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Objective To compare the antimicrobial susceptibility patterns of three common mastitis pathogens ( Staphylococcus aureus, Streptococcus uberis and Str. dysgalactiae) isolated from milk samples from New Zealand and the USA. Methods A total of 182 S. aureus, 126 Str. uberis and 89 Str. dysgalactiae isolates from New Zealand (107, 106 and 41, respectively) and the USA (75, 20 and 48, respectively) were assessed using the disk diffusion test. Results Susceptibility varied among the bacterial species. All isolates were susceptible to the amoxicillin-clavulanic acid combination. Resistance to lincomycin was most frequent (susceptibility of 8.6%) across all species. Non-susceptible (i.e. resistant or intermediate) isolates of S. aureus were identified for the three non-isoxazolyl penicillins (amoxicillin, ampicillin and penicillin: 20.6% and 36.0%) and lincomycin (99.9% and 94.6%) for NZ and the USA, respectively. Resistance to erythromycin (5.3%) and tetracyclines (6.7%) was detected only in isolates from the USA. There were differences in susceptibility between Str. uberis and Str. dysgalactiae; all streptococcal isolates demonstrated resistance to aminoglycosides (neomycin 52.4% and streptomycin 27.9%) and enrofloxacin (28%). Resistance of Str. dysgalactiae to tetracycline was almost 100.0% and to oxytetracycline 89.9%. Conclusion and clinical relevance Most of the isolates tested were susceptible to most of the antimicrobials commonly used for treatment of bovine mastitis, with the exception of the lincosamides. Susceptibility to a selected class-representative antimicrobial and at the genus level should be interpreted with caution. Differences between NZ and the USA confirm the value of national surveys to determine the susceptibility patterns of mastitis pathogens. [ABSTRACT FROM AUTHOR]

The article discusses a study of the causes of abortion in cows in dairy farms in New Zealand which have a history of abortion associated with Neospora caninum. The researchers enrolled a total of 2,246 cows and primiparous heifers and monitored them for abortion. They performed histopathological examinations and collected maternal paired sera when abortion was found. They observed that organism was the most prevalent cause of abortion.

The mode of action of bismuth subnitrate in teat sealant formulations as a preventative for intramammary infections during the dry period is unknown. Although previous studies proposed an action mechanism-creating a physical barrier in the teat canal to prevent bacterial invasion-it has not been proven experimentally. We hypothesized that bismuth subnitrate has an inhibitory effect on bacterial growth, in addition to its barrier effect. The objective of this study was to assess the effect of bismuth subnitrate on bacterial growth of major mastitis-causing agents. A strain of Streptococcus uberis (SR115), 2 strains of Staphylococcus aureus (SA3971/59 and SA1), and a strain of Escherichia coli (P17.14291) were tested in vitro for their ability to grow in the presence or absence of bismuth subnitrate. Disk diffusion testing, impedance measurement, and evaluation of bacterial growth in shaking conditions were the methods used to test this hypothesis. A reduction of growth in the presence of bismuth subnitrate occurred for all the strains tested. However, we observed strain and species variations in the extent of growth inhibition. These results suggest that an inhibitory effect on bacterial growth by bismuth subnitrate could partially explain the efficacy of bismuth-based formulations for preventing intramammary infections over the dry period. Further research is required to test the effect of teat sealant formulations on bacterial growth., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

The aim of this study was to assess the effect of bismuth subnitrate and micronized keratin on bovine neutrophils in vitro. We hypothesized that recruitment and activation of neutrophils into the teat canal and sinus are the mechanisms of action of bismuth subnitrate and keratin-based teat sealant formulations. To test this, a chemotaxis assay (Experiment 1) and a myeloperoxidase (MPO) assay (Experiment 2) were conducted in vitro. Blood was sampled from 12 mid-lactation dairy cows of variable ages. Neutrophils were extracted and diluted to obtain cell suspensions of approximately 10 6 cells/mL. In Experiment 1, test substances were placed in a 96-well plate, separated from the cell suspension by a 3 µm pore membrane and incubated for 3 h to allow neutrophils to migrate through the membrane. In Experiment 2, neutrophils were exposed to the test products and the amount of MPO released was measured by optical density. Results showed that neutrophils were not activated by bismuth or keratin products ( p < 0.05) in all of the tests performed. These results suggest that the mechanisms of action of bismuth subnitrate and keratin-based teat sealants do not rely on neutrophil recruitment and activation in the teat canal and sinus after treatment.

Biofilm formation is of growing concern in human and animal health. However, it is still unclear how biofilms are related to mastitis infections in dairy cattle. In this study, a comparison between two tests for biofilm formation and the association between biofilm and the presence of genes associated with biofilm formation were investigated for 92 Staphylococcus aureus isolates from clinical mastitis cases. Congo red agar (CRA) and microtitre test assay (MTA) in vitro phenotypic tests were used to evaluate biofilm formation. The presence of icaA , icaD , and bap genes associated with biofilm formation was confirmed using the polymerase chain reaction. Results show that most of the S. aureus isolates, though not possessing one of the biofilm-forming genes, were able to produce biofilms. MTA was more frequently positive in identifying biofilm-forming isolates than CRA., Competing Interests: The authors declare no conflict of interest.

