Yersinia enterocolitica of porcine origin: carriage of virulence genes and genotypic diversity.

Yersinia enterocolitica is an important foodborne pathogen, and pigs are recognized as a major reservoir and potential source of pathogenic strains to humans. A total of 172 Y. enterocolitica recovered from conventional and antimicrobial-free pig production systems from different geographic regions (North Carolina, Ohio, Michigan, Wisconsin, and Iowa) were investigated to determine their pathogenic significance to humans. Phenotypic and genotypic diversity of the isolates was assessed using antibiogram, serogrouping, and amplified fragment length polymorphism (AFLP). Carriage of chromosomal and plasmid-borne virulence genes were investigated using polymerase chain reaction. A total of 12 antimicrobial resistance patterns were identified. More than two-thirds (67.4%) of Y. enterocolitica were pan-susceptible, and 27.9% were resistant against ß-lactams. The most predominant serogroup was O:3 (43%), followed by O:5 (25.6%) and O:9 (4.1%). Twenty-two of 172 (12.8%) isolates were found to carry Yersinia adhesion A (yadA), a virulence gene encoded on the Yersinia virulence plasmid. Sixty-nine (40.1%) isolates were found to carry ail gene. The ystA and ystB genes were detected in 77% and 26.2% of the strains, respectively. AFLP genotyping of isolates showed wide genotypic diversity and were grouped into nine clades with an overall genotypic similarity of 66.8-99.3%. AFLP analysis revealed that isolates from the same production system showed clonal relatedness, while more than one genotype of Y. enterocolitica circulates within a farm.

Evaluation of intra-abdominal pressure in horses that crib.

OBJECTIVE: To measure intra-abdominal pressure (IAP) in horses that crib and compare it with IAP in horses that do not have this vice. STUDY DESIGN: Cohort study. ANIMALS: Healthy cribbing horses (cribbing cohort, n = 8) and 8 healthy noncribbing horses (noncribbing cohort). METHODS: A microsensor catheter was introduced into the peritoneal cavity through the right paralumbar fossa, using local anesthesia, for measurement of IAP. These pressures were recorded in 1-minute intervals for 2 hours, while the horses were standing tied in a stall. IAPs of cribbing horses were compared to the noncribbing cohort. RESULTS: Baseline IAPs were not significantly different between cribbing and noncribbing cohorts (P = .076); however, IAPs in the cribbing cohort were significantly increased when compared with the noncribbing cohort, during active cribbing behavior (P = .0016). Frequency of cribbing was not associated with increased IAP (P = .35). IAPs in the cribbing cohort remained significantly elevated compared with the noncribbing cohort, even after the behavior had ceased (P = .0002). CONCLUSION: Cribbing is associated with increased IAP in the horse, both during and after the behavior.

Milk yield and somatic cell count during the following lactation after selective treatment of cows at dry-off.

Selective dry cow therapy (SDCT) has received increasing attention in recent years owing to global concerns over agricultural use of antimicrobial drugs and development of antimicrobial resistance. The objective of this study was to evaluate the effect of SDCT on milk yield and somatic cell count (SCC) in dairy herds in the USA. Cows in four Ohio dairy herds were categorized into two groups (low-SCC and high-SCC) at dry-off based on their SCC and clinical mastitis (CM) history during the lactation preceding the dry-off. Low-SCC cows were randomly assigned to receive or not to receive intramammary antibiotics at dry-off. Milk yield and SCC of these cows during the following lactation were compared using linear mixed effects models, adjusting for parity, calving season, stage of lactation, previous lactation milk yield and herd. Milk yield of untreated and treated low-SCC cows at dry-off did not differ significantly during the following lactation. Overall, treated low-SCC cows had 16% lower SCC (approximately 35 000 cells/ml, P = 0·0267) than the untreated cows during the following lactation; however, the effect was variable in different herds. Moreover the impact of treatment, or the lack thereof, on milk yield varied considerably between herds. The results suggested that in some herds treating all cows at dry-off may be beneficial while in other herds leaving healthy cows without antibiotic dry cow treatment has no negative impact on milk yield or milk quality (SCC), and in fact, may be beneficial. Further studies are needed to identify characteristics of herds where treating all cows routinely at dry-off may be needed for maintaining good udder health and where switching to selective treatment of cows at dry-off would be the optimal approach to achieve best results.

