Hygiene and Sanitation in the Boar Stud
Hygiene is defined as the science of the establishment and maintenance of health, or the conditions or practices as of cleanliness conducive to health (Merriam-Webster Collegiate Dictionary: www.m-w.com). Providing a clean, disease free environment for the boars will enhance sperm production and prevent disease spread to the sow units it serves. Bacterial contamination of extended boar semen can originate from the ejaculate itself as well as from the environment. While contamination of the ejaculate from the boar is considered normal, it is not desirable and may reduce fertility in sows (Althouse, et. al., 2000; Rillo, et. al., 1998). Indications of bacterial contamination of extended boar semen range from sperm cell agglutination and decreased storage life to reduced fertility and vaginal discharges in mated sows (Althouse, et. al., 1998). In extreme cases with high levels of contamination, endometritis may occur in mated sows, which could result in culling and even sow death (Payne et al., 2008). Find out more about hygiene and sanitation in the board stud in this Factsheet.
Biosecurity is an important component for providing protection to animals from diseases that are not present on the farm. When considering hygiene and external contamination of semen doses, it is common to associate these with bacterial contamination, while biosecurity generally invokes thoughts of prevention of viral infection of the boars themselves. However, boar stud managers should strive to create a culture within all employees that embraces biosecurity in such a way that it is taken into consideration for all forms of disease and all processes and activities within and outside of the stud. With an average ratio of 150 to 200 sows per boar on stud, the potential impact of disease in a boar stud contaminating the sow units to which it supplies semen is enormous, and the liability to the stud could be immense.
Biosecurity encompasses a large number of processes from rodent control and bird-proofing to human traffic control, personnel showering, and provision of clean clothing and boots for workers. Animal products from outside the stud, which could potentially transmit disease to the boars in stud, must never be allowed in the boar diets. It is possible for contaminants to enter the boar stud from outside sources via a number of routes, including laboratory supplies, personnel, boars, air, water, and feed. Conducting periodic audits of biosecurity practices and monitoring of feed will identify potential breaches, assist the manager in maintaining proper protocols throughout the system, and educate the employees in areas that need improvement.
The National Pork Board has published a series of documents on biosecurity that can be accessed either directly from them at P.O. Box 9114, Des Moines, IA 50306, (515) 223-2600 or at www.porkboard.org (http://www.pork.org/Producers/ Security.aspx?c=Security). These documents include a guide for general principals of biosecurity and a series of questions for those purchasing semen to ask their semen supplier.
In 2003, the American Association of Swine Veterinarians (AASV) published a document titled Health, Hygiene, and Sanitation Guidelines for Boar Studs Providing Semen to the Domestic Market. This document contains a section on Hygiene and Sanitation Requirements for Semen Collection, Processing, and Storage and is available to all AASV members and can be provided to the boar stud manager by their veterinarian.
Once biosecurity has become a culture with the employees, the risk of disease introduction to the stud will be significantly reduced. It is critical to the success of the stud that this culture be instilled in every new employee as quickly as possible. Having the entire team understand the importance will expedite this process with new hires. The best biosecurity protocols are useless without monitoring and meaningful punishment for breaches. Ultimately, the manager’s actions and attitude towards biosecurity will set the tone for the entire team.
Routine cleaning of the boar barn includes daily scraping of manure from alleys and power washing and disinfecting of the semen collection areas. With the exception of the very largest boar studs that have multiple rooms, it is unlikely that most studs would replace boars on an all-in, all-out basis, making regular cleaning and disinfecting of an empty barn or room difficult. Ambient temperature and boar comfort must be taken into consideration when routine power washing is performed with boars present in the barn. When boars are present during washing, supplemental heat and increased air movement may be required to dry the boars and the barn quickly. Further, care must be taken to not spray boars directly with the power washer. Considerable variation exists in the ability to remove contaminants with a power washer, depending on the type of surface being cleaned (Amass, S. F. 2004). Surfaces that are rough will give the boars more traction and result in fewer injuries due to slipping, but may result in more residual bacteria following cleaning (Madec, et. al., 1999). Therefore, a compromise must be reached between abrasiveness for traction and smoothness to enhance cleaning ability. Slat quality in the boar barn is particularly important to ensure injury is not caused in normal day to day housing and moving within the unit. A thick rubber mat (3/8 inch) with large, 1 inch diameter holes, is always recommended for the semen collection pen. The mat must be removable to facilitate complete washing, disinfection, and drying on a daily basis (Figure 1). Rotation of disinfectants is recommended in conjunction with thorough testing to ensure that resistant bacteria are not present.