Curriculum managers of the Bachelor of Veterinary Science program at Massey University have undertaken major curricular review every 5-10 years and also made adjustments to the program as a result of student and other stakeholder feedback. New curricula introduced in 2003 and 2013 aimed to address specific stakeholder requirements in the veterinary, agricultural, and allied industries. The new curricula initially sought to strengthen clinical skills but more recently focused on the core professional skill of client communication, the integration of knowledge and clinical skills, and a better understanding of the effects of herd health interventions on farm economics. The need for greater emphasis on the veterinarian's role in One Health at the intersection of humans, animals, and the environment was also recognized. The most recent curricular review was preceded by faculty enlightenment and discussion about innovative models of medical education with a focus on student-centered and integrated learning. A new curriculum was introduced from 2013 that presented more material in its clinical context, attempted to manage curriculum overload through a focus on Day One Competences, implemented vertical and horizontal integration of subjects, and introduced more problem-based and student-centered learning. Regular reviews of student workload were needed to ensure that the objectives were achieved, but student feedback has generally been positive.

To determine if toltrazuril was effective in eliminating Neospora caninum infection from congenitally infected lambs. Twenty-eight ewes were allocated to 3 groups where animals in Groups A and B were inoculated with 1 × 10(7)N. caninum tachyzoites on Day 120 of gestation and Group C was maintained as a negative control group. Lambs born from ewes in Group A were treated with toltrazuril (20mg/kg) on Days 0, 7, 14 and 21 after birth. Lambs in Groups B and C were untreated. All lambs in Groups A and B were seropositive at 12 weeks of age. At 12 weeks of age, no differences between lambs in Group A and Group B were observed in serological results (ELISA and western blot), presence of N. caninum-related brain histopathological lesions or the number of organisms detected by qPCR. Group C remained negative for serology, detection of N. caninum DNA as well as histopathology throughout the study. Results indicate that N. caninum congenitally-infected lambs had a continuing infection with N. caninum despite being treated with toltrazuril., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Recent reports indicate N. caninum has a possible role in causing abortions in sheep in New Zealand. Knowledge about the mode of transmission of neosporosis in sheep in New Zealand is limited. This study aimed to determine the rate of vertical transmission that would occur in lambs born from experimentally inoculated ewes and to determine if previous inoculation would protect the lambs from N. caninum infection. A group of 50 ewes was divided into 2 groups with one group being inoculated with 5×10(6) N. caninum tachyzoites prior to pregnancy in Year 1. In Year 2, each of these groups was subdivided into 2 groups with one from each original group being inoculated with 1×10(7) N. caninum tachyzoites on Day 120 of gestation. Inoculation of N. caninum tachyzoites into ewes prior to mating resulted in no congenital transmission in lambs born in Year 1 but without further inoculation, 7 out of 11 lambs in Year 2 were positive for N. caninum infection. Ewes that were inoculated in both years resulted in all 12 lambs born in Year 2 being positive for N. caninum infection. This indicates that previous inoculation in Year 1 did not result in any vertical transmission in that year but did not provide any protection against vertical transmission in Year 2. These results suggest that vertical transmission occurs readily once the ewe is infected., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Recent reports indicate Neospora caninum has a possible role in causing abortions in sheep in New Zealand. Knowledge about the epidemiology of neosporosis in sheep is limited. This study aimed to adapt and validate a commercially available ELISA assay as an IgG avidity assay to discriminate between acute (primary and re-inoculated) and chronic N. caninum infections in sheep. In addition, it was used to compare the antibody avidity values between lambs from ewes inoculated with N. caninum either during the pregnancy or in the previous year. The avidity assay was undertaken by using 6M urea for the first wash after incubation with the primary antibody in the commercial ELISA (Chekit* Neospora antibody test kit, IDEXX Laboratories, Australia). Sequential serum samples were obtained from naïve ewes (n=16) experimentally inoculated with live N. caninum tachyzoites. All ewes were seropositive by two weeks post-inoculation and remained seropositive for 20 weeks post-inoculation. There was a linear relationship between time after inoculation and avidity values (p<0.05) over the first 24 weeks. In Week 4, all animals had avidity values <35% and by Week 8, 8/16 animals had avidity values of >35%. These results suggest that an avidity value of <35% indicates a recent primary infection while a value of >35% is indicative of a chronic infection. The assay was then validated using samples from other groups of experimentally inoculated sheep as well as samples from naturally infected ewes. When comparing sample to positive ratio (S/P) and avidity values from lambs born from recently inoculated ewes with those from ewes inoculated the previous year and re-inoculated in the current year, it was possible to differentiate the lambs at 2 weeks of age. Lambs from recently inoculated