol. Furthermore, the percentage of false positives is higher in heifers that did not ovulate to 1st TAI protocol.

Biofilm formation under high shear stress increases resilience to chemical and mechanical challenges.

The effect that the hydrodynamic conditions under which biofilms are formed has on their persistence is still unknown. This study assessed the behaviour of Pseudomonas fluorescens biofilms, formed on stainless steel under different shear stress (τw) conditions (1, 2 and 4 Pa), to chemical (benzalkonium chloride - BAC, glutaraldehyde - GLUT and sodium hypochlorite - SHC) and mechanical (20 Pa) treatments (alone and combined). The biofilms formed under different τw  showed different structural characteristics. Those formed under a higher τw were invariably more tolerant to chemical and mechanical stresses. SHC was the biocide which caused the highest biofilm killing and removal, followed by BAC. The sequential exposure to biocides and mechanical stress was found to be insufficient for effective biofilm control. A basal layer containing biofilm cells mostly in a viable state remained on the surface of the cylinders, particularly for the 2 and 4 Pa-generated biofilms.

Effects of eCG and FSH in timed artificial insemination treatment regimens on estrous expression and pregnancy rates in primiparous and multiparous Bos indicus cows.

Effects were evaluated in Bos indicus cows of eCG and FSH on follicular growth, estrous expression, and pregnancy per artificial insemination (P/AI) as a result of fixed-time artificial insemination (TAI). In Experiment 1, extent of timing-of-ovulation synchronization among cows was evaluated after imposing an estrogen/progesterone-based treatment regimen. At progesterone device removal (D8), cows were administered: eCG, or FSH or served as untreated Controls. In Experiment 2, percentage of cows P/AI was evaluated when the Experiment 1-treatment regimen was imposed. On D10, all cows were artificially inseminated. In Experiment 3, cows were assigned to two treatment groups (Control and eCG) on D8 to evaluate percentage of cows P/AI and estrous expression. In Experiment 1, follicular dynamics were similar among treatment groups. In Experiment 2, follicular growth was greater (P = 0.0001) with the eCG treatment. There was an interaction of treatment × parity (P = 0.007) on percentage of cows P/AI. There was a greater percentage of primiparous cows P/AI in the eCG-treated than Control and FSH-treated cows. There was a greater percentage of eCG-treated multiparous cows pregnant as a result of TAI than Control cows. There was an interaction of treatment × parity (P = 0.005) on P/AI in Experiment 3, in which the eCG effect was more pronounced in primiparous cows. Treatment with FSH, therefore, was not as effective as eCG in stimulation of follicular growth or enhancing percentage of cows pregnant as a result of TAI. Physiological effects of eCG, however, were also more evident in primiparous cows.

Probiotic properties of yeasts isolated from Brazilian fermented table olives.

AIMS: To investigate the probiotic potential of yeasts isolated from naturally fermented Brazilian table olives. METHODS AND RESULTS: Eighteen yeast strains were tested in terms of: safety; survival of gastrointestinal and digestion conditions; antimicrobial activity; cellular hydrophobicity; autoaggregation ability and adhesion to epithelial cells; coaggregation and inhibition of pathogenic bacteria adhesion. Six yeasts showed favourable results for all probiotic attributes: Saccharomyces cerevisiae CCMA 1746, Pichia guilliermondii CCMA 1753, Candida orthopsilosis CCMA 1748, Candida tropicalis CCMA 1751, Meyerozyma caribbica CCMA 1758 and Debaryomyces hansenii CCMA 1761. These yeasts demonstrated resistance to 37°C, pH 2·0 and bile salts, and survived in vitro digestion (≥106  CFU per ml). Furthermore, the yeasts exhibited a hydrophobic cell surface (42·5-92·2%), autoaggregation capacity (41·0-91·0%) and adhesion to Caco-2 (62·0-82·8%) and HT-29 (57·6-87·3%) epithelial cell lines. Also, the strains showed antimicrobial activity against Salmonella Enteritidis as well as the ability to coaggregate and reduce the adhesion of this pathogen to intestinal cells. CONCLUSIONS: Autochthonous yeasts from naturally fermented Brazilian table olives have probiotic properties, with potential for development of new probiotic food products. SIGNIFICANCE AND IMPACT OF STUDY: These data are important and contribute to the knowledge of new potential probiotic yeasts capable of surviving gastrointestinal tract conditions and inhibiting pathogenic bacteria.