The effect of inoculum volume on the microbiologic detection of naturally occurring Staphylococcus aureus intramammary infections.

Currently no standard definitions for the diagnosis of Staphylococcus aureus intramammary infection (IMI) exist. As a result, criteria applied in research to diagnose S. aureus IMIs have varied making comparisons between published works difficult. The goal of the current study was to define the optimal inoculum volume used in the diagnosis of naturally occurring S. aureus IMIs. Microbiologic results from 2 field studies examining S. aureus IMIs were used to examine the effects of inoculum volume on the microbiologic detection of S. aureus. A total of 1,583 milk samples were included in the analysis, and the results of using a 0.01-ml and a 0.1-ml inoculum are presented. Using a 0.01-ml inoculum resulted in a sensitivity of 91% (95% confidence interval [CI]: 88.6-93%) and a specificity of 99.4% (95% CI: 98.6-99.8%). Using the larger 0.1-ml inoculum resulted in a sensitivity of 96.8% (95% CI: 95.2-97.9%) and a specificity of 99.3% (95% CI: 98.4-99.7%). All false-positive samples were from S. aureus-negative quarters in S. aureus-positive cows. There were no false-positive cultures from S. aureus-negative cows. Of the false-negative samples, the majority (77%) were from 6 of the 34 S. aureus-positive quarters. Results from the current study of naturally occurring S. aureus IMIs support the hypothesis that, when using quarter level milk samples, a S. aureus IMI is most accurately diagnosed using a 0.1-ml inoculum. Regardless of inoculum volume, a single quarter sample culture that is positive with S. aureus (>or=1 colony-forming unit) is sufficient to diagnose a S. aureus IMI.

Association of milk yield and infection status at dry-off with intramammary infections at subsequent calving.

The dry period plays an important role in maintenance of udder health. Cows are most susceptible to intramammary infections (IMI) after dry-off and near parturition and drying-off procedures may affect the likelihood of IMI at calving. The objective of this study was to evaluate the association of milk yield and infection status at dry-off with the likelihood of IMI at calving by examining different drying-off methods. Cows (n=112) at the Ohio State University Waterman Dairy Teaching and Research Herd were randomly assigned to either an intermittent or a standard, twice-daily milking group 1 week prior to dry-off. All quarters of all cows in the herd were treated with an antibiotic dry-cow product after the last milking. Milk samples were collected 1 week prior to dry-off (pre-dry), on the day of dry-off, and within 3 d of parturition to determine infection status of the quarters. Association between IMI at calving and cumulative milk yield for the final week of lactation and drying-off method was examined using generalized estimation equations with logic link, accounting for potential confounders, such as pre-dry and dry-off infection status, and for the correlated data structure due to quarters clustered within cows. Intermittent milking significantly reduced milk yield at the end of lactation. Increasing cumulative milk yield during the last week of lactation was significantly associated with a greater probability of IMI at calving for quarters that were uninfected prior to dry-off: uninfected quarters of cows producing more than 115 kg during the last week of lactation were 7.1-times more likely to be infected at calving (P=0.0081) than uninfected quarters of cows producing less than 75 kg. Even though the overall cure rate over the dry period was relatively high at 84%, the odds of a quarter being infected at calving was 7.6- and 3.3-times higher if it was infected at dry-off with major pathogens (P<0.0001) or minor pathogens (P=0.028), respectively, compared with an uninfected quarter at dry-off. The results suggest that decreasing milk yield prior to dry-off may serve as an effective means to maintain good udder health in a herd.

Comparison of speed of onset and analgesic effect of 2% mepivacaine hydrochloride deposited within or outside the neurovascular bundle at the level of the proximal sesamoid bones in horses with naturally occurring forefoot-related lameness.

