Abstract
Full text: Infection with Mycoplasma spp, typically M. bovis, is an important disease complex of dairy cattle. Mycoplasma spp can cause mastitis, arthritis, metrititis, pneumonia, septicemia, and death of cattle. Standard microbial cultures of milk samples do not isolate Mycoplasma spp; special methods are necessary. Mycoplasma infections have been reported as contagious in nature, primarily by milking machines and respiratory spread. Bulk tank milk samples (n = 5 samples per tank) were collected from all bulk tanks on most dairy farms in Utah, USA (n = 222 farms, 292 tanks) at 3-4 day intervals, resulting in a sensitivity of 97% for Mycoplasma spp. Mycoplasma was detected on 16/222 dairy farms in Utah (7%), a relatively high prevalence compared to the rest of the USA. After initial surveillance, follow up was conducted on positive farms. One farm milking approximately 4500 Holstein cows in dry lot and free stall housing experienced an outbreak of clinical mastitis (CM) caused by Mycoplasma spp., affecting 35 cows per month vs. the endemic rate of approximately 3 CM cases per month (aseptic milk samples from all CM cases were cultured from this herd). Bedding sand was used following a recycling and manure separation process on the
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Wilson, D J;
Trujillo, J;
Justice-Allen, A. , E-mail: David.Wilson@usu.edu;
[1]
Goodell, G
[2]
- Utah State University, Logan, UT (United States)
- Dairy Authority, Greeley, CO (United States)
Citation Formats
Wilson, D J, Trujillo, J, Justice-Allen, A. , E-mail: David.Wilson@usu.edu, and Goodell, G.
Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals.
IAEA: N. p.,
2009.
Web.
Wilson, D J, Trujillo, J, Justice-Allen, A. , E-mail: David.Wilson@usu.edu, & Goodell, G.
Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals.
IAEA.
Wilson, D J, Trujillo, J, Justice-Allen, A. , E-mail: David.Wilson@usu.edu, and Goodell, G.
2009.
"Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals."
IAEA.
@misc{etde_21287167,
title = {Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals}
author = {Wilson, D J, Trujillo, J, Justice-Allen, A. , E-mail: David.Wilson@usu.edu, and Goodell, G}
abstractNote = {Full text: Infection with Mycoplasma spp, typically M. bovis, is an important disease complex of dairy cattle. Mycoplasma spp can cause mastitis, arthritis, metrititis, pneumonia, septicemia, and death of cattle. Standard microbial cultures of milk samples do not isolate Mycoplasma spp; special methods are necessary. Mycoplasma infections have been reported as contagious in nature, primarily by milking machines and respiratory spread. Bulk tank milk samples (n = 5 samples per tank) were collected from all bulk tanks on most dairy farms in Utah, USA (n = 222 farms, 292 tanks) at 3-4 day intervals, resulting in a sensitivity of 97% for Mycoplasma spp. Mycoplasma was detected on 16/222 dairy farms in Utah (7%), a relatively high prevalence compared to the rest of the USA. After initial surveillance, follow up was conducted on positive farms. One farm milking approximately 4500 Holstein cows in dry lot and free stall housing experienced an outbreak of clinical mastitis (CM) caused by Mycoplasma spp., affecting 35 cows per month vs. the endemic rate of approximately 3 CM cases per month (aseptic milk samples from all CM cases were cultured from this herd). Bedding sand was used following a recycling and manure separation process on the farm; sand samples were cultured for mycoplasmas and other bacteria during the outbreak. Acholeplasma laidlawii was found in one sample, 2 samples were positive for M. bovis by PCR, and one month later 14/20 cow pens' and bedding samples tested Modified Hayflick medium culture-positive for Mycoplasma spp. (testing by 3 different laboratories). During the same month, one recycled bedding sand sample and one cow pen sand sample tested PCR-positive at the Utah Veterinary Diagnostic Laboratory; amplicon sequencing of both isolates showed 99% homology with M. bovis. Positive bedding sand (18,000 kg) was transported from the farm to Utah State University and stored in a pile outdoors. As the weather progressed from late winter (March) to summer (May), the colony forming units/gram (cfu/g) of Mycoplasma spp. decreased. From 770,000 and 720,000 cfu/g on the surface and 2,130,000 and 990,000 cfu/g deep in the pile during the first 2 weeks, surface counts all were negative and the deep counts decreased steadily, below 150,000/g during April and to 2,200 -7,800 cfu/g during May and June, then negative through July. Controlled incubation temperatures for 24 -72 hr showed the following: - 20 deg. C - negative (n = 4); 4 deg. C - 64,000, 59,400, 6,000 cfu/g and negative; 37 deg. C - 25,400 cfu/g in wet slurry, all 4 other (dry) samples negative; 450 C - negative (n = 1); 60 deg. C - negative (n = 2). Together with the weather temperature data (not shown), results suggested that frozen or warm conditions did not support mycoplasmal growth in sand as well as cool temperatures, approximately 4 deg. C. Mycoplasma also grew readily deep within the sand pile, but not as much on the surface. During July, 3 recycled sand bedding piles tested on the initial farm and one each from 2 other project follow up farms with mycoplasma-positive cows were culture-positive for Mycoplasma spp. Chemical disinfection experiments were unsuccessful except for use of 10% bleach (0.5% sodium hypochlorite) solution in a pan of sand for 10 minutes at room temperature. A mycoplasma-negative closed herd was identified and tested until 32/32 calves were negative for mycoplasmas on nasal and ear swabs, resulting in probability > 99% that the herd was negative. Bull calves (n = 12) were then obtained from the negative herd and 6 were exposed to Mycoplasma spp.