In vitro culture and morphology of Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae) collected from the intestine of tambaqui (Colossoma macropomum) farmed in the Brazilian Amazon.

The tambaqui (Colossoma macropomum) is a species of great economic importance for fish farming in the Brazilian Amazon, and acanthocephaliasis caused by Neoechinorhynchus buttnerae (Golvan 1956) represents an obstacle to its production due to it causing severe morphological damage to the intestinal mucosa, thus impairing the absorption of nutrients and causing weight loss in the fish. Therefore, the establishment of in vitro protocols for evaluation of anthelmintic drugs is the first step to development of effective measures for in vivo control of this endoparasite. The present study evaluated the in vitro survival of N. buttnerae maintained in Eagle's minimum essential medium under different culture conditions. Three assays were carried out to evaluate whether temperature, supplementation with the antibiotics penicillin and streptomycin, and culture medium replacement or no replacement would influence the motility and morphology of the acanthocephalans. The results of the Kaplan-Meier analysis indicated that the use of culture in minimum essential medium together with penicillin and streptomycin prolonged the parasite's survival when kept at temperatures of 24 °C or 28 °C. We describe herein for first time an alternative protocol that is ideal for the in vitro culture of N. buttnerae. As such, this protocol ensures greater reliability in further in vitro studies with N. buttnerae.

Acute toxicity of essential oils of Aloysia triphylla (L'Hér.) Britton, Lippia gracilis Schauer, and Piper aduncum L. in Colossoma macropomum (Cuvier, 1818).

The aim of this study was to determine the acute toxicity of the essential oils (EOs) of Aloysia triphylla, Lippia gracilis and Piper aduncum in juvenile tambaqui (Colossoma macropomum), and evaluate the possible histopathological alterations in their gills. For the acute toxicity tests, juvenile tambaqui (n=24/treatment) were distributed in six treatments with three replicates, which comprised the control and five EO concentrations of A. triphylla (60, 80, 100, 120 and 140 mg L-1), L. gracilis (35, 40, 45, 50 and 55 mg L-1) and P. aduncum (42.5, 45, 47.5, 50 and 52.5 mg L-1), with an exposure period of 4 h. The mortality rate and severity of damage to the tambaqui gills were proportional to the increase in the concentration of the EO, with LC50-4 h values estimated at 109.57 mg L -1 for A. triphylla, 41.63 mg L -1 for L. gracilis and 48.17 mg L -1 for P. aduncum. The main morphological damages observed in the gills of the tambaqui exposed to the three EOs, were Grade I: hypertrophy and hyperplasia of lamellar epithelial cells, lamellar fusion, epithelial detachment, capillary dilation and constriction, proliferation of chloride cells and mucosal cells and edema; in low frequency Grade II damage as epithelial rupture and lamellar aneurysm. Necrosis (Grade III damage) was observed only in gill lamellae exposed to P. aduncum EO (47.5, 50.0 and 52.5 mg L-1). Concentrations of EOs below LC50-4 h can be used sparingly, for short periods of exposure for the treatment of diseases in tambaqui breeding.

Review: Non-surgical artificial insemination and embryo recovery as safe tools for genetic preservation in small ruminants.

Artificial insemination (AI) and in vivo embryo production (or multiple ovulation and embryo transfer, MOET) programs are both instrumental in accelerating the propagation of genetically and economically superior goats and sheep. The aim of this review was to present the current gestalt of non-surgical AI and embryo recovery (NSER) procedures in small ruminants. Small body size, precluding rectal palpation, and highly limited penetrability of the uterine cervix in ewes are the major reasons for the scarce use of non-surgical assisted reproduction techniques in this species. As a result, AI and embryo recovery techniques in sheep mainly involve laparoscopy or laparotomy (LAP). In does, however, the Embrapa method of AI allows for successful intrauterine deposition of semen, resulting in pregnancy rates from 50 to 80% under field conditions (>3 000 goats inseminated) when frozen-thawed semen is used. After the administration of prostaglandin F2α (PGF2α), non-surgical (transcervical) embryo recovery is also feasible in goats, with the cervical penetration rate approaching 100%. There is a paucity of information on the efficacy of non-surgical AI using frozen semen in sheep, but the results are satisfactory with fresh, cooled, or chilled ram semen. An application of the NSER technique in ewes has greatly improved over the last decade, and cervical penetration rates of ∼90% can be achieved when a hormonal cervical dilation protocol using PGF2α, oxytocin, and/or estradiol ester (e.g., estradiol benzoate) is applied. In some genotypes of sheep, sufficient cervical dilation can be induced without estradiol ester included in the protocol. Several studies indicated that recovery of transferable quality ovine embryos using NSER is comparable to that employing a ventral midline laparotomy, and NSER is evidently a method of choice when animal welfare is concerned. Considering both the number of retrievable embryos and animal well-being, the NSER is a viable alternative for surgical procedures. With further developments, it has the makings of a primary, if not exclusive, embryo recovery technique in small ruminants worldwide.

