Sperm subpopulations influence the pregnancy rates in cattle

This study aimed to study the characteristics and subpopulations of spermatozoa from bulls with low and high reproductive performance based on pregnancy rates. Based on historical records of pregnancy rate from four farms, 24 bulls were selected. Two groups were established, with low pregnancy rates (n = 12; LOW), including bulls that presented pregnancy rates <52.27% (33.33% to 51.81%); and a group with high pregnancy rates (n = 12; HIGH), with pregnancy rates >52.27% (52.27% to 69.64%), after fixed-time artificial insemination (FTAI). The thawed sperm straws were analysed to sperm kinetics, morphology, plasma membrane integrity and sperm subpopulations. The LOW group exhibited a higher proportion of static cells (p < .05). In contrast, the HIGH group showed greater percentages for membrane integrity and total and progressive motility, and cells with fast and medium velocity (p < .05). In the cluster procedures, four sperm subpopulations were established. The low-fertility bulls presented the highest percentage of subpopulation 2 (41.46%), characterized by slow and progressive spermatozoa. The high-fertility bulls exhibited the highest percentage of subpopulation 3 (37.17%), characterized by fast and nonlinear spermatozoa. Results from this study indicated that bulls with greater percentages of fast and nonlinear spermatozoa seem to have greater fertilization capacity and the subpopulations analysis can be considered a tool to identify ejaculates with high fertility.     

Effects of Azoxystrobin on Mycotoxin Production in a Carbendazim-Resistant Strain ofFusarium sporotrichioides

Carbendazim-resistant (RS) and control (CS) strains ofFusarium sporotrichioides Sherb., previously developed in our laboratory, were exposed to graded concentrations of azoxystrob in in broth media under shake-culture conditions for 2, 3, 4 and 8 days. Azoxystrobin concentrations were 0, 1, 10 and 100 mg 1^-1 broth and cultures were incubated at a constant 25°C. Mycelial growth was significantly affected by strain (P<0.01), azoxystrobin concentration (P<0.001) and incubation time (P<0.001). Combined results for the four incubation times showed that CS yielded higher mycelial mass than RS (P<0.01) only in the absence of azoxystrobin. At fungicide additions of 1, 10 and 100 mg P^-1 mycelial growth was reduced (P<0.001) with minimal strain differences (P>0.05) at all three doses of azoxystrobin. Significant (P<0.05 or better) strain-fungicide interactions were recorded in trichothecene production following exposure to azoxystrobin. At 4 and 8 days of incubation, the 10 mg 1^-1 addition of azoxystrobin stimulated T-2 toxin synthesis (P<0.05) only in RS cultures. In contrast, T-2 toxin enhancement in CS cultures occurred only on day 8 but at a lower level of azoxystrobin (1 mg1^-1). Thus, the stimulation of T-2 toxin synthesis depended upon strain and azoxystrobin level. Production of diocetoxyscirpenol (DAS) was affected by a more complex set of interactions. Overall means showed that, in comparison with initial values (on day 2 or 3), DAS output maximized significantly(P<0.05) on day 4 in RS cultures and on day 8 in CS. Marked strain effects were observed on exposure to the 10 mg 1^-1 level of azoxystrobin. At this level, DAS production was enhanced in RS only after 4 (P<0.01 ) and 8 (P<0.05) days of incubation, while in contrast, CS reduced DAS production. As with T-2 toxin, DAS production in CS was stimulated (P<0.05 or better) only at low exposure levels of azoxystrobin. In the case of neosolaniol (NEO), however, the main effect of strain was significant (P<0.05), with CS producing consistently more of the mycotoxin than RS on day 4 of the experiment. At this point, the NEO:T-2 toxin ratio was also higher in CS (0.63) than in RS (0.12), a feature reported by us previously. In conclusion, the present investigation has shown for the first time that the development of resistance to one fungicide can affect trichothecene production inF. sporotrichioides on exposure to a second fungicide. These results have been incorporated into a new classification scheme for fungicide efficacy which is also presented in this paper. http://www.phytoparasitica.org posting Oct. 7,2001.     

Some observations on the morphology of the pigeon's seminiferous epithelium cells

A light microscopic examination of the seminiferous epithelium of the pigeon was carried out. Comparisons are made between the spermiogenic stages of the pigeon and that of the domestic fowl and mammals.     