There is limited information regarding the presence of bacteria in showers at swine barns, but evidence reported to date suggests that the risk of such contamination is low, despite the fact that most farm showers are not visibly kept clean (Amass, et. al., 2005). Simple hand washing has been demonstrated to be linked with a reduction in Salmonella prevalence on swine farms (Wong, et. al., 2004). This suggests the minimum requirements of showering into the barn and wearing unit clothing and boots will prevent most contamination problems that could potentially be introduced by employees. There are several areas that can help reduce disease movement into a boar stud and these include: 1) observing minimum “down” times between exposure with other swine and entry into the boar stud, 2) off-site isolation with daily chores performed by non-stud personnel, 3) washing, disinfecting, and drying of vehicles before returning to the boar stud after a visit to another swine operation (particularly for semen delivery vehicles), and 4) fogging of packages with disinfectant prior to their entry in to the unit. Enforcing a policy that discourages personnel with active influenza infections from reporting for work will protect the boars from possible cross-contamination (low risk) and prevent infection of other workers at the unit (high risk).
Under normal circumstances, and provided good hygiene practices are followed, the limited numbers of bacteria that are typically present in an ejaculate do not pose a significant problem to semen storage and fertility. If, however, the bacteria level is significant and/or there are bacteria present that are resistant to the antibiotic(s) in the semen extender used, semen quality can deteriorate rapidly in storage and/or fertility may be compromised. Often, this is manifested by an increase in the number of sows that recycle following breeding and an increased incidence of vulvar discharges in sows three weeks post mating.
Follow These Five Methods To Minimize The Bacterial Load In Semen Collections:
- Maintain a high level of hygiene in the barn, including daily power washing and disinfection of the warm-up pens, semen collection area and dummy sows. With the proper type of flooring and ventilation, the warm-up and semen collection areas will dry between periods of use. Bacteria, which are present on the skin and hair and in the feces of the boar, are more easily washed from a metal dummy sow with a rubberized covering, or no covering at all, compared to one covered with an absorbent material such as cloth, carpet or canvas. Periodic washing of the semen collection pen and equipment between collections will reduce the environmental load of bacteria present in the semen collection area.
- Keep all semen collection materials as clean as possible prior to use. Single use disposable collection bags and/ or insulated containers that can be prepared ahead of time and kept in clean plastic bags until immediately prior to use are preferred over reusable equipment. Automated semen collection systems that take the place of the technician in the actual semen collection process use disposable products that can be kept clean and sterile prior to use.
- Use clean, disposable collection gloves for each boar. A second plastic glove over the collection glove should be worn until the boar penis is actually exposed and the boar is ready to fully extend. Many boars will attempt several mounts and extensions (full or partial) prior to their final extension and ejaculation. This may necessitate the wearing of two or more collection gloves. Keeping additional gloves in a pocket to put on if the collection glove gets contaminated (although not sterile) is a good alternative to using a glove contaminated by contact with the boar. Some technicians also have a habit of placing a hand on the back of the boar as he mounts and extends to help balance and support the boar. This may result in direct contamination of the collection glove and should be avoided unless the plastic glove worn over the collection glove is kept on during this process.
- Collect only the ejaculate fractions with the most sperm cells and least amount of bacterial contamination. The first fraction, or pre-sperm portion, and the fourth, or gel plug fraction, each contain higher levels of bacteria and contain minimal numbers of sperm cells and, therefore, should not be collected into the container (Gall, et. al., 1998). The first, pre-sperm fraction is easily distinguished as it is clear and ends with the first jets of the darker, creamy appearing spermrich fraction (the first fraction to save). The fourth, gel fraction is accompanied by pellets of sticky “gel” that make it easily distinguishable from the second fraction or the sperm rich, and the third fraction of seminal fluid that are to be saved. A disadvantage of the automated semen collection equipment is that this gel fraction cannot be diverted from the semen collection container, although the filter should prevent it from passing into the collection bag. Preputial fluids have been shown to be a major contributor to bacterial contamination of ejaculates in the boar (Rillo, et. al., 1998). Surgical removal of the preputial diverticulum is an effective method of eliminating this source of contamination (Althouse and Evans, 1994). While this intervention is not practical for most studs, there may be a limited number of applications where it could be advantageous.