Effect of the addition of antifreeze protein type I on the quality of post-thawed domestic cat epididymal sperm.

Cryopreservation of domestic cat semen is mainly performed as a model for the establishment of endangered wild feline protocols. The supplementation of antifreeze protein type I (AFP I) to cryopreservation medium has shown improvement in frozen-thawed sperm quality in other species, but its effect on cat semen has not yet been tested. This study aimed to assess the addition of AFP I to cryopreservation medium in domestic cats. Sperm was obtained from the cauda epididymis of orchiectomized cats; sperm was then pooled in Tris buffer and allocated into three treatments, according to AFP I final concentration: 0 (control), 0.1, and 0.5 µg/ml. Nine replicates were cryopreserved in a two-step protocol and subsequently thawed at 37°C for 30 s. There was no difference (P > 0.05) among the control, 0.1 and 0.5 µg/ml groups for parameters such as motility, vitality, functional membrane integrity, mature chromatin, normal morphology, and sperm binding to egg perivitelline membrane. In the 0.5 µg/ml group only, percentages of live sperm with intact acrosome and of sperm with most inactive mitochondria (DAB III) showed a significant reduction, along with a tendency (P = 0.053) to an increase in the percentage of sperm with most active mitochondria (DAB II). In conclusion, the supplementation of 0.1 and 0.5 µg/ml of AFP I did not promote consistent beneficial effects on the overall sperm cryotolerance in domestic cats.

Antimicrobial resistance profile of Aeromonas spp. isolated from asymptomatic Colossoma macropomum cultured in the Amazonas State, Brazil.

Bacterial diseases are important factors that limit productivity in aquaculture. To reduce negative economic impacts, fish farmers use antimicrobials, often indiscriminately, and this action has led to bacterial resistance to drugs. The objectives of this study were to isolate and identify the main putative pathogenic bacterial species in tambaqui (Colossoma macropomum), establish the profile of resistance to antimicrobials by the methods of disc diffusion, and determine the minimum inhibitory concentration (MIC) values. Two hundred and ninety asymptomatic fish were collected between March and November 2015 from ten fish farms in the Amazonas state (Brazil). Of the total strains recovered from tambaqui, seven were identified as Aeromonas spp. by sequencing the 16S rRNA gene. These seven isolates showed resistance to ampicillin, 28% to erythromycin, and 28% to sulfonamide. Additionally, the seven isolates showed a MIC higher than the range evaluated for amoxicillin, penicillin, novobiocin, tylosin tartrate, and clindamycin, and 85% showed resistance to erythromycin. The results of this study indicate the need to increase the awareness of fish farmers and, most importantly, the government, about the lack of drug regulations for use in aquaculture, and good management practices, so the indiscriminate prophylactic and systemic use of antimicrobials be inhibited.

Ovarian steroids modulate the systemic inflammatory response OF COWS challenged with lipopolysaccharide (LPS) intrauterine infusion.

Postpartum uterine infections of dairy cows promote a local and systemic inflammation and interfere with reproductive efficiency. The aim of this study was to evaluate the effect of steroid hormones including progesterone (P4) and estradiol (E2) on the systemic inflammatory response of cows after being challenged with an intrauterine infusion of lipopolysaccharide (LPS). For this, a hemogram and serum dosage of haptoglobin (Hp) in eight primiparous Gir cows ovariectomized were performed on day (day 0) and after 24 h (day +1). Four cows (n = 4) were challenged (day 0) with 20 mL of 0.9% NaCl + 12.5 µg/kg LPS, and four cows (n = 4) were challenged (day 0) with 20 mL of 0.9% NaCl. For this, the study was divided in four experimental groups as: (1) Control group: without any hormonal treatment before day 0; (2) Group 24 h - E2: 1 mg of estradiol benzoate 24 h before (day -1); (3) Group 24 h - P4: 2.0 g of P4 device 24 h before (day -1); (4) Group 14 d - P4: 2.0 g of P4 device 14 days before (day -14). In the systemic response to LPS, there was an increase in Hp (control group; 24 h - P4 group; 14 d - P4 group), and on day +1 the Hp of 14 d - P4 group was higher when compared to the other groups. On day 0, the 14 d - P4 group had an increase in circulating leukocytes and lymphocytes cells than the control group (P < 0.01). On day +1 after LPS-challenge the 14 d - P4 group showed a decrease in circulating lymphocytes, eosinophils, and monocytes (P < 0.05). A neutrophilia with left shift in the two treatments with P4 (day +1), in addition to a thrombocytopenia and lower platelets compared to the 24 h - E2 group (P < 0.05) (day 0) were recorded. It was concluded that ovariectomized cows challenged with LPS, previously submitted to steroid hormones induce a systemic inflammatory response. Also, the systemic response is more intense after previous prolonged exposure to P4 and less intense after exposure to E2. This study provided important information relating the effect of ovarian steroids on the systemic inflammatory response of cows challenged with intrauterine LPS.