Nuclear maturation kinetics and in vitro fertilization of immature bovine oocytes injected into pre-ovulatory follicles.

The maturation kinetics and in vitro fertilization of immature bovine oocytes injected by the intra-follicular oocyte injection (IFOT) technique into pre-ovulatory follicles of previously synchronized cows were evaluated. In Experiment 1, grade I, II and III cumulus-oocyte complexes (COCs) were randomly distributed to one of three Groups: Matvitro22 (COCs matured in vitro for 22 h), MatFol20 and MatFol28 (COCs matured in vivo after being injected into a pre-ovulatory follicle of previously synchronized cows for 19.8 ± 0.1 h and 28.3 ± 0.1 h, respectively). Cows received 12.5 mg of LH (Lutropin, Bioniche, Canada) at the time of IFOT in the MatFol20 Group or 10 h after IFOT in the MatFol28 Group. MatFol20 and MatFol28 COCs were aspirated approximately 20 h after the LH injection for nuclear maturation kinetics and recovery rate assessment. In Experiment 2, grade I, II, and III COCs were randomly distributed into two Groups: Matvitro22 Group, COCs were matured and fertilized in vitro, and MatFol20 Group, COCs were matured as in the MatFol20 Group in Experiment 1, but COCs were fertilized in vitro. Putative zygotes were classified as fertilized, unfertilized or polyspermic. In Experiment 1, the recovery rate was lower (P < 0.001) in the MatFol20 Group (52.9%, 91/172) compared with MatFol28 (72.9%, 113/155). Rate of oocytes in germinal vesicle stage, metaphase I, anaphase I and telophase I were similar among Groups. However, oocytes matured in vivo for 28.3 h had lower rate of metaphase II (P = 0.001) and greater rates of degenerated (P = 0.001) and parthenogenetically activated (P = 0.001) oocytes. In experiment 2, the rates of polyspermy and degenerated were similar between Groups. However, the rate of fertilized oocytes was greater (P = 0.05) in oocytes in the MatFol20 Group. It is concluded that oocyte in vivo maturation for 19.8 h after IFOT does not compromise the nuclear maturation kinetics and increases in vitro fertilization rates. However, the extra 10 h of intra-follicular incubation time decreased oocyte viability.

Braylin induces a potent vasorelaxation, involving distinct mechanisms in superior mesenteric and iliac arteries of rats.

Arterial hypertension is a risk factor for various cardiovascular and renal diseases, representing a major public health challenge. Although a wide range of treatment options are available for blood pressure control, many hypertensive individuals remain with uncontrolled hypertension. Thus, the search for new substances with antihypertensive potential becomes necessary. Coumarins, a group of polyphenolic compounds derived from plants, have attracted intense interest due to their diverse pharmacological properties, like potent antihypertensive activities. Braylin (6-methoxyseselin) is a coumarin identified in the Zanthoxylum tingoassuiba species, described as a phosphodiesterase-4 (PDE4) inhibitor. Although different coumarin compounds have been described as potent antihypertensive agents, the activity of braylin on the cardiovascular system has yet to be investigated. To investigate the vasorelaxation properties of braylin and its possible mechanisms of action, we performed in vitro studies using superior mesenteric arteries and the iliac arteries isolated from rats. In this study, we demonstrated, for the first time, that braylin induces potent vasorelaxation, involving distinct mechanisms from two different arteries, isolated from rats. A possible inhibition of phosphodiesterase, altering the cyclic adenosine monophosphate (cAMP)/cAMP-dependent protein kinase (PKA) pathway, may be correlated with the biological action of braylin in the mesenteric vessel, while in the iliac artery, the biological action of braylin may be correlated with increase of cyclic guanosine monophosphate (cGMP), followed by BKCa, Kir, and Kv channel activation. Together, these results provide evidence that braylin can represent a potential therapeutic use in preventing and treating cardiovascular diseases.

