Age-related reproductive performance of the Adélie penguin, a long-lived seabird exhibiting similar outcomes regardless of individual life-history strategy.

Age-related variation in reproductive performance in long-lived iteroparous vertebrate species is common, with performance being influenced by within-individual processes, such as improvement and senescence, in combination with among-individual processes, such as selective appearance and disappearance. Few studies of age-related reproductive performance have compared the role of these drivers within a metapopulation, subject to varying degrees of resource competition. We accounted for within- and among-individual changes among known-aged Adélie penguins Pygoscelis adeliae during 17 years (1997-2013), at three clustered colonies of disparate size, to understand patterns in age-related reproductive success during early and late adulthood. Age at first reproduction (AFR) was lowest, and number of breeding attempts highest, at the largest colony. Regardless of AFR, success improved with early post-recruitment experience. For both oldest and youngest recruitment groups, peak performance occurred at the end of their reproductive life span indicating a possible cost of reproduction. Intermediate recruitment groups reached peak performance in their mid-reproductive life span and with intermediate breeding experience, before decreasing. Breeding success was lowest for the initial breeding attempt regardless of AFR, but we observed subsequent variation relative to recruitment age. Gaining experience by delaying recruitment positively influenced reproductive performance early in the reproductive life span and was most evident for the youngest breeders. Oldest recruits had the highest initial and peak breeding success. Differences in AFR resulted in trade-offs in reproductive life span or timing of senescence but not in the overall number of breeding attempts. Patterns differed as a function of colony size, and thus competition for resources. Early life improvement in performance at the larger colonies was primarily due to within-individual factors and at the largest colony, AFR. Regardless of colony size late-life performance was positively related to the age at last reproduction, indicating selective disappearance of lower performing individuals. These results highlight that different life-history strategies were equally successful, indicating that individuals can overcome potential trade-offs associated with early- and late-life performance. These results have important implications for understanding the evolution of life-history strategies responsible for driving population change.

Of mast and mean: differential-temperature cue makes mast seeding insensitive to climate change.

Mast-seeding plants often produce high seed crops the year after a warm spring or summer, but the warm-temperature model has inconsistent predictive ability. Here, we show for 26 long-term data sets from five plant families that the temperature difference between the two previous summers (ΔT) better predicts seed crops. This discovery explains how masting species tailor their flowering patterns to sites across altitudinal temperature gradients; predicts that masting will be unaffected by increasing mean temperatures under climate change; improves prediction of impacts on seed consumers; demonstrates that strongly masting species are hypersensitive to climate; explains the rarity of consecutive high-seed years without invoking resource constraints; and generates hypotheses about physiological mechanisms in plants and insect seed predators. For plants, ΔT has many attributes of an ideal cue. This temperature-difference model clarifies our understanding of mast seeding under environmental change, and could also be applied to other cues, such as rainfall.

Preoperative gabapentin for acute post-thoracotomy analgesia: a randomized, double-blinded, active placebo-controlled study.

BACKGROUND: The role of preoperative gabapentin in postoperative pain management is not clear, particularly in patients receiving regional blockade. Patients undergoing thoracotomy benefit from epidural analgesia but still may experience significant postoperative pain. We examined the effect of preoperative gabapentin in thoracotomy patients. METHODS: Adults undergoing elective thoracotomy were enrolled in this prospective, randomized, double-blinded, placebo-controlled study, and randomly assigned to receive 600 mg gabapentin or active placebo (12.5 mg diphenhydramine) orally within 2 hours preoperatively. Standardized management included thoracic epidural infusion, intravenous patient-controlled opioid analgesia, acetaminophen and ketorolac. Pain scores, opioid use and side effects were recorded for 48 hours. Pain was also assessed at 3 months. RESULTS: One hundred twenty patients (63 placebo and 57 gabapentin) were studied. Pain scores did not significantly differ at any time point (P = 0.53). Parenteral and oral opioid consumption was not significantly different between groups on postoperative day 1 or 2 (P > 0.05 in both cases). The frequency of side effects such as nausea and vomiting or respiratory depression was not significantly different between groups, but gabapentin was associated with decreased frequency of pruritus requiring nalbuphine (14% gabapentin vs. 43% control group, P < 0.001). The frequency of patients experiencing pain at 3 months post-thoracotomy was also comparable between groups (70% gabapentin vs. 66% placebo group, P = 0.72). CONCLUSIONS: A single preoperative oral dose of gabapentin (600 mg) did not reduce pain scores or opioid consumption following elective thoracotomy, and did not confer any analgesic benefit in the setting of effective multimodal analgesia that included thoracic epidural infusion.

