Buhr R J, Musgrove M T, Richardson L J, Cox N A, Wilson J L, Bailey J S, Cosby D E, Bourassa D V
Poultry Microbiological Safety Research Unit, USDA, ARS, Richard B. Russell Agricultural Research Center, Athens, GA 30604-5677, USA.
Avian Dis. 2005 Dec;49(4):577-81. doi: 10.1637/7337-020205R.1.
We previously reported the recovery of Campylobacter (naturally colonized) from the ductus deferens of 5 of 101 broiler breeder roosters, and four of those five positive roosters had previously produced Campylobacter-positive semen samples. Those results prompted further evaluation to determine if inoculation route influenced the prevalence or level of Campylobacter contamination of semen, the digestive tract, or reproductive organs. Individually caged roosters, confirmed to be feces and semen negative for Campylobacter, were challenged with a marker strain of Campylobacter jejuni either orally using 1.0 ml of a diluted cell suspension (log(10)4.3 to 6.0 cells), by dropping 0.1 ml of suspension (log(10)5.3 to 7.0 cells) on the everted phallus immediately after semen collection or by dip coating an ultrasound probe in the diluted cell suspension (log(10)4.3 to 6.0 cells) and then inserting the probe through the vent into the colon. Six days postinoculation, individual feces and semen samples were again collected and cultured for Campylobacter. Seven days postinoculation, roosters were killed, the abdomen aseptically opened to expose the viscera, and one cecum, one testis, and both ductus deferens were collected. The samples were then suspended 1:3 (weight/volume) in Bolton enrichment broth for the culture of Campylobacter. Samples were also directly plated onto Cefex agar to enumerate Campylobacter. Campylobacter was recovered 6 days after challenge from feces in 82% of samples (log(10)4.1 colony-forming units [CFU]/g sample), 85% of semen samples (log(10)2.9 CFU/ml), and on the seventh day postchallenge from 88% of cecal samples (log(10)5.8 CFU/g sample). Campylobacter was not directly isolated from any testis sample but was detected following enrichment from 9% (3/33) of ductus deferens samples. Roosters challenged with Campylobacter orally, on the phallus, or by insertion of a Campylobacter dip-coated ultrasound probe were all readily colonized in the ceca and produced Campylobacter-positive semen and feces on day 6 after challenge. The low prevalence of recovery of Campylobacter from the ductus deferens samples and failure to recover from any testis sample suggests that semen may become Campylobacter positive while traversing the cloaca upon the everted phallus. The production of Campylobacter-positive semen could provide a route in addition to fecal-oral for the horizontal transmission of Campylobacter from the rooster to the reproductive tract of the hen.
我们之前报告过,在101只肉种鸡公鸡中,有5只的输精管中检测到弯曲杆菌(自然定植),这5只呈阳性的公鸡中有4只此前精液样本检测出弯曲杆菌阳性。这些结果促使我们进一步评估接种途径是否会影响精液、消化道或生殖器官中弯曲杆菌污染的发生率或水平。单独笼养的公鸡,经确认粪便和精液中弯曲杆菌呈阴性,用空肠弯曲杆菌标记菌株进行攻毒,攻毒途径分别为:口服1.0 ml稀释的细胞悬液(10的4.3至6.0次方个细胞),采精后立即在翻出的阴茎上滴加0.1 ml悬液(10的5.3至7.0次方个细胞),或将超声探头浸入稀释的细胞悬液(10的4.3至6.0次方个细胞)后经泄殖腔插入结肠。接种后6天,再次采集个体粪便和精液样本并培养弯曲杆菌。接种后7天,处死公鸡,无菌打开腹腔暴露内脏,采集一段盲肠、一个睾丸和双侧输精管。然后将样本按1:3(重量/体积)悬浮于博尔顿增菌肉汤中培养弯曲杆菌。样本也直接接种于头孢克肟琼脂平板上以计数弯曲杆菌。攻毒后6天,82%的粪便样本(10的4.1次方个菌落形成单位[CFU]/g样本)、85%的精液样本(10的2.9次方个CFU/ml)中检测到弯曲杆菌,攻毒后第7天,88%的盲肠样本(10的5.8次方个CFU/g样本)中检测到弯曲杆菌。未从任何睾丸样本中直接分离出弯曲杆菌,但从9%(3/33)的输精管样本增菌后检测到弯曲杆菌。经口服、阴茎或插入浸有弯曲杆菌的超声探头攻毒的公鸡,盲肠均易被定植,攻毒后第6天精液和粪便检测出弯曲杆菌阳性。从输精管样本中分离出弯曲杆菌的发生率较低,且未从任何睾丸样本中分离出弯曲杆菌,这表明精液在经翻出的阴茎穿过泄殖腔时可能会被弯曲杆菌污染。弯曲杆菌阳性精液的产生可能为弯曲杆菌从公鸡水平传播至母鸡生殖道提供了一条除粪-口途径之外的传播途径。