ewes had low S/P and avidity values at 2 weeks of age which increased by 12 weeks of age. In comparison, lambs from re-inoculated ewes had high S/P and avidity values at 2 weeks of age, due to maternal antibody influence but values were similar to those from lambs that were born from recently inoculated ewes at 12 weeks of age. Avidity values for four naturally infected ewes were all >60% indicating chronic infection. These results suggest that the assay is able to discriminate between recent and chronic infection in sheep as well as able to differentiate lambs with maternal immunity compared to their own de novo immunity. As such it can be utilized to understand the kinetics of N. caninum infection in sheep., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Dairy herd managers have attempted to increase and maintain profits by selectively breeding dairy cattle for high production. Selection for milk production may have resulted in a tendency for greater liveweight (LW) loss postpartum. This study aimed to: (1) determine if excessive LW loss and milk yield in the first 50 days in milk (DIM) was associated with the development of lameness after 50 DIM, and (2) estimate the incidence risk of lameness in this herd attributable to excessive liveweight loss. The dataset comprised details from 564 mixed age cows from a single, seasonally calving, pasture fed dairy herd in New Zealand. After adjusting for the confounding effects of parity, LW at calving, breed, the presence of specified disease events in the first 50 DIM and milk yield, LW loss in the first 50 DIM increased the risk of lameness after 50 DIM by a factor of 1.80 (95% CI 1.00-3.17). The risk of lameness was greatest for high yielding cows that lost excessive LW (risk ratio 4.36, 95% CI 4.21-8.19), but the effect LW loss on lameness risk at the herd level was relatively small. Based on data accumulated during the study we estimate that for this herd, there would be a 3% (95% CI 1-6%) reduction in the incidence risk of lameness if excessive LW loss was prevented. Twenty three percent of the incidence of lameness in this herd was attributable to excessive LW loss. We conclude that policies and interventions to reduce the rate and amount of LW loss in the first 50 DIM will have a non-negligible impact on the incidence risk of lameness in this herd., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Recent reports from New Zealand indicate Neospora caninum has a possible role in causing abortions in sheep. Transmission of N. caninum via semen has been documented in cattle. This study aimed to investigate if horizontal transmission through semen was also possible in sheep. Initially, 6-month old crossbred ram lambs (n=32), seronegative to N. caninum, were divided into 4 equal groups. Group 1 remained uninoculated whilst the remainder were inoculated with N. caninum tachyzoites intravenously as follows: Group 2 - 50 tachyzoites; Group 3 - 10(3) tachyzoites; Group 4 - 10(7) tachyzoites. Semen samples were collected weekly for 8 weeks for the detection of N. caninum DNA and quantified using quantitative PCR (qPCR). Plasma collected 1 month post-inoculation was subjected to ELISA (IDEXX Chekit) and Western blot. At 2 weeks post-infection, three rams from Group 1 (uninoculated) and three rams from Group 4 (10(7)tachyzoites/ml) were mated with two groups of 16 ewes over two oestrus cycles. Ewe sera collected 1 and 2 months post-mating were tested for seroconversion by ELISA and Western blot. All experimentally infected rams seroconverted by 1 month with ELISA S/P% values ranging from 11% to 36.5% in Group 2, 12-39.5% in Group 3 and 40-81% in Group 4. However, none of the ewes mated with the experimentally infected rams seroconverted. For the Western blot, responses towards immunodominant antigens (IDAs) were observed in ram sera directed against proteins at 10, 17, 21, 25-29, 30, 31, 33 and 37 kDa. Rams in Group 2, 3 and 4 were noted to have at least 3 IDAs present. None of the ewes showed any of the 8 prominent IDAs except for the one at 21 kDa which was seen in 30 out of 32 ewes in both groups. N. caninum DNA was detected intermittently in the ram's semen up to 5 weeks post-inoculation with the concentrations ranging from that equivalent to 1-889 tachyzoites per ml of semen. Low concentrations of N. caninum DNA were also detected in the brain tissue of two rams (Groups 1 and 4). These results suggest that although N. caninum DNA can be found in the semen of experimentally infected rams, the transmission of N. caninum via natural mating is an unlikely event., (Copyright © 2013 Elsevier B.V. All rights reserved.)

This study investigated the effect of lameness on liveweight (LW) in pasture-fed dairy cattle. The data comprised 222,446 averaged daily LW measurements from 828 lactations of 542 mixed-age cows in a seasonally calving, pasture-fed New Zealand dairy herd. The LW measurements for individual cows were aggregated into weekly averages and analyses conducted to evaluate the effect of a diagnosis of lameness on LW change after controlling for the effect of week in milk, parity, LW at calving, breed, calendar month, and season. In lame cows, LW decreased for up to 3 wk before lameness was diagnosed and for up to 4 wk after treatment. Total LW loss arising from a single lameness episode was, on average, 61 kg (95% confidence interval: 47 to 74 kg). The results from this study demonstrate how LW records for individual animals can be used to enhance a herd manager's ability to detect lame cows and present them for treatment. The methods presented here show how daily LW monitoring might be used as a tool for early detection of lameness in dairy cattle., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