OBJECTIVE: To compare the speed of onset and analgesic effect of mepivacaine deposited within or immediately outside the neurovascular bundle at the base of the proximal sesamoid bones in horses. ANIMALS: 6 horses with naturally occurring forefoot-related lameness. PROCEDURES: In a crossover study design, horses were randomly assigned to receive 1 of 2 treatments first, with the second treatment administered 3 to 7 days later. Trotting gait was analyzed with an inertial sensor-based motion analysis system immediately before treatment to determine degree of lameness. Afterward, ultrasound guidance was used to inject 2% mepivacaine hydrochloride around the palmar digital nerves of the affected forelimb at the level of the base of the proximal sesamoid bones either within the subcircumneural space or outside the circumneural sheath. After injection, gait was reevaluated at 5-minute intervals for 45 minutes. RESULTS: Mepivacaine deposition outside the circumneural sheath did not resolve lameness in any horse; for 3 horses, the mean time to 70% reduction of initial vertical head movement was 13.3 minutes, and the remaining 3 horses had no such reduction at any point. Mepivacaine deposition within the subcircumneural space resulted in a mean time to 70% reduction of initial vertical head movement of 6.7 minutes and mean time to resolution of lameness of 21.7 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that when peripheral nerves of horses lie within a sheath, local anesthetic solution should be deposited within the sheath for an effective nerve block. If local anesthetic solution is deposited outside the sheath, the nerve block may yield erroneous results.

Effect of 3% chloroprocaine hydrochloride when used for median and ulnar regional nerve blocks in lame horses.

OBJECTIVE: To assess onset of analgesia for 3% chloroprocaine hydrochloride and 2% mepivacaine hydrochloride when used for median and ulnar nerve blocks in lame horses. ANIMALS: 6 naturally lame horses. PROCEDURES: A crossover experiment was conducted. Horses were assigned to 1 of 2 treatment groups (3% chloroprocaine or 2% mepivacaine first). Median and ulnar nerve blocks were performed in the lame limb with the assigned treatment. Lameness was objectively evaluated before treatment administration and at various points for 120 minutes after treatment with a wireless inertial sensor-based motion analysis system. Following a 7-day washout period, horses then received the other treatment and lameness evaluations were repeated. RESULTS: Median and ulnar nerve blocks performed with 3% chloroprocaine resulted in more consistent, rapid, and profound amelioration of lameness than did blocks performed with 2% mepivacaine. Lameness decreased more between 20 and 40 minutes after injection when 3% chloroprocaine was used than when 2% mepivacaine was used. Complete resolution of lameness was detected a mean of 9 minutes after injection when median and ulnar nerve blocks were performed with 3% chloroprocaine and a mean of 28 minutes after injection when performed with 2% mepivacaine. CONCLUSIONS AND CLINICAL RELEVANCE: 3% chloroprocaine had a more rapid onset and provided better analgesia for median and ulnar nerve blocks in horses with naturally occurring lameness, compared with 2% mepivacaine. These favorable properties suggest that 3% chloroprocaine would be useful for performance of median and ulnar regional nerve blocks during complicated lameness evaluations.

Diagnosis of intramammary infections at dry-off based on sampling strategy, epidemiology of pathogens, and agreement beyond chance.

Isolation of pathogens from duplicate or multiple milk samples is currently considered the gold standard in diagnosis of bovine intramammary infections (IMI). However, in large field studies and especially in normal dairy production conditions, collection of single samples is often the most practical option to determine the causal agents of mastitis in a herd. The objective of the present study was to determine how well results between the first and the second sample in pairs of duplicate and successive quarter milk samples agree, using 5 different IMI definitions, based on the number of colony forming units (CFU) per milliliter of milk and epidemiology of the pathogens isolated. Agreement between microbiologic results from the first and the second sample of a pair was assessed by calculating the percentage of agreement and kappa coefficient. Milk samples collected at dry-off from 561 Holstein cows in 4 Ohio dairy herds were included in the analyses. Results of the study indicate that the agreement between the first and the second sample of a duplicate pair was high when criteria to call a sample positive was adjusted for the number of CFU/ml of milk by considering the epidemiology of different mastitis organisms. This finding suggests that an IMI can be accurately diagnosed with single samples. For contagious pathogens (Staphylococcus aureus) a cutoff of 100 CFU/ml and a cutoff of 1,000 CFU/ml for major environmental and minor pathogens will serve as a sensible approach to diagnose bovine IMI with single milk samples.

