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1.  Efficacy and Pharmacokinetics of Bacteriophage Therapy in Treatment of Subclinical Staphylococcus aureus Mastitis in Lactating Dairy Cattle†  
Bovine mastitis is an inflammation of the udder caused by microbial infection. Mastitis caused by Staphylococcus aureus is a major concern to the dairy industry due to its resistance to antibiotic treatment and its propensity to recur chronically. Growing concerns surrounding antibiotic resistance have spurred research into alternative treatment methods. The ability of lytic S. aureus bacteriophage K to eliminate bovine S. aureus intramammary infection during lactation was evaluated in a placebo-controlled, multisite trial. Twenty-four lactating Holstein cows with preexisting subclinical S. aureus mastitis were treated. Treatment consisted of 10-ml intramammary infusions of either 1.25 × 1011 PFU of phage K or saline, administered once per day for 5 days. The cure rate was established by the assessment of four serial samples collected following treatment. The cure rate was 3 of 18 quarters (16.7%) in the phage-treated group, while none of the 20 saline-treated quarters were cured. This difference was not statistically significant. The effects of phage intramammary infusion on the bovine mammary gland were also studied. In healthy lactating cows, a single infusion of either filter-sterilized broth lysate or a CsCl gradient-purified phage preparation elicited a large increase in the milk somatic cell count. This response was not observed when phage was infused into quarters which were already infected with S. aureus. Phage-infused healthy quarters continued to shed viable bacteriophage into the milk for up to 36 h postinfusion. The phage concentration in the milk suggested that there was significant degradation or inactivation of the infused phage within the gland.
doi:10.1128/AAC.01630-05
PMCID: PMC1563511  PMID: 16940081
2.  Distribution and characteristics of verocytotoxigenic Escherichia coli isolated from Ontario dairy cattle. 
Epidemiology and Infection  1992;108(3):423-439.
Faecal swabs obtained from a random sample of 1131 cows and 659 calves on 100 southern Ontario dairy farms were examined for verocytotoxigenic Escherichia coli (VTEC) using a Vero cell assay. Five isolates from each positive culture were tested similarly. Positive colonies were examined with DNA probes for Shiga-like toxin I (SLT-I) and SLT-II sequences. Probe-negative colonies were tested for neutralization of verocytotoxicity using anti-SLT-I and anti-SLT-IIv antisera. Colonies showing no neutralization response were examined in a polymerase chain reaction procedure. Colonies positive by any test were confirmed to be E. coli biochemically, serotyped, biotyped and tested for antimicrobial resistance. Faecal culture supernatants which were positive in the Vero cell assay, but culture negative, were examined using the neutralization assay. Animals were classified positive by faecal culture supernatant or by positive VTEC isolate. The prevalence rates of VTEC infection in cows and calves were estimated to be 9.5 and 24.7%, respectively. The proportion of animals infected on each farm ranged from 0 to 60% for cows and 0 to 100% for calves. Of 206 VTEC isolates identified, few were of serotypes which have been isolated from humans and none were E. coli O 157.H7.