A clinical trial was undertaken to assess the efficacy of Bovilis(®) Neoguard, a killed Neospora caninum tachyzoite vaccine on 5 commercial dairy farms in New Zealand with a history of Neospora-associated abortion. Cattle were enrolled in the trial at 30-60 days of gestation and randomly allocated to treatment or control groups. Treatment consisted of 5 mL doses of Bovilis Neoguard administered subcutaneously at enrolment then 4 weeks later. Isotonic saline was administered to the control group. Of 2246 cattle enrolled in the trial, 10.7% of cows and 12.6% of heifers were seropositive to N. caninum. Sampling of a randomly selected proportion of enrolled animals 6 weeks after the second treatment showed that 188/232 (81.0%) vaccinated with Bovilis(®) Neoguard had seroconverted, while 11/130 (8.5%) cows and 10/36 (27.8%) heifers in the control group had seroconverted. Forty-eight vaccinated and 63 control animals aborted. On one farm 12.5% of control animals and 6.1% of vaccinated animals aborted (vaccine efficacy 0.61; p=0.03). On another farm with a high level of abortion 8.4% of control animals and 8.7% of vaccinates aborted. On the remaining 3 farms fewer abortions occurred than expected. A modified Poisson regression approach was used to calculate relative risks for abortion and vertical transmission. Overall vaccine efficacy was 0.25 (p=0.12). Heifer replacement calves from the animals enrolled in the trial were sampled for antibodies to N. caninum at 6-9 months of age. Fourteen of 17 calves from vaccinated, seropositive cows were seropositive as were 13/23 calves from seropositive cows in the control group. The interaction between dam serostatus and treatment group was significant (p=0.05) with vaccination increasing the risk of vertical transmission. It was concluded that vaccination after conception prevented 61% abortions in one of five herds and that vaccination may have increased the risk of early embryonic death., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Daily walkover liveweight (WoLW) records (n=79,697) from 463 pasture-fed dairy cows from a single dairy herd in the lower North Island of New Zealand were recorded over the first 100 d of lactation. The aims of this study were to (1) describe LW records retrieved by a standalone automatic Wo daily weighing system; (2) describe the frequency and nature of outlier LW records measured by the system and develop an approach for excluding identified outlier LW records; (3) quantify the agreement between cow LW measured using the Wo system and those measured statically; and (4) describe the autocorrelation between daily LW measurements to provide an indication of how frequently management decisions need to be reviewed to effectively monitor cow LW change in the early-lactation period. The standard deviation of daily LW measurements across parities was 17 kg, on average. A near perfect association between LW measured statically and WoLW (concordance correlation coefficient 0.99, 95% CI 0.99-1.0) was observed. After controlling for the effect of LW at calving and long-term LW change using a mixed-effects linear regression model, the autocorrelation between WoLW recorded on successive days was 0.21, decaying to zero by 8 d. This study shows that by using a standalone automatic Wo weighing system positioned in the exit race of a rotary milking parlor, it was possible to record LW of individual cows on a daily basis and, with controlled cow flow over the weighing platform (allowing for sufficient succession distance to prevent congestion), results were similar to those recorded using conventional, static weighing techniques using the same scales. Based on the autocorrelation analyses, we recommend that LW are recorded on a daily basis to allow changes in physiological status such as the onset of acute illness or estrus to be detected. For managerial purposes, such as using LW change as a guide for adjusting the herd feeding program, we recommend a 7-d decision interval to effectively monitor significant changes in cows' recorded daily LW measurements., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The incidence of clinical mastitis and infection status at calving was assessed in quarters treated with 1 of 2 internal teat sealants at the time of dry off. Two contralateral quarters per cow (n=63 cows) were treated with a sealant that contained 0.5% chlorhexidine; the other quarters were treated with a commercial teat sealant. Ten cows were untreated (controls). On d 2, 4, and 16 after dry off, cows were challenged with Streptococcus uberis S210 strain. Cows were examined daily for 34 d after drying off and cases of clinical mastitis were recorded. Milk samples were collected for culture from any quarters that developed clinical mastitis during the first 34 d after drying-off and from all quarters on d -5 and 0 relative to treatment and at the first and twentieth milking after calving. The incidence of clinical mastitis during the examination period was lower in treated quarters (n=7/252; 1.5%; lower incidence for those treated with chlorhexidine-containing teat sealant n=3/126; 1.2%) than in untreated quarters (n=13/40; 26.8%). The protection against intramammary infection after calving, adjusted for the effect of cow, was higher in quarters treated with the novel teat sealant (89/105; 15.2%; 95% CI=9.6-23.4) than in those treated with the commercial teat sealant (71/104; 31.7%; 95% CI=23.5-41.3) and untreated controls (6/28; 78.6%; 95% CI=59.8-90.0), respectively. Quarters treated with teat sealants were less likely to have an intramammary infection after calving and had a lower incidence of clinical mastitis during the early dry period than did untreated controls in this challenge study., (Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Pregnant ewe lambs (7-8 months old at breeding) were inoculated intravenously at 90 days after joining with the ram with 50, 5 x 10(3), 10(6), or 10(8)Neospora caninum tachyzoites and outcomes were compared to a control group. Seroconversion was measured by indirect fluorescent antibody test (IFAT) at fortnightly intervals and by enzyme-linked immunosorbent assay (ELISA) at 28 days post-challenge. Seroconversion (by IFAT) occurred in all animals except for 2 sheep in the lowest dose group, including the 9 sheep in the control group. IFAT antibody titres rose and fell rapidly, peaking at 1:800 and in 8 animals was negative within 7 days of abortion. ELISA results more closely reflected infection dose and outcome of pregnancy than IFAT serology did. All 10 ewes in each of the 2 highest dose groups aborted, 5 of 10 ewes in the mid-dose group aborted and no ewes in the lowest dose or control groups aborted. Histological lesions consistent with N. caninum infection were seen in the brains of all 25 aborted lambs, as well as in 2 live-born premature lambs from the group receiving 5 x 10(3) tachyzoites and 2 clinically normal lambs (one from the control group). Polymerase chain reaction (PCR) detected N. caninum DNA in a variety of tissues from lambs, aborted foetuses and dams including 3 of 9 ewe-lamb pairs from the control group. The results from this study showed a strong relationship between challenge dose of N. caninum tachyzoites, the sample to positive (S/P) percentage in an ELISA test 28 days after challenge and pregnancy outcome. IFAT results did not correlate well with the level of challenge or the outcome of pregnancy and their relevance in studies of this kind should be questioned.