Metástase de seminoma em região orbital em cão - relato de caso / Metastatic seminoma in the orbital region in a dog - case report

O seminoma é uma neoformação testicular originária de células germinativas de ocorrência comum em cães, com maior prevalência em animais senis. Em geral, o comportamento biológico do seminoma canino é benigno. Relata-se neste trabalho um caso de seminoma com metástase em região orbital em um cão com 14 anos de idade. O animal foi atendido com queixa de aumento de volume em órbita esquerda, com posterior detecção de nódulo testicular. A punção aspirativa por agulha fina da massa orbital sugeriu tratar-se de linfoma de alto grau, contudo o diagnóstico definitivo de seminoma difuso foi estabelecido pela avaliação histopatológica, a qual revelou tratar-se de neoplasia maligna pouco diferenciada, sendo o diagnóstico de seminoma difuso confirmado pelo exame imunoistoquímico. Relatos de seminomas metastáticos em cães são incomuns. Objetivou-se com este trabalho relatar um caso de seminoma anaplásico difuso em cão cujo foco principal de metástase ocorreu em região orbital, além de descrever e discutir as dificuldades diagnósticas encontradas.(AU)

Seminoma is a testicular neoformation originating from germ cells, commonly occurring in dogs. With higher prevalence in senile animals, the biological behavior of canine seminomas generally benign. This case reports seminoma with mestastasis in the orbital region in a 14-year-old dog. The animal was treated with a complaint of increased volume in the left orbit, and later a nodule in the testicle was discovered. Fine-needle aspiration of the orbit mass initially indicated a high-grade lymphoma. The definitive diagnosis of diffused seminoma was established by histopathological examination, resulting in poorly differentiated malignant neoplasia. Finally, the diagnosis was confirmed through immunohistochemistry, being the result compatible with diffused seminoma. Metastatic seminomas reported in dogs are quite uncommon. In this work we report a case of diffused anaplastic seminoma in dogs, where the main focus of metastasis was observed in the orbital region, and we also describe and discuss the difficulties encountered in the diagnostic.(AU)

Influence of surface copper content on Stenotrophomonas maltophilia biofilm control using chlorine and mechanical stress.

This work aimed to evaluate the action of materials with different copper content (0, 57, 96 and 100%) on biofilm formation and control by chlorination and mechanical stress. Stenotrophomonas maltophilia isolated from drinking water was used as a model microorganism and biofilms were developed in a rotating cylinder reactor using realism-based shear stress conditions. Biofilms were characterized phenotypically and exposed to three control strategies: 10 mg l-1 of free chlorine for 10 min, an increased shear stress (a fluid velocity of 1.5 m s-1 for 30s), and a combination of both treatments. These shock treatments were not effective in biofilm control. The benefits from the use of copper surfaces was found essentially in reducing the numbers of non-damaged cells. Copper materials demonstrated better performance in biofilm prevention than chlorine. In general, copper alloys may have a positive public health impact by reducing the number of non-damaged cells in the water delivered after chlorine exposure.

Prolonged exposure of Stenotrophomonas maltophilia biofilms to trace levels of clofibric acid alters antimicrobial tolerance and virulence.