PMCID: PMC2272218  PMID: 1601076
3.  In vitro exposure of bovine morulae to Ureaplasma diversum. 
Ureaplasma diversum has been associated with infertility in the cow experimentally and in naturally occurring cases. However, the pathogenic mechanism is undetermined. The purpose of this study was to determine whether ureaplasmas are pathogenic for bovine morulae in vitro. Twenty-one morulae were recovered from three superovulated, mature, Holstein cows six or seven days postestrus. The embryos were divided into three groups (A,B,C) and incubated for 16 hours at 37 degrees C in humidified air with 10% CO2. Group A was incubated in embryo culture medium alone, Group B was incubated in culture medium with sterile ureaplasma broth added and Group C was incubated in culture medium containing 1.7 X 10(6) colony forming units Ureaplasma diversum strain 2312. After incubation, the morulae were examined using an electron microscope. Structures morphologically identical to U. diversum were present on the outer surface of the zonae pellucidae of all the morulae exposed to the organism and none were present on the unexposed control embryos. No other morphological differences were observed in either the ureaplasma-exposed embryos or the two groups of control embryos. Ureaplasma diversum was isolated from three of the five embryos incubated in culture medium with sterile ureaplasma broth added. These three embryos were recovered from one donor cow which cultured positive for U. diversum from the vulva and flush fluid. This finding suggests that the contaminating organisms entered the embryo culture wells either in the embryo collection medium or attached to the embryos.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMCID: PMC1255303  PMID: 3607652
4.  The effects of gonadotrophin releasing hormone administration four days after insemination on first-service conception rates and corpus luteum function in dairy cows. 
One hundred and eighty-five Holstein-Friesian dairy cows received either sterile water or 250 micrograms of gonadotrophin releasing hormone intramuscularly on the fourth day after the first service postpartum. Heparinized blood samples were taken immediately prior to treatment (day 4) and on day 8 postinsemination for analysis of plasma progesterone concentration. Pregnancy diagnosis was carried out by rectal palpation at 42 days postinsemination and reconfirmed after 60 days postbreeding. The pregnancy rates after first, second or third service were not significantly different between gonadotrophin releasing hormone-treated and control cows. Plasma progesterone concentrations on day 4 and day 8 postinsemination, as well as the change in plasma progesterone concentration from day 4 to day 8, were similar for gonadotrophin releasing hormone-treated and control cows. The plasma progesterone concentrations on day 8 postbreeding were significantly higher (p less than 0.005) and the change in progesterone concentrations between days 4 and 8 were significantly greater (p less than 0.002) in pregnant cows compared to nonpregnant cows.
PMCID: PMC1255187  PMID: 3530415
5.  Observations on Intramammary Infections in First Calf Heifers in Early Lactation 
The Canadian Veterinary Journal  1986;27(3):112-115.
Intramammary infections and mastitis were monitored on four occasions at three-week intervals in 61 first calf heifers in five dairy herds during the first ten weeks of lactation. Of 940 quarter milk samples examined 65% were classed as negative, 10.4% as positive for mastitis (mainly subclinical), 1.8% as group 3 (infection present but no elevation in somatic cell count) and 22.8% as group 4 (elevated somatic cell count). Seventy-seven percent of the infections detected and identified were those due to coagulase-negative staphylococci, the main species being S. hyicus, S. epidermidis, S. simulans and S. hominis. Other infections detected with Corynebacterium pyogenes (three samples), Escherichia coli (one sample), Micrococcus spp. (one sample), S. aureus (two samples) and Streptococci (non-agalactiae) (seven samples).
The geometric mean somatic cell count for 23 quarters infected with coagulase-negative staphylococci was 311 × 103 cells / mL compared to 134 × 103 cells / mL in noninfected adjacent contralateral quarters. The respective figures for% cell volume in Channel 8 (mainly neutrophils) were 10.6% and 3.5%. There was a highly significant association between herd and the proportion of quarter milk samples in the four mastitis categories.