Lactoferrin concentrations in bovine milk prior to dry-off.

Concentration of natural protective factors (NPFs) which have the ability to inhibit growth of mastitis-causing pathogens increase rapidly following the cessation of milking of dairy cows. One such NPF is lactoferrin, an iron-binding protein present in high concentrations in dry-cow secretions. Earlier studies have demonstrated that intermittent milking at the end of lactation increases levels of NPFs in milk and may decrease prevalence of intramammary infections at calving; however, most of these studies date back several decades and may not apply to current high-producing cows. The objective of this study was to assess whether an intermittent milking schedule prior to dry-off increases the concentration of lactoferrin in mammary secretions at the end of lactation and what other factors influence lactoferrin concentration at dry-off. One week prior to dry-off (pre-dry), cows were randomly assigned to an intermittent milking schedule or they continued to be milked twice daily. Duplicate quarter milk samples for microbiological culture were taken at pre-dry and at dry-off to determine infection status of quarters. Quarter somatic cell counts (SCC) were measured on the day of dry-off. Lactoferrin concentrations were quantified by ELISA. Intermittent milking, mean SCC for the last three months prior to dry-off, SCC at dry-off, lactoferrin concentration at pre-dry, quarter infection status at pre-dry and dry-off, days in milk at dry-off, breed, parity, cumulative milk yield for the final week of lactation and season were considered as potential explanatory variables. Their effect on lactoferrin concentration at dry-off was assessed using a mixed-effects linear regression model. Lactoferrin concentration increased significantly during the final week of lactation for cows on an intermittent milking schedule and was significantly associated with initial lactoferrin concentration and infection status at dry-off.

A new approach to teaching veterinary public health at the Ohio State University.

Public-health practitioners with expertise in the area of veterinary public health are expected to understand the prevention and control of zoonotic infectious diseases in both human and animal populations. This focus on multiple species is what makes the veterinary public health (VPH) official unique. The development of a new VPH specialization within the existing Master of Public Health (MPH) degree program at the Ohio State University represents a significant new collaboration between the College of Veterinary Medicine and the College of Public Health. The main objective of the VPH specialization is to educate and train professionals to provide them with the skills, knowledge, and resources necessary to protect and improve human health using a One Medicine approach. The program targets a population of students who will likely enter the professional veterinary medicine curriculum but have one year available to enhance their preparatory training in health sciences before beginning the program. A core series of VPH courses was initiated to complement the existing MPH course requirements. The program has been successful in attracting students from the primary target population, but it has also attracted students wanting the MPH as a terminal degree and veterinarians returning to school to expand their career options.

Effect of the addition of epinephrine to a lidocaine solution on the efficacy and duration of palmar digital nerve blocks in horses with naturally occurring forefoot lameness.

OBJECTIVE To determine whether addition of epinephrine to a lidocaine solution would prolong and potentiate the efficacy of a palmar digital nerve block (PDNB) in horses. ANIMALS 6 adult horses with naturally occurring forefoot lameness. PROCEDURES Initially, a PDNB with a 2% lidocaine solution was performed on the affected foot of each horse. Three days later, the PDNB was repeated with a 1% lidocaine solution or a 1% lidocaine solution containing epinephrine (dilution, 1:200,000). After another 3-day washout period, the PDNB was repeated with the treatment opposite that administered for the second PDNB. Gait was analyzed with a computerized lameness analysis system and heart rate and extent of skin sensation between the heel bulbs of the blocked foot were evaluated at predetermined times for 2 hours after each PDNB. RESULTS Efficacy and duration of the PDNB did not differ significantly between the 2% and 1% lidocaine treatments. The addition of epinephrine to the 1% lidocaine solution improved the efficacy and prolonged the duration of the PDNB. It also resulted in a positive correlation between skin desensitization and amelioration of lameness. Median heart rate remained unchanged throughout the observation period for all 3 treatments. No adverse effects associated with the PDNBs were observed. CONCLUSIONS AND CLINICAL RELEVANCE Addition of epinephrine (dilution, 1:200,000) to a 1% lidocaine solution improved the efficacy and prolonged the duration of a PDNB in horses with naturally occurring lameness and might be clinically useful for lameness evaluations and standing surgery of the forefoot of horses.