The presence of pharmaceuticals in water sources, including in drinking water (DW), is increasingly being recognized as an emerging and global concern for the environment and public health. Based on the principles of the "One Health" initiative, the present work aims to understand the effects of clofibric acid (CA), a lipid regulator, on the behavior of a selected bacterium isolated from drinking water (DW). Biofilms of the opportunistic pathogen Stenotrophomonas maltophilia were exposed to CA for 12 weeks at 170 and 17000 ng/L. The effects of CA were evaluated on planktonic S. maltophilia susceptibility to chlorine and antibiotics (amoxicillin, ciprofloxacin, erythromycin, kanamycin, levofloxacin, oxacillin, spectinomycin, tetracycline and trimethoprim-sulfamethoxazole), biofilm formation, motility, siderophores production and on the adhesion and internalization of the human colon adenocarcinoma cell line (HT-29). It was found that CA did not affect planktonic S. maltophilia tolerance to chlorine exposure. Additionally, no effects were observed on biofilm formation, motility and siderophores production. However, biofilms formed after CA exposure were more tolerant to chlorine disinfection and lower CFU reductions were obtained. Of additional concern was the effect of CA exposure on S. maltophilia increased tolerance to erythromycin. CA exposure also slightly reduced S. maltophilia ability to invade HT-29 cells. In conclusion, this work reinforces the importance of studying the effects of non-antibiotic contaminants on the behavior of environmental microorganisms, particularly their role as drivers affecting resistance evolution and selection.

Morfologia do pâncreas da paca (Cuniculus paca, Linnaeus, 1766) / Morphology of the pancreas of pacas (Cuniculus paca, Linnaeus, 1766)

Este trabalho se baseou na descrição morfológica da paca (Cuniculus paca), espécie selvagem pertencente à ordem dos roedores, típica de regiões tropicais, sendo o seu conhecimento anatômico detalhado muito importante para sua conservação, experimentação e comercialização, pois ainda são escassos na literatura os trabalhos sobre morfologia de espécies selvagens. Macroscopicamente, o pâncreas da paca é formado por lóbulos, organizados em ramificações difusas, e, microscopicamente, tal órgão possui uma camada de tecido conjuntivo frouxo, a partir da qual partem septos, que dividem a glândula em lóbulos, compostos de ácinos e ilhotas pancreáticas.(AU)

The present study aimed to describe the morphology of pacas (Cuniculus paca), a wild species belonging to the order Rodentia, found in tropical areas. The detailed anatomical knowledge of pacas is very important for conservation, experimental studies and commercialization because studies in literature about wild species are scarce. The pancreas of paca has lobes organized in diffuse branches at the macroscopic level and microscopically this organ has a loose connective tissue layer, originating septa that divide the gland into lobes composed of acini and pancreatic islets.(AU)

The role of surface copper content on biofilm formation by drinking water bacteria.

Copper pipes are conventionally used to supply tap water. Their role in biofilm prevention remains to be understood. This study evaluates the ability of selected surface materials with different copper contents (0, 57, 79, 87, 96, 100% of copper) to control biofilm formation and regrowth. Further experiments were performed to assess copper leaching and corrosion under conditions mimicking real plumbing systems. Acinetobacter calcoaceticus and Stenotrophomonas maltophilia isolated from a drinking water distribution system were used as model bacteria. All the copper materials showed positive results on the control of single and dual species biofilms presenting high reductions of bacterial culturability > 4 log CFU per cm2. The antimicrobial action of the selected materials seem not to be related to copper leaching or to the formation of reactive oxygen species. However, bacterial-copper contact demonstrated damage to bacterial membranes. The alloy containing 96% copper was the most promising surface in reducing biofilm culturability and viability, and was the only surface able to avoid the regrowth of single species biofilms when in contact with high nutrient concentrations. The alloy with 87% copper was shown to be unsuitable for use in chlorinated systems due to the high copper leaching observed when exposed to free chlorine. The presence of viable but non-culturable bacteria was remarkable, particularly in dual species biofilms. The overall results provide novel data on the role of copper alloys for use under chlorinated and unchlorinated conditions. Copper alloys demonstrated comparable or even higher biofilm control effects than elemental copper surfaces.