This study quantified Neospora caninum DNA in the blood and brain of pregnant and aborted heifers by monitoring PCR product formation in real-time using SYBR Green I, a double-stranded DNA-binding dye. Primers were designed to amplify a 188 bp product specific to N. caninum from the Nc-5 gene fragment of N. caninum. Similarly, a 71 bp product was amplified from the 28S rRNA gene of bovine genomic DNA that served as a control. Agarose gel electrophoresis and analysis of the melting curve for PCR products showed that both primer pairs were specific to their targets. Standard curves were generated for both bovine and N. caninum genomic DNA, and were used to compute the relative concentration of parasite to bovine DNA in the test samples. The concentration of N. caninum DNA in 1 ng of bovine genomic DNA obtained from blood ranged between 0.097 ng at the 1st week of the observation and 0 ng at the 15th week in aborted cows. In pregnant cows, the values ranged between 0.080 ng at the 1st week and 0.155 ng at the 15th week of observation. There was a sustained decrease of DNA concentration in the aborted group after abortion and an increase in DNA concentration in the pregnant group. Comparison of parasite DNA in blood and brain of infected heifers showed a higher DNA concentration in brain than in blood. This study shows that N. caninum DNA can be quantified to obtain the relative concentration of parasite DNA to host genomic DNA in blood. This technique allows testing of a large number of samples at one time and can be done without the need for slaughter of tested animals.

Aim: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates., Methods: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent antibody test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice., Results: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3-7%., Conclusion: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.

Twelve 2-year old heifers in their fifth month of gestation when pregnancy tested were used in this study. Six heifers aborted at approximately 4 months of gestation and had blood samples drawn less than 6 weeks after the abortions were identified. Blood samples were also drawn from three sero-positive pregnant and three sero-negative pregnant heifers. DNA was isolated from the samples and a 350 bp fragment of the Nc-5 gene was PCR amplified using primer pair Np21+ and Np6+. Also, the Internal Transcribed Spacer 1 (ITS1) was PCR amplified using Tim 3 and Tim 11 primer pair. The Nc-5 gene fragment was cloned, sequenced and the sequence BLAST-tested. Similarly, the ITS1 product was sequenced and BLAST-tested. The BLAST test results revealed that Neospora caninum DNA was present in these blood samples indicating that polymerase chain reaction can be used in the detection of N. caninum DNA in the blood of sero-positive cows.

This study describes qualitative and quantitative antibody response in cows naturally infected with Neospora caninum. The study was carried out with 269 serum samples obtained from 24 cows over a period of 15 weeks. Prior to sample collection, the cows were tested with ELISA. The 269 samples were screened with IFAT and categorized into seven IFAT titre groups (< 1 : 80, 1 : 80, 1 : 200, 1 : 600, 1 : 1000, 1 : 2000, > 1 : 2000). The samples were finally analysed by Western blotting. Seven immunodominant antigens (approximately 18-, approximately 25-, approximately 33-, approximately 35-36-, approximately 45-46-, approximately 47-, approximately 60-62 kDa) and five minor antigens (approximately 25, approximately 51, approximately 64, approximately 77, approximately 116 kDa) were recognized by cow sera. The recognition of approximately 46 kDa antigen by cow sera was common to samples with IFAT titre 1 : 80 and above. Another common antigen was the approximately 18 kDa antigen, which was recognized by samples with IFAT titre 1 : 200 and above. The most remarkable observation was the presence of the 45-46 kDa, the 77 kDa, and absence of the 18 kDa antigenic bands in samples with IFAT titre 1 : 80. This observation was consistent even in the face of fluctuating antibody titre where serum antibody titres from an animal exceeded then failed to reach 1 : 80. Antibody fluctuation was observed across all cows (pregnant and aborted) with no discernible fluctuation pattern. However, the fluctuation in antibody titre observed appeared to be most remarkable in initially ELISA-negative pregnant cows, and to a lesser extent in ELISA-positive pregnant cows, and ELISA-positive aborted cows. Although there was fluctuation in antibody titre, the banding patterns of N. caninum tachyzoite antigens by cows within the same IFAT titre group remained similar.