-positive sand bedding for 28-34 d, until the sand became culture negative as freezing temperatures developed. Four calves were exposed from 24-30 d old to 52-64 d old, and 2 calves were exposed from a few days old to 15 -21 d old. The remaining 6 calves were controls, bedded with mycoplasma-negative sand. Preliminary results are: 86 nasal and ear swabs collected every 7 d and 17 tracheal swabs collected at 35 d (n = 9) and 70 d (n = 8) after exposure to either mycoplasmal or controls and have all been negative for mycoplasmas. (author)}
place = {IAEA}
year = {2009}
month = {Jul}
}
title = {Mycoplasma diagnosis by PCR from bedding of mycoplasmal dairy herds and association with disease in dairy animals}
author = {Wilson, D J, Trujillo, J, Justice-Allen, A. , E-mail: David.Wilson@usu.edu, and Goodell, G}
abstractNote = {Full text: Infection with Mycoplasma spp, typically M. bovis, is an important disease complex of dairy cattle. Mycoplasma spp can cause mastitis, arthritis, metrititis, pneumonia, septicemia, and death of cattle. Standard microbial cultures of milk samples do not isolate Mycoplasma spp; special methods are necessary. Mycoplasma infections have been reported as contagious in nature, primarily by milking machines and respiratory spread. Bulk tank milk samples (n = 5 samples per tank) were collected from all bulk tanks on most dairy farms in Utah, USA (n = 222 farms, 292 tanks) at 3-4 day intervals, resulting in a sensitivity of 97% for Mycoplasma spp. Mycoplasma was detected on 16/222 dairy farms in Utah (7%), a relatively high prevalence compared to the rest of the USA. After initial surveillance, follow up was conducted on positive farms. One farm milking approximately 4500 Holstein cows in dry lot and free stall housing experienced an outbreak of clinical mastitis (CM) caused by Mycoplasma spp., affecting 35 cows per month vs. the endemic rate of approximately 3 CM cases per month (aseptic milk samples from all CM cases were cultured from this herd). Bedding sand was used following a recycling and manure separation process on the farm; sand samples were cultured for mycoplasmas and other bacteria during the outbreak. Acholeplasma laidlawii was found in one sample, 2 samples were positive for M. bovis by PCR, and one month later 14/20 cow pens' and bedding samples tested Modified Hayflick medium culture-positive for Mycoplasma spp. (testing by 3 different laboratories). During the same month, one recycled bedding sand sample and one cow pen sand sample tested PCR-positive at the Utah Veterinary Diagnostic Laboratory; amplicon sequencing of both isolates showed 99% homology with M. bovis. Positive bedding sand (18,000 kg) was transported from the farm to Utah State University and stored in a pile outdoors. As the weather progressed from late winter (March) to summer (May), the colony forming units/gram (cfu/g) of Mycoplasma spp. decreased. From 770,000 and 720,000 cfu/g on the surface and 2,130,000 and 990,000 cfu/g deep in the pile during the first 2 weeks, surface counts all were negative and the deep counts decreased steadily, below 150,000/g during April and to 2,200 -7,800 cfu/g during May and June, then negative through July. Controlled incubation temperatures for 24 -72 hr showed the following: - 20 deg. C - negative (n = 4); 4 deg. C - 64,000, 59,400, 6,000 cfu/g and negative; 37 deg. C - 25,400 cfu/g in wet slurry, all 4 other (dry) samples negative; 450 C - negative (n = 1); 60 deg. C - negative (n = 2). Together with the weather temperature data (not shown), results suggested that frozen or warm conditions did not support mycoplasmal growth in sand as well as cool temperatures, approximately 4 deg. C. Mycoplasma also grew readily deep within the sand pile, but not as much on the surface. During July, 3 recycled sand bedding piles tested on the initial farm and one each from 2 other project follow up farms with mycoplasma-positive cows were culture-positive for Mycoplasma spp. Chemical disinfection experiments were unsuccessful except for use of 10% bleach (0.5% sodium hypochlorite) solution in a pan of sand for 10 minutes at room temperature. A mycoplasma-negative closed herd was identified and tested until 32/32 calves were negative for mycoplasmas on nasal and ear swabs, resulting in probability > 99% that the herd was negative. Bull calves (n = 12) were then obtained from the negative herd and 6 were exposed to Mycoplasma spp.-positive sand bedding for 28-34 d, until the sand became culture negative as freezing temperatures developed. Four calves were exposed from 24-30 d old to 52-64 d old, and 2 calves were exposed from a few days old to 15 -21 d old. The remaining 6 calves were controls, bedded with mycoplasma-negative sand. Preliminary results are: 86 nasal and ear swabs collected every 7 d and 17 tracheal swabs collected at 35 d (n = 9) and 70 d (n = 8) after exposure to either mycoplasmal or controls and have all been negative for mycoplasmas. (author)}
place = {IAEA}
year = {2009}
month = {Jul}
}