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Purpose: Non-obstructive azoospermia (NOA) is a common, but complex problem, with multiple therapeutic options and a lack of clear guidelines. Hence, there is considerable controversy and marked variation in the management of NOA. This survey evaluates contemporary global practices related to medical and surgical management for patients with NOA. Materials and Methods: A 56-question online survey covering various aspects of the evaluation and management of NOA was sent to specialists around the globe. This paper analyzes the results of the second half of the survey dealing with the management of NOA. Results have been compared to current guidelines, and expert recommendations have been provided using a Delphi process. Results: Participants from 49 countries submitted 336 valid responses. Hormonal therapy for 3 to 6 months was suggested before surgical sperm retrieval (SSR) by 29.6% and 23.6% of participants for normogonadotropic hypogonadism and hypergonadotropic hypogonadism respectively. The SSR rate was reported as 50.0% by 26.0% to 50.0% of participants. Interestingly, 46.0% reported successful SSR in

Purpose: Non-obstructive azoospermia (NOA) represents the persistent absence of sperm in ejaculate without obstruction, stemming from diverse disease processes. This survey explores global practices in NOA diagnosis, comparing them with guidelines and offering expert recommendations. Materials and Methods:Materials and Methods: A 56-item questionnaire survey on NOA diagnosis and management was conducted globally from July to September 2022. This paper focuses on part 1, evaluating NOA diagnosis. Data from 367 participants across 49 coun-tries were analyzed descriptively, with a Delphi process used for expert recommendations. Results:Results: Of 336 eligible responses, most participants were experienced attending physicians (70.93%). To diagnose azoosper-mia definitively, 81.7% requested two semen samples. Commonly ordered hormone tests included serum follicle-stimulating hormone (FSH) (97.0%), total testosterone (92.9%), and luteinizing hormone (86.9%). Genetic testing was requested by 66.6%, with karyotype analysis (86.2%) and Y chromosome microdeletions (88.3%) prevalent. Diagnostic testicular biopsy, distinguishing obstructive azoospermia (OA) from NOA, was not performed by 45.1%, while 34.6% did it selectively. Differ-entiation relied on physical examination (76.1%), serum hormone profiles (69.6%), and semen tests (68.1%). Expectations of finding sperm surgically were higher in men with normal FSH, larger testes, and a history of sperm in ejaculate. Conclusions:Conclusions: This expert survey, encompassing 367 participants from 49 countries, unveils congruence with recommended guidelines in NOA diagnosis. However, noteworthy disparities in practices suggest a need for evidence-based, international consensus guidelines to standardize NOA evaluation, addressing existing gaps in professional recommendations.

Lately, nickel oxide nanoparticles (NiO NPs) have been employed in different industrial and biomedical fields. Several studies have reported that NiO NPs may affect the development of reproductive organs inducing oxidative stress and, resulting in male infertility. We investigated the in vitro effects of NiO NPs on porcine pre-pubertal Sertoli cells (SCs) which undergone acute (24 h) and chronic (from 1 up to 3 weeks) exposure at two subtoxic doses of NiO NPs of 1 μg/ml and 5 μg/ml. After NiO NPs exposure we performed the following analysis: (a) SCs morphological analysis (Light Microscopy); (b) ROS production and oxidative DNA damage, gene expression of antioxidant enzymes (c) SCs functionality (AMH, inhibin B Real-time PCR analysis and ELISA test); (d) apoptosis (WB analysis); (e) pro-inflammatory cytokines (Real-time PCR analysis), and (f) MAPK kinase signaling pathway (WB analysis). We found that the SCs exposed to both subtoxic doses of NiO NPs didn’t sustain substantial morphological changes. NiO NPs exposure, at each concentration, reported a marked increase of intracellular ROS at the third week of treatment and DNA damage at all exposure times. We demonstrated, un up-regulation of SOD and HO-1 gene expression, at both concentrations tested. The both subtoxic doses of NiO NPs detected a down-regulation of AMH and inhibin B gene expression and secreted proteins. Only the 5 μg/ml dose induced the activation of caspase-3 at the third week. At the two subtoxic doses of NiO NPs a clear pro-inflammatory response was resulted in an up-regulation of TNF-α and IL-6 in terms of mRNA. Finally, an increased phosphorylation ratio of p-ERK1/2, p-38 and p-AKT was observed up to the third week, at both concentrations. Our results show the negative impact of subtoxic doses NiO NPs chronic exposure on porcine SCs functionality and viability.

