Higher Fertilization Rate Research Articles

Influence of Sludge Incineration Ash on Ryegrass Growth and Soil Phosphorus Status

Phosphorus (P) is a limited resource that could be depleted. Consequently, recycling the P contained in sewage sludge, including sewage sludge incineration ash (SIA), from wastewater treatment plants is a possibility to be explored. A greenhouse experiment using annual ryegrass (Lolium multiflorum L.) was performed with an experimental design of three completely randomized blocks of two soils and 29 treatments: one control without P and two levels of 9 and 26 kg total P ha−1 from 14 different sources: twelve SIAs (not contaminated by trace metals) from the US and Canada, one commercial synthetic fertilizer (triple superphosphate (TSP)), and one commercial rock phosphate (RP). Higher ryegrass biomass levels were achieved at the higher fertilization rate (26 kg total P ha−1) and when using the SIAs with the highest P solubility percentage (PSP) (≥ 54% of total P). The biomass increases following SIA application were as high as 29% and 59% more than the control for the sandy loam and clayey soil, respectively, but 40% less than in TSP for both soils. A similar behavior was observed for P uptake, with a maximum increase of 26% for the clayey soil, and 165% for the sandy loam soil. The ryegrass biomass and P uptake increases due to SIA application were larger than those due to RP application in the clayey soil, but similar to those in the sandy loam soil. The SIAs with a PSP of ≥ 54% significantly increased soil available P stocks and saturation. According to our findings, we conclude that the SIAs from municipal and agrifood industries have a potential for P agricultural recycling, but their efficiencies vary.

Динаміка росту, розвитку та відтворної здатності телиць різного походження знам’янського типу поліської м’ясної породи

The article presents the results of the study of the growth and development and reproductive abilities of heifers of different linear origin of the Znamyansky type Poliska meat breed Experimental research was carried out in the Agrofirma Kolos Znamyansky district, Kirovograd region. The object of research was the heifers of the Znamyansky type of different lines: I - control - animals, which are not included in the experimental lines, II – Mazun 6; III – Radyst 113; IV – Darovanyy 400. As a result of the carried out research, we found out that heifers of control and experimental groups at identical conditions of feeding and maintenance, have shown various intensity of growth. Heifers of experimental groups by live weight indicators in all age periods exceeded peers of the control group with a true difference. Among the experimental groups there was an obvious advantage for the animals of the third experimental group. The same tendency is observed in the indicators of the daily average and relative increments. High daily average increment of the animals of the third experimental group was 828,8 g per day during the period of 12-15 months. Comparing the body measurements of heifers of different experimental groups, we can note the advantage of animals of the III experimental group in all altitude and latitude parameters. The other three groups had lower and rather similar indicators. In general, the animals are quite large, harmoniously built, with a strong, but not massive skeleton, which is confirmed by the indicators of animal physique. By physique indexes the highest marks were given to heifers of III and IV experimental groups. They had higher indexes of physique such as breast, index of blockiness, which characterizes their better meat forms. The growth energy of animals in meat cattle breeding is an indicator that has a direct impact on reproduction ability, because a fully developed animal is able to reach puberty quite early and give a healthy offspring every year. By the indexes of reproductive ability of heifers it was found out that there are certain differences between heifers of different origins. Animals of different genotypes had quite different indicators of live weight during the first hunting. The animals of the third experimental group had the highest weight 333,9 kg, the animals of the first control group had the lowest weight 230,4 kg. Heifers of the third experimental group were also characterized by a high rate of fertilization from the first mating in comparison with other groups with a true difference. By the indicators of duration of pregnancy, there is no significant difference between the genotypes.

Early production of offspring by in vitro fertilization using first-wave spermatozoa from prepubertal male mice.