Jamides wananga, a new species from Papua New Guinea and Australia (Lepidoptera: Lycaenidae).

Jamides wananga sp. n. is described and illustrated from Madang Province of Papua New Guinea and Dauan Island in the Torres Strait (Queensland, Australia). The new species is similar in appearance to several other Jamides Hübner species in the bochus-group. DNA sequence data and morphology were used to distinguish the nominotypical subspecies found on the New Guinea mainland from J. w. roxina subsp. n. on Dauan Island. Notes on the habitat and behaviour are also provided.

Working less to gain more: when breeding quality relates to foraging efficiency.

In animal populations, a minority of individuals consistently achieves the highest breeding success and therefore contributes the most recruits to future generations. On average, foraging performance is important in determining breeding success at the population level, but evidence is scarce to show that more successful breeders (better breeders) forage differently than less successful ones (poorer breeders). To test this hypothesis, we used a 10-year, three-colony, individual-based longitudinal data set on breeding success and foraging parameters of a long-lived bird, the Adélie Penguin, Pygoscelis adeliae. Better breeders foraged more efficiently than poorer breeders under harsh environmental conditions and when offspring needs were higher, therefore gaining higher net energy profit to be allocated to reproduction and survival. These results imply that adverse "extrinsic" conditions might select breeding individuals on the basis of their foraging ability. Adélie Penguins show sufficient phenotypic plasticity that at least a portion of the population is capable of surviving and successfully reproducing despite extreme variability in their physical and biological environment, variability that is likely to be associated with climate change and, ultimately, with the species' evolution. This study is the first to demonstrate the importance of "extrinsic" conditions (in terms of environmental conditions and offspring needs) on the relationship between foraging behavior and individual quality.

Interlayer Hybridization of Virgin Carbon, Recycled Carbon and Natural Fiber Laminates.

To meet sustainability objectives in the transport sector, natural fiber (NF) and recycled carbon fiber (RCF) have been developed, although they have been typically limited to low to medium performance components. This work has considered the effect of interlayer hybridization of woven NF and non-woven RCF with woven virgin carbon fibers (VCF) on the mechanical and damping performance of hybrid laminates, produced using double bag vacuum infusion (DBVI). The mean damping ratio of the pure laminates showed a trend of NF>RCF>VCF, which was inversely proportional to their modulus. The tensile, flexural and damping properties of hybrid laminates were dominated by the outermost ply. The VCF-RCF and VCF-NF hybrid laminates showed a comparatively greater mean damping ratio. The results of this work demonstrate a method for the uptake of alternative materials with a minimal impact on the mechanical properties and improved damping performance.

Estimation of the burden of leptospirosis in New Zealand.

BACKGROUND: Human leptospirosis mainly affects people in close occupational contact with domestic livestock and their products in New Zealand. The disease has an unquantified impact on both human health and animal production in the country. This study aimed to estimate the burden of leptospirosis in terms of disability-adjusted life years (DALYs) and cost associated with loss due to absence from work, treatment of disease, animal production loss and cost of vaccination. METHODS: Previously published studies of abattoir workers farmers, and veterinarians, reporting annual risks of influenza-like illness attributable to Leptospira infection, were used to estimate the expected number of cases in a year. The cost of lost animal production was based on results of observational studies in beef cattle, sheep and deer conducted in New Zealand. RESULTS: Expected median annual number of severe and mild cases of human leptospirosis was 2,025 (95% probability interval [95% PI] 1,138-3,422). Median annual DALYs were 0.42 (95% PI: 0.06-2.40) per 100,000 people for the entire population, and 15.82 (95% PI: 2.09-90.80) per 100,000 people working in at-risk occupations (i.e. abattoir workers, farmers and veterinarians). Human infection resulted in a median cost of 4.42 (95% PI: 2.04-8.62) million US dollars (USD) due to absence from work and disease treatment. Median production loss cost in beef cattle, sheep and deer was USD 7.92 (95% PI: 3.75-15.48) million, while median vaccination cost in cattle, (including dairy), sheep and deer was USD 6.15 (95% PI: 5.30-7.03) million. Total annual cost of leptospirosis plus vaccination was USD 18.80 (95% PI: 13.47-27.15) million, equivalent to USD 440,000 (95% PI: 320,000-640,000) per 100,000 people. CONCLUSION: This study provides an estimate of the disease burden and cost of leptospirosis in New Zealand that could support occupational health authorities and livestock industries in assessing interventions for this disease.