A 600-cow New Zealand dairy herd experienced an abortion storm in 1997 and was monitored (blood sampling at about 3-month intervals) from May 1997 until January 1999. Abortion risk reached 9% in 1997 and was highest in heifers at 19%. The abortion risk decreased in 1998 to 3.2% (still somewhat higher than during the years prior to the outbreak). The serological reaction pattern for Neospora caninum showed an association with abortion risk only around the time of the 1997 outbreak when seropositive cows were 4.2 times more likely to abort than negative ones. Over the whole study period, only 27% of cows that were sampled on all nine visits always tested negative. Offspring from dams which had positive tests for Neospora caninum were 2.4 times more likely to abort than those from dams testing consistently negative. Controlling for age and breed, seropositive cows produced more milk than those that were consistently negative. Infection might have been present endemically within this herd prior to the epidemic, but in 1997 an additional factor appeared to have triggered the outbreak.

Introduction: Neospora caninum is the most frequently diagnosed cause of cattle abortion in New Zealand and is an important pathogen worldwide. The parasite has been found in aborted bovine foetuses and in puppies with neurological disease. Recent developments have provided new insights into the epidemiology of the disease that are reviewed in this article., Transmission: Transplacental infection is of major importance in the spread of N. caninum overseas. Most congenital infections result in the birth of apparently healthy calves. Seroepidemiological studies indicate introduced point-source infections are also likely, as do investigations of abortion outbreaks in cattle herds attributed to N. caninum. Horizontal transmission is an important determinant of the stability of infection prevalence in a herd. Two potential sources of horizontal transmission that have recently been reported are: via a canine definitive host and; cow-to-calf transmission through pooled colostrum or milk. Transmission parameters for the organism have been estimated using mathematical modeling, to explore the epidemiology and options for control of N. caninum infection in dairy cattle in the absence of precise epidemiological data. LIFE-CYCLE: Seroepidemiologic studies, combined with the knowledge that dogs can be a definitive host, provide evidence supporting a dog-cattle life-cycle. The importance of dogs in the epidemiology of bovine neosporosis is not yet clear. It is likely that N. caninum oocysts in dog faeces could serve as a source of infection for cattle and recent studies have shown that the canine and bovine isolatesflare the same organism. Infection of cattle orally exposed to oocysts has been demonstrated experimentally but not in nature. It is not known if other wild carnivores may serve as definitive hosts of N. caninum., Conclusion: Despite active research for over a decade, very little is known about the mechanisms of transmission of N. caninum. Vertical transmission is well documented but horizontal spread and the pathogenesis of abortion from this disease need further investigation. It is evident that, even with a very high probability of vertical transmission, some form of horizontal transmission is required for the disease to persist in a herd.

Three hundred and forty-two Streptococcus uberis isolates were cultured from milk samples from subclinical and clinical cases of dairy cattle mastitis. The samples were collected from 15 different New Zealand farming regions, including eight specific farms, during field research trials and veterinary diagnostic investigations. Pulsed-field gel electrophoresis was used to determine and compare the degree of genetic dissimilarity between the restriction endonuclease fragment pattern of the 342 New Zealand and a single United States S. uberis isolate. The 343 isolates exhibited 330 different restriction endonuclease fragment patterns. The United States isolate had a pattern unlike any of the New Zealand isolates. Most of the isolates were genetically different strains (pattern deferred by at least 33%), but identical patterns were noted within the same or different quarters of an individual cow, different cows within the same farm, and from different cows from the same or different districts, farming regions or islands. Seven of the eight selected farms had at most only one pair of isolates with banding patterns, which differed by less than 33%. A high degree of dissimilarity was noted in individual herds in which all the samples were collected on the same day or over a 2-year period. The high degree of dissimilar isolates is an indication that S. uberis infections in New Zealand dairy cattle are largely due to the opportunistic nature of the organism in the cows' environment. Prevention and treatment of S. uberis mastitis will therefore need to be directed at a multitude of different strains present throughout the country as well as in individual herds.