Mature male mice (aged 10–12 weeks or older) are conventionally used for in vitro fertilization (IVF) in order to achieve high fertilization rates (e.g., > 70%). Here, we sought to determine the earliest age at which male mice (C57BL/6J strain) can be used efficiently for producing offspring via IVF. Because we noted that the addition of reduced glutathione (GSH) to the IVF medium significantly increased the fertilizing ability of spermatozoa from prepubertal males, we used this IVF protocol for all experiments. Spermatozoa first reached the caudal region of the epididymides at day 35; however, they were unable to fertilize oocytes. Caudal epididymal spermatozoa first became competent for oocyte fertilization at day 37, albeit at a low rate (2.9%). A high fertilization rate (72.0%) was obtained at day 40, and 52.4% of the embryos thus obtained developed into offspring after embryo transfer. Moreover, we found that corpus epididymal spermatozoa in prepubertal mice could fertilize oocytes; however, the fertilization rates were always < 50%, regardless of the age of the males. Caput epididymal spermatozoa failed to fertilize oocytes irrespective of the age of the males. Therefore, we propose that caudal epididymal spermatozoa from male mice aged 40 days can be efficiently used for IVF, to obtain offspring in the shortest attainable time. This protocol will reduce the turnover time required for the generation of mice by ~1 month compared with that of the conventional IVF protocol.

The Outcome of Intracytoplasmic Sperm Injection with Fresh and Cryopreserved Ejaculated Sperm: A Comparative Study at the Yaounde Gynecological Endoscopic Surgery and Human Reproductive Teaching Hospital

Objective: To evaluate fertilization rate, embryo quality and pregnancy rate in patients undergoing Intracytoplasmic Sperm Injection (ICSI) using fresh and cryopreserved ejaculated sperm. Methods: We carried out an analytical cross-sectional study with retrospective data collection from December 1st, 2017 to May 1st, 2018. We compared two groups of patients undergoing ICSI using ejaculated sperm: group I comprised of those in whom fresh semen was used and group II was comprised of those in whom cryopreserved semen was used. We collected the fertilization rate, embryo quality, and pregnancy rate. The Odds Ratio was calculated to determine the association between the variables. A p-value of <0.05 was considered significant. A logistic regression analysis was done. Results: when OATS sperm was used, we had higher fertilization rate 67, 5% vs. 46, 9%, p=0,042 and better embryo quality on day 3, 95% vs. 5%, p=0.008 with freshly ejaculated sperm against the cryopreserved semen. After logistic regression, the fertilization rate, embryo quality and pregnancy rate in the fresh and cryopreserved ejaculated semen were similar. Conclusion: The semen with OATS may be more susceptible to damages caused by cryopreservation, resulting in lower fertilization rate and embryo quality but similar pregnancy rates.

Development and application of a twin open-top chambers method to measure soil HONO emission in the North China Plain

HONO (nitrous acid) is a crucial precursor for tropospheric OH radicals, and its sources are not well understood. In the past decade, soil was proven to be a potential source for HONO. However, more field measurements of soil HONO emission flux are needed to explore the mechanism and its impact on regional air quality. Here, we developed a system based on twin open-top chambers (OTCs) and wet chemical methods to measure HONO emission flux from agricultural soil in the North China Plain (NCP). The performance of the OTC system was tested under laboratory and field measurement conditions. The results showed that the system could reflect the strength (>90%) and variation of gas emission with an average residence time of 4–5 min. The greenhouse effect and chemical reaction interference in the chamber was proven to have no significant influence on the HONO flux measurement. Field measurement revealed that agricultural soil before fertilization was an important source of HONO. The emission flux showed radiation-dependent or temperature-dependent variation, with a peak of 3.21 ng m−2 s−1 at noontime that could account for approximately 67 pptv h−1 of the missing HONO source under an assumed mixing layer height of 300 m. Fertilization substantially accelerated HONO emission, which was rationally attributed to biological processes including nitrification. Considering the high fertilization rate in the NCP and other similar regions in China, HONO emission from agricultural soil likely has enormous impact on regional photochemistry and air quality, suggesting that more research should be conducted on this aspect.

Revisiting the impact of ejaculatory abstinence on semen quality and intracytoplasmic sperm injection outcomes.