Epidemiological investigation of Leptospira spp. in a dairy farming enterprise after the occurrence of three human leptospirosis cases.

An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one- (R1, N = 125) and rising two-year-old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy-five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty-five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non-vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira.

Effect of core material and restoration design on strength of endodontically treated bovine teeth: a laboratory study.

PURPOSE: The purpose of this study was to evaluate the fracture resistance of bovine teeth restored with one-piece cast core/crowns and no ferrule, compared to teeth restored with amalgam cores and full coverage crowns, with and without a dentine ferrule. MATERIALS AND METHOD: Thirty bovine incisors were selected and modified to ensure all teeth had axial dentine walls of similar size. The teeth were then randomly allocated to one of the three groups: control group restored with amalgam core and cast crown without ferrule; ferrule group restored with amalgam core and cast crown with a 2-mm dentine ferrule; one-piece group restored with one-piece cast core/crown without ferrule. Each tooth was loaded to the point of fracture. RESULTS: The mean load resisted by the control group, the ferrule group, and the one-piece group were 1092.5, 1843.5, and 1463.1 N, respectively. The mean load resisted by the ferrule group was significantly greater than the control group (p < 0.001) and the one-piece cast core/crown group (p= 0.04). The mean load resisted by the one-piece cast core/crown group was significantly greater than the control group (p= 0.04). CONCLUSIONS: The maximum load resistance was significantly enhanced by a 2-mm ferrule compared with teeth with no ferrule and teeth restored with one-piece cast core/crowns. Teeth restored with one-piece cast core/crowns were significantly more resistant to loading than teeth restored with amalgam cores and crowns without a ferrule.

Effects of natural infection by L. borgpetersenii serovar Hardjo type Hardjo-bovis and L. interrogans serovar Pomona, and leptospiral vaccination, on sheep growth.

In New Zealand, up to 97% of NZ sheep flocks are seropositive to Leptospira borgpetersenii serovar Hardjo and/or Leptospira interrogans Pomona, yet vaccination is rare. This study evaluated the impact of exposure to these serovars and of vaccination on sheep growth. One third of 2260 ewe lambs on eight farms were randomly selected and vaccinated with a primary and booster bivalent Hardjo and Pomona vaccine starting at one month of age on seven farms and at around five months of age on one farm. Repeated blood samples were taken over one (n = 6 farms, bred as ewe lambs at 7-8 months of age) or two (n = 2 farms, bred as rising 2-year-old ewes) years and tested by microscopic agglutination test to assess exposure to Hardjo and Pomona. Individual weights were recorded at the same time and modelled using a multilevel linear model accounting for within-farm clustering and repeated measures. Predicted average weights were computed and compared based on the vaccination status and within the control group based on exposure status (positive for Hardjo only, Pomona only, Hardjo and Pomona and negative) for each combination of farm and weighing episode. Statistical significance of the comparison was evaluated after adjustment for multiple comparisons. There was no difference in average weight between vaccinated and control sheep before or after vaccination in any of the flocks. The comparison between sheep seropositive for either or both serovars and seronegative sheep was inconclusive, with variations of direction and magnitude of the difference between farms and weighing episodes. In the absence of an overall growth response to vaccination, widespread adoption of vaccination would unlikely yield an economic response at the industry level. However, the inconsistency observed when comparing animals based on their exposure status suggests that the actual effect of leptospirosis on growth is difficult to predict. A study of the effect on sheep reproduction is needed to fully assess the effect of vaccination on sheep production.

Effects of natural infection by L. borgpetersenii serovar Hardjo type Hardjo-bovis, L. interrogans serovar Pomona and leptospiral vaccination on sheep reproduction.

