Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality

Martínez‐Pastor, Felipe; Lacalle, Estíbaliz; Martínez-Martínez, Sonia; Fernández‐Alegre, Estela; Álvarez-Fernández, Laura; Martinez-Alborcia, María J.; Bolarín, Alfonso; Morrell, Jane M.

doi:10.1016/j.theriogenology.2021.02.009

Cited by 14 publications

(9 citation statements)

References 38 publications

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“…To this end, two low-density Porcicoll (P20 and P30) has been evaluated, and a considerable reduction or complete elimination of some bacteria by both colloids, without affecting sperm quality, was observed. Furthermore, the recovery rates are 86% for P20 and 81% for P30, presenting a very efficient alternative to the use of antibiotics [ 68 ]. In addition, spermatozoa can be separated from bacteria in semen via single-layer centrifugation (SLC).…”

Section: Resultsmentioning

confidence: 99%

“…However, there are also more difficult microorganisms to remove; for example, flagellated motile bacteria are more challenging to eliminate than non-flagellated ones [ 70 ]. Martínez-Pastor et al [ 68 ] evaluated SLC on modified low-density Porcicoll. A sterile inner tube is included inside some 50 mL centrifuge tubes to facilitate harvesting of the sperm pellet.…”

Section: Resultsmentioning

confidence: 99%

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Bacteria and Boar Semen Storage: Progress and Challenges

et al. 2022

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Porcine breeding today is based on artificial insemination with chilled semen. This is stored at 5 °C with antibiotic supplementation to avoid bacteriospermia. There are many negative consequences on sperm quality and functionality as a result of bacterial contamination, as well as on the health of the sow. Nowadays, various techniques are being developed to reduce the indiscriminate use of antibiotics and thus avoid the generation of antibiotic resistance genes. This review aims to inform about the bacterial contamination consequences of storing liquid semen from boar and to provide an update on current methods and alternatives to antibiotic use in cold storage.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Bacteria and Boar Semen Storage: Progress and Challenges

et al. 2022

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“…The special challenge in pig breeding results from the relatively high semen storage temperature (15–18 °C) favouring bacterial growth, the high volume of insemination doses (50 to 100 mL, two to three semen doses per cycle), and the environmental pollution due to seminal backflow into manure (approximately 85% of the inseminated volume; [ 2 , 3 ]), thus providing a source and reservoir of resistance genes in soil and water [ 4 , 5 ]. Attempts to replace conventional antibiotics for boar semen preservation range from the use of alternative antimicrobial substances such as peptides [ 6 , 7 , 8 , 9 ], phytoextracts [ 10 , 11 ] or nanoparticles [ 12 ], to the physical removal of bacteria through single layer centrifugation [ 13 , 14 ] and microfiltration [ 15 ]. However, until now, none of the concepts have been feasible for AI practice because they lack antimicrobial efficiency, are sperm-damaging or impracticable and costly.…”

Section: Introductionmentioning

confidence: 99%

Antimicrobially Active Semen Extenders Allow the Reduction of Antibiotic Use in Pig Insemination

et al. 2021

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Antibiotic use in semen extenders for livestock may contribute to the development and spreading of multi-drug resistance. Antimicrobial control in semen doses for artificial insemination of pigs is indispensable due to the relatively high storage temperature (17 °C). The objectives of this study were first, to examine whether the antimicrobial capacity differs between antibiotic-free extenders and second, to determine whether an antimicrobial active extender provides the possibility to reduce antibiotics. Antibiotic-free semen extenders Beltsville Thawing Solution (BTS) and Androstar Premium were inoculated at 103 to 104 CFU/mL with four pure bacterial strains isolated from boar ejaculates or a mixture thereof, and then stored for 144 h at 17 °C. Bacterial counts after aerobic culture decreased in BTS up to one log level and decreased in Androstar Premium by 2 to 3.5 log levels (p < 0.05). In semen samples from nine boars stored in the inoculated Androstar Premium extender containing half of the standard concentration of gentamicin, bacteria counts were below 101 CFU/mL. Likewise, half of the standard dose of apramycin and ampicillin was fully antimicrobially active and sperm quality was maintained. In conclusion, semen extenders with intrinsic antimicrobial activity allow a reduction in antibiotic use in pig insemination.

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“…Clearly, boar semen may contain many other bacterial species than studied here, either from animal, environmental, or human sources. Among those, Enterococcus species , Pseudomonas , Staphylococcus , Proteus , and E. col i, occur most frequently [ 16 , 17 , 20 , 32 , 33 ]. In the present field data analysis, however, none of these bacteria were identified in low-quality semen samples, indicating that they either were absent or sensitive to the antibiotic in the semen extender or grew only slowly, so that spermicidal effects were not reached.…”

Section: Discussionmentioning

confidence: 99%

Growth Dynamic and Threshold Values for Spermicidal Effects of Multidrug-Resistant Bacteria in Extended Boar Semen

et al. 2023

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The aim of this study was first to examine the prevalence of bacteria-associated loss of sperm quality in samples from insemination centers during a seven-year semen monitoring program and, second, to investigate the growth dynamic of four different multidrug-resistant bacterial species and their impact on sperm quality during semen storage. A reduced sperm quality associated with bacterial contamination was found in 0.5% of 3219 of the samples from insemination centers. In samples spiked with Serratia marcescens and Klebsiella oxytoca, bacterial growth by six log levels was seen during storage at 17 °C, causing loss of sperm motility, membrane integrity, membrane fluidity, and mitochondrial membrane potential at >107 CFU/mL (p < 0.05). Storage at 5 °C in the Androstar Premium extender efficiently inhibited their growth. Achromobacter xylosoxidans and Burkholderia cepacia showed limited growth up to two log levels at 17 °C and did not impair sperm quality. In conclusion, spermatozoa tolerate moderate loads of multidrug-resistant bacteria, and hypothermic, antibiotic-free semen storage effectively limits bacterial growth. The constant use of antibiotics in semen extenders should be reconsidered.

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Low density Porcicoll separates spermatozoa from bacteria and retains sperm quality

Cited by 14 publications

References 38 publications

Bacteria and Boar Semen Storage: Progress and Challenges

Bacteria and Boar Semen Storage: Progress and Challenges

Antimicrobially Active Semen Extenders Allow the Reduction of Antibiotic Use in Pig Insemination

Growth Dynamic and Threshold Values for Spermicidal Effects of Multidrug-Resistant Bacteria in Extended Boar Semen

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