Regulatory bodies recommend inconsistent ejaculatory abstinence lengths before semen analysis. The literature exploring the effect of ejaculatory abstinence length on the outcomes of intracytoplasmic sperm injection is scarce. To study the influence of ejaculatory abstinence length on semen quality and intracytoplasmic sperm injection outcomes. This prospective cohort study included 818 patients undergoing conventional semen analysis from October 2015 to October 2016, in a private university-affiliated IVF centre. Generalized linear models adjusted for potential confounders were used to investigate the associations between ejaculatory abstinence length and seminal parameters and intracytoplasmic sperm injection outcomes. Increasing ejaculatory abstinence length was positively correlated with semen volume, sperm concentration, total sperm count, total motile sperm count and sperm DNA fragmentation index. Significant inverse correlations were observed between ejaculatory abstinence length and fertilization rate, blastocyst formation rate, implantation rate and pregnancy rate. A discriminant analysis showed a mean ejaculatory abstinence length in the positive pregnancy group of 3.14±1.64days and 4.83±3.66days in the negative pregnancy group. A cut-off point was established halfway between ejaculatory abstinence length averages, at 4days. The ejaculatory abstinence ≤4days group showed significant lower semen volume, sperm concentration, total sperm count and total motile sperm count compared to ejaculatory abstinence>4days group. The ejaculatory abstinence≤4days group showed significant lower sperm DNA fragmentation index, and higher rates of fertilization, high-quality embryos on day 3, blastocyst development, implantation and pregnancy compared to ejaculatory abstinence>4days group. The implantation rate was significantly higher and the pregnancy rate tended to be higher with one day of ejaculatory abstinence, compared to 2-4days of ejaculatory abstinence. Ejaculatory abstinence periods of >4days have a detrimental effect on sperm DNA and intracytoplasmic sperm injection outcomes. One day of ejaculatory abstinence significantly improves implantation rate and tends to increase pregnancy rate, compared to 2, 3 and 4days of ejaculatory abstinence.

The overlapping reproductive traits of the two male mating types of the oval squid Sepioteuthis lessoniana

The oval squid Sepioteuthis lessoniana is polyandrous, and a female mates with several males during a spawning season. This has resulted in two alternative male mating tactics in this species. The larger consort males usually use the male-parallel (MP) mating posture to put spermatophores at the opening of the oviduct; the smaller sneaker males predominantly use the male-upturned (MU) mating posture to place spermatophores around the female’s buccal membrane. Since squid eggs are released from the oviduct, MP mating is expected to have a higher fertilization rate than MU mating. Given the fact that these two male mating tactics are largely dependent on the body size of the male relative to the female, it is unclear how the smaller males are able to reproductively compete with the larger males. On analyzing spermatophore and sperm morphology from populations of adult male squids, spermatophore size was found to be positively correlated with mantle length of these squids, but sperm length was negatively correlated with squid mantle length. More importantly, no distinct size dimorphism exists in relation to either spermatophore morphology or sperm morphology. The overlap in size distributions of the spermatophores and sperms of the male squid thus implies that medium-sized males may have flexible reproductive strategies, switching between MP and MU mating tactics depending on the context of the mating event and/or female choice. This presumed behavioral plasticity might increase the reproductive success of these males and maintain the dynamic stability of two alternative mating types in this polyandrous species.