- Remove the semen collection filter from the top of the collection container prior to transport of the ejaculate into the laboratory. This step reduces potential for bacterial contamination and load in the lab.
In the collection barn, having sterile, disposable products available does not guarantee they will be clean at the time of use. If the bag of collection supplies is left open or a technician uses a dirty hand to remove the items from the bag, contamination can compromise the entire bag’s contents (Figure 2). Placing a sterile collection container on the dirty floor, or even placing it in a pocket or under an arm may also result in contamination. An open box of collection gloves kept in the barn will become contaminated with dust and bacteria circulating in the air and should therefore be kept in a closed plastic bag. The goal is to use the best practices that result in the least amount of bacterial exposure during these processes.
Everything that enters the semen processing laboratory, including water, air, people, and ejaculates, has the potential to introduce bacterial contamination into the processed semen doses. The ideal semen processing laboratory is physically separated from the barn with independent showers, break rooms, and other areas. If it is not possible to provide showers for the lab personnel, then at minimum, technicians should change clothing and shoes and wash hands. In smaller units where barn personnel may also spend time working in the lab, they should change clothing and wash hands prior to entering the lab if it is not possible for them to shower between duties. Unfortunately, having showers does not guarantee every employee will take a complete shower every time they enter the unit. Developing a “culture” among all employees where taking a shower or changing clothing and washing hands (depending on the facilities) is considered “normal” should be the goal of every boar stud manager. For all hand washing in the unit, the type of soap used is not as important as the washing of hands itself. Antibacterial soap does not remove more bacteria than ordinary soap when hands are washed for 20 seconds, although E2 rated soaps (those containing 50 parts per million of chlorine) are significantly more effective in removing bacteria than either antibacterial or ordinary soap. Instant hand sanitizers are not as effective in removing bacteria as washing with soap and water (Miller, et. al., 1994).
For cleaning laboratory surfaces, just as with washing hands, cleaning appears to be more important than “disinfecting”. In 1995, disinfection protocols were tested in a boar stud comparing a cleaning procedure used at a hospital to a commercial cleaning solution. The hospital protocol incorporated wiping all surfaces with a solution of 10% bleach (5.25% sodium hypochlorite) in water, which was compared to use of only a commercially available detergent cleaning solution and wiping with disposable towels before and after daily work. In all tests run at the boar stud, there were fewer bacterial colonies grown from swabs of counters using the commercially available detergent compared to those wiped with the bleach solution (Gall, T.J., unpublished data). Chlorine bleach solutions are routinely used for disinfection but are not effective as cleaning agents for removal of organic matter or dirt (Emswiler, et. al., 1978; Northcutt, et. al., 2007). When selecting cleaning and disinfecting agents for the boar stud, first decide whether you want a cleaning agent alone, a combination cleaning and disinfecting solution, or a disinfecting solution alone. The label of each agent should clearly state which type it is and all disinfectants must be registered with the Environmental Protection Agency (EPA). Each manufacturer is required to submit to the EPA proof that the product kills the organisms listed on the label. The label should also state which types of surfaces the product is safe to use on. Ten % bleach solution was once considered the “Gold Standard” for disinfection in hospitals, but many hospitals, as well as food service and medical care facilities have switched from bleach to quaternary ammonia chloride solutions (Quats) for disinfection of surfaces. Quats provide broad spectrum bacteria, virus and mold control, are less corrosive than bleach and have cleaning properties that bleach solutions do not provide (McFadden, R., 2005). The active ingredient in Quats is benzalkonum chloride which can be formulated with a variety of ingredients to provide a neutral pH disinfectant. Since Quats provide both cleaning and disinfectant properties, they provide an economical one-step process for use in the lab. There are several other types of disinfectants that may be considered for use in the lab but may be less practical. One of these is the phenols, which are flammable and corrosive, can leave a residue that eventually has to be removed, and must be mixed up fresh every day.