Most New Zealand sheep flocks are seropositive to Leptospira serovars Hardjo and/or Pomona, yet vaccination is rare. This study evaluated the impact of exposure to these serovars and of vaccination, on primiparous one- (P1) and two-year-old (P2) sheep reproduction outcomes. The study was designed as a split-flock vaccination trial, with a third of the animals vaccinated starting at one month of age. Reproduction outcomes were the proportion of bred P1 (7 months old) and as P2 (19 months old) scanned pregnant, the proportion of pregnant ewes rearing a lamb to tail docking and the proportion of docked lambs that were weaned. Odds ratios and their 95% confidence intervals were calculated to compare reproductive performance between vaccinated and control sheep, and within the control group, between seropositive and seronegative sheep. Odds ratios (OR) were also calculated to assess the relationship between vaccination and loss to follow-up. There was no difference in pregnancy and docking rates between vaccinated and control sheep, or between seropositive and seronegative sheep. P1 with a Hardjo titre ≥1536 were significantly less likely (OR = 0.41, 95%CI = 0.19-0.93) to keep a lamb between docking and weaning than P1 with both Hardjo and Pomona titres <1536, for an observed difference in weaning rate of up to 22.6% points on one farm. A reduction of weaning rates in 2-tooths seropositive for Pomona alone and both Hardjo and Pomona was observed but this was non-significant, possibly because of a lack of power. No difference in weaning rate was observed between vaccinated and control P1 or P2. On one farm vaccinated P1 were less likely to be lost to follow-up (OR = 0.27, 95% CI 0.08 to 0.95) between breeding and weaning. Comparing reproductive performance of vaccinated and control sheep revealed no significant difference. However, comparing exposed and non-exposed ewes revealed a possible adverse effect of Leptospira on weaning rates. This suggests that a full vaccination program may result in an improvement of reproductive outcomes, possibly by providing herd immunity.

Effectiveness of a commercial leptospiral vaccine on urinary shedding in naturally exposed sheep in New Zealand.

L. borgpetersenii serovar Hardjo and L. interrogans serovar Pomona are endemic in New Zealand sheep. An effective vaccine and vaccination strategy would protect both humans and livestock. Four to 12 lambs were selected from each of eight farms (total=84, vaccinated group), while four to 16 lambs (total=98) served as unvaccinated controls. A commercial Hardjo/Pomona vaccine was given at 1-6 weeks of age, 5-11 weeks later and 33-67 weeks later on seven farms and at 18 weeks of age and 5 weeks later on the eighth farm. Vaccinates and controls were grazed together. Blood was regularly collected from the control group to assess flock exposure. Urine was collected from both groups 26-82 weeks after the second vaccination and tested by quantitative PCR. Seroprevalence in controls at the time of urine sampling ranged from 2.7 to 98.2% for Hardjo and from 0 to 54.1% for Pomona with seroconversion occurring 13 to 67 weeks after the second vaccination in all but one farm where exposure had happened by the time of vaccination. The shedding prevalence adjusted for clustering in farms was 45.1% [95% CI 17.6-72.7] (for an observed number of 50/98) in the control animals and 1.8% [95% CI 0.0-10.1] (for an observed number of 5/84) in the vaccinated animals. The vaccine was 100% effective on five farms where animals were vaccinated before 12 weeks of age and before natural exposure occurred, but the effectiveness was 80% [0-97] on one farm where the lambs were exposed before vaccination and 65% [9-87] to 80% [0-97] on one farm where the animals were fully vaccinated by 24 weeks of age. The overall vaccine effectiveness was 86.3% [63.6-94.8%] despite maternal antibodies in some flocks at first vaccination. Vaccination timing seemed to be crucial in achieving optimum reduction in shedding in urine in vaccinated sheep.

Pressure transmission across dentine in response to an external pressure: a laboratory study.

PURPOSE: To determine whether transducers could provide a convenient method of measuring dentine permeability and to investigate the effects of removal of pulp tissue and perfusion time on the pressure increase across human dentine after low-pressure perfusion. METHOD: Human premolar teeth that had been stored for 1-2 months were prepared for full crown preparations. The pulp tissue was removed from half the samples and all the coronal segments perfused with saline at 1.3kPa for varying times. An external pressure was then applied and the response within the pulp chambers recorded with transducers. RESULTS: Extirpation of pulp tissue reduced the rate of rise of pressure inside the pulp chamber by approximately 50%. Perfusion at 1.3kPa for up to 1h had no effect on pressure rise, but 6h of perfusion produced a very significant increase. CONCLUSION: Transducers can provide a relatively simple, convenient and clinically relevant measurement of dentine permeability. For stored coronal segments that have been perfused at physiological pressures, extirpation of pulp tissue and perfusion time have significant effects on pressure transmission across dentine.