32 Vitrification of in vitro-matured bovine oocytes in triacetate cellulose hollow fibres

A prospective method of vitrification in triacetate cellulose hollow fibres (HF) introduced by Matsunari et al. (2012 J. Reprod. Dev. 58, 599-608) allowed significant simplification and standardization of vitrification/warming procedures and was successfully used for group cryopreservation of various pre-implantation mammalian embryos. The goal of the current work was to evaluate the effectiveness of the HF vitrification method for cryopreservation of in vitro-matured bovine oocytes. The base medium for all the vitrification and rewarming solutions was calcium-free TBP-like protein-HEPES supplied with 20% of fetal bovine serum. Groups of 15 morphologically normal in vitro-matured bovine oocytes were equilibrated with 3% (vol/vol) ethylene glycol for 15min, loaded into HF, and transferred into vitrification solution containing 30% ethylene glycol and either 0.5M (Group 1) or 1.0M (Group 2) sucrose. Hollow fibres were incubated for either 60s (Group 1) or 30s (Group 2) and immediately plunged into LN. Rewarming was conducted at 39°C. Oocytes within HF were placed in decreasing concentrations of sucrose solutions to remove cryoprotectants. Then, oocytes were subjected to IVF. Non-vitrified denuded oocytes were used as a control. Survival rates were evaluated at 21h post-rewarming. Part of the presumptive zygotes were fixed and stained with acetolacmoid for fertilization rate. The remaining zygotes were cultured for 10 days. Developmental rates were evaluated at 44h and 7 and 10 days post-IVF. All results are presented as mean percentage±standard deviation. Data were analysed using Mann-Whitney U test. Significance was set at P&amp;lt;0.05. Survival rate was significantly lower in Group 1 (79.0±8.0%) and Group 2 (75.0±5.0%) compared with the control group (97.0±4.0%). Fertilization rate in Group 1 differed significantly from the control (80.5±18.3% v. 95.5±9.1%). Cleavage rates in Groups 1 and 2 did not differ significantly from the control (42.5±15.7% v. 60.7±11.1% v. 63.0±15.8%, respectively). Blastocyst yields at 7 days post-IVF were 0.9±2.3 (1/116) and 9.6±5.4% (6/65) in Groups 1 and 2, respectively. The former was significantly lower than in the control group (17.0±10.3%, 23/154). It should be noted that hatching in the control group started at 8 days post-IVF and was delayed in Groups 1 and 2 for at least 24h. Day 10 blastocyst yields were 3.0±3.3 (P&amp;lt;0.05), 20.9±13.8, and 30.4±9.6% in Groups 1 and 2 and the control group, respectively. All obtained Day 10 blastocysts (3/116) in Group 1 hatched. Hatching rate in Group 2 was significantly lower than in the control group. Both Groups 1 and 2 showed relatively high survival and fertilization rates, but embryo development rates in both groups had a tendency to be lower than in the control. However, the obtained results indicate that the modifications of the protocol may increase the effectiveness of HF vitrification. The HF vitrification method remains a prospective option for simultaneous cryopreservation of a group of bovine oocytes.

Acrosome reaction and chromatin integrity as additional parameters of semen analysis to predict fertilization and blastocyst rates

BackgroundTraditional semen parameters have shown little to none predictive value for fertilization and blastocyst viability for a successful pregnancy. Therefore, the purpose of this study was to explore the usefulness of incorporating the acrosome reaction (AR) and chromatin integrity to conventional semen analysis to individually predict the fertile potential of sperm samples.MethodsA cross-sectional study was conducted in 69 participants undergoing IVF using oocyte donation. Semen samples were collected and evaluated for: AR [spontaneous (sAR) and induced (iAR)] by flow cytometry using anti-CD46-FITC, Acrosome Response to an Ionophore Challenge (ARIC), chromatin integrity by Sperm Chromatin Structure Assay (DNA Fragmentation Index-%DFI and High DNA Stainability-%HDS), WHO semen analysis, fertilization and blastocyst rates.ResultsThe participant age was 40.0 ± 6.1 years (66% were normozoospermic). Sperm morphology, sAR, iAR, and ARIC were associated with the fertilization (β = 3.56, R2 = 0.054; β = − 5.92, R2 = 0.276; β = 1.83, R2 = 0.150; and β = 2.10, R2 = 0.270, respectively, p < 0.05). A logit model was developed to calculate the probability of fertilization (≥ 60%) for each participant, using the sperm morphology and ARIC as independent variables, followed by ROC analysis to determine a cutoff probability of 0.65 (specificity = 80.6%, sensitivity = 63.2%). %DFI was inversely associated with the viable blastocyst rate (β = − 1.77, R2 = 0.057, p = 0.003), by the logit model and ROC analysis, a cutoff probability of 0.70 (specificity = 80.6%, sensitivity = 72.3%) was obtained to predict blastocyst viability (≥ 40%). There was no difference in the results with normozoospermic samples (n = 46).ConclusionsThe incorporation of ARIC and %DFI allowed to obtain predictive models for high fertilization and blastocyst rates in an individualized way, being promising tools to improve the diagnosis of male fertility potential for research or assisted reproduction, even in men with unknown infertility.