IntroductionAmong substances released into the environment by anthropogenic activities, the heavy metal cadmium (Cd) is known to induce severe testicular injury causing male subfertility/infertility. Zinc (Zn) is another heavy metal that, unlike Cd, is physiologically present in the testis, being essential for spermatogenesis. We aimed to examine the possibility that 50 µM ZnCl2 could counteract the toxic effects induced by Cd in an in vitro model of porcine prepubertal Sertoli cells (SCs) exposed to both subtoxic (5 μM) and toxic (10 μM) concentrations of CdCl2 for 48 h.Materials and MethodsApoptosis, cell cycle, and cell functionality were assessed. The gene expression of Nrf2 and its downstream antioxidant enzymes, ERK1/2, and AKT kinase signaling pathways were evaluated.Materials and ResultsWe found that Zn, in co-treatment with subtoxic and toxic Cd concentration, increased the number of metabolically active SCs compared to Cd exposure alone but restored SC functionality only in co-treatment with subtoxic Cd concentration with respect to subtoxic Cd alone. Exposure of Cd disrupted cell cycle in SCs, and Zn co-treatment was not able to counteract this effect. Cd alone induced SC death through apoptosis and necrosis in a dose-dependent manner, and co-treatment with Zn increased the pro-apoptotic effect of Cd. Subtoxic and toxic Cd exposures activated the Nrf2 signaling pathway by increasing gene expression of Nrf2 and its downstream genes (SOD, HO-1, and GSHPx). Zn co-treatment with subtoxic Cd attenuated upregulation on the Nrf2 system, while with toxic Cd, the effect was more erratic. Studying ERK1/2 and AKT pathways as a target, we found that the phosphorylation ratio of p-ERK1/2 and p-AKT was upregulated by both subtoxic and toxic Cd exposure alone and in co-treatment with Zn.DiscussionOur results suggest that Zn could counteract Cd effects by increasing the number of metabolically active SCs, fully or partially restoring their functionality by modulating Nrf2, ERK1/2, and AKT pathways. Our SC model could be useful to study the effects of early Cd exposure on immature testis, evaluating the possible protective effects of Zn.

Fertility preservation for prepubertal male patients undergoing gonadotoxic therapies, potentially depleting spermatogonial cells, is an expanding necessity, yet most of the feasible options are still in the experimental phase. We present our experience and a summary of current and novel possibilities regarding the different strategies to protect or restore fertility in young male patients, before proceeding with chemotherapy or radiotherapy for malignances or other diseases. Adult oncological patients should always be counselled to cryopreserve the semen before starting treatment, however this approach is not suitable for prepubertal boys, who aren’t capable to produce sperm yet. Fortunately, since the survival rate of pediatric cancer patients has skyrocketed in the last decade and it’s over 84%, safeguarding their future fertility is becoming a major concern for reproductive medicine. Surgical and medical approaches to personalize treatment or protect the gonads could be a valid first step to take. Testicular tissue autologous grafting or xenografting, and spermatogonial stem cells (SSCs) transplantation, are the main experimental options available, but spermatogenesis in vitro is becoming an intriguing alternative. All of these methods feature both strong and weak prospects. There is also relevant controversy regarding the type of testicular material to preserve and the cryopreservation methods. Since transplanted cells are bound to survive based on SSCs number, many ways to enrich their population in cultures have been proposed, as well as different sites of injection inside the testis. Testicular tissue graft has been experimented on mice, rabbits, rhesus macaques and porcine, allowing the birth of live offspring after performing intracytoplasmic sperm injection (ICSI), however it has never been performed on human males yet. In vitro spermatogenesis remains a mirage, although many steps in the right direction have been performed. The manufacturing of 3D scaffolds and artificial spermatogenetic niche, providing support to stem cells in cultures, seems like the best way to further advance in this field.