Complex regional pain syndrome.

Complex regional pain syndrome (CRPS), formerly known as reflex sympathetic dystrophy, is a regional, posttraumatic, neuropathic pain problem that most often affects 1 or more limbs. Like most medical conditions, early diagnosis and treatment increase the likelihood of a successful outcome. Accordingly, patients with clinical signs and symptoms of CRPS after an injury should be referred immediately to a physician with expertise in evaluating and treating this condition. Physical therapy is the cornerstone and first-line treatment for CRPS. Mild cases respond to physical therapy and physical modalities. Mild to moderate cases may require adjuvant analgesics, such as anticonvulsants and/or antidepressants. An opioid should be added to the treatment regimen if these medications do not provide sufficient analgesia to allow the patient to participate in physical therapy. Patients with moderate to severe pain and/or sympathetic dysfunction require regional anesthetic blockade to participate in physical therapy. A small percentage of patients develop refractory, chronic pain and require long-term multidisciplinary treatment, including physical therapy, psychological support, and pain-relieving measures. Pain-relieving measures include medications, sympathetic/somatic blockade, spinal cord stimulation, and spinal analgesia.

Comparison of lidocaine and compression for velvet antler analgesia in wapiti.

This research compared ring block lidocaine anesthesia (L) and compression (C) for velvet antler removal in elk. Thirty-two wapiti were given 1 mg/kg body weight of zuclopenthixol acetate. The next day, they were restrained in a hydraulic chute and given either a compression device or a lidocaine ring block on the antler pedicle. Behavioral and physiological responses to treatment application and antler removal were recorded, and blood was collected for cortisol analysis. During application of L and C, increases in mean heart rate and systolic arterial blood pressure were greater in the C treatment group (P < 0.05, and P = 0.05, respectively). When antler was removed, more behavioral responses occurred in the C treatment group (P = 0.02) and its median behavior score was higher (P = 0.03). Mean heart rates increased for both treatment groups when antlers were removed (P < 0.01). It was concluded that application of C may be painful, and that C was not as effective as L for analgesia for velvet antler removal.

Interlaboratory and between-specimen comparisons of diagnostic tests for leptospirosis in sheep and cattle.

A study was performed to investigate interlaboratory test agreement between a research and a commercial veterinary diagnostic laboratory on blood and urine samples, and to investigate test agreement between blood, urine, and kidney samples (research laboratory) for leptospirosis diagnosis. Samples were sourced from 399 sheep and 146 beef cattle from a local abattoir. Interlaboratory agreement for real-time quantitative polymerase chain reaction (qPCR) results on urine samples was almost perfect (kappa = 0.90), despite the use of different amplification targets (DNA gyrase subunit B gene vs. 16s ribosomal RNA gene), chemistries (SYTO9 vs. TaqMan probe), and pre-PCR processing. Interlaboratory agreement for microscopic agglutination test (MAT) positivity was almost perfect (kappa = 0.93) for Leptospira borgpetersenii serovar Hardjo subtype Hardjobovis (Hardjobovis) but moderate (kappa = 0.53) for Leptospira interrogans serovar Pomona (Pomona). Among animals that had different titers recorded, higher Hardjobovis and lower Pomona titers were reported by the commercial laboratory than by the research laboratory (P < 0.005). These interlaboratory comparisons can assist researchers and diagnosticians in interpreting the sometimes discrepant test results. Within the research laboratory, the comparison of qPCR results on urine and kidney showed almost perfect agreement (kappa = 0.84), suggesting that the qPCR on these 2 specimens can be used interchangeably. The agreement between MAT positivity and urine and kidney qPCR results was fair (kappa = 0.32 and kappa = 0.33, respectively). However, the prevalence ratio of urine and kidney qPCR positivity in Hardjobovis-seropositive versus Hardjobovis-seronegative sheep indicated that Hardjobovis seropositivity found in sheep may be able to predict shedding or renal carriage.

Trends in the breeding population of Adélie penguins in the Ross Sea, 1981-2012: a coincidence of climate and resource extraction effects.