Presynchronization by induction of a largest follicle using a progesterone device in GnRH-based-ovulation synchronization protocol in crossbred dairy cows.

The objective was to compare the fertility of dairy cows using a presynchronization protocol by induction of a largest follicle using a progesterone intravaginal device prior to an Ovsynch protocol (P4synch) with the Double-Ovsynch in lactating dairy cows. Lactating Bos indicus x Bos taurus crossbred cows (n = 440) were randomly allocated to one of two treatments: (I) Double-Ov (n = 228), GnRH (D-17), PGF2α 7 days later (D-10) and GnRH 3 days later (D-7) followed by an Ovsynch protocol 7 days later (GnRH on D0, PGF on D7, GnRH on D9); (II) P4synch (n = 212), insertion of a sustained release progesterone intravaginal device (D-10), 10 days later (D0) an Ovsynch protocol was initiated (GnRH on D0, PGF on D7, GnRH on D9) with progesterone device withdrawal on Day 7. All cows were artificially inseminated (TAI) 16 h after the second GnRH of the Ovsynch protocol and pregnancy diagnosis was performed by transrectal ultrasonography 30 and 60 days after TAI. A subset of cows (n = 52 for Double-Ov and n = 50 for P4synch) ultrasonography was performed on days 0, 7, 9 and 24 of the experimental period. There were no differences among treatments on presynchronization rate [presence of a follicle>12 mm on D0, Double-Ov 94.2% (49/52) and P4synch 92.0% (46/50); P = 0.66], follicular diameter on the 1st GnRH (Double-Ov 17.2 ±â€¯0.7 mm e P4synch 18.6 ±â€¯0.9 mm; P = 0.28), ovulation rate to the 1st GnRH [Double-Ov 86.3% (44/51) and P4synch 81.2% (39/48); P = 0.50], synchronization rate [presence of a follicle>12 mm on D9; Double-Ov 84.6% (44/52) and P4synch 86.0% (43/50); P = 0.84], follicular diameter on the 2nd GnRH (Double-Ov 17.5 ±â€¯0.6 mm and P4synch 18.0 ±â€¯0.5 mm; P = 0.48), ovulation rate to the 2nd GnRH [Double-Ov 90.9% (40/44) and P4synch 86.0% (37/43); P = 0.48] and CL diameter on D24 (Double-Ov 27.9 ±â€¯0.7 mm and P4synch 29.4 ±â€¯0.9 mm; P = 0.19). Corpus luteum presence on D0 was different (P = 0.03) among treatments [Double-Ov 57.7% (30/52) and P4synch 36.0% (18/50)]. There was no difference (P = 0.85) among the pregnancy per AI on day 30 [Double-Ov 39.0% (89/228) and P4synch 40.1% (85/212)], on day 60 [Double-Ov 34.8% (79/227) and P4synch 38.7% (82/212); P = 0.41] and pregnancy loss [Double-Ov 10.2% (9/88) and P4synch 3.5% (3/85); P = 0.08]. The presynchronization by induction of a largest follicle using a sustained release progesterone device prior to Ovsynch yielded similar results compared with the Double Ovsynch protocol on follicular development patterns and on the fertility of lactating dairy cows.

Subclinical mastitis interferes with ovulation, oocyte and granulosa cell quality in dairy cows.