Effect of exercise and environmental terrain on development of the digital cushion and bony structures of the bovine foot.

OBJECTIVE: To determine whether exercise on alternative terrain affects the development of the digital cushion and bony structures of the bovine foot. ANIMALS: 20 weaned bull calves. PROCEDURES: Two-month-old calves were randomly allocated to an exercise or control group. For 4 months, the control group was maintained in grass paddocks, and the exercise group was maintained in a 0.8-km lane with a mixed terrain of dirt, stones (0.32- to 0.95-cm pea gravel and 5-cm crusher run), and grass. Water and food for the exercise group were located at opposite ends of the lane; calves were fed twice daily, which ensured they walked 3.2 km/d. Pedometers were applied to all calves to measure distance traveled. All calves were slaughtered at 6 months of age. The right forefeet and hind feet were harvested for MRI and CT evaluation. RESULTS: Control calves walked a mean of 1.1 km daily, whereas the exercised calves walked a mean of 3.2 km daily. Mean digital cushion volume and surface area were 25,335 mm(3) and 15,647 mm(2), respectively, for the exercised calves and 17,026 mm(3) and 12,745 mm(2), respectively, for the control calves. When weight was controlled, mean digital cushion volume and surface area for the exercise group were increased by 37.10% and 18.25%, respectively, from those for the control group. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that exercise on alternative terrain increased the volume and surface area of the digital cushion of the feet of dairy calves, which should make them less susceptible to lameness.

High-sensitivity quantitative PCR platform.

Real-time PCR methods have become widely used within the past few years. However, real-time PCR is rarely used to study chronic diseases with low pathogen loads, presumably because of insufficient sensitivity. In this report, we developed an integrated nucleic acid isolation and real-time PCR platform that vastly improved the sensitivity of the quantitative detection of the intracellular bacterium, Chlamydia spp., by fluorescence resonance energy transfer real-time PCR. Determinants of the overall detection sensitivity were analyzed by extracting nucleic acids from bovine milk specimens spiked with low amounts of chlamydial organisms. Nucleic acids were optimally preserved and recovered by collection in guanidinium stabilization buffer, binding to a matrix of glass fiber fleece, and elution in low volume. Step-down thermal cycling and an excess of hot-start Taq polymerase vastly improved the robustness and sensitivity of the real-time PCR while essentially maintaining 100% specificity. The amplification of Chlamydia 23S rRNA allowed for the differentiation of chlamydial species and was more robust at low target numbers than amplification of the omp1 gene. The best combined method detected single targets per a 100-microL specimen equivalent in a 5-microL real-time PCR input. In an initial application, this high-sensitivity real-time PCR platform demonstrated a high prevalence of chlamydial infection in cattle.

Mastitis therapy and pharmacology.

Mastitis remains the most frequent cause of antibacterial use on dairy farms and contributes to a substantial portion of total drug and veterinary costs incurred by the dairy industry. Ultimately, the best outcome of mastitis therapy is a positive effect on the amount of marketed milk harvested and long-term survival of the cow. This article describes the strategies of therapy for bovine mastitis, with an emphasis on antibacterial and anti-inflammatory therapy.

Staphylococcus aureus shedding pattern throughout lactation in dairy cows with naturally occurring intramammary infection.

