Study on the use of toltrazuril to eliminate Neospora caninum in congenitally infected lambs born from experimentally infected ewes.

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.

Vertical transmission in experimentally infected sheep despite previous inoculation with Neospora caninum NcNZ1 isolate.

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.

Epidemiology of vaginal prolapse in mixed-age ewes in New Zealand.

AIMS: Identify environmental, animal, and management factors associated with risk of vaginal prolapse in ewes, to enable farmers and advisors to make pragmatic decisions based on empirical observations for control of the condition. METHODS: Two longitudinal studies conducted over 2 years to identify factors associated with incidence of prolapse in (i) cohorts of 200 individually identified mixed-age (MA) ewes, and (ii) all MA ewes, on voluntarily participating sheep-breeding farms in Hawkes Bay (HB) and Southland regions of New Zealand. RESULTS: The overall annual incidences of prolapse on 113 farms in 2000 and 88 in 2001 were 1.21 and 0.82 per 100 MA ewes, respectively, and 1.05 for both years combined. A total of 406 prolapses were recorded among 36,695 individually identified cohort ewes. Individual farm incidences for both years varied from 0-5.9 (mean=1.56, median=1.39) on Southland and 0-3.9 (mean=0.75, median=0.54) per 100 ewes on HB farms. The crude relative risk of a prolapse occurring in a MA ewe was 5.31 times higher for ewes carrying twins and 11.3 times higher for ewes carrying triplets, than single lambs. Flocks made up of predominantly pure or crossbred Perendale ewes appeared to be at lower risk than flocks with other breeds. Shearing in the 3 months leading up to mating appeared to be protective, as was shearing in the second half of pregnancy. The risk was higher on farms with moderate to steep terrain than on farms with flat terrain. The identified risk factors in the individually identified cohorts were: access to salt and feeding of swedes in the latter part of pregnancy, moderate to steep lambing paddocks, multiple lambs detected at scanning, and weight gain between start of mating and scanning. The condition recurred in 2001 in six (35%) of 17 study ewes that had prolapsed during 2000. Culling policies for female offspring of affected ewes did not influence incidence at the farm level; nor did feeding hay or grain in late pregnancy. Furthermore, there was no association between incidence and body condition scores measured prior to and after mating, at scanning, or at time of set stocking. CLINICAL SIGNIFICANCE: Vaginal prolapse is an inevitable consequence of sheep reproduction and its incidence is expected to increase as reproductive rates increase. This study provides some firm leads as to the relative importance of risk factors and gives guidance for risk reduction, e.g. by identification and separate management of ewes carrying twins or triplets, using flat paddocks for lambing, and guarding against gain in weight between the start of mating and scanning.

Adaptation of a commercial ELISA to determine the IgG avidity in sheep experimentally and naturally infected with Neospora caninum.

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.

Liver biopsy histopathology for diagnosis of Johne's disease in sheep.

Sheep with Johne's disease develop epithelioid macrophage microgranulomas, specific to Mycobacterium avium subsp. paratuberculosis (Map) infection, in the terminal ileum, mesenteric lymph nodes, and organs distant to the alimentary tract such as the liver. The objectives of this study were to determine whether liver pathology was present in ewes affected by Map and whether liver cores provide adequate tissue for this potential diagnostic marker. One hundred and twenty-six adult, low body condition ewes were euthanized, necropsied, and underwent simulated liver biopsy. Ileal lesions typical of Map were found in 60 ewes. Hepatic epithelioid microgranulomas were observed in all ewes with Type 3b (n = 40) and 82% (n = 11) with Type 3c ileal lesions. None were found in ewes unaffected by Map or with Type 1, 2, or 3a ileal lesions. Liver biopsy core samples provided adequate tissue for histopathology with a sensitivity and specificity of 96% (95% confidence interval [CI], 0.87-0.99) and 100% (95% CI, 0.95-1), respectively for detection of types 3b and 3c ileal lesions.

Seroconversion and semen shedding in rams experimentally infected with Brucella ovis.