Aim: To determine the effect of calcium supplementation of dairy cattle after calving on serum concentrations of calcium, magnesium, and phosphorous after calving, and on reproductive and productive performance in the following lactation., Methods: This was a double-blind randomised clinical trial using 356 dairy cows of parity 3 or greater from six seasonally calving commercial dairy herds. Cows were randomly assigned to a treatment and control group. The treatment group received two doses of 380 ml of calcium chloride in oil (49 g of elemental calcium); the first dose was given as soon as practical after calving and the second 12 hours later. Control cows received 380 ml of a placebo using the same treatment regime. Treatment and control cows had blood samples taken during the first 7 days after calving to determine serum calcium, magnesium and phosphorous levels. Treatment and control cows were monitored throughout the lactation to measure production and reproductive performance., Results: Treatment had no effect on PSM to first service intervals. Treatment significantly reduced PSM to conception intervals in only one of the four herds with data available for analysis (p = 0.09). There was no difference between treatment groups for adjusted corrected milk yield calculated from the first herd test milk yield (p = 0.93) or 305-day milk solids production (p = 0.19)., Conclusion: Further studies are required to identify reasons for the between-herd effectiveness of calcium chloride treatment in improving fertility.

The objectives of this experiment were to determine the effects of 0.5 mg estradiol benzoate, administered intramuscularly 24 h after removal of CIDR-B progesterone containing intravaginal devices, on the time to estrus, ovulation and peak LH concentration in dairy heifers. Ovulatory responses and plasma LH concentrations were examined using 14 Friesian dairy heifers in 2 separate treatment periods. All heifers received a CIDR-B progesterone-containing intravaginal device with an attached 10-mg estradiol benzoate capsule for 12 d. Within each period, 24 h after CIDR-B removal, 7 heifers received an intramuscular injection of 0.5 mg estradiol benzoate while the remaining 7 heifers received an intramuscular injection of a placebo. Blood samples for LH assay were collected at 0, 6 and 12 h, and then every 4 h for 60 h after estradiol injection. Detection of estrus was conducted at 4-h intervals, and ultrasonographical examination to detect ovulation was conducted every 8 h for 88 h after removal of the CIDR-B device. Treatment with estradiol benzoate tended to reduce the time from device removal to the LH peak in Period 1 (median time to LH peak 40.1 vs 63.9 h; P = 6.07). In Period 2, treatment with estradiol had no significant effect on the time to the LH peak, standing estrus or ovulation. We hypothesize that the period effect was due to the stage of cycle at the time of treatment. For heifers treated in Period 1, the stage of cycle was random. However, because of the prior synchronization of estrus, which was implicit in the experimental design, heifers in Period 2 tended to be in late diestrus. The administration of estradiol benzoate after treatment with exogenous progesterone appears to overcome the variability in timing of LH peaks typically occurring in a herd of synchronized heifers due to different stages of follicular development.

Recommendations for oestrus synchronisation of dairy heifers using progesterone-containing intravaginal devices suggest re-insertion of used devices 16 days after first insemination for a period of 5 days to allow a second opportunity for artificial insemination. Controlled studies on the effectiveness of re-using intravaginal devices to synchronise returns to oestrus in non-pregnant dairy heifers are lacking. A clinical trial was conducted involving 750 Friesian heifers in 13 herds. After an initial synchronisation programme, the used intravaginal devices were re-inserted 14 or 16 days after first insemination into half of the heifers in each herd for a period of 5 days. After the first synchronisation programme, 47.5% of heifers remained non-pregnant. Re-insertion of used intravaginal devices for 5 days significantly increased the number of non-pregnant heifers detected in oestrus and inseminated by 48 hours after device removal compared to heifers in which devices were not re-inserted (45.2% v. 27.3%, p < 0.05, in herds where intravaginal devices were re-inserted on day 14; 48.8% v. 13.6%, p < 0.05, in herds where intravaginal devices were re-inserted on day 16). Re-insertion at 14 or 16 days after first insemination was equally effective in increasing visible returns to service. However, the number of non-pregnant heifers synchronised for a second round of artificial insemination was less than expected. Conception rate to the re-synchronised oestrus was unaffected by the treatment. It is concluded that the additional procedures of CIDR re-insertion, removal, tailpainting and insemination involved in there-synchrony programme, and the relatively low in-calf rate to the re-synchronised round of insemination, reduced the potential benefits of re-synchronisation.

The effects of Se and I supplementation on growth rate and on thyroid and somatotropic function were examined for heifer calves from two herds fed pasture. Supplementation of calves with intraruminal Se pellets increased the basal plasma concentration of 3,5,3'-triiodothyronine and reduced the basal plasma concentration of thyroxine for both herds. For one herd, supplementation with Se increased the triiodothyronine response to challenge with thyrotropin-releasing hormone, increased BW gain, and tended to increase the plasma concentration of IGF-I. The plasma concentration of growth hormone was unaffected by Se supplementation. Supplementation with I increased the response of thyroid hormones to thyrotropin-releasing hormone but did not increase BW gain. Interaction between Se and I treatment within the herds was not apparent for any outcome variable. These data suggest that the effects of Se deficiency in grazing calves may be mediated by alterations in thyroid hormone metabolism but apparently are not mediated through modulation of the peripheral concentration of growth hormone.