Heterotopic autotransplantation of equine ovarian tissue using intramuscular versus subvulvar grafting sites: Preliminary results.

Ovarian tissue transplantation (OTT) is a technique well established and successfully applied in humans using mainly orthotopic or heterotopic transplantation sites. In livestock, OTT is still in its infancy and, therefore, different aspects of the technique, including the efficiency of different heterotopic OTT sites as well as the potential effect of age (i.e., young vs. old mares) in the ovarian graft quality, need to be investigated. The present study investigated the efficacy of the intramuscular (IM) or the novel subvulvar mucosa (SV) heterotopic autotransplantation sites to maintain the survivability of the grafts for 3 and 7 days post-OTT. Ovarian biopsy fragments were obtained in vivo and distributed to the following treatments: Fresh control group (ovarian fragments immediately fixed), SV-3, IM-3, SV-7, and IM-7. During and after graft harvesting, the macroscopic characteristics of the grafts (i.e., adherence, morphology, and bleeding) were scored, and the percentages of morphologically normal and developing preantral follicles as well as the follicular and stromal cell densities of the grafts were evaluated. The results were that similar (P > 0.05) macroscopic scores were observed between both transplantation sites 7 days post-OTT, with positive correlations (P < 0.01) found among adherence, morphology, and bleeding of the grafts. A lower (P < 0.05) percentage of morphologically normal follicles was found 7 days post-OTT in the SV site (82%) compared with the Fresh control group (99%) and IM site (95%); however, the percentages of developing follicles were similar (P > 0.05) between both transplantation sites 7 days post-OTT (30-43%). Although similar (P > 0.05) follicular densities were found in both transplantation sites in young and old mares at 3 and 7 days post-OTT, large individual variation in the follicular depletion rate was observed regardless of transplantation site. The Fresh control group and SV-7 treatments had higher (P < 0.05) stromal cell densities in young and old mares compared with both IM-7 treatments. When comparing transplant sites between young and old mares, the follicular density in old mares and the stromal cell density in young mares were greater (P < 0.05) in the SV than in the IM site. In conclusion, even though the transplantation sites differentially affected some end points, overall comparable findings of the OTT technique using both heterotopic autotransplantation sites (i.e., IM and SV) for equine ovarian tissue were observed.

Oviduct fluid during IVF moderately modulates polyspermy in in vitro-produced goat embryos during the non-breeding season.

The present study determined i) the presence of proteins (oviduct-specific glycoprotein, OVGP1; heat shock protein-70A, HSPA1A; heat shock protein-A8, HSPA8; annexin A1, ANXA1; annexin A5, ANXA5; and myosin-9, MYH9) known to be involved in early reproduction in the oviduct fluid (OF) of anestrous goats; and ii) the functional effect of during IVF on polyspermy modulation and embryonic development. In vitro-matured oocytes were co-cultured with spermatozoa (1.0, 2.0, or 4.0 x 106 cells/mL) for 18 h in SOF medium supplemented with 5 µg/mL of heparin, 4 µg/mL gentamicin, and 10% estrus sheep serum (CTRL1, CTRL2, and CTRL4 groups) or the same medium plus 10% OF (OF1, OF2, and OF4 groups) obtained from anestrus goats. The analysis of OF by western blotting confirmed the presence of the six proteins tested for. The increase in sperm concentration had no effect (P > 0.05) on the penetration rate in any group; however, monospermy rate decreased as sperm concentration was increased in both OF and CTRL. Regardless of the concentration used, when data were pooled, OF supplementation improved (P < 0.05) monospermy and tended (P = 0.057) to enhance IVF efficiency. Additionally, IVF efficiency was higher (P < 0.05) in OF1 than in OF4 [60 ± 13 vs 37 ± 5%). The development capacity was not affected (P > 0.05) by the sperm concentration and OF treatment, and the average values were cleavage (72 ± 2.6%), blastocyst (37 ± 3.0%), blastocyst in relation to the cleaved (51 ± 4.8%), hatched (62 ± 1.2%), and number of cells per blastocyst (174 ± 1.8%). In conclusion, the six proteins analyzed are present in the OF of anestrous goats, and the supplementation of this OF during IVF may modulate the polyspermy incidence and enhance IVF efficiency, especially when 1x106 sperm per mL is used.