AIM: To determine the time taken for rams to develop antibodies to Brucella ovis in serum, shed B. ovis in semen and develop lesions of epididymitis following infection with B. ovis. METHODS: Fifteen 19-month-old rams were artificially infected with B. ovis by inoculation of infected semen onto the nasal and rectal mucus membranes (Day 0). Serum was collected from each ram at 2 to 8-day intervals and tested at commercial laboratories using a complement fixation test (CFT) and an ELISA. Cut-off values for the CFT were 0-4/4 negative; 1/8-3/8 suspicious and 4/8-4/128 positive, and for the ELISA were <10% negative; ≥10 to <50% suspicious and ≥50% positive. Selected serum samples were also tested using a gel diffusion test (GDT). At 7 to 8-day intervals semen was collected for bacterial culture and the scrotal contents were palpated to identify lesions of epididymitis. The study was terminated after 56 days. RESULTS: On Day 28 B. ovis was isolated from the semen of one ram and by Day 49 it was isolated from the semen of 10 rams. All 10 rams had suspicious or positive ELISA or CFT titres by Day 36 and 56, respectively. The GDT results were all negative on Day 36 and in general did not become positive in individual rams until 7-28 days after semen shedding commenced. Epididymitis was detected in one ram on Day 36; by Day 56 eight rams had epididymitis detectable by scrotal palpation. CONCLUSIONS: The B. ovis ELISA test identified infected rams at an earlier stage than the CFT; this was at 19-36 days after exposure. Rams can begin shedding B. ovis in semen as early as 28 days after exposure and lesions of epididymitis develop as early as 36 days after exposure. CLINICAL RELEVANCE: During a test and slaughter campaign for the control of B. ovis, the most appropriate serological re-testing interval is likely to be around 28 days (4 weeks) using the ELISA with or without the CFT, although caution is required in interpretation of "suspicious" ELISA results. Following a B. ovis breakdown, two negative CFT or ELISA tests 60 days apart are recommended to confirm freedom from infection, supporting current guidelines.

Detection of Neospora caninum DNA in semen of experimental infected rams with no evidence of horizontal transmission in ewes.

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.

The impact of highly concentrated Mo and Cu dietary supplements, fed as a bolus, on the efficacy of chelated versus inorganic Cu in cattle on a low-Cu diet.

AIM: To compare the efficacy of chelated versus inorganic forms of dietary Cu supplements, fed as a bolus, when challenged by a daily bolus of dietary Mo in cattle on a low-Cu diet. METHODS: Forty non-lactating, Friesian dairy cows of adequate Cu status were assigned to four groups and fed a basal diet of baled silage containing 5.3 mg Cu and 0.4 mg Mo/kg DM. The experimental design was a factorial of two chemical forms of supplemental Cu and two levels of Mo intake, provided as pelleted grain supplements made from crushed barley/molasses plus Cu and Mo. The supplements contained 140 mg Cu/kg as Cu sulphate pentahydrate (CS), 140 mg Cu/kg as Cu glycinate (CG), CS plus 38 mg Mo/kg as sodium molybdate (CS+Mo), or CG plus 38 mg Mo/kg (CG+Mo). Commencing on Day 0, supplements were fed once daily (offered 1-1.2 kg/cow) and were completely consumed within 5-10 minutes, which constitutes a bolus type of administration. Liver samples were collected by biopsy at Days -24, 13, 41 or 47, and 69 for Cu determinations. RESULTS: The diets fed to the Cu+Mo groups were roughly equivalent to 25 mg Cu and 5.7 mg Mo/kg DM. Mean initial concentration of Cu in liver for all groups was 516 (SE 54) µmol Cu/kg fresh tissue. In cows supplemented with CS and CG, the final (Day 69) concentrations increased (p<0.01) to 939 (SE 166) and 853 (SE 163) µmol Cu/kg, respectively. These values were not different (p=0.72). For groups CS+Mo and CG+Mo, the final concentrations of 535 (SE 122) and 453 (SE 102) µmol Cu/kg were not different from initial values or from each other (p>0.25). The rate of accumulation of Cu in liver following bolus Cu and Mo intake was highly variable but was not affected by initial concentration of Cu in liver (p>0.9) or by the form of Cu (p>0.6). Mean rates of accumulation of Cu in liver were 4.0 (SD 3.8) and 0.65 (SD 2.0) µmol Cu/kg fresh tissue/day for the Cu-only treatments and the Cu+Mo treatments, respectively. CONCLUSIONS: When fed together as a bolus, high Mo intake negated the effect of supplemental Cu but it did not reduce liver Cu stores. There was no difference in the reaction of dietary Mo with chelated Cu (as glycinate) versus inorganic Cu (as sulphate) dietary supplements.