OBJECTIVE: To evaluate shedding patterns of Staphylococcus aureus, specifically the association between clonal relatedness and shedding patterns of S aureus for cows with naturally occurring S aureus intramammary infection. DESIGN: Longitudinal field study. SAMPLE: Milk samples from 22 lactating cows (29 mammary glands) of varied numbers of lactations on 2 dairies. Procedures-Foremilk samples were collected weekly for 26 to 44 weeks during lactation from individual mammary glands. Milk samples were cultured bacteriologically with a 0.01-mL inoculum. Samples were considered culture positive for S aureus if ≥ 1 colony-forming units were obtained. Milk samples from known S aureus-positive mammary glands that were culture negative for S aureus or culture positive with a single colony of S aureus were cultured bacteriologically a second time with a 0.1-mL inoculum. Longitudinal shedding patterns of S aureus and the effect of strain type on ln(colony forming unit count) were examined. RESULTS: With the 0.01-mL inoculum, 914 of 1,070 (85%) samples were culture positive. After reculturing of negative samples with a 0.1-mL inoculum, 1,011 (95%) of the samples were culture positive. There was no significant difference in the detection of S aureus between genotypic clusters when either the 0.01- or 0.1-mL inoculum was used. There was no significant difference in the amount of shedding between mammary glands infected with isolates in genotypic cluster 1 or 2. No consistent shedding patterns were identified among or within cows. There was a significant difference in mammary gland linear score and test day (composite) linear score between mammary glands infected with isolates in genotypic clusters 1 and 2. CONCLUSIONS AND CLINICAL RELEVANCE: S aureus was shed consistently in cows with naturally occurring intramammary infection in cows, and regardless of the pulsotype, variations in the amount of S aureus shedding had no significant effect on the ability to detect S aureus with a 0.1-mL inoculum.

Variation in daily shedding patterns of Staphylococcus aureus in naturally occurring intramammary infections.

The goal of the current prospective field study was to examine the shedding patterns of naturally occurring Staphylococcus aureus intramammary infections and the association of pulsed field gel electrophoresis pulsotype with shedding. Milk samples from 5 multiparous and 2 primiparous cows identified with S. aureus intramammary infections were collected for 21 consecutive days, 3 times throughout the lactation (63 days total). Cyclicity of each quarter was evaluated using a locally weighted regression. Pulsed field gel electrophoresis was used for genotypic cluster comparisons to evaluate the association of strain type and shedding patterns. Although the amount of shedding varied greatly, 97.5% of the samples were culture positive. There were notable differences in S. aureus shedding patterns among cows as well as within cows; however, no consistent cyclic pattern was identified. Quarters infected with S. aureus isolates grouped in genotypic cluster 1 appeared to shed at consistently higher levels with a median cfu/0.01 ml of 154 (ln[cfu] = 5.0). In comparing ln(cfu)/0.01 ml between genotypic clusters over the first 21-day sample period, accounting for the effect of sample day, samples collected from quarters infected with S. aureus in genotypic cluster 1 had a 1.5 times greater ln(cfu) than those collected from quarters infected with strains in genotypic cluster 2. The ability to detect S. aureus from day to day was very consistent. The current study examining naturally occurring intramammary infections would support the conclusions of other studies suggesting that a single quarter sample would be adequate in determining S. aureus intramammary infections status.

Using dairy herd improvement records and clinical mastitis history to identify subclinical mastitis infections at dry-off.

Interest in selective dry cow therapy (SDCT) has been increasing owing to concerns over development of antimicrobial resistance. Implementation of SDCT, however, requires a quick and cost-effective on-farm method for identifying cows for treatment and cows that can be left without treatment. The objective of the present study was to evaluate the use of clinical mastitis (CM) history and somatic cell counts (SCC) from monthly Dairy Herd Improvement (DHI) records in identification of infected and uninfected cows at dry-off. A total of 647 Holstein cows were classified as uninfected or infected at dry-off based on CM history and varying number of monthly SCC records (with three different SCC cut-offs). Cows were considered uninfected based on the following criteria: (1) SCC <100,000 cells/ml and no CM during the lactation; (2) SCC <200,000 cells/ml and no CM during the lactation; (3) as criterion two, but additionally a cow was also considered uninfected if it experienced a case of CM during the first 3 months of the lactation and the SCC was <100,000 cells/ml for the rest of the lactation; (4) SCC <300,000 cells/ml and no CM during the lactation; otherwise they were considered infected. Infected and uninfected cows at dry-off were most efficiently identified using three months' SCC records with a threshold of 200,000 cells/ml for cows without CM during the lactation and a threshold of 100,000 cells/ml during the rest of lactation for cows with CM during the first 90 days in milk. Moreover, this criterion also most efficiently identified cows infected with major pathogens only at dry-off. The success of the criteria used for identifying infected and uninfected cows will, however, depend on herd characteristics, such as prevalence of infection and type of pathogens present in the herd.