Exogenous progestogens differentially alter gene expression of immature cumulus-oocyte complexes in sheep.

This study evaluated the role of progesterone (P4) and medroxyprogesterone acetate (MAP) on the molecular status of immature cumulus-oocyte complexes (COCs) and the implications for oocyte quality in sheep. The number of viable COCs per ewe and the rate of COCs screened for developmental competence by brilliant cresyl blue positive (BCB+) were similar (P > 0.05), respectively, across treatments (P4: 7.7 ± 0.7 and 4.7 ± 1.2; MAP: 5.7 ± 1.0 and 3.5 ± 2.3; and control: 5.7 ± 1.1 and 3.6 ± 2.4). The COCs' gene expression was altered by exogenous progestogens compared with the control group: markers of steroidogenic pathway (FSH receptor [FSHr], LH receptor [LHr], and estradiol receptor α) and of quality (zygote arrest 1, growth differentiation factor 9, and B-cell lymphoma 2) were in abundance in P4 (P < 0.05). In addition, reelin protein (RELN) was downregulated, and Bcl-2 was upregulated in MAP (P < 0.05). In the P4 vs MAP comparison, FSHr, LHr, and RELN genes were upregulated (P < 0.05) in the P4 group. In conclusion, P4 and MAP promoted dissimilar effects on transcriptome profiling of immature BCB-selected COCs, possibly due to the differences in the chemical structure of progestogens and concentrations of serum P4. Exogenous P4 impacted positively on the profile of genes related to oocyte quality.

Administration of a single dose of 300 IU of human chorionic gonadotropin seven days after the onset of estrus improves pregnancy rate in dairy goats by an unknown mechanism.

This study examined the effects of exogenous hCG administration on ovarian function and pregnancy rates in estrous-induced dairy goats during the transition into the breeding season. Eighty-six Toggenburg does received 60 mg of medroxyprogesterone acetate intravaginal sponge for 6 d plus 200 IU of equine chorionic gonadotropin and 30 µg of d-cloprostenol i.m. 24 h before sponge removal, and were then bred for 96 h. Seven days (D7) after first mating the does received either 1 mL of saline (the control group, n = 43) or 300 IU of hCG (the hCG-treated group, n = 43) i.m. Transrectal ovarian ultrasonography (B-mode and color Doppler) was performed on D7, D13, D17, and D21 and ultrasonographic pregnancy detection on D30. Pregnancy rate was higher (P < 0.05) in hCG-treated goats (90.7%; 39/43) than that in control animals (74.4%; 32/43). Accessory luteal structures (ALSs) were detected in 46.5% (20/43) of hCG-treated does. All hCG-treated does that had ALSs and 82.6% of goats without ALS post-treatment remained pregnant. The total luteal area increased (P < 0.05) from D7 to D13 in pregnant animals of both groups, whereas mean vascular area declined (P < 0.05) by D21 in all nonpregnant does. Serum progesterone concentrations increased (P < 0.05) on D21 in pregnant goats of both groups, but they were related to changes in luteal tissue content only in control does throughout the present study. Mean daily numbers of small- and medium-sized antral follicles decreased (P < 0.05) only in pregnant animals of both groups with a decline in medium follicle numbers occurring earlier in hCG-treated (D13) compared with control does (D17). To summarize, a single dose of hCG given on D7 after estrus was followed by a decrease in the number of medium-sized antral follicles in gestating hCG-treated does, induced the formation of ALSs in ~47% of all hCG-treated does, and significantly increased the pregnancy rate in estrous-induced Toggenburg goats in the transition to the breeding season.

Complete pathological response (pCR) in gastroesophageal cancer: Correlation with metabolic response.

PURPOSE: Neoadjuvant chemoradiotherapy (nCRT) followed by surgery in patients with resectable esophageal or esophagogastric junctional (GEJ) (Siewert I) cancer is associated with long term overall survival benefits. Up to one third of all patients submitted to nCRT present pathological complete response (pCR). 18F-fluorodeoxyglucose positron emission tomography with CT (18F-FDG PET-CT) is an important tool for assessing treatment response. Purpose was to assess retrospectively the power of 18F-FDG PET-CT in predicting pCR to evaluate the feasibility of a "watch and wait" approach. PATIENTS AND METHODS: Retrospective analysis of a prospective database with esophageal or GEJ submitted to pre-operative chemoradiation. Pre and pos treatment 18F-FDG PET-CT were reviewed and classified using visual assessment and PERCIST criteria and the values of maximum standard uptake value were also recorded. Patients were classified as pCR or non-PCR. 18F-FDG PET-CT and pathological findings were compared against each other. RESULTS: Forty-three patients were included. The median age was 67 years and 90.7% were male. All patients underwent preoperative CRT and were evaluated with 18F-FDG PET-CT pre and post treatment. Transthoracic surgery was performed in all patients. Histological type was adenocarcinoma in 37% and squamous cell carcinoma in 58%. pCR was achieved in 56% of cases. Visual assessment of 18F-FDG PET-CT showed overall sensitivity 57.9%, specificity 62.5% and PERCIST criteria had 100% sensibility and 16.7% specificity. CONCLUSIONS: 18F-FDG PET-CT is not an ideal predictor of pCR but if we use the PERCIST criteria we will have a high sensitivity and negative predictive value, avoiding false negative scans.