The prevalence of disseminated Mycobacterium avium subsp. paratuberculosis infection in tissues of healthy ewes from a New Zealand farm with Johne's disease present.

AIM: To determine the prevalence of disseminated Mycobacterium avium subsp. paratuberculosis (Map) infection in healthy ewes in a flock with a history of clinical Johne's disease. METHODS: Twenty-four healthy ewes, from a large sheep and cattle farm with a history of clinical Johne's disease in the ewe flock, were randomly selected, euthanased, blood sampled, and examined at necropsy. BACTEC™ radiometric culture for Map was performed on samples of faeces, ileum, mesenteric lymph node, biceps femoris muscle and mononuclear cells in peripheral blood. Serum antibody ELISA tests were performed. Histological sections and Ziehl Neelsen (ZN) stains of impression smears of ileum and mesenteric lymph node were examined for pathological lesions characteristic of Johne's disease and acid fast organisms (AFO). Indirect quantification of Map was performed, using BACTEC radiometric growth indices measuring the time taken for the production of (14)CO(2.) RESULTS: No histological evidence of Johne's disease or AFO was found in the ileum and mesenteric lymph nodes. Twelve of the 24 ewes (50%) had Map cultured from the ileum (n=6) and/or mesenteric lymph nodes (n=8) while none had Map cultured from the faeces, biceps femoris muscle or blood mononuclear cells. One of the 12 Map culture positive ewes was serum ELISA positive. The culture growth rates in liquid medium suggest low numbers of Map were present in the tissues of the culture positive ewes. CONCLUSION: Fifty per cent of clinically healthy ewes exposed to Map within a Johne's infected flock were Map culture positive in the ileum and/or mesenteric lymph node(s), while the ELISA was positive in 8% of those animals (n=1). There was no faecal shedding of Map and no Map was cultured from skeletal muscle or from blood mononuclear cells suggesting that systemic Map infection, defined as positive culture of Map from skeletal muscle and/or blood, may be uncommon in healthy mixed age ewes without clinical Johne's disease. CLINICAL RELEVANCE: ELISA serology detected 1 of 12 ewes infected with Map whilst none were detected from faecal BACTEC radiometric culture, suggesting biosecurity measures used to control the spread of Map may be of limited use. Map was not cultured from blood mononuclear cells or skeletal muscle, indicating that meat from healthy ewes, from farms where Johne's disease is present, is an unlikely source of Map exposure for humans. Further research is warranted to establish the prevalence and dissemination of Map in tissues outside the alimentary tract of healthy ewes from farms throughout New Zealand where Map is present.

The role of liver Cu kinetics in the depletion of reserves of Cu in dairy cows fed a Cu-deficient diet.

AIM: To determine how the concentration of Cu in liver affects the rate of depletion of that Cu when cows are fed a Cu-deficient diet under experimental conditions, and to mathematically model the rate of depletion of Cu over time. METHODS: In June 2010, 25 non-lactating Friesian cows were assigned to three groups such that initial mean concentrations of Cu in liver were 265, 534 and 1,486 µmol Cu/kg fresh tissue (Day 0). All cows were managed as a single group and fed a Cu-deficient diet of primarily baled silage. No mineral Cu supplements were given. Liver biopsies were collected from cows on Days 0, 53, 98 and 161 to determine concentrations of Cu. At about the same time, samples of silage and pasture herbage were collected to determine Cu, Mo and S concentrations. RESULTS: Median concentration of Cu in silage was 6.5 (min 6, max 9) mg/kg DM. Concentration of Cu in liver decreased in all groups (p<0.001), over the duration of the study. The amount of Cu depleted from liver was greater in groups that started the study with higher initial concentrations of Cu in liver. The rate of decline followed exponential first-order kinetics with an elimination rate constant k of 0.0057 (CI 95%=0.0039-0.0074), meaning that about 0.57% of liver Cu reserves were depleted each day. For individual cows this loss amounted to 0.1-14 µmol Cu/kg liver/day. CONCLUSIONS AND CLINICAL RELEVANCE: Depletion of Cu from liver was dependent on initial concentration of Cu. These results can be used to predict how long an unsupplemented herd will remain in adequate Cu status, which adds confidence to decisions about when Cu supplementation should be withdrawn or reinstated. Cows with high concentrations of Cu in liver can maintain adequate Cu status for months without supplements. Intake of less Cu and more Mo would increase the rate of depletion, and seasonal factors would also have some influence.