PMCID: PMC1680159  PMID: 17422635
Heifers; mastitis; somatic cell counts
6.  The sensitivity and specificity of postbreeding plasma progesterone levels as a pregnancy test for dairy cows. 
Plasma progesterone levels on day 4 and day 8 postbreeding were measured for one hundred and eighty-four dairy cows. These two parameters (PPD4, PPD8), their absolute difference (PPDIFF) and their ratio (PPRATIO) were assessed for their ability to identify cows not conceiving, using the principles of sensitivity and specificity. PPD4 was significantly higher (p less than 0.10) and PPD8, PPDIFF and PPRATIO were significantly lower (p less than 0.01) in cows remaining open than in pregnant cows. Evaluating each parameter separately, PPDIFF greater than 3.00 units had the highest specificity, 85.7%, but a low sensitivity (27.0%). Combining two parameters using series interpretation to increase specificity resulted in the best combination of specificity (87%) and sensitivity (27%). Maximum specificity was 97% for PPD4 less than or equal to 1.00 units and PPD8 greater than 4.00 units, and also for PPD4 less than or equal to 1.00 units and PPDIFF greater than 3.00 units, but sensitivity was very low (7% and 10% respectively). Predictive values of the test results with the best specificity were evaluated; given the population pregnancy rate of 54%, none exceeded 50%, indicating that the plasma progesterone parameters were not very useful for identifying open dairy cows.
PMCID: PMC1236186  PMID: 4041979
7.  Effects of gonadotrophin releasing hormone on reproductive performance of dairy cows with retained placenta. 
The effects of gonadotrophin releasing hormone (GnRH) on the reproductive performance of dairy cows with retained placenta were studied. Three hundred and seventy-eight cows diagnosed as having retained placenta received intramuscular injections of either 2 mL sterile water or 200 micrograms of GnRH in 2 mL sterile water between day 8 and day 14 postpartum. Rectal palpation was performed at the time of treatment and ten to 20 days after treatment in order to determine the rate of uterine involution. Thereafter, monthly rectal examinations were carried out until insemination. Pregnancy diagnosis was made by rectal palpation at 40 days or more after breeding. Using the entire experimental population, there were no significant differences between GnRH-treated and control cows for the rate of uterine involution, the occurrence of reproductive problems, the interval from parturition to first observed estrus, the interval from parturition to first insemination, the interval from parturition to conception, the number of services per conception, the total number of services per cow regardless of conception and the incidence of culling for infertility. When the data for herds in which breeding began earlier in the postpartum period (herds having a mean less than or equal to 80 days from parturition to first service for retained placenta cows) were considered, the GnRH treatment resulted in a significantly shorter (p less than or equal to 0.01) calving to conception interval as compared to control cows. Also, there was a significant reduction (p less than or equal to 0.05) in the total number of services per cow regardless of conception and a significant reduction in the interval from parturition to first service.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID: PMC1236082  PMID: 6391640
8.  Reproductive performance in dairy cows following postpartum treatment with gonadotrophin releasing hormone and/or prostaglandin: a field trial. 
Three hundred and five Holstein Friesian cows were given either 250 micrograms gonadotrophin releasing hormone (GnRH) or saline on day 15 postpartum followed by 500 micrograms cloprostenol or saline on day 24 postpartum. Four treatment groups were formed using random allocation: Group I -- placebo (Day 15)/placebo (Day 24), Group II -- GnRH (Day 15)/placebo (Day 24), Group III -- placebo (Day 15)/cloprostenol (Day 24), Group IV -- GnRH (Day 15)/cloprostenol (Day 24). Double blind techniques were used during the follow-up period. Rectal palpation, to assess uterine involution and ovarian activity was performed just prior to each treatment and again at 28 days postpartum. In addition blood samples were collected at 15, 24 and 28 days postpartum for measurement of plasma progesterone. There were no significant differences among treatment groups with respect to services per conception, number of heats detected before first service and culling for infertility. Cows treated only with GnRH had an increased calving to first estrus and calving to first breeding interval, and tended to have an increased calving to conception interval. Treatment with cloprostenol significantly decreased calving to conception and calving to first observed estrus intervals. Treatment with GnRH on day 15 postpartum resulted in a significant increase in the subsequent incidence of pyometra and prebreeding anestrus. On the other hand, cloprostenol treatment on day 24 postpartum resulted in a decreased incidence of pyometra, regardless of GnRH treatment and a decreased incidence of prebreeding anestrus in GnRH treated cows compared to cows receiving only GnRH at day 15 postpartum.(ABSTRACT TRUNCATED AT 250 WORDS)
PMCID: PMC1236054  PMID: 6383577
9.  Plasma Progesterone Concentrations in Dairy Cows with Cystic Ovaries and Clinical Responses Following Treatment with Fenprostalene 
The Canadian Veterinary Journal  1983;24(11):352-356.