Инвагинация оолеммы в нативных и криоконсервированных ооцитах человека при их оплодотворении in vitro методом ICSI

Development of novel prognostic criteria for evaluation in vitrofertilization of fresh and cryopreserved oocytes is an important component of modern assisted reproductive technologies. There was compared a degree of oolemma invagination during intracytoplasmic sperm injection into fresh and cryopreserved oocytes. The classification of the gamete turgor state based on oolemma degree invagination during intracytoplasmic sperm injection was developed. This behavior of oocyte membrane is a prognostic characteristic of zygotic formation rate. Oocytes with normal turgor were characterized by a high fertilization rate higher than 80%. Immediate oolemma puncture during oocyte micromanipulation showed structural and functional damage of membrane and led to further oocyte degeneration. After cryopreservation by a standard method the ratio of different types of oolemma invagination has been changed. A relative number of cells with normal turgor was decreased down to 54% due to an increase of oocyte number with higher membrane elasticity. The four-stage method of cryoprotective media removing after oocyte freeze-thawing is more effective for preserving gametes with normal turgor contrary to the standard two-stage one. Probl Cryobiol Cryomed 2018; 28(3): 258 – 265

Application of a low-cost camera on a UAV to estimate maize nitrogen-related variables

The development of small unmanned aerial vehicles and advances in sensor technology have made consumer digital cameras suitable for the remote sensing of vegetation. In this context, monitoring the in-field variability of maize (Zea mays L.), characterized by high nitrogen fertilization rates, with a low-cost color-infrared airborne system could be the basis for a site-specific nitrogen (N) fertilization support system. An experimental field with different N treatments applied to silage maize was monitored during the years 2014 and 2015. Images of the field and reference destructive measurements of above ground biomass, its N concentration and N uptake were taken at V6 and V9 development stages. Classical normalized difference vegetation indices (NDVI) and the indices adjusted by crop ground cover were calculated and regressed against the measured variables. Finally, image colorgrams were used to explore the potential of band-related information in variable estimation. A colorgram is a linear signal that summarizes the color content of each digital image. It is composed of a sequence of the frequency distribution curves of the camera bands, of their related parameters and of results of the principal components analysis applied to each image. The best predictors were found to be the ground cover and the adjusted green-based NDVI: regression equation at V9 resulted in R2 of 0.7 and RRMSE < 25% in external validation. Colorgrams did not improve prediction performance due to the spectral limitations of the camera. Therefore, the feasibility of the method should be tested in future research. In spite of limitations of sensor setup, the modified camera was able to estimate maize biomass due to the very high spatial resolution. Since the above ground biomass is a robust proxy of N status, the modified camera could be a promising tool for a low-cost N fertilization support system.

Effect of glutathione on sperm quality after short-term cryopreservation in seurukan fish Osteochilus vittatus (Cyprinidae)

The objective of this present study is to determine the optimum concentration for glutathione in the cryopreservation of seurukan fish (Osteochilus vittatus) spermatozoa. A completely randomized design (CRD) was used with 6 treatments and 3 replications. The Seurukan fish sperm was diluted in extender with a ratio of 1:20 (sperm: extender), then glutathione was added at a concentration of 0, 10, 20, 30, 40 and 50 mg L−1. Next, the sperm thawed at 39–40 °C for 3 min and mixed with 100 eggs which were randomly selected. The success of the fertilized egg was observed 6 h after fertilization, while the hatching rate was examined 60 h after fertilization. The ANOVA test showed that the addition of glutathione significant affected the sperms motility, fertility and hatching rate of seurukan fish Osteochilus vittatus eggs (P < 0.05). By segregation the fresh sperm, a higher sperm motility rate was recorded with an addition of 30 mgL−1 of glutathione (63.00 ± 5.89), however, this value was not significantly different from using concentration of 10 and 20 mg L−1. A higher fertilization rate was produced using glutathione concentration of 50 mg L−1 (51.33 ± 17.01); however, it was also not significantly different from the concentration of 20, 30 and 40 mgL−1. In addition, a higher hatching rate was also recorded using glutathione concentration of 50 mg L−1 (40.33 ± 12.89), this value was not significantly different from using 40 mgL−1 glutathione. Hence, a conclusion was drawn that the optimum concentration of glutathione is 40 mg L−1 of diluent.