Measurements of the size of Adélie penguin (Pygoscelis adeliae) colonies of the southern Ross Sea are among the longest biologic time series in the Antarctic. We present an assessment of recent annual variation and trends in abundance and growth rates of these colonies, adding to the published record not updated for more than two decades. High angle oblique aerial photographic surveys of colonies were acquired and penguins counted for the breeding seasons 1981-2012. In the last four years the numbers of Adélie penguins in the Ross and Beaufort Island colonies (southern Ross Sea metapopulation) reached their highest levels since aerial counts began in 1981. Results indicated that 855,625 pairs of Adélie penguins established breeding territories in the western Ross Sea, with just over a quarter (28%) of those in the southern portion, constituting a semi-isolated metapopulation (three colonies on Ross Island, one on nearby Beaufort Island). The southern population had a negative per capita growth rate of -0.019 during 1981-2000, followed by a positive per capita growth rate of 0.067 for 2001-2012. Colony growth rates for this metapopulation showed striking synchrony through time, indicating that large-scale factors influenced their annual growth. In contrast to the increased colony sizes in the southern population, the patterns of change among colonies of the northern Ross Sea were difficult to characterize. Trends were similar to southern colonies until the mid-1990s, after which the signal was lost owing to significantly reduced frequency of surveys. Both climate factors and recovery of whale populations likely played roles in the trends among southern colonies until 2000, after which depletion of another trophic competitor, the Antarctic toothfish (Dissostichus mawsoni), may explain the sharp increasing trend evident since then.

Bayesian estimation of the sensitivity and specificity of individual fecal culture and Paralisa to detect Mycobacterium avium subspecies paratuberculosis infection in young farmed deer.

A Bayesian latent class model was used to estimate the sensitivity and specificity of an immunoglobulin G1 serum enzyme-linked immunosorbent assay (Paralisa) and individual fecal culture to detect young deer infected with Mycobacterium avium subsp. paratuberculosis. Paired fecal and serum samples were collected, between July 2009 and April 2010, from 20 individual yearling (12-24-month-old) deer in each of 20 South Island and 18 North Island herds in New Zealand and subjected to culture and Paralisa, respectively. Two fecal samples and 16 serum samples from 356 North Island deer, and 55 fecal and 37 serum samples from 401 South Island deer, were positive. The estimate of individual fecal culture sensitivity was 77% (95% credible interval [CI] = 61-92%) with specificity of 99% (95% CI = 98-99.7%). The Paralisa sensitivity estimate was 19% (95% CI = 10-30%), with specificity of 94% (95% CI = 93-96%). All estimates were robust to variation of priors and assumptions tested in a sensitivity analysis. The data informs the use of the tests in determining infection status at the individual and herd level.

Evaluation of a SYTO9 real-time polymerase chain reaction assay to detect and identify pathogenic Leptospira species in kidney tissue and urine of New Zealand farmed deer.

A SYTO9 real-time polymerase chain reaction assay for detection of pathogenic Leptospira spp. based on amplification of DNA gyrase subunit B (gyrB) gene has been optimized and evaluated for sensitivity and specificity on kidney and urine samples of New Zealand farmed deer. The detection limit was 10(3) cells/ml (2-10 copies/reaction). Comparison of the assay on deer kidneys (n = 268) with culture as the gold standard revealed a sensitivity and specificity of 85% and 99.2%, respectively. For deer urine (n = 113), the assay was compared with known inoculated samples and revealed a sensitivity and specificity of 96.7% and 100%, respectively. The assay was applied for quantifying pathogenic leptospires shed naturally in deer urine and revealed a detectable concentration of 3.7 × 10(3) to 1.7 × 10(6) cells/ml. To assess the assay's capability for identifying pathogenic Leptospira spp., 14 field isolates of L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona were amplified for polymerase chain reaction (PCR) product, purified, and sequenced. When compared with the National Center for Biotechnology Information database, sequence data matched with L. borgpetersenii serovar Hardjo-bovis in 13 samples and L. interrogans serovar Pomona in 1 sample, which was consistent with the microscopic agglutination test (MAT). Sequence analysis of purified PCR product amplified directly from kidney and urine samples also yielded serovar-comparable MAT results. Results suggest that the assay is rapid, sensitive, and specific for detection of pathogenic leptospires in deer clinical samples. The developed assay can also be used for estimating the concentration of leptospires and identifying Leptospira spp. in combination with DNA sequencing.