Idade à puberdade e características reprodutivas de novilhas mestiças F1 Holandês x Gir com fenótipos divergentes para consumo alimentar residual / [Age at puberty and reproductive characteristics of F1 Holstein x Gyr crossbred heifers with divergent phenotypes for residual feed intake]

A relação da eficiência nutricional com a idade e o peso à puberdade e as características reprodutivas foram estudadas em 35 novilhas F1 Holandês x Gir, em ensaios usando o consumo alimentar residual (CAR) nos períodos pós-desaleitamento (PD) e pós-puberdade (PP). Novilhas PD (P<0,0001) e PP (P<0,001) foram ranqueadas como de alta, intermediária e baixa eficiências e apresentaram CAR de -0,259 -0,014 e 0,346kg/dia e -0,848 -0,096 e 0,842kg/dia, respectivamente. Novilhas PD diferiram (P<0,05) na idade à puberdade entre ranques de alta, intermediária e baixa eficiências, em média, aos 339,7; 346,4; 315,3 dias, respectivamente. Os pesos à puberdade de novilhas PD com alta, intermediária e baixa eficiências foram, em média, de 321±28,1kg e não diferiram (P>0,05) entre CAR. Não foi verificada correlação entre diâmetro folicular, taxa de crescimento do folículo, número de ondas foliculares no ciclo estral e taxa de gestação à primeira IA com o CARPD ou CARPP. Novilhas F1HG selecionadas para CAR com baixa eficiência podem ter idade à puberdade 10 dias a mais, quando comparadas às novilhas de alta eficiência para CAR. Não foi verificado que novilhas selecionadas para alta ou baixa eficiências alimentares, usando CAR, tivessem algum impacto negativo sobre características reprodutivas avaliadas.(AU)

The relationship between nutritional efficiency and age and weight at puberty and reproductive traits was studied in 35 F1 Holstein x Gyr heifers, in trials using residual food consumption (CAR) in the post-weaning (PW) and post-pubertal (PP) periods. PD heifers (P<0.0001) and PP (P<0.001) were classified as high, intermediate and low efficiencies and presented CAR of -0.259 -0.014 and 0.346kg/day and -0.848 -0.096 and 0.842kg/day, respectively. PD heifers differed (P<0.05) in age at puberty between high, intermediate and low efficiencies, on average, at 339.7, 346.4 and 315.3 days, respectively. The weight at puberty of PD heifers with high, intermediate and low efficiencies were, on average, 321±28.1kg and did not differ (P>0.05) between CAR. No correlation was found between follicular diameter, follicle growth rate, number of follicular waves in the estrous cycle and pregnancy rate at 1st AI with the CARPD or CARPP. F1HG heifers selected for CAR with low efficiency can be aged at puberty 10 days longer when compared to high-efficiency heifers for CAR. These results did not show that heifers selected for high or low food efficiency, using CAR, had negative impact on the reproductive characteristics evaluated.(AU)

Repeated trans-cervical embryo recoveries in Santa inês ewes subjected to short- or long-term superovulatory treatment regimens.

Outcomes of short- (6.5 days) and long-term (14.5 days) estrous synchronization for 6.5 d (G-6.5d) or 14.5 d (G-14.5d) and followed by the 4-day or 3-day declining-dose follicle-stimulating hormone superovulatory regimen, respectively, were compared using 16 estrous-cycling Santa Inês ewes. Non-surgical embryo recovery (NSER) procedures were performed 60 d apart starting 6 or 7 d after the onset of estrus; an i.m. injection of estradiol benzoate and of d-cloprostenol at 16 h was followed by an i.v. oxytocin injection administered 20 min before NSER. There was a longer (P < 0.05) period before estrous onset in ewes during the second (September) compared with the first study replicate (July) by approximately 14 h. The NSER could be performed in 11 of 15 ewes that were in estrus, with an average of three viable-embryos/donor and the mean duration of the procedure being 29 min. There were no differences in superovulatory responses between the two groups of ewes, but there were only degenerated embryos in ewes of the G-6.5d group. In summary: i. the duration of progestin-priming and of multiple-dose pFSH treatment had a limited effect on superovulatory responses in estrous-cycling Santa Inês ewes; ii. NSER is a safe and repeatable method of embryo collection in ewes subsequent to superovulation; and iii. duration of the superovulatory treatment regimen may alter the effects of endogenous steroids on oocyte/embryo quality in ewes.