The objective was to evaluate the effect of mastitis by somatic cell count (SCC) on follicular growth, ovulation, oocytes and cumulus cells quality and on the concentration and size of exosomes in follicular fluid of dairy cows. In the study, crossbred cows (Bos taurus - Holstein x Bos indicus - Gir) were classified for analysis as Control (SCC<200.000 cells/mL) and Mastitis (SCC>400.000 cells/mL) groups. In experiment 1 (follicular dynamics), cows (n = 57) were submitted to ultrasound evaluations every 24 h, from progesterone-releasing-intravaginal-device (PRID) removal (D8) until 48 h later (D10). Thereafter, evaluations were performed every 12 h, until ovulation or up to 96 h after PRID removal. In experiment 2 (oocyte, cumulus complexes, and follicular fluid evaluation), cows (n = 26) were submitted to follicular aspiration (OPU) for oocyte quality and cumulus cells transcript evaluation. The amount of cumulus complexes transcripts (BCL2, BAX, PI3K, PTEN, FOXO3) was determined by Real-Time Polymerase Chain Reaction. Moreover, seven days after the OPU session, the dominant follicle was aspirated. Exosomes were isolated from the follicular fluid for evaluation of particle size and concentration. Ovulation rate [Control 77.4% (24/31) and Mastitis 57.7% (15/26); P = 0.09] and viable oocytes rate [Control 59.1% (130/220) and Mastitis 41.9% (125/298); P = 0.01] were higher in Control animals. Additionally, there was a greater number of degenerate oocytes [Control 6.7 ±â€¯1.2 and Mastitis 13.3 ±â€¯5.5; (P = 0.001)] in subclinical mastitis cows. There was greater abundance (P = 0.003) of BAX cumulus cell transcripts and exosome mean (P = 0.03) and mode (P = 0.02) was smaller in subclinical mastitis cows. In conclusion, ovulation rate, oocyte quality, and exosome diameter were smaller in cows with SCC>400.000 cells/mL.

Exposure to progesterone previous to the protocol of ovulation synchronization increases the follicular diameter and the fertility of suckled Bos indicus cows.

The objective was to evaluate the effect of injectable progesterone previous to the timed artificial insemination (TAI) protocol on follicular growth, ovulation and pregnancy rate of suckled Bos indicus cows. In experiment 1, 10 days before the beginning to TAI protocol (D-10), 431 suckled-anestrus Nelore cows (249 multiparous and 182 primiparous), were allocated to one of three treatments groups (control, P4i and P4iGnRH). At this moment, cows in the P4i and P4iGnRH group received 150 mg of injectable progesterone intramuscularly (Sincrogest injetável®, Ouro Fino, Brazil). On Day 0 (D0), all cows were synchronized using an estradiol/progesterone-based TAI protocol. Simultaneously, in the P4iGnRH group, cows received 10 µg of Busereline intramuscularly (Sincroforte®, Ouro Fino, Brazil). Ultrasound exams were performed to evaluate the diameter of the largest follicle (D0, D8 and D10), ovulation rate and diameter of the corpus luteum (D24). In experiment 2, 681 suckled Nelore cows (356 multiparous and 325 primiparous) were synchronized using an estradiol/progesterone-based TAI protocol and received treatments similar to experiment 1. TAI was performed 48 h after removal of the progesterone (P4) device. Pregnancy diagnosis was 30 d after TAI. In experiment 3, blood samples were collected to evaluated the progesterone concentration for 168 h after administration of injectable progesterone intramuscularly. Statistical analyses were performed by GLIMMIX procedure of SAS. In experiment 1, the diameter of the largest follicle (LF) on D10 (P = 0.21), follicular growth rate (P = 0.34) and ovulation rate (P = 0.62) were similar among experimental groups. However, there was difference among groups for the LF on D0 [Control (10.9 ±â€¯0.2 mm)b, P4i (12.7 ±â€¯0.3 mm)a and P4iGnRH (12.6 ±â€¯0.3 mm)a; P = 0.001], LF on D8 [Control (9.7 ±â€¯0.2 mm)b, P4i (10.4 ±â€¯0.2 mm)a and P4iGnRH (9.9 ±â€¯0.2 mm)ab; P = 0.05], presence of the CL on D8 [Control 0% (0/136)b, P4i 0% (0/140)band P4iGnRH 26.4% (38/144)a; P = 0.001], diameter of the CL on D24 [Control (19.7 ±â€¯0.4 mm)ab, P4i (20.1 ±â€¯0.4 mm)a and P4iGnRH (18.5 ±â€¯0.4 mm)b; P = 0.001] and pregnancy rate [Control 35.0% (78/223)b, P4i 45.9% (105/229)a and P4iGnRH 40.6% (93/229)ab; P = 0.01]. The circulating concentration of P4 remained above 1.5 ng/mL until 168 h after the P4i treatment. In conclusion, the injectable progesterone previous to the TAI protocol increased diameter of the LF on D0 and D8 without interfering on the ovulation rate. Furthermore, such exposure increases the pregnancy rate in suckled Nelore cows.