Pathology of Brucella ovis infection in red deer stags (Cervus elaphus).

AIM: To describe the pathology of the reproductive tract of red deer stags with active Brucella ovis infection and in stags in which B. ovis infection had resolved. METHODS: Twenty-three red deer stags of varying history were slaughtered and their epididymides and accessory sex glands examined grossly and by histopathology. At the time of slaughter five of the stags had an active B. ovis infection of 24-55 days duration following exposure to infected rams, 10 stags had been experimentally infected with B. ovis by intravenous inoculation 649 days previously and had developed an active infection but the bacterial infection had resolved at least 308 days prior to slaughter, and eight stags had not been exposed to B. ovis at any time. RESULTS: Of the five stags with an active infection, one had gross enlargement of the epididymides that could be detected by scrotal palpation. Histological lesions in all five stags included mild to severe, predominantly non-suppurative epididymitis, vesiculitis, prostatitis and ampullitis, with neutrophil exudation in associated glandular ducts. Additional lesions in the epididymides were spermatic granulomas and epithelial hyperplasia with intra-epithelial cyst formation. Of the 10 stags in which the bacterial infection had resolved, two had gross enlargement of the epididymides. The histological lesions were similar to those in stags with active infection but were generally milder, with increased periductal scar tissue in the epididymides. The lesions seen in stags resembled those seen in rams with B. ovis infection but they were usually less florid and had fewer plasma cells. No gross abnormalities or histopathological lesions were detected in the non-infected stags. CONCLUSIONS: Only a small percentage of red deer stags infected with B. ovis develop lesions of epididymitis that can be detected by scrotal palpation. Gross and histological lesions of the genital tract of stags associated with B. ovis infection are similar to the lesions seen in rams. Lesions in stags persist for >300 days after the bacterial infection has resolved. CLINICAL RELEVANCE: Brucella ovis infection should be considered when there are gross lesions of epididymitis or histological evidence of inflammation in the epididymides or accessory sex glands of red deer stags. Retrospective diagnosis of B. ovis in stags could be achieved by histological examination of the reproductive organs.

Ram and buck management.

Careful management is necessary to ensure the reproductive success in any small ruminant farm and to maximise the productive longevity of rams and bucks. Rams and bucks are frequently overlooked outside of the breeding period, but year-round attention to nutrition, parasite control and general disease control is important in keeping them healthy and sound for breeding. Pre-mating soundness examinations are an inexpensive and relatively easy way to assess the potential ability of a ram or buck to perform during the breeding period and should be incorporated annually into the management of any flock. During the breeding period, careful thought should be given to the appropriate use of males, and the effectiveness of mating should be monitored.

Potential involvement of Neospora caninum in naturally occurring ovine abortions in New Zealand.