Does paternal age influence the fertilization rate of egg donation cycles with male factor?

Evaluate the impact of paternal age in egg donation cycles with male factor. Observational, analytical, retrospective, transversal. In vitro fertilization cycles from the egg donation program in a private fertility center from January 2012 through December 2016 were reviewed. The cycle outcomes were assessed in relation to semen samples to analyze the impact of male factor and paternal age. Cycles were divided into two groups: Group 1, those who had male factor (oligo, terato, asthenospermia of any degree) and Group 2, those who had normal seminal parameters. Male age, fertilization rate, embryo development, and pregnancy rate were compared between groups. From 472 samples, 213 (45.13%) had male factor (Group 1), and 259 (54.87%) had normal seminal parameters (group 2). Overall, male patients from group 2 were younger (mean 38.2 yr, ± 5.8, P = 0.002) and had a higher fertilization rate (68.22%, P = 0.005). In this group, fertilization rates didn’t fluctuate with age (r2 = 0.010). However, patients from group 1 were older (mean age 40.5 a, ± 4.85, P = 0.002) and paternal age was slightly correlated with fertilization rate (r2=-0.126) with lower fertilization rates (62.51%, P=0.005) and a low probability of pregnancy when normal forms were less than 6% (AUROC = 0.677, SEN 57%, SPE 76.7%). Male age mildly reduces fertilization rates in IVF cycles with male factor. In cycles without male factor, age did not correlate with fertilization rates or any seminal parameter.

Standardized cryopreservation protocol of European perch (Perca fluviatilis) semen allows to obtain high fertilization rates with the use of frozen/thawed semen

The general aim of this study was to develop an efficient and standardized cryopreservation procedure for European perch semen. The specific aims of this study were: to test the effects of (i) final glucose concentration, (ii) final sperm concentration in the extended semen and (iii) the storage time of post-thaw semen on sperm motility parameters. Moreover, the effects of cryopreservation of semen on fertilization rates were tested at sperm:egg ratios ranging between 50,000:1 and 500,000:1 (iv), as well as the fertilization of 25 g portions of eggs (v). The range of optimal sperm concentrations in straws with high post-thaw sperm motility (66–73%) was quite wide, from 1.0 to 6.0 × 109 spermatozoa ml−1. The final glucose concentration of 0.30–0.34 M in 7.5% methanol produced the highest results of sperm motility (76 ± 5%) after cryopreservation. Storage time of post-thaw semen negatively influenced sperm motility parameters. The fertilization rates while using cryopreserved semen were high (79%) and did not differ from fresh semen (85%). Similar results (73–77%) were obtained between sperm:egg ratio ranged from 50,000:1 to 500,000:1. Moreover, cryopreserved semen was successfully used for fertilization as much as 12,500 eggs (25 g). However, semen has to be used for fertilization within 2 min after thawing. In our opinion, standardizing the procedure of semen cryopreservation presented in this study is a prerequisite for the development of repeatable procedures and the future implementation of cryopreserved semen in hatchery practice. Further studies should focus on extension of time of post-thaw sperm motility, determination of optimal sperm:egg ratio at the conditions of large-scale fertilization and scaling up procedure to fertilize 50–150 g of eggs (one ribbon) at single application.

High-quality spontaneous spawning in greater amberjack (Seriola dumerili, Risso 1810) and its comparison with GnRHa implants or injections

Production of sufficient high-quality eggs of greater amberjack (Seriola dumerili) still constitutes the main bottleneck for commercial production of this species. The main objective of this study was to compare the quality of spontaneous spawn of greater amberjack with those obtained by either GnRHa injection or GnRHa implant protocols. Captive amberjack broodstock were distributed in three circular tanks of 40 m3. Broodstock from Tank 1 were not hormonally induced and spawned spontaneously, whereas those of Tank 2 were intramuscularly injected with GnRHa (20 µg/kg body weight) and those of Tank 3 were given EV-500 µg GnRHa implants. The number of eggs per spawn obtained in the broodstock without hormonal treatment was larger than in those obtained with injections or implants. Egg quality was best in broodstock with spontaneous spawn, followed by GnRHa-injected fish and then GnRHa implants. Besides, size of larvae from control and injected broodstock was similar between them and significantly higher (p < 0.01) than those from GnRHa implant spawn. Overall, this study showed that it is possible to obtain very high-quality spontaneous spawn in greater amberjack, providing the adequate conditions. Furthermore, GnRHa weekly injections lead to similar egg viability and hatching rates than spontaneous spawn and higher fertilization rates than GnRHa hormonal implants, which is better than in previous studies.

Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa

High quality of sperm is essential to a high fertilization rate, especially post- cryopreservation. Assessment of sperm integrity, motility and energy reserves before cryopreservation is necessary for selection of milt with optimal fertilizing potential. We describe the effect of cryopreservation on the quality of black-lip pearl oyster, Pinctada margaritifera var. cumingii sperm. Evaluated quality indices of fresh and frozen/thawed P. margaritifera spermatozoa, included morphology, ultrastructure and motility characteristics relative to the energy content (ATP) and its capacity to be sustained by mitochondrial respiration. Morphology and ultrastructure were quantitatively evaluated using images obtained by optical microscopy assisted by the Image J software and TEM, respectively. Sperm motility was assessed using Image J software combined with a computer assisted sperm analysis plugin adapted for assessing P. margaritifera spermatozoa. Other sperm quality parameters evaluated included O2 consumption, ATP content, and creatine kinase activity. Frozen/thawed spermatozoa exhibited damage to the head but retained a compact spherical shape. Sperm motility indicators showed a significant decrease in quality resulting from the freeze/thaw process. The percent of motile cells was 54% compared to 84% in fresh sperm, O2 consumption was 4.8 compared to 44 nanomol min−1, ATP content was 0.72 nmol/109 spermatozoa in the activating medium compared to 4.54 nmol/109 spermatozoa, and creatine kinase activity was 9.06 × 10−5 IU mg−1 protein compared to 12.5 × 10−5 IU mg−1 protein.The cryopreservation protocol allowed obtaining an acceptable motility rate after thawing, confirming the predictive value of sperm motility measurements before cryopreservation in terms of their ability to withstand freezing process.

Plant and Microbial Responses to Repeated Cu(OH)2 Nanopesticide Exposures Under Different Fertilization Levels in an Agro-Ecosystem.

The environmental fate and potential impacts of nanopesticides on agroecosystems under realistic agricultural conditions are poorly understood. As a result, the benefits and risks of these novel formulations compared to the conventional products are currently unclear. Here, we examined the effects of repeated realistic exposures of the Cu(OH)2 nanopesticide, Kocide 3000, on simulated agricultural pastureland in an outdoor mesocosm experiment over 1 year. The Kocide applications were performed alongside three different mineral fertilization levels (Ambient, Low, and High) to assess the environmental impacts of this nanopesticide under low-input or conventional farming scenarios. The effects of Kocide over time were monitored on forage biomass, plant mineral nutrient content, plant-associated non-target microorganisms (i.e., N-fixing bacteria or mycorrhizal fungi) and six soil microbial enzyme activities. We observed that three sequential Kocide applications had no negative effects on forage biomass, root mycorrhizal colonization or soil nitrogen fixation rates. In the Low and High fertilization treatments, we observed a significant increase in aboveground plant biomass after the second Kocide exposure (+14% and +27%, respectively). Soil microbial enzyme activities were significantly reduced in the short-term after the first exposure (day 15) in the Ambient (-28% to -82%) and Low fertilization (-25% to -47%) but not in the High fertilization treatment. However, 2 months later, enzyme activities were similar across treatments and were either unresponsive or responded positively to subsequent Kocide additions. There appeared to be some long-term effects of Kocide exposure, as 6 months after the last Kocide exposure (day 365), both beta-glucosidase (-57% in Ambient and -40% in High fertilization) and phosphatase activities (-47% in Ambient fertilization) were significantly reduced in the mesocosms exposed to the nanopesticide. These results suggest that when used in conventional farming with high fertilization rates, Kocide applications did not lead to marked adverse effects on forage biomass production and key plant–microorganism interactions over a growing season. However, in the context of low-input organic farming for which this nanopesticide is approved, Kocide applications may have some unintended detrimental effects on microbially mediated soil processes involved in carbon and phosphorus cycling.