Sixty-two dairy cows diagnosed as having cystic ovarian degeneration were used to study the correlation between rectal palpation findings and plasma progesterone concentrations and the response of cysts to treatment using fenprostalene, a luteolytic agent. Rectal palpation accurately determined the presence of luteal cysts as confirmed by plasma progesterone concentrations of 3 ng/mL or more. Treatment with fenprostalene was very effective for luteal cysts: a high percentage of treated cows exhibited estrus within seven days after treatment. The conception rate following artifical insemination during the induced estrus was 87.5% (21/24). Rectal palpation was much less accurate for the diagnosis of follicular cysts. Cows diagnosed as having follicular cysts had wide variations in plasma progesterone concentrations. Response to fenprostalene treatment was poor in cows with nonluteinized cystic follicles associated with low progesterone concentrations. However, cows diagnosed as having follicular cysts, but with progesterone concentrations of 1 ng/mL or more, responded better to fenprostalene treatment than cows with low progesterone concentrations.
It was concluded that, if correctly diagnosed, luteal cysts can be successfully treated with fenprostalene, and conception rates following treatment can be expected to be normal.
PMCID: PMC1790459  PMID: 17422330
10.  The Effects of Gonadotrophin Releasing Hormone Administration in Early Postpartum Dairy Cows on Hormone Concentrations, Ovarian Activity and Reproductive Performance: A Review 
The Canadian Veterinary Journal  1983;24(4):116-122.
Gonadotrophin releasing hormones have become widely used hormonal compounds in veterinary medicine, particularly with respect to bovine reproduction. The character and physiological actions of gonadotrophin releasing hormone are briefly reviewed and its clinical applications are summarized. The endocrinological research concerned with the use of gonadotrophin releasing hormone in the early postpartum period is discussed. Field trials which have been conducted to assess the effects of postpartum gonadotrophin releasing hormone administration on reproductive performance have varied widely in both design and interpretation of results. These experiments are reviewed, including the clinical trials using normal cows as well as those on cows with retained placenta.
PMCID: PMC1790294  PMID: 17422245
11.  The Events of Normal and Abnormal Postpartum Reproductive Endocrinology and Uterine Involution in Dairy Cows: A Review 
The results of numerous hormonal profile studies in the dairy cow are reviewed with respect to specific hormone changes in the early postpartum period, and factors contributing to the onset of cyclic reproductive activity. Variations from the normal pattern of postpartum hormonal activity or uterine involution are reviewed also. The normal involutionary changes of the bovine uterus are discussed.
PMCID: PMC1790317  PMID: 17422232
12.  An evaluation of protein/fat ratio in first DHI test milk for prediction of subsequent displaced abomasum in dairy cows. 
First DHI test milk that was sampled prior to displaced abomasum (DA) diagnosis was used to evaluate milk protein/fat ratio (PFR) for prediction of subsequent DA in dairy cows. Odds ratio, sensitivity, specificity, and likelihood ratio were determined. Twenty-seven DA cases were matched to 3 controls per case by herd and calving date. Milk was tested at a median of 19 d after calving, which was 8 d prior to the median time of DA diagnosis. Adjusted for parity and days in milk, a protein/fat ratio < or = 0.72 was 8.2 times more likely to come from a cow subsequently diagnosed with DA than a protein/fat ratio > 0.72. Using the cut off value of 0.72, the sensitivity of PFR for DA was 80% and the specificity was 68%. A receiver operating characteristics curve indicated that the minimum sum of false negative and false positive results was at a PFR cut off value of 0.72. The likelihood ratio indicated that protein/fat ratios < or = 0.62 are 3.8 times more likely to come from cows that are diagnosed subsequently with DA than from cows without DA. The protein/fat ratio in 1st DHI test milk may predict subsequent DA in dairy cows.