Neospora caninum is an obligate intracellular parasite and is recognised as the leading cause of bovine abortion worldwide. Natural infection with N. caninum has been described in sheep but it has generally not been regarded as a significant cause of abortion. Recently, there have been several New Zealand cases of foetal abortions where N. caninum was detected which strongly suggested the involvement of Neospora in these abortions. However, there is minimal information about the prevalence of N. caninum infection naturally occurring in New Zealand sheep flocks and particularly its impact on reproduction success. Thus, this present study provides preliminary data on the role that Neospora is playing in ovine reproductive failure by establishing the prevalence of N. caninum antibodies and DNA in ewe blood and foetal material present in 21 New Zealand sheep farms with ongoing unexplained abortion problems and 10 farms with consistently high fertility levels. The results of this study demonstrated an overall seroprevalence of 1.4% which varied between Aborting/non-pregnant (1.8%), age-matched pregnant controls (0.6%) and high fertility (2.1%) ewes. However, despite the variation observed, there was no statistical difference between the three groups. In addition, Neospora DNA was detected by PCR in 13% of submitted foetal brains and in ewe blood from aborting/non-pregnant (6.9%), age-matched pregnant controls (3.6%) and high fertility pregnant (2.1%) ewes. When the PCR results were considered with the IFAT and IDEXX ELISA results, there was no correlation between serology positive and PCR positive blood samples. Taken together, these results reveal that reliance on ELISA-based serology or PCR alone may underestimate the involvement of Neospora. Furthermore, determining the involvement of Neospora appears to require a multi-facetted approach where diagnostic methods and serological cut-off values may need to be adjusted as further information about the effect of natural infections with N. caninum in the ovine host is elucidated.

Detection of Mycobacterium avium subsp. paratuberculosis in skeletal muscle and blood of ewes from a sheep farm in New Zealand.

AIM: To determine whether viable Mycobacterium avium subsp. paratuberculosis (Map) is present in skeletal muscle and blood in ewes with and without Johne's disease confirmed histologically. METHODS: A total of 51 mixed-aged ewes in poor body condition from a farm with a history of clinical Johne's disease were culled and examined at necropsy. BACTEC radiometric culture was performed on samples of skeletal muscle from the biceps femoris, mononuclear cells in peripheral blood (hereafter referred to as blood), and ileum. Histological sections and Ziehl-Neelsen (ZN)-stained impression smears of terminal ileum and mesenteric lymph nodes were examined. Ewes were defined as having confirmed Johne's disease if there was histopathological evidence typical of the disease within the ileum and adjacent lymph nodes. RESULTS: Eighteen of 21 (86%) ewes with confirmed clinical Johne's disease were culture-positive for Map from sites peripheral to the alimentary tract, comprising 15 from skeletal muscle and 13 from blood. Five of 30 (17%) ewes that did not have Johne's disease were culture-positive, with four from skeletal muscle and one from blood. The likelihood that ewes with confirmed Johne's disease had systemic Map infection compared with ewes without was determined as OR=30 (95% CI=6.3-142.0; p<0.001). CONCLUSION: The prevalence of Map infection of skeletal muscle and blood in ewes with confirmed Johne's disease was 71% and 62% respectively, and in unaffected ewes was 13% for muscle and 3% for blood. CLINICAL RELEVANCE: Skeletal muscle and blood are potential sources of exposure of humans to Map, and the risk appears higher from sheep with Johne's disease.

Histopathology of grossly normal mesenteric lymph nodes of New Zealand farmed red deer (Cervus elaphus) including identification of lipopigment.

This article describes the histopathology of grossly normal mesenteric lymph nodes (MLNs) of New Zealand farmed red deer (Cervus elaphus). Eighty MLNs were sourced from 10 deer from 5 North Island herds and 5 South Island herds classified as low risk and high risk of Mycobacterium avium subspecies paratuberculosis (MAP) infection, respectively. Fixed sections were stained with hematoxylin and eosin; Ziehl-Neelsen; and, selectively, periodic acid-Schiff, Perl's, and Sudan black. Positive Ziehl-Neelsen stain, follicular hyperplasia, capsular eosinophil infiltration, focal granulomas, foci of macrophages containing lipopigment, parasitic granulomas, and calcified foci are described and severity graded where appropriate. Animal age, sex, and herd of origin are variably associated with the presence of one or more features. Trabecular fibrosis and dilated edema-filled sinusoids are described. These observations allow differentiation between likely nonpathologic histologic features in deer MLNs and features possibly attributable to infection with a pathogen such as MAP.

Assessment of occupational exposure to leptospirosis in a sheep-only abattoir.

