title: Critical sources of bacterial contamination and adoption of standard sanitary protocol during semen collection and processing in Semen Station.
abstract: OBJECTIVE
  The present investigation was conducted to locate the critical sources of bacterial contamination and to evaluate the standard sanitation protocol so as to improve the hygienic conditions during collection, evaluation, and processing of bull semen in the Semen Station.
  
  
  METHODS
  The study compared two different hygienic procedures during the collection, evaluation and processing of semen in Central Semen Station, Anjora, Durg. Routinely used materials including artificial vagina (AV) inner liner, cone, semen collection tube, buffer, extender/diluter, straws; and the laboratory environment like processing lab, pass box and laminar air flow (LAF) cabinet of extender preparation lab, processing lab, sealing filling machine, and bacteriological lab were subjected to bacteriological examination in two phases of study using two different sanitary protocols. Bacterial load in above items/environment was measured using standard plate count method and expressed as colony forming unit (CFU).
  
  
  RESULTS
  Bacterial load in a laboratory environment and AV equipments during two different sanitary protocol in present investigation differed highly significantly (p<0.001). Potential sources of bacterial contamination during semen collection and processing included laboratory environment like processing lab, pass box, and LAF cabinets; AV equipments, including AV Liner and cone. Bacterial load was reduced highly significantly (p<0.001) in AV liner (from 2.33±0.67 to 0.50±0.52), cone (from 4.16±1.20 to 1.91±0.55), and extender (from 1.33±0.38 to 0) after application of improved practices of packaging, handling, and sterilization in Phase II of study. Glasswares, buffers, and straws showed nil bacterial contamination in both the phases of study. With slight modification in fumigation protocol (formalin @600 ml/1000 ft(3)), bacterial load was significantly decreased (p<0.001) up to 0-6 CFU in processing lab (from 6.43±1.34 to 2.86±0.59), pass box (from 12.13±2.53 to 3.78±0.79), and nil bacterial load was reported in LAFs.
  
  
  CONCLUSIONS
  Appropriate and careful management considering critical points step by step starting right from collection of semen to their processing can significantly minimize bacterial contamination.
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science & A.H., Chhattisgarh Kamdhenu Vishwavidyalaya, Anjora, Durg, Chhattisgarh, India.
authors.forename: Chandrahas
authors.initials: C
authors.lastname: Sannat
authors.affiliation: Central Semen Station, Livestock Development Department, Government of Chhattisgarh, Anjora, Durg, Chhattisgarh, India.
authors.forename: Ajit
authors.initials: A
authors.lastname: Nair
authors.affiliation: Central Semen Station, Livestock Development Department, Government of Chhattisgarh, Anjora, Durg, Chhattisgarh, India.
authors.forename: S B
authors.initials: SB
authors.lastname: Sahu
authors.affiliation: Central Semen Station, Livestock Development Department, Government of Chhattisgarh, Anjora, Durg, Chhattisgarh, India.
authors.forename: S A
authors.initials: SA
authors.lastname: Sahasrabudhe
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science & A.H., Chhattisgarh Kamdhenu Vishwavidyalaya, Anjora, Durg, Chhattisgarh, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science & A.H., Chhattisgarh Kamdhenu Vishwavidyalaya, Anjora, Durg, Chhattisgarh, India.
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Gupta
authors.affiliation: Department of Veterinary Microbiology, College of Veterinary Science & A.H., Chhattisgarh Kamdhenu Vishwavidyalaya, Anjora, Durg, Chhattisgarh, India.
authors.forename: R K
authors.initials: RK
authors.lastname: Shende
keywords: Semen Station; bacterial contamination; critical sources; environment; laboratory equipments