Exogenous progestogen does not affect first-wave follicle populations and oocyte quality during ovarian stimulation with FSH in sheep.

The effect of short-term administration of medroxyprogesterone acetate (MPA) or natural progesterone (P4) during ovarian stimulation with FSH on oocyte recovery was investigated in Santa Inês ewes. Ewes were treated with an intravaginal sponge containing MPA for 6 d; GnRH was applied 36 h after sponge removal and FSH was given in 3 injections (40, 24, and 16 mg, respectively) every 12 h after (D0, approximate ovulation time). At the first FSH dose, the ewes received either a new MPA sponge (n = 10) or a controlled device for internal release impregnated with P4 (n = 10) or did not receive any device (n = 10). Ovarian dynamics were assessed every 12 h by transrectal ultrasonography from D-3 to D2. Oocytes were recovered by laparoscopic ovum pick-up (LOPU) on D2 and graded by morphologic quality. The number of small, medium, and large follicles at D0 and D2 (ultrasound examinations), number of both follicles aspirated and oocytes recovered at LOPU, recovery rate, and oocyte grade did not differ (P > 0.05) among treatments. Thus, the short-term use of MPA or P4 during ovarian stimulation did not affect the first-wave follicle population or morphologic quality of oocytes. We would suggest that, in this protocol, the use of exogenous progestin is unnecessary.

Influência da dificuldade de inseminação, temperamento e cortisol plasmático sobre a taxa de concepção de vacas e novilhas da raça Nelore inseminadas em tempo fixo / Influence of difficulty of insemination, temperament and plasma cortisol on conception rate of Nelore cows and heifers in timed AI

O objetivo do presente experimento foi avaliar efeito do estresse e da dificuldade de inseminação (DifIA) sobre a taxa de concepção (TC) de vacas (n=93) e novilhas (n= 72) Nelore submetidas à IATF. No D9, anotou-se nota de temperamento (NTe) e tempo da saída do brete (TSB) de todos os animais e coletou-se sangue das novilhas. No dia da IATF (D11), anotou-se NTe, TSB, DifIA e tempo de IA. A TC foi 36% para vacas e 46% para novilhas (P>0,05). Não foi observado efeito de Nte sobre TC (P>0,05). Porém, houve tendência para maior (P<0,10) TC nos animais que não apresentaram dificuldade de inseminação (DifIA1; TP=42%) em comparação aos animais com moderada ou alta dificuldade (DifIA2+DifIA3; TP=27%). Foi observado menor (P<0,05) tempo de IA para animais DifIA1 (17:31±06:02s) que animais DifIA2-3 (30:10±15:45s). Novilhas com maiores (P<0,05) níveis de cortisol apresentaram maior NTe (P<0,05). Entretanto, TC (59%) das novilhas menos reativas (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) não diferiu da TC (41%; P>0,05) das mais agitadas (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). Concluiu-se que avaliações de temperamento se relacionaram com nível de estresse, embora esses parâmetros não tenham afetado a TC deste trabalho. A maior dificuldade e/ou tempo necessário para se completar a IA demonstrou ser um potencial fator para a redução da fertilidade na IATF.(AU)

The objective was to evaluate the influence of stress and difficulty of insemination (DifIA) on conception rate (CR) of Nellore cows (n= 93) and heifers (n= 72) in Timed-AI. On D9, temperament (NTe) and time for chute exit (TSB) were recorded for all animals, and blood samples were colected from heifers. On the day of Timed-AI (D11), NTe, TSB, DifIA and time for AI were recorded. For cows, CR was 36% and for heifers 46% (P> 0.05). No effect (P> 0.05) of NTe was observed on CR. However, a tendency (P< 0,10) for higher CR was observed in animals with no difficulty for insemination (DifIA1; CR=42%) compared to animals that presented moderate or high difficulty (DifIA2+DifIA3, CR=27%). Time required for AI was lower (P< 0.05) in animals DifIA1 (17:31±06:02sec) than in animals DifIA2-3 (30:10±15:45sec). Heifers with greater (P< 0.05) cortisol levels presented higher Nte (P< 0.05). However, CR (59%) of less reactive heifers (cortisol=4,12±1,12ng/mL; NTe=3,2±0,6) did not differ from CR (41%; P> 0.05) of stressed animals (cortisol=7,76±1,33ng/mL; NTe=3,82±0,79). It was concluded that assessments of temperament were related to stress level, although these parameters did not affect the CR of this study. However, the higher difficulty and/or time to complete AI showed to be a potential factor for reducing fertility after timed-AI.(AU)