Another type of disinfectant is iodine solution (iodophors) which are slow to kill organisms, making exposure time critical. They are also corrosive to metal and rubber and stain many types of surfaces. Iodophors must also be mixed fresh daily and may be inactivated by hard water. Ethyl alcohol diluted to 70% (with water) is effective against most organisms, but is flammable and requires that exposure time be monitored (Favero, M.S., 1985). Ethanol is very effective as a disinfectant for skin and can be used on hands prior to handling clean supplies and equipment. The importance of cleaning organic material to achieve disinfection has been shown for common surfaces used in the swine industry, such as rubber boots (Amass, et. al., 2001). For washing lab surfaces, disposable cleaning cloths are ideal, but reusable clothes that are washed daily are an acceptable alternative. Sponges should be avoided for cleaning, unless they are replaced regularly (daily, or at least weekly). Ultraviolet (UV) light (253.7nm wavelength) is effective against airborne and surface organisms but will not penetrate solid materials. Bulbs must be checked for intensity and kept clean to be effective. UV lights should never be turned on while people are present in the lab.
Protocols for minimizing contamination within the lab begin with thorough cleaning on a daily basis. Surfaces that are used routinely should be cleaned and disinfected frequently throughout the processing period. Technicians should wash hands frequently and use disposable towels as much as possible. All surfaces that are touched need frequent cleaning, including telephones, the computer keyboard and mouse, doorknobs, drawer pulls, and all other items. A plastic cover can be ordered for the most popular computer keyboards which allows for effective cleaning while providing for unimpeded operation. Plastic kitchen wrap makes for an ideal cover for the computer mouse and telephone, and can be replaced easily and frequently at low cost. Cell phones should not be allowed into the lab or past the dirty side of the showers at any time, since they cannot be readily cleaned and disinfected.
Water is the single largest component of semen extender and is used for routine cleaning and rinsing of lab equipment. Regardless of the source, water should be periodically tested for contaminants. Low cost in-line UV light systems are available that will kill bacteria present in water used in the lab.
Use of disposables in the lab is preferable to using equipment that must be washed and sterilized. Similar to use of supplies in the barn, you cannot assume that disposable items taken from an open bag are sterile, so following best hygiene practices will help minimize contamination. Thoroughly washing hands before handling supplies is the best way to decrease cross contamination. Once containers of bulk supplies are opened, their contents should be divided into re-sealable bags or boxes (Figure 4). Bulk semen extender should be repackaged in individual aliquots for use when the bulk package is initially opened to prevent repeated opening and possible contamination of the extender powder. Semen extenders should only be purchased from reputable suppliers who can verify quality control (QC) processes and routinely perform these processes for the ingredients used in the final product.
Antibiotics in Semen Extenders
By design, semen extenders contain ingredients that make them well suited as culture media for the bacteria that may be present in neat semen at the time of collection or picked up from the environment during processing. Antibiotics are either present or are added to semen extenders to control bacteria growth and allow for increased storage times (Poolperm, et. al., 1998). Cooling of semen to storage temperatures from 59°F to 64°F (15°C to 18°C) also aids in suppression of bacterial growth. Breakthroughs in semen extender formulations that would allow storage at or near 32°F (0°C), or improvements in semen freezing technology could further prolong storage times. Microscopic evaluation of extended samples throughout the useful storage life of the semen should be performed and will often reveal problems with bacterial contamination. This process should be carried out at least to the recommended maximum shelf-life of the extender or longer if a problem is suspected. Visual analysis of sperm cells, with and without stain, can be coupled with culturing of samples of extended semen on agar plates with incubation for 48 hours at 96°F to 100°F (36°C to 38°C) to confirm contamination by observation for bacterial colony growth. Keep in mind that problems will often only be revealed after the semen in question has been used at the farm. When a bacterial problem is discovered in the lab, follow-up of fertility results for each affected pool or ejaculate must be performed at all farms that utilized the affected Important considerations for choosing the proper antibiotics for a semen extender are spermicidal activity, cost, and sensitivity and resistance information of the target bacteria to a specific antibiotic. The first step is to determine which bacteria are normally present and what antibiotics provide control. Table 1 lists antibiotic combinations and the levels that are available in various semen extenders from some of the suppliers in the U.S.A.