Screening the whole genome of a pathogen in vivo for individual protective antigens.

We report the results of a general protocol that was used to screen the whole genome of Chlamydophila abortus, type strain B577 (formerly Chlamydia psittaci strain B577), in a mouse pneumonia model. Genetic immunization was used to functionally test the genes of C. abortus as vaccines in a mouse challenge system. Nine gene fragments were isolated that conferred protection, with five protecting as effectively as the live-vaccine positive control. Bioinformatics approaches were unable to reconstruct isolation of these antigens. These results suggest that pathogen genomes can be functionally screened for vaccine candidate antigens in a mouse model to reveal new classes of vaccine candidate antigens that may have therapeutic efficacy across host species, disease manifestations, and delivery platforms.

High prevalence of natural Chlamydophila species infection in calves.

We investigated the acquisition and prevalence of Chlamydophila sp. infection in calves. Specimens were collected at weekly intervals from birth to week 12 postpartum from 40 female Holstein calf-dam pairs in a dairy herd. Real-time PCR detected, quantified, and differentiated Chlamydophila 23S rRNA gene DNA from vaginal cytobrush swabs and milk samples. Chemiluminescence enzyme-linked immunosorbent assay with lysed Chlamydophila abortus or Chlamydophila pecorum elementary body antigens quantified antibodies against Chlamydophila spp. in sera. Chlamydophila sp. DNA was found in 61% of calves and 20% of dams in at least one positive quantitative PCR. In calves, clinically inapparent C. pecorum infection with low organism loads was fivefold more prevalent than C. abortus infection and was most frequently detected by vaginal swabs compared to rectal or nasal swabs. In dams, C. abortus dominated in milk and C. pecorum dominated in the vagina. The group size of calves correlated positively (P < 0.01) with Chlamydophila infection in quadratic, but not linear, regression. Thus, a doubling of the group size was associated with a fourfold increase in frequency and intensity of Chlamydophila infection. For groups of 14 or 28 calves, respectively, logistic regression predicted a 9 or 52% probability of infection of an individual calf and a 52 or 99.99% probability of infection of the group. Anti-Chlamydophila immunoglobulin M antibodies in Chlamydophila PCR-positive calves and dams and in dams that gave birth to calves that later became positive were significantly higher than in PCR-negative animals (P

Reinfection with Chlamydophila abortus by uterine and indirect cohort routes reduces fertility in cattle preexposed to Chlamydophila.

This study investigated the effects of controlled reinfection on fertility of cattle naturally preexposed to Chlamydophila abortus. All animals had high prechallenge levels of immunoglobulin M (IgM), IgG, IgG1, and IgG2 serum antibodies against ruminant C. abortus in a chemiluminescent enzyme-linked immunosorbent assay. Twenty virgin heifers were estrus synchronized with prostaglandin F2, artificially inseminated 2 to 3 days later, and challenged immediately by intrauterine administration of 0, 10(4), 10(5), 10(6), or 10(8) inclusion-forming units (IFU) of C. abortus. Ten heifers were estrus synchronized, inseminated, and uterine challenged 2 weeks later. These animals were also indirectly exposed to C. abortus infection (cohort challenged) by contact with their previously challenged cohorts. Pregnancy was determined by rectal palpation 42 days after insemination. All anti-C. abortus antibody isotypes increased in heifers following uterine challenge with 10(8) IFU. A total of 11, 83, 50, 66, and 0% of heifers were pregnant after uterine challenge with 0, 10(4), 10(5), 10(6), and 10(8) IFU of C. abortus, respectively. A total of 50 and 65% of heifers were pregnant with and without cohort challenge, respectively. Uterine inoculum dose and cohort challenge (or, alternatively, a negative pregnancy outcome [infertility]) correlated highly significantly with a rise in postchallenge anti-C. abortus IgM levels over prechallenge levels. Logistic regression modeled fertility, with uterine challenge dose and cohort challenge or prechallenge IgM as predictors (P < 0.05). The models predict that the uterine C. abortus inoculum causing infertility is 8.5-fold higher for heifers without cohort exposure and 17-fold higher for heifers with high IgM levels than for heifers with cohort exposure or with low IgM levels.