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Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

ABSTRACT: This study examined the effect of Fe3O4 nanoparticles on boar semen. Beltsville thawing solution without antibiotics was used to extend ejaculates from 5 boars (4 ejaculates/boar). Semen samples of control group (C) and group with Fe3O4 (Fe; 0.192 mg/mL semen) were incubated under routine boar semen storage temperature (17 °C) for 0.5 h and nanoparticles were removed by a magnetic field. Before and after treatment, aliquots of all groups were cultured using standard microbiological methods. The samples after treatment were stored (17 °C) for 48 h and sperm parameters (computer-assisted sperm analyzer (CASA) variables; morphology; viability; hypo-osmotic swelling test (HOST); DNA integrity) were evaluated at storage times 0, 24, 48 h. Semen data were analyzed by a repeated measures mixed model and microbial data with Student's t-test for paired samples. Regarding CASA parameters, Fe group did not differ from C at any time point. In group C, total motility after 24 h and progressive motility after 48 h of storage decreased significantly compared to 0 h. In group Fe, linearity (LIN) after 48 h and head abnormalities after 24 h of storage increased significantly compared to 0 h. The microbiological results revealed a significant reduction of the bacterial load in group Fe compared to control at both 24 and 48 h. In conclusion, the use of Fe3O4 nanoparticles during semen processing provided a slight anti-microbiological effect with no adverse effects on sperm characteristics.

SUBMITTER: Tsakmakidis IA

PROVIDER: S-EPMC7466471 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

Tsakmakidis Ioannis A IA Tsakmakidis Ioannis A IA Samaras Theodoros T Anastasiadou Sofia S Basioura Athina A Ntemka Aikaterini A Michos Ilias I Simeonidis Konstantinos K Karagiannis Isidoros I Tsousis Georgios G Angelakeris Mavroeidis M Boscos Constantin M CM

Nanomaterials (Basel, Switzerland) 20200810 8

This study examined the effect of Fe3O4 nanoparticles on boar semen. Beltsville thawing solution without antibiotics was used to extend ejaculates from 5 boars (4 ejaculates/boar). Semen samples of control group (C) and group with Fe3O4 (Fe; 0.192 mg/mL semen) were incubated under routine boar semen storage temperature (17 °C) for 0.5 h and nanoparticles were removed by a magnetic field. Before and after treatment, aliquots of all groups were cultured ...[more]

PMID: 32784995

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Project description:Artificial insemination (AI) is the preferred method for reproduction in the majority of the intensive pig production systems Worldwide. To this end, fresh extended ready-to-use semen doses are either purchased from AI-centres or produced by boars kept on-farm. For profitable semen production, it is necessary to obtain a maximum amount of high quality semen from each boar. This paper reviews current knowledge on factors that may affect semen quality by influencing the boar or the semen during processing. Genetic markers could be used for early detection of boars with the highest fertility potential. Genetic selection for fast growth might jeopardize semen quality. Early detection of boars no longer fit for semen production might be possible by ultrasonography of the testes. Seasonal variation in sperm quality could be associated with changes in photoperiod and heat stress during summer. Comfortable housing, with appropiate bedding material to avoid locomotion problems is essential. In some areas, cooling systems may be necessary to avoid heat stress. The sperm quality can be manipulated by feeding strategies aiming, for instance, to increase sperm resistance to oxidative stress and extend storage duration. High collection frequency will negatively influence sperm quality. Also, if collection is not hygienically performed it will result in bacterial contamination of the semen doses. The concern over bacterial contamination has risen not only because of its negative effect on semen quality but also due to the detection of antimicrobial resistance in isolates from extended semen. Moreover, bacterial and viral pathogens must be monitored because they affect semen production and quality and constitute a risk of herd infection. During processing, boar sperm are submitted to many stress factors that can cause oxidative stress and capacitation-like changes potentially reducing their fertility potential. Dilution rate or dilution temperature affects the quality of the semen doses. Some packaging might preserve semen better than others and some plastic components might be toxic for sperm. Standard operation procedures and quality assurance systems in AI centres are needed.