Mini-percoll gradient may be used for frozen-thawed sperm selection in sheep / Gradiente de mini-Percoll pode ser utilizado na seleção espermática de sêmen congelado de ovinos

This study evaluated the effect of increasing centrifugal force and reducing centrifugation time and volume in Percoll protocols on ram sperm parameters. Commercial semen of Santa Inês rams were used and five treatments were performed: traditional Percoll and mini-Percoll (MP) techniques (I- 5000 x g, 5min; II- 2500 x g, 5min; III- 1250 x g, 5min; IV- 700 x g, 10min). At post-thawing (PT) and post-selection protocols (0h), samples were assessed for spermatozoa recovery rate, motility, plasma membrane (PM) integrity, sperm capacitation and morphology and incubated at 37 C for 1, 2 and 3h. The sperm recovery rate averaged 9.1±1.4%, and most motility parameters were similar (P> 0.05) among protocols. VCL (µm/s) was higher (P< 0.05) after MP-II, III and IV (66.1±4.5) than traditional Percoll (46.3±4.9). Capacitation status and PM integrity were similar (P> 0.05) among treatments. For the first time, we have demonstrated the reduction of the gradient volume and centrifugation time associated with an increase on centrifugation force at Percoll can be successfully used for frozen-thawed ram sperm selection. MP may be used instead of traditional Percoll, decreasing costs and semen handling time.(AU)

O presente estudo avaliou o efeito do aumento da força de centrifugação, bem como da redução do tempo de centrifugação e do volume do gradiente de Percoll em diferentes protocolos nos parâmetros espermáticos de ovinos. Foi utilizado sêmen comercial de carneiros da raça Santa Inês, e cinco tratamentos foram realizados: Percoll tradicional e quatro técnicas de mini-Percoll (I- 5000 x g, 5min; II- 2500 x g, 5min; III- 1250 x g, 5min; IV- 700 x g, 10min). Após o descongelamento e a seleção espermática em cada técnica utilizada (0h), amostras foram avaliadas quanto à taxa de recuperação espermática, motilidade, integridade de membrana plasmática, capacitação e morfologia. Ao final, foram incubadas a 37 ºC por uma, duas e três horas. A taxa de recuperação média (9,1±1,4%) e a maioria dos parâmetros de motilidade foram similares (P>0,05) entre os tratamentos. VCL foi maior (P<0,05) após MP-II, III e IV (66,1±4,5) quando comparados ao Percoll tradicional (46,3±4,9). O status da capacitação e a integridade de membrana foram similares (P>0,05) entre os tratamentos. Pela primeira vez, foi demonstrado que a redução do volume do gradiente utilizado e do tempo de centrifugação, associada com o aumento da força de centrifugação nos protocolos de Percoll, pode ser usada com sucesso na seleção espermática de sêmen congelado de ovinos. O mini-Percoll pode ser utilizado em alternativa à técnica de Percoll tradicional, diminuindo custos e tempo de manipulação do sêmen durante a técnica.(AU)

Use of human intravaginal tampon embedded with natural progesterone induces synchronous estrus in Santa Inês ewes / Uso de tampão intravaginal humano embebido com progesterona humana para indução e sincronização de estro em ovelhas Santa Inês

Este estudo avaliou a eficiência de três protocolos de indução de estro síncronizado em ovelhas da raça Santa Inês. Vinte e quatro ovelhas adultas foram equitativamente distribuídas em três grupos, de acordo com ordem de parto, peso corporal (kg) e escore da condição corporal. As ovelhas receberam implante vaginal de progesterona natural por seis dias mais 37,5µg de d-cloprostenol laterovulvar e 300UI de eCG i.m., 24 horas antes da remoção do dispositivo. Ovelhas controle receberam CIDR330mg de progesterona, e as demais receberam dispositivo absorvente intravaginal humano, tamanho mini, embebido com 200 (OB200) ou 400mg (OB400) de progesterona. Coletas de sangue foram feitas nos momentos D0 (antes da inserção dos dispositivos), D0+6h e diariamente, até um dia após retirada do dispositivo (D7). A progesterona (ng/mL) foi semelhante (P>0,05) em todos os tratamentos ao longo do período experimental, exceto no dia da remoção do dispositivo, quando as ovelhas controle (2,5±0,3) tiveram progesterona superior (P<0,05) às ovelhas OB200 (0,6±0,1) e OB400 (1,2±0,4). O percentual de animais em estro (100,0, 62,5 e 100,0%) foi semelhante (P>0,05) e o intervalo para o estro (46,3±3,9a, 26,4±4,5b e 31,2±5,8a,b) foi diferente (P<0,05) entre os animais dos grupos controle, OB200 e OB400, respectivamente. A taxa de concepção das ovelhas foi de 50,0% (4/8) para cada grupo (P>0,05). Dispositivos humanos e fonte de progesterona podem ser usados para induzir o estro sincronizado em ovelhas Santa Inês.(AU)

Environmental effects on collared peccaries (Pecari tajacu) serum testosterone, testicular morphology, and semen quality in the Caatinga biome.