PMCID: PMC1189462  PMID: 9553715
13.  Clinical mastitis in dairy cattle in Ontario: frequency of occurrence and bacteriological isolates. 
The objective of this study was to describe the frequency of occurrence of clinical mastitis in dairy herds in Ontario. The study group consisted of 65 dairy farms involved in a 2-year observational study, which included recording all clinical mastitis cases and milk sampling of quarters with clinical mastitis. Lactational incidence risks of 9.8% for abnormal milk only, 8.2% for abnormal milk with a hard or swollen udder, and 4.4% for abnormal milk plus systemic signs of illness related to mastitis were calculated for 2840 cows and heifers. Overall, 19.8% of cows experienced one or more cases of clinical mastitis during location. Teat injuries occurred in 2.1% of lactations. Standard bacteriology was performed on pretreatment milk samples from 834 cows with clinical mastitis. The bacteria isolated were Staphylococcus aureus (6.7%), Streptococcus agalactiae (0.7%), other Streptococcus spp. (14.1%), coliforms (17.2%), gram-positive bacilli (5.5%), Corynebacterium bovis (1.7%), and other Staphylococcus spp. (28.7%). There was no growth in 17.7% of samples, and 8.3% of samples were contaminated. Clinical mastitis is a common disease in dairy cows in Ontario; approximately 1 in 5 cow lactations have at lease one episode of clinical mastitis. There is, however, considerable variation in the incidence of clinical mastitis among farms. The majority of 1st cases of clinical mastitis occur early in lactation, and the risk of clinical mastitis increases with increasing parity. Environmental, contagious, and minor pathogens were all associated with cases of clinical mastitis.
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PMCID: PMC1539829  PMID: 9442950
14.  Use of test day milk fat and milk protein to detect subclinical ketosis in dairy cattle in Ontario. 
The Canadian Veterinary Journal  1997;38(11):713-718.
Serum beta-hydroxybutyrate (BHB) levels were determined for 1333 dairy cows in various stages of lactation and parity on 93 dairy farms in Ontario. The data were collected in a cross-sectional manner, as part of the 1992 Ontario Dairy Monitoring and Analysis Program. The median serum BHB was 536 mumol/L for all cows, with a range of 0 to 5801 mumol/L. When subclinical ketosis was defined as a serum BHB level of 1200 mumol/L or higher, the prevalence of ketosis for cows in early lactation (< 65 days in milk (DIM)) was 14.1%. Prevalences for mid lactation (65-149 DIM), late lactation (> 149 DIM), and dry cows were 5.3%, 3.2%, and 1.6%, respectively. The mean serum BHB was significantly higher in the early group compared with each of the other 3 groups (P < 0.05). There was a trend of increasing prevalence with increasing parity across all stages of lactation. Only the difference between the parity-1 group and the parity-4 and greater group was statistically significant (P < 0.05). Both test-day fat percent and test-day protein percent were significantly associated with subclinical ketosis. However, test-day fat percent and test-day protein percent, used alone or in combination, were not useful screening tests for identifying cows with subclinical ketosis.
PMCID: PMC1576823  PMID: 9360791
15.  Dynamics and regulation of bulk milk somatic cell counts. 
Somatic cell count (SCC) in milk is inversely related to dairy cow productivity and milk quality. In an effort to improve product quality, and indirectly farm productivity, regulatory limits on somatic cell counts have been established by many of the major dairy producing countries. The purpose of this paper was to assess the impact of regulations on bulk milk somatic cell counts in Ontario and to assist producers in meeting regulatory limits through development of prediction models. Through the use of a transfer function model, provincial SCC was found to have dropped by approximately 60,000 as a result of the reduction program. Limits of the regulatory program, seasonality and herd characteristics were found through time series cross-sectional models to have an impact on prediction of SCC at the farm level, but the major influence was historical SCC levels.