γ-Amino Butyric Acid Control of Arginine Vasopressin Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol

The present study aims to ascertain the influence of gamma-amino butyric acid (GABA)(A or B) receptors on arginine vasopressin (AVP) release in vitro and determine whether E(2) modulates GABA-AVP interaction. Within 10 min of ewe killing, saggital midline hypothalamic slices (from the anterior preoptic area to the mediobasal hypothalamus along with the median eminence, 2-mm thick, two per ewe) were dissected, placed in oxygenated minimum essential media (MEM)-alpha at 4 degrees C and within 2 h were singly perifused at 37 degrees C with oxygenated MEM-alpha (pH 7.4; flow rate 0.15 ml/min), either with or without E(2) (24 pg/ml). After 4-h equilibration, 10-min fractions were collected for 4 h interposed with a 10-min exposure at 60 min to a specific GABA(A or B) receptor agonist or antagonist at various doses (0.1-10 mm). GABA(A) (muscimol; no E(2), n = 7 perifusion chambers, with E(2), n = 11) or GABA(B) (baclofen; no E(2), n = 8, with E(2), n = 15) agonists (10 mm) did not influence AVP concentrations. However, AVP release increased (p < 0.05) 20-30 min after exposure to 10 mm GABA(A or B) antagonists (bicuculline, no E(2), n = 7: from 4.6 +/- 0.7 to 33.0 +/- 0.4, with E(2), n = 17: from 11.9 +/- 1.4 to 32.8 +/- 6.0; CGP52432, with E(2), n = 14: from 14.0 +/- 2.6 to 28.8 +/- 3.9 pg/ml). At the end of the collection period, hypothalamic slices responded to KCl (100 mm) with AVP efflux (p < 0.05). GABA(B) but not GABA(A) antagonist-stimulated AVP release was enhanced in the presence of E(2). In summary, AVP release is under the inhibitory influence of GABA input with further potentiation by E(2) through GABA(B) receptors in vitro.     

Immunolocalization of Aquaporins 1 and 9 in the Ram Efferent Ducts and Epididymis

Aquaporins (AQPs) are essential membrane protein channels for the transport of water across membranes. Fluid movement in the epididymis is important for modulation of the luminal environment, in which sperm mature and reside. This study was designed to understand the morphology and localization of AQPs in ram efferent ducts (ED) and epididymis. For this purpose, the epididymis of seven animals were removed for histologic and immunohistochemical analyses. AQP1 immunoreactivity was observed in the apex of the ED, and AQP9 was found adjacent to the nuclei of the epithelial cells of the ED. The epithelial lining of ram epididymis is pseudostratified columnar and presents principal, basal, apical and narrow cells. In the initial segment (IS), a moderate reaction for AQP1 was observed in the apical cytoplasm of epithelial cells. An intense reactivity for AQP1 was noted over the microvilli of principal cells and in spermatozoa in the caput. In the corpus and cauda, AQP1 was noted only over the endothelial cells of vascular channels located in intertubular spaces. A weak‐to‐moderate reaction for AQP9 was observed in the nuclei of epithelial cells in the IS, caput and corpus of the epididymis. In the cauda, an intense reaction to AQP9 was observed in the epithelial border. In the IS, caput and corpus, the reactivity for AQP9 differed from those observed in domestic animals. The cauda showed a pattern similar to that previously described. These results indicate that AQPs 1 and 9 have reversed locations and roles in rams, suggesting activity variations related with fluid and solute absorption throughout the epididymis.     

Detection of retinoid receptors in non-neoplastic canine lymph nodes and in lymphoma

This study evaluated the difference in retinoid receptor expression between non-neoplastic lymph nodes and nodal lymphoma in dogs. Retinoid receptor expression was evaluated by immunohistochemistry in 32 canine lymph nodes. The lymph nodes had been previously diagnosed as non-neoplastic (6 normal and 7 hyperplastic lymph nodes) and B- and T-cell lymphoma (19 cases). Immunohistochemistry for retinoic acid receptors and retinoid-X receptors (and their subtypes α, β, and γ) was performed in all cases. In addition, immunohistochemistry for CD3 and CD79a was performed in all lymphoma cases. Non-neoplastic lymphocytes were negative for all retinoid receptors. Retinoic acid receptor-γ was detected in 100% of B-cell lymphoma and 78% of T-cell lymphoma, while retinoid X receptor-γ was positive in 78% of T-cell lymphoma cases. When normal lymph node architecture was still present, a contrast between retinoid-negative benign cells and retinoid-positive malignant cells was clear. Retinoid receptors were expressed in neoplastic, but not in benign lymphocytes, suggesting their value for both diagnosis and treatment of canine lymphoma.     