As wild fish are a finite resource, the role of aquaculture in providing the necessary fish per capita is becoming increasingly important. Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss) are species of interest to Romanians, rich in omega-3 polyunsaturated fats and a good source of protein, while being low in calories and saturated fats. The purpose of this study was to examine and compare the biochemical composition, respectively the nutritional value, of Atlantic salmon and rainbow trout obtained from Romanian aquaculture and from import, that can be found on the Romanian market. The results of this study confirm similarities regarding the biochemical composition of salmon obtained in Romanian aquaculture with wild Atlantic salmon. Rainbow trout bred in trout farms in different areas in our country, can be considered a valuable food source for human consumption.

Temporal variations in the biochemical and haematological parameters of blood must be taken into account as it is a useful indicator for assessing the nutritional status, non-specific immunity and health of fish in general. The aim of this study was to analyse changes in the biochemical and haematological profile of blood in common carp (Cyprinus carpio) depending on the environmental changes characteristic for each season. The lower mean haematocrit, identified in the spring, which correlates with the lower erythrocyte count, implies a more pronounced anaemia in carp after the winter period. The specimens analysed in spring and summer show an increase in the percentage of neutrophils by 60%, compared to the specimens analysed in autumn. Lower values for albumin in spring (0.65±0.22 g/dL) suggest a disruption of protein synthesis function in liver, which is not true for biological material during summer (0.90±0.60 g/dL), and autumn (0.95±0.25 g/dL). CK and CRE values are within normal limits throughout the study period, indicating no injury or muscle atrophy for the analysed specimens. These results suggest that seasonal changes in the environment cause temporary changes in the indicators of the biochemical and haematological profile of the blood of Cyprinus carpio.

To determine the macronutrient variations in fish meat at the end of the winter period compared to the start of wintering, 7 species from the Danube River were fished (Abramis brama, Ballerus sapa, Barbus barbus, Blicca bjoerkna, Gymnocephalus schraetser, Sander lucioperca, Vimba vimba). Biochemical analyses of fish meat were performed for the winter of 2020-2021 and 2021-2022. The biochemical profile followed the same trends for the 7 fish species as well as the 2 wintering periods considered. At the start of the winter period, in the meat of all analysed species, the percentage of water was lower compared to the end of the winter period. Proteins and lipids varied inversely with the amount of water in the fish meat for both wintering periods. In 2020-2021, the percentages of proteins and lipids were lower than in 2021-2022, for all 7 species analysed. The variations of macronutrients between the start and the end of the winter period are explained by the hibernation behaviour of the fish, which is directly influenced and dependent on temperature variations. The thermal regime recorded higher values in the winter of 2020-2021, interrupting the hibernation of fish, which led to a higher consumption of energy reserves in the meat of the considered species.

The use of probiotics in aquaculture has become a necessity to increase disease resistance. Probiotics are also capable of increasing feed digestion and conversion, decreasing sensitivity to stress, and improving the health of the fish. The aim of this study was to analyse the effect of probiotics on survival, welfare, growth indices and blood composition in Siberian sturgeon (Acipenser baerii), reared in a recirculating system. Diets were supplemented with Lactobacillus acidophilus (FLa) and Saccharomyces boulardii (FSb), separately and combined (FLa + Sb). The effect of probiotics was investigated on 2000 fish with a mean weight of 8.82 ± 0.29 g/specimen. Fish were fed for eight weeks with four different diets: a control without probiotics, one with the addition of lactic acid bacteria (La), one with the addition of yeast (Sb), and one with a mixture of bacteria and yeast in equal proportions (50% La + 50% Sb). FLa + Sb showed a better condition factor (Fulton coefficient, K = 0.39%) and significantly higher growth performance in terms of individual growth (WGi = 35.56 g), total growth gain (WGt = 15.30 g), specific growth rate (SGR, 2.70%/day), and feed conversion ratio (FCR = 1.58), compared to the control. The concentration of lymphocytes, monocytes, and neutrophils was higher in the tanks fed with probiotics compared to the control. The presence of probiotics caused a decrease in cholesterol and glucose. There were significant differences between the Immunoglobulin M values identified in the tank fed with FLa + Sb and the control tank. The results showed that the addition of lactic acid bacteria and yeasts, introduced as probiotics in the diets of Acipenser baerii, reared in a recirculating system, improved the growth indicators, survival, and welfare of the fish.