This study estimated the frequency of exposure of meat workers to carcasses infected with Leptospira serovars Hardjobovis or Pomona in a sheep-only abattoir in New Zealand. A stochastic spreadsheet model was developed to assess the daily risk of exposure of eviscerators, meat inspectors and offal handlers to live leptospires in sheep carcasses from May to November 2004 (high-risk period), and from December 2004 to June 2005 (low-risk period). The average sheep processed per day were 225 for an eviscerator, 374 for a meat inspector, and 1123 for an offal handler. The median daily exposures during high- and low-risk periods were 11 [95% distribution interval (DI) 5-19] and three (95% DI 1-8) infected carcasses/day for eviscerators, 18 (95% DI 9-29) and six (95% DI 2-12) for meat inspectors, and 54 (95% DI 32-83) and 18 (95% DI 8-31) for offal handlers, respectively. Stochastic risk modelling provided evidence that processing of sheep carcasses exposed meat workers regularly to live leptospires with substantial seasonal variation.

Accumulation and depletion of liver copper stores in dairy cows challenged with a Cu-deficient diet and oral and injectable forms of Cu supplementation.

AIM: To quantify the capacity of the liver to respond to a Cu-deficient diet and various forms of Cu supplementation in dairy cows with high Cu status. METHODS: Sixty non-pregnant, non lactating mixed-age Friesian dairy cows were fed baled silage for 116 days, containing 5.8 mg Cu/kg dry matter (DM), that provided an inadequate Cu intake. They were either unsupplemented (Control); or treated thrice weekly orally with solutions containing the equivalent of 150 mg Cu/day as copper sulphate pentahydrate (CS), copper amino chelate (CAC), or copper glycinate (CG); 16 g Cu as CuO wire particles, administered in an intra-ruminal bolus on Day 0; or 100 mg Cu as calcium copper edetate, administered by S/C injection on Days 0 and 58. On Days -5, 14, 28, 58, 86 and 116, liver biopsies and blood samples were obtained for determination of Cu. RESULTS: Mean initial concentrations of Cu in liver for all groups was 827 (SE 42) micromol/kg fresh tissue. In control cows, this decreased to 552 micromol/kg on Day 116, and averaged 670 micromol/kg over the length of the trial. Oral forms of Cu supplementation increased overall mean concentrations in liver to 960 (SE 79), 1,050 (SE 81) and 1,100 (SE 84) micromol/kg for CS, CAC and CG, respectively, but there was no difference between form of supplement. Mean concentrations were significantly increased by bolus treatment, but not by injection. Concentrations of Cu in serum in all groups decreased from 12.1 (SE 0.3) to 10.4 (SE 0.6) micromol/L by Day 116, with no differences due to treatments. The initial concentration of Cu in liver significantly affected the rate of accumulation of Cu among cows supplemented orally. In cows with an initial concentration <1,100 micromol/kg, the average increase was 4.1 micromol/kg fresh tissue/day, whereas rates were variable, even negative, when initial concentrations were >1,100 micromol/kg. Release of Cu over 116 days in cows given the bolus was calculated to be equivalent to an oral intake of CS of 106 mg Cu/day. CONCLUSIONS: In dairy cows with a high Cu status fed a Cu-deficient diet, responses to supplementation with two chelated forms of Cu administrated orally were similar to those due to CS. Copper as CuO was about 77% as effective as oral supplementation with chemically similar inorganic Cu. Initial concentrations of Cu in liver influenced the rate of accumulation following oral supplementation. CLINICAL RELEVANCE: Changes in concentrations of Cu in liver were readily observed following oral supplementation over a wide range of initial concentrations, whereas these changes could not be detected in serum. The liver is thus a good index of the Cu status of dairy cows.

The impact of high zinc intake on the copper status of dairy cows in New Zealand.