Project description:The optimum storage temperature for liquid-preserved boar semen has been empirically determined to be between 15 and 20°C. Lower temperatures provide an advantage to inhibit bacterial growth, but are regarded as critical due to the high sensitivity of boar spermatozoa to chilling injury. Higher storage temperatures are supposed to induce energy deficiency due to an insufficient depression of metabolic cell activity. However, experimental evidence for alterations of the sperm's energy status in relation to storage temperature and duration is missing. Therefore, we aimed to revisit the upper and lower storage temperature limits for liquid-preserved boar semen from the perspective of the sperm's energy metabolism. Ejaculates (n = 7 boars) were cooled down in Beltsville Thawing Solution (BTS) to 25, 17, 10, or 5°C and stored for up to 120 h. ATP and adenylate energy charge (EC) levels were assessed at storage temperature (24, 72, and 120 h storage) and after subsequent re-warming (38°C). Sperm quality and energy status remained at a stable level in samples stored at 25 and 17°C. Chilling to and storage at 10 or 5°C in BTS provoked cold shock in a subset of sperm as shown by a loss in viability and motility (P < 0.05), which was accompanied by a significant release of adenine nucleotides into the semen extender. Prolonged storage for 120 h resulted in significantly lower mean ATP concentrations in viable spermatozoa at 5 or 10°C compared to 17°C (P < 0.05). Cluster analysis revealed that the main sperm subpopulation, i.e., sperm with moderate speed and linearity, decreased from 50 to 30% (P < 0.05) in favor of slow-moving spermatozoa (5°C) or spermatozoa with a hyperactivation-like motility pattern (10°C). The results point to a sublethal imbalance in available ATP in a subset of the surviving sperm population, rather than a general decrease in available ATP in all spermatozoa. In conclusion, storing diluted boar semen at a stable temperature between 17 and 25°C is a safe procedure concerning the spermatozoa's energy status. Future concepts for hypothermic boar semen preservation below 17°C require measures which ameliorate the imbalanced energy status in viable spermatozoa.

Project description:IntroductionFacing the global threat of antimicrobial resistance, the reduction of antibiotic use in semen extenders is a main goal in artificial insemination (AI) of pigs. The aim of this study was to investigate the potential of a commercial extender containing an organic bactericidal supplement in the absence of conventional antibiotics to control bacterial growth and to maintain the quality of boar spermatozoa during long-term semen storage for up to 144 h at 17°C.MethodsSemen from 233 boars housed at 16 European AI centers was split and diluted in the long-term extender "Androstar Plus without antibiotics + organic bactericidal supplement" (APlus) and in the control extender Beltsville Thawing Solution (BTS) with gentamicin, which is routinely used in many AI centers. Sperm motility was assessed with computer-assisted semen analysis (CASA) and membrane integrity was evaluated with flow cytometry. The number of bacteria was determined by counting colonies on agar plates.ResultsAt the end of storage, bacterial counts were ≥ 106 CFU/mL in 10.7% of the APlus and in 0.4% of the BTS samples. At the same time, bacterial counts were only weakly correlated with sperm motility (r = -0.23, p < 0.05), and there was no correlation with sperm membrane integrity (p > 0.05). Among the 12 identified bacterial species in APlus samples, loss of sperm quality was exclusively observed in the presence of >106 CFU/mL Serratia marcescens and Klebsiella oxytoca. Both these bacterial species, despite their known multi-drug resistance and the continuous use of gentamicin in Europe, proved sensitive to this antibiotic, thus indicating an efficient quality assurance program and responsible antibiotic use.ConclusionLong-term storage of boar semen at 17°C without conventional antibiotics in an extender containing an organic bactericidal supplement is an option if semen samples are regularly tested for the presence of S. marcescens and K. oxytoca, and the source of contamination is eliminated.

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Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

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Iron Oxide Nanoparticles as an Alternative to Antibiotics Additive on Extended Boar Semen.

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