The action of chemical and mechanical stresses on single and dual species biofilm removal of drinking water bacteria.

The presence of biofilms in drinking water distribution systems (DWDS) is a global public health concern as they can harbor pathogenic microorganisms. Sodium hypochlorite (NaOCl) is the most commonly used disinfectant for microbial growth control in DWDS. However, its effect on biofilm removal is still unclear. This work aims to evaluate the effects of the combination of chemical (NaOCl) and mechanical stresses on the removal of single and dual species biofilms of two bacteria isolated from DWDS and considered opportunistic, Acinectobacter calcoaceticus and Stenotrophomonas maltophilia. A rotating cylinder reactor was successfully used for the first time in drinking water biofilm studies with polyvinyl chloride as substratum. The single and dual species biofilms presented different characteristics in terms of metabolic activity, mass, density, thickness and content of proteins and polysaccharides. Their complete removal was not achieved even when a high NaOCl concentrations and an increasing series of shear stresses (from 2 to 23Pa) were applied. In general, NaOCl pre-treatment did not improve the impact of mechanical stress on biofilm removal. Dual species biofilms were colonized mostly by S. maltophilia and were more susceptible to chemical and mechanical stresses than these single species. The most efficient treatment (93% biofilm removal) was the combination of NaOCl at 175mg·l-1 with mechanical stress against dual species biofilms. Of concern was the high tolerance of S. maltophilia to chemical and mechanical stresses in both single and dual species biofilms. The overall results demonstrate the inefficacy of NaOCl on biofilm removal even when combined with high shear stresses.

Molecular and endocrine factors involved in future dominant follicle dynamics during the induction of luteolysis in Bos indicus cows.

The growth profiles of the future dominant follicle (DF) and subordinate follicle (SF) and the gene expression of the granulosa cells during luteolysis induction in Bos indicus cows were evaluated. Forty cows were synchronized with a progesterone and estradiol based protocol. After synchronization, cows with a corpus luteum (CL) were evaluated by ultrasonography every 12 h, beginning at eight days post ovulation. Cows identified with a follicle of at least 6.0 mm in diameter in the second wave were split into two groups (BD-before follicular deviation and AD-after follicular deviation. In the BD group cows received 500 µg of cloprostenol (a synthetic analogue of prostaglandin F2α) when the DF reached a mean diameter of 7.0 mm (6.5-7.5 mm). In the AD group, cows received 500 µg of cloprostenol when the DF reached a mean diameter of 8.0 mm (7.5-8.5 mm). Cows in both groups were submitted to aspiration of the DF at 96 and 72 h after prostaglandin was given. Follicular aspirations were performed to quantify IGF1R, LHR and PAPPA transcripts in the granulosa cells. The diameter of the DF at the moment of prostaglandin administration (P = 0.001) and the growth rate of the SF (P = 0.05) were greater in the AD group. There was greater abundance of LHR transcripts in BD cows (P = 0.04). The remaining variables tested were similar between the experimental groups (P > 0.05). In conclusion, the induction of luteolysis before follicular deviation does not interfere with dominant follicle dynamics. However, it causes granulosa cell LHR down regulation.