The objective of the study was to understand the influence of climatic variations in a semiarid environment on serum testosterone, testicular morphology and semen quality in collared peccaries (Pecari tajacu). Reproductive metrics (semen quality, testicular morphometry and testosterone serum profiles) of 10 mature males were measured monthly for 18 months. Meteorological data (rainfall, air temperature, relative humidity, wind speed and radiant heat load) also were recorded during the same period. Rainfall regimes were classified in different classes (Class 1: months with no rain; Class 2: months with up to 50 mm of rain; and Class 3: months with >50 mm of rain). Among rainfall classes, average air temperature (°C) and relative humidity (%) were different. Climatic changes between rainfall classes did not lead to overall variations of testicular size, testosterone production, and semen metrics. However, relative humidity recorded before semen collection (one day, one week, or over 51-55 days) was positively correlated (P < 0.05) with semen motility metrics (total motility, beat cross frequency and straightness) and sperm subpopulations (medium and static sperm), as well as with volume. Negative correlations (P < 0.05) were revealed between air temperature and the same semen motility patterns and volume. Additionally, radiant head load measured on the day of semen collection negatively influenced (P < 0.05) sperm straightness. This study demonstrates for the first time that no seasonal changes could be detected overt the 18-month period on the serum testosterone, testicular morphology and semen quality of collared peccaries raised in the Caatinga biome; however, it is expected that long term environmental changes will influence the reproductive physiology of species leaving in that habitat.

Hydrometra in dairy goats: Ultrasonic variables and therapeutic protocols evaluated during the reproductive season.

Hydrometra is characterized by the accumulation of fluid within the uterus due to the persistence of corpus luteum. The diagnosis of this disorder occurs with an ultrasonic exam. This study evaluated uterine drainage and fertility rates in goats after the use of d-cloprostenol in association or not with Gonadotropin-releasing hormone (GnRH) treatment. Twenty Saanen goats, diagnosed with hydrometra, received three 37.5-µg doses of d-cloprostenol laterovulvarly at 10-day intervals. On D5, the goats were assigned into two groups receiving 1 mL of GnRH or saline solution intramuscularly. Ultrasonography (US) was performed from D0 to D25. An US approach was used to rank hydrometra in scores. The pregnancy rate was assessed 45 and 90 days after the end of treatment. The uterine fluid was totally drained after the first and second administration of d-cloprostenol in 50% and 95% of the goats, respectively. In one female, full emptying of the uterus occurred only after D20. US performed at 45 and 90 days after the end of treatment indicated there was a pregnancy rate of 45.0% and 55.0%, respectively. Fertility did not differ between the GnRH-treated and control goats. Those goats not pregnant at 45 days had a follicular cyst, hydrosalpinx or hydrometra. At 90 days, no change was observed in the hydrosalpinx, and four goats had hydrometra. The use of three doses of d-cloprostenol 10 days apart was efficient for induction of draining the contents of the uterus, resulting in a relatively acceptable pregnancy rate. This treatment associated with the US approach can be important when applied in the field.

Leptospirosis is strongly associated to estrus repetition on cattle.

Although prevalent, the exact impact of infectious diseases on reproductive failures remains to be determined. Among them, leptospirosis has commonly been reported as cause of abortions on outbreaks. Nevertheless, the majority of the animals present a chronic, silent form of the disease, which is characterized by low reproductive efficiency and is frequently neglected. In that context, we conducted a study that aims to estimate the impact of chronic leptospirosis on reproductive disorders on cattle. A total of 25 different dairy herds with history of reproductive losses from Rio de Janeiro, Brazil, were selected. From each herd, a questionnaire was applied and sera from 20 cows were randomly tested for leptospirosis (totaling 500 cows). Chi-square was performed to estimate the association of seroreactivity with reproductive disorders. A total of 32% of the herds were positive, all of them against serogroup Sejroe. Estrus repetition was the most important reported reproductive problem and it was strongly associated to seroreactivity against leptospirosis. Besides, specific vaccination against leptospirosis was an important protection factor against that disorder. In conclusion, control programs including, but not limited to, vaccines must be implemented on those herds in order to reduce reproductive losses, particularly estrus repetition.