Pre-breeding in Passiflora subrotunda Mast.: Morphological and Reproductive Characterization at Different Light Levels

Passiflora are ornamental plants that are appreciated as part of outdoor decor, composing pergolas and gardens, as well as in interior ornamentation where species tolerant to environments with less light availability are used. The objective of this study was to evaluate the influence of different levels of light and pot types on morphological and reproductive characteristics in Passiflora subrotunda and to support genetic breeding programs of ornamental passifloras. The conditions of 75% and 100% light favored vegetative morphological characteristics through the time (105 days). Floral characteristics also presented higher values along increasing light levels. All qualitative characters related to flower and plant coloration did not vary among genotypes. The species possesses diurnal anthesis and flowers throughout the year. Plants cultivated in concrete pots showed greater growth and flower production. Pollen grains (PGs) are large, with an isopolar form, a small polar area, and a long aperture, and amylaceous. Percentage of viable PG was high: above 97% using Alexander solution and reaching up to 91% with fluorescein diacetate. Stigmas were partially receptive during the flower’s opening period. Percentage of self-compatibility was lower, based on the higher fertilization rate through cross-pollination. These information will be used in planning of ornamental Passiflora-breeding programs, assisting in the selection of characteristics and breeding methods.

Is semen analysis without strict criteria misleading decisions in IVF? A prospective systematic study

Sperm morphology has been strongly linked to fertilization. This makes it an important component in semen analysis. They are usually assessed by world health organization (WHO) standard or Kruger strict criteria in in-vitro fertilization (IVF) centers all over the world. Sperm count, motility, and morphology together form the basis by which patients are allocated into IVF or intra-cytoplasmic sperm injection. This study aimed to compare fertilization rates in standard IVF from patients with normal sperm count and motility with and without morphological assessment by WHO guideline. In this prospective cohort study, sperm count, motility, and morphology of 504 men candidate for IVF program over a three years period in our center were evaluated in two groups: Group A (case group) included men with normal sperm count and motility but with a poor morphology and group B (control group) included men with normal sperm count, motility and morphology evaluated by WHO criteria. Fertilization rate in both groups were then analyzed after 16-18 hr post insemination. Fertilization rate was higher in group B (p=0.028). Participants in group B, apart from having a normal sperm count (32.9±7.2) and motility (62.4±8.9), have a strict morphology of ≥30%. Our result has shown that spermatozoa in group B had a higher fertilization rate (71.4%). Though the sperm count (36.4±6.7) and motility (66.3±7.4) in group A were slightly higher (p=0.058 and p=0.060 respectively) than group B, the fertilization rate was lower. Our study showed that sperm morphology could be a very important consideration before decisions towards allocation of patients into IVF or intra-cytoplasmic sperm injection.

EFFECT OF MINERAL NPK AND ORGANIC FERTILIZATION ON GROWTH, YIELD, ESSENTIAL OIL AND CHEMICAL COMPOSITION OF CORIANDER PLANTS

A field trial was conducted at Esna, Luxor Governorate, Egypt during 2012/2013 and 2013/2014 seasons to study the effect of mineral NPK and organic fertilization on growth, yield, essential oil and herb N, P and K % of Coriandrum sativum, L. plants. The obtained results revealed that all studied characters of growth, yield, essential oil and herb % of NPK were significantly augmented due to the use of both low and high NPK fertilization rates with the high one giving the highest values. Concerning organic fertilization, the different growth traits (plant height, stem diameter and herb dry weight), yield and yield component characters (number of umbels/plant and fruit yield per plant and per fed), essential oil parameters (percent and yield per plant and per fed) and NPK % in the herb were gradually increased parallel to the gradual increase in compost level. The combined treatment between the low NPK rate (50 g kristalon/10.8 m2 plot)and medium compost level (10 ton/fed) resulted in better fruit and essential oil yield than that obtained from the high NPK rate. So, the possibility of substituting one half of the mineral NPK dose by the medium compost level is justified.