The objective of this study was to investigate the effects of selenium (Se) and alpha-tocopherol supplementation on uterine involution and ovarian function in dairy heifers fed a prepartum diet containing low concentrations of Se and alpha-tocopherol. Twenty-four pregnant Friesian heifers were randomly allocated to one of four experimental groups in a 2 x 2 design balanced for age and body weight. Prepartum treatments consisted of supplementation with either 2 intraruminal Se pellets or 3600 mg of alpha-tocopherol p.o. 4 times per wk, or both. Control animals received no supplementation. For 8 wk before calving, the heifers were fed exclusively on pasture hay which contained less than 10 microg/kg of Se and 19 mg/kg of alpha-tocopherol. After calving, the heifers grazed perennial ryegrass and white clover pasture. Concentrations of Se and alpha-tocopherol in serum for the prepartum heifers of the control group were 10 ng/ml and 1.3 microg/ml, respectively, indicating deficiencies of these nutrients. Treatment with Se and alpha-tocopherol increased prepartum serum concentrations of Se and alpha-tocopherol to 74 ng/ml and 5 microg/ml, respectively (P < 0.001). However, treatment with Se, alpha-tocopherol, or both, failed to enhance uterine involution, hasten resumption of postpartum ovarian activity or reduce the incidence of clinical postpartum abnormalities. These findings suggest that postpartum reproductive dysfunction is not a primary feature of moderate Se or vitamin E deficiency of cattle at pasture.

The objective of this study was to determine if serum glutathione peroxidase activity reflects short-term changes in the selenium status of goats. Angora goat kids (n=14) were fed pelleted luceme containing 20 microg/kg of selenium, and treated orally with either selenium (0.1 mg/kg of liveweight weekly, as sodium selenate) or de-ionised water. Serum activity of glutathione peroxidase was increased in response to supplementation and differed from that of controls within 24 hours of supplementation. The change in serum glutathione peroxidase activity during the 21 days after the start of weekly supplementation closely followed changes in serum selenium concentration. The results of this study suggest that serum glutathione peroxidase activity reflects the short-term improvement in the selenium status of Angora goat kids following oral supplementation with sodium selenate.

Cows from three herds calving in spring that were fed on pasture were paired by herd, production index, predicted calving date, and age. One cow from each pair was allocated randomly to the treatment group and received a controlled-release capsule of monensin 1 mo prior to the start of AI. A random sample of 20 matched pairs of cows was selected within each herd to monitor BW and metabolic parameters. Treated cows produced more fat, protein, and liters of milk per day during the 2nd mo of monensin activity and produced a greater total milk volume than did control cows. Treated cows gained less BW during the 1st mo after treatment. No significant differences were identified in submission rates, overall pregnancy rates, or pregnancy rate to first or second AI. Blood urea nitrogen was elevated in the treatment group as a main effect, and NEFA were elevated in the 2nd and 3rd mo after treatment. There was no significant effect on blood concentrations of glucose, albumin, or BHBA.

Anecdotal reports suggest that abortion occurs in New Zealand cattle after ingestion of macrocarpa (Cupressus macrocarpa). The clinical signs observed are similar to those seen in abortions caused by ponderosa pine (Pinus ponderosa Laws). Isocupressic acid has been identified as an abortifacient component of the needles of ponderosa pine. Macrocarpa collected from the Palmerston North area contained 0.89% and 1.24% isocupressic acid, compared to 1.70% isocupressic acid in Ponderosa pine needles from Oregon in the United States as analysed by the USDA-ARS Poisonous Plant Research Laboratory. These findings support observations that macrocarpa causes abortion in late gestation cattle in New Zealand and suggest the causal agent is probably isocupressic acid.

The objectives of this trial were to determine the effects of 0.5 mg estradiol benzoate administered intramuscularly 24 h after the removal of progesterone-containing intravaginal devices on the occurrence and timing of estrus, synchronized pregnancy rate and synchronized conception rate in dairy heifers. A clinical trial was conducted involving 750 dairy heifers in 13 herds. Within each herd heifers were randomly allocated to 1 of 2 estrus synchronization treatments. All heifers received a CIDR-B progesterone-containing intravaginal device containing a 10 mg estradiol benzoate capsule for 12 d. Twenty-four hours after CIDR-B removal one group received 0.5 mg, im estradiol benzoate while the other group received an intramuscular injection of a placebo. Estrus detection was performed at 48 and 72 h after intravaginal device removal, and heifers detected in estrus at those times were inseminated. Administration of estradiol benzoate 24 h after removal of CIDR-B devices significantly increased the number of heifers exhibiting estrus within the observation period (96.1 vs 90.5%; P < 0.01). It also altered the onset of estrus so that significantly more heifers were in estrus (86.6 vs 72.3%; P < 0.01) and conceived (47.1 vs 37.5%; P < 0.05) by 48 h after CIDR-B device removal. The synchronized conception rate was unaffected by treatment. The distribution of estrus was such that fixed-time insemination after estrus synchronization with this treatment program may be recommended.