Study of the Antibiotics Effect on Sperm Parameters in the Breeding Rooster

Abstract

Antibiotics have become an integral part of modern medical treatments in humans and animals, but their potential effects on male reproductive health remain an area of concern. The present thesis aims to investigate and elucidate the impact of commonly prescribed antibiotics on sperm parameters in breeding roosters, that to our knowledge, no similar studies have been conducted before; focusing on both in-vitro and in-vivo settings. This study sheds light on the effects of oxytetracycline, enrofloxacin, erythromycin, ampicillin, colistin, tylosin, and sulfonamides on sperm movement characteristics and viability. Semen from forty Cobb 500 (n = 40) reproductive roosters, 45-week-old, weighing 5-6 kg was collected (using Burrows and Quin method) and evaluated for different parameters. Neat semen samples were handled immediately after collection. The sperm volume and PH were evaluated by graduating tube and PH Test paper strips respectively. Whereas sperm count, viability, and all motile parameters (total and progressive motility (TM% and PM%) in addition to all sperm cells kinematics including velocities. [Curvilinear velocity (VCL μm/s), straight-line velocity (VSL μm/s), average path velocity (VAP μm/s), the amplitude of the lateral head displacement (ALH μm), linearity (LIN% = (VSL / VCL) x 100) and frequency to which the sperm head crosses the mean trajectory (beat-cross frequency [BCF]/Hz] were assessed by CASA system (Computer-Aided Sperm Analyzer) after dilution. In the in-vitro phase of the study, only ejaculates with good quality (volume, mass motility, sperm concentration) were sampled and used. The semen donors were collected at a 3-day interval. The pooled sperm was immediately diluted 1/4 ratio (1 part of semen and 3 parts of physiological solution NaCl 0.9%) to protect and preserve the fresh semen quality and the motility of spermatozoa, then transferred to the laboratory where it was diluted again (1/16 ratio) and divided into eight groups. A control group without antibiotics diluted using 0.9% NaCl and treated groups using varying concentrations of oxytetracycline, erythromycin, tylosin, ampicillin, enrofloxacin, colistin, and sulfonamide solutions (30 µg/ml, 15 µg/ml, 100 µg/ml, 30 µg/ml, 5 µg/ml, 10 µg/ml and 300 µg /ml of 0.9% NaCl respectively). Motility parameters were assessed through advanced analyses after 0, 1, 2, 3, 4, 5, 6, and 24 hours of incubation at 37ºC. Moving to the in-vivo investigation and based on the treatment, the breeding roosters were divided into seven groups with five (n=5) individuals each. They were respectively received therapeutic doses of oxytetracycline (40 mg/kg/bwt), erythromycin (20 mg/kg/bwt), tylosin (20 mg/kg/bwt), ampicillin (30 mg/kg/bwt), enrofloxacin (10 mg/kg/bwt), colistin (2.5 mg/kg/bwt) and sulfonamides (140 mg/kg/bwt) for 9 consecutive days via drinking water that was prepared each day just before its administration. Each group was subjected to 3 semen collections: before treatment (control T0), 3rd day (T3), and 9th day (T9) of treatment. Our in-vitro results revealed that spermatozoa were particularly sensitive to almost all antibiotics except for enrofloxacin, which showed a positive relationship with sperm kinematics. In the in-vivo study, enrofloxacin, colistin, tylosin, and oxytetracycline caused a significant negative effect on motile parameters including velocities compared to ampicillin-treated samples. Consequently, in-vitro, most antibiotics appear not suitable in cryopreservation extenders contrary to enrofloxacin. The drug was highly tolerated by the rooster's sperm cells by enhancing their motility and prolonging their survival by up to 6 hours compared to the other antibiotics. The negative effect of tylosin on sperm motile parameters was greater than erythromycin although both drugs are macrolides. The results highlighted the in-vivo effectiveness of sulfonamides and erythromycin on sperm movement characteristics. Both antibiotics considerably improve total and progressive motility, viability as well as all kinematics.