PMCID: PMC1263607  PMID: 8490807
16.  A case-control study of selected pathogens including verocytotoxigenic Escherichia coli in calf diarrhea on an Ontario veal farm. 
A case-control study of diarrheal disease in veal calves was conducted over a three month period on a single large veal farm in southern Ontario. One hundred diarrheic calves (cases) were identified by visual examination of their feces. Each case was matched to two nondiarrhetic controls from the same room on the same day, and a fecal sample was obtained from each animal. Fecal consistency of cases and controls was observed daily for one week following sample collection. Control calves which developed diarrhea during that period were excluded from the study. Breed, sex and the date and nature of antimicrobial drugs administered to each calf were recorded. Moisture content of fecal samples was measured by weighing samples before and after oven drying. Samples were screened for verocytotoxigenic Escherichia coli (VTEC) using a Vero cell assay, for enterotoxigenic E. coli (ETEC) using an immunoblot procedure with anti-K99 monoclonal antibodies, and for Salmonella species using modified semi-solid Rappaport-Vassiliadis medium. A latex agglutination test was used to detect rotaviruses, and samples were examined for cryptosporidia using sucrose wet mounts. No VTEC were identified in cases or controls. One calf was positive for Salmonella and three were positive for ETEC. Rotaviruses were detected in four cases and four controls. A significant positive association was found between diarrhea and infection with Cryptosporidium. This study thus provided no evidence of an association between diarrhea and infection with either VTEC, ETEC, Salmonella spp. or rotaviruses in the population examined. On the other hand our results do suggest that Cryptosporidium infection may promote transient diarrheal disease in veal calves in Ontario.
PMCID: PMC1263535  PMID: 1330276
17.  Observations on intramammary infection and somatic cell counts in cows treated with recombinant bovine somatotropin. 
Data were collected on udder health variables as part of a study of the effects of recombinant bovine somatotropin on production in lactating dairy cows. Milk samples, obtained at three intervals during the study, were assessed for their somatic cell count and bacteriological culture result. There was an increase in the prevalence of infection at mid-lactation in the 20.6 and 41.2 mg per day treatment groups as compared to the controls. There was no difference detected in the mean cell count between groups from the samples collected pretrial, mid-lactation, or late lactation. However, analysis of the individual cow Dairy Herd Improvement somatic cell count data showed a difference between groups which was most evident in mid-lactation.
PMCID: PMC1263443  PMID: 1884302
18.  Effects of progesterone and human chorionic gonadotrophin administration five days postinsemination on plasma and milk concentrations of progesterone and pregnancy rates of normal and repeat breeder dairy cows. 
Treatment with a progesterone-releasing intravaginal device between days 5 and 12 after estrus elevated (p less than 0.05) plasma progesterone concentrations between days 6 and 8 in comparison with controls. Treatment with injectable progesterone (200 mg) on days 5, 7, 9 and 11 postestrus did not increase plasma progesterone concentrations over controls. The administration of 1500 IU human chorionic gonadotrophin (hCG) on day 5 after estrus resulted in a sustained increase (p less than 0.01) in plasma progesterone concentrations from day 8 until day 20 when measurements ceased. Pregnancy rates, as a result of artificial insemination (AI) at the pretreatment estrus, in these treatments (n = 12-14 each), were unaffected by any of the treatments and ranged from 57.1 to 75.0% at 45-60 days post-AI. In a field trial, of 36 repeat breeder cows treated with 1500 IU hCG 5.5 days after insemination, 47.2% were pregnant at 60 days, whereas 39.5% of saline-treated controls were diagnosed pregnant. Treatment with hCG significantly (p less than 0.05) increased milk progesterone concentrations over controls on days 14 and 20 after insemination.
PMCID: PMC1255660  PMID: 2379109

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