Table 1. Antibiotic combinations available in semen extenders in the U.S.A. by supplier.
|Supplier||Antibiotic (quantity per Liter)|
|A||Ampicillin (not given) A Ceftiofur (not given)|
|A||Gentamycin (not given)|
|A||Neomycin (not given)|
|B||Apramycin (250mg) + Ampicillin (250mg)|
|B||Gentamycin (250mg or 400mg)|
|B||Gentamycin (400mg) + Ampicillin (200mg)|
|B||Gentamycin (400mg) + Ampicillin (200mg) + Cefoperazone (50mg)|
|B||Gentamycin (400mg) + Neomycin (600mg)|
|B||Gentamycin (400mg) + Spectinomycin (270mg) + Lincomycin (120mg)|
|C||Ampicillin (100 to 200mg)|
|C||Gentamycin (250 to 500mg)|
|C||Neomycin (250 to 600mg)|
|D||Amikacin (per customer request)|
|D||Gentamycin (250 to 400mg)|
|D||Penicillin (100,000 IU)|
|E||Gentamycin + Amoxicillin + Tylosin|
|F||Colistine (30mg) + Neomycin (80mg)|
|F||Colistine (30mg) + Enrofloxacine (30mg) + Gentamycin (60mg)|
|F||Colistine (30mg) + Gentamycin (60mg) + Lincospectin (30mg)|
|F||Colistine (33mg) + Enrofloxacine (33mg) + Gentamycin (83mg)|
|F||Colistine (30mg) + Gentamycin (60mg) + Lincospectin (30mg)|
|F||Colistine (60mg) + Gentamycin (60mg) + Enrofloxacine (30mg)|
Samples of extended semen should be routinely submitted for bacterial culture to an independent laboratory. This service is provided by many veterinary clinics, diagnostic laboratories and artificial insemination (AI) supply companies. Samples should be stored at the boar stud at the proper temperature until the sample is tested. Information desired from the diagnostic laboratory would include types and relative levels of bacteria present and a list of antibiotics to which they are both sensitive and resistant. This is an on-going process as the micro-flora of the boars and environment can change over time. These processes are extremely important in order to provide a high quality product that will maximize fertility for the consumer as well as maintain customer confidence that the boar stud is monitoring their own production. Customers should rely on the boar stud to provide the QC for the semen they provide. Sow farms rarely have the equipment or expertise to properly evaluate semen they have received, so it is best to keep this process in the hands of the boar stud personnel. Placing an expiration date on semen does not necessarily result in proper semen use and storage times and can result in shortages if doses are discarded prior to the next shipment arriving at the farm, particularly in the event of an unforeseen delivery delay. Semen does not die on its appointed expiration date, but farm employees will perceive that to be the case if such a date exists. Training sow farm employees in the proper techniques for semen handling, storage and timeliness of use is the responsibility of the boar stud that is supplying it and must be done if expiration dates are used on semen doses.
Three distinct levels of risk associated with bacterial contamination of extended boar semen have been defined for the boar semen industry (Kuster, C.E., 2004). Risk Level I is the lowest and is characterized by no significant growth of bacteria from extended semen samples after 48 hours of incubation on agar. Level II is defined as medium risk with growth of non-spermicidal bacteria that do not affect shelf life of the sperm cells. However, Level II indicates a breakdown in hygiene in the lab and/or barn which needs to be addressed immediately. Level III is the highest risk level and is characterized by the presence of spermicidal bacteria at a significant level. Intervention should involve the unit’s veterinarian to determine the source and the most expedient means of correcting the problem. Level III may also require a temporary change in extender antibiotic until the problem is corrected.
Biosecurity and hygiene in the boar stud include processes and developing a culture within the employees to support these goals. Providing written protocols will enhance training of new employees and serve as a guide for addressing contamination problems, should they occur. Having the boar stud certified by the International Organization of Standards (ISO) will require the documentation of every process in the unit, resulting in detailed protocols, and offer an avenue for tracking down problems that have occurred. Although ISO certification can be an expensive process, going though the steps to be certified without actually completing the certification itself can be a great place to start. Finally, boar studs that rely on the regular use of outside consultants, veterinarians and laboratories for input on QC and processes will generally maintain a higher level of hygiene and biosecurity than those that do not.
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