Kisspeptin,c‐Fos and CRFR Type 2 Co‐expression in the Hypothalamus after Insulin‐Induced Hypoglycaemia

Normal reproductive function is dependent upon availability of glucose and insulin‐induced hypoglycaemia is a metabolic stressor known to disrupt the ovine oestrous cycle. We have recently shown that IIH has the ability to delay the LH surge of intact ewes. In the present study, we examined brain tissue to determine: (i) which hypothalamic regions are activated with respect to IIH and (ii) the effect of IIH on kisspeptin cell activation and CRFR type 2 immunoreactivity, all of which may be involved in disruptive mechanisms. Follicular phases were synchronized with progesterone vaginal pessaries and at 28 h after progesterone withdrawal (PW), animals received saline (n = 6) or insulin (4 IU/kg; n = 5) and were subsequently killed at 31 h after PW (i.e., 3 h after insulin administration). Peripheral hormone concentrations were evaluated, and hypothalamic sections were immunostained for either kisspeptin and c‐Fos (a marker of neuronal activation) or CRFR type 2. Within 3 h of treatment, cortisol concentrations had increased whereas plasma oestradiol concentrations decreased in peripheral plasma (p < 0.05 for both). In the arcuate nucleus (ARC), insulin‐treated ewes had an increased expression of c‐Fos. Furthermore, the percentage of kisspeptin cells co‐expressing c‐Fos increased in the ARC (from 11 to 51%; p < 0.05), but there was no change in the medial pre‐optic area (mPOA; 14 vs 19%). CRFR type 2 expression in the lower part of the ARC and the median eminence was not altered by insulin treatment. Thus, disruption of the LH surge after IIH in the follicular phase is not associated with decreased kisspeptin cell activation or an increase in CRFR type 2 in the ARC but may involve other cell types located in the ARC nucleus which are activated in response to IIH.     

Growth and Nutritional Disorders of Eggplant Cultivated in Nutrients Solutions with Suppressed Macronutrients

The objective was to evaluate the effects of omitting macronutrients in the nutrients solution on growth characteristics and nutritional status of eggplants. The treatments were complete nutrients solution and solutions with nutrient omission: nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sulfur (S). The experiment was carried out under greenhouse conditions with three replicates in a completely random design. Plant height, number of leaves per plant, leaf area, relative chlorophyll index, photosynthesis rate, stomatal conductance, dry matter, concentration levels of macronutrients in plant aerial part and root system, and nutritional disorders were evaluated. Omitting elements interfered in the concentration of elements in the various plant tissues and this had as consequences limited vegetative growth, reduced dry matter and led to the development of the typical deficiency symptoms of each element. Although potassium was the most demanded of all elements, nitrogen and calcium were the most growth limiting ones.     

DRIS norms and limiting nutrients in banana cultivation in the South of Ecuador

Abstract

Foliar analysis is an effective method to diagnose the nutritional status of plants. However, the mineral concentration in foliar tissue has traditionally been evaluated by assessing the activity of each element, without considering the interactions between them. To address this, dual interactions were calculated using the Diagnosis and Recommendation Integrated System (DRIS) to identify which crop nutrients are most influential in nutrient imbalances and which are the most limiting nutrients for the nutritional status of banana crops in Ecuador. To achieve this, a regional survey of the nutritional status and its productivity levels was conducted for 188 different sites during the crop season in 2017–2018, involving banana cultivars ‘Vallery’ and ‘Williams’, from the Cavendish subgroup. The DRIS calculation method was combined with Beaufils and Jones functions. From the initial 188 foliar samples, 83 samples (representing 44% of the population) were considered to represent the high-yield reference population, with yields of 38–60 t ha^?1. The DRIS method defined the mean nutritional balance index, which was not found to be statistically correlated (p?>?0.05) with productivity, revealing that there was no significant association with the nutritional status of the plants. Specific DRIS norms were obtained and indicated that deficiencies in K, N, Ca, and Fe, and excesses in Mn, B, Cl, Zn, S, Cu, and Mg were the most limiting nutrients for banana cultivars in the south of Ecuador.