AIM: To determine the effect of a high Zn intake on the concentration of Cu in the liver of dairy cows. METHODS: Sixty non-pregnant, non-lactating, mixed-age Friesian dairy cows were randomised into four groups: unsupplemented control; thrice weekly oral treatment with 350 mg Cu (equivalent to 150 mg Cu/day) as CuSO(4) x 5H(2)O; a ZnO bolus releasing 6.628 g Zn/day given on Days 0 and 28; and oral treatment with Cu and ZnO boluses. The cows were fed baled silage for the duration of the study (56 days). On Days -12, 14, 28, 42 and 56 liver biopsies were collected for determination of concentrations of Cu, and blood samples for measurement of Cu and Zn in serum. RESULTS: Mean concentrations of Cu in the liver of unsupplemented control cows decreased from an initial 717 (SE 108.5) to 396 (SE 88.0) micromol/kg fresh tissue on Day 28, and then increased to 542 (SE 105.3) micromol/kg fresh tissue on Day 42, before decreasing to 434 (SE 81.6) micromol/kg fresh tissue on Day 56. Administration of ZnO resulted in no change in the concentration of Cu in liver compared with controls. Supplementing with 150 mg Cu/day significantly increased the concentration of Cu in liver, compared with controls, to 1,074 (SE 105.2) micromol/ kg fresh tissue on Day 56. Treatment with ZnO significantly reduced concentrations of Cu in liver on Day 56, compared with Cu-supplemented cows, to 786 (SE 107.7) micromol/kg fresh tissue. The treatments had no effect on concentrations of Cu in serum. In cows given the ZnO, mean concentrations of Zn in serum increased from 17 (SE 0.6) on Day 0 to 30 (SE 1.7) micromol/L on Day 14, then decreased to 20 (SE 1.3) micromol/L on Day 28, increased to 40 (SE 2.9) micromol/L on Day 42, then decreased to 24 (1.16) micromol/L on Day 56. In cows given ZnO plus Cu, concentrations of Zn in serum were significantly greater than in cows only given ZnO on Day 41 (46 (SE 2.71) vs 35 (SE 3.23) micromol/L, respectively). Mean concentrations of Cu, Mo and Zn in the baled silage were 4.4, 0.5 and 21 mg/kg dry matter (DM), respectively. CONCLUSIONS: High Zn intakes used in the prevention of facial eczema in dairy cows had little effect on concentrations of Cu in liver when the Cu intake was low, but decreased the efficacy of a Cu supplement by about 50%. CLINICAL RELEVANCE: When using Zn as a prophylactic treatment for facial eczema the level of Cu supplementation should be adjusted, which may mean that in some situations it is recommended not to supplement cows with Cu.

The effects of pasture availability for twin- and triplet-bearing ewes in mid and late pregnancy on ewe and lamb behaviour 12 to 24 h after birth.

This experiment was conducted to evaluate the effect of offering ewes two different feeding levels, during mid and late pregnancy, on ewe and lamb behaviour 12 to 24 h after birth. Romney ewes, bearing twin (n = 80) or triplet foetuses (n = 56), were allocated to a pasture sward height of 2 or 4 cm between 70 and 107 days of pregnancy. In late pregnancy (day 107 to 147), half of the ewes were reallocated the alternate sward height, which produced four treatments: 2-2, 2-4, 4-2 and 4-4. Ewes were weighed on days 65, 92, 107 and 130 of pregnancy and lamb live weights were recorded 12 to 24 h after birth. Twelve to 24 h after birth the maternal behaviour score (MBS) of the ewes were determined, whilst their lambs were tagged. After the lambs were released, the behaviour of each ewe and her lambs was observed for 5 min. Ewe treatment and litter size had no effect on ewe MBS. However, as MBS increased (ewes stayed closer to lambs during tagging), ewes bleated less in a high-pitch and were quicker to make contact with their lamb. During the observation period, ewes in the 4-4 treatment had a greater percentage of their bleats in a low pitch (P < 0.05) than ewes in the 2-2 and 4-2 treatment (61.3% v. 41.3% and 38.8% low bleats, respectively) and more lambs born to 4-4 ewes (95%) bleated than lambs born to 2-2 ewes (84%; P < 0.05). However, lambs born to ewes in the 2-2 treatment bleated earlier than lambs in all other treatments (P < 0.05). Lambs born to 4-4 ewes were less likely (P < 0.05) to move towards their dam in order to make contact than lambs born to 2-2 or 4-2 ewes (3.1% v. 16.9% and 16.7%, respectively). These findings suggest that under the conditions of the present study, ewe nutrition had little effect on maternal behaviour. However, lambs born to ewes offered 2 cm pasture sward heights during mid and/or late pregnancy (2-2, 2-4 and 4-2 treatments) displayed behaviour that demonstrated greater 'need' whereas lambs born to ewes offered 4 cm during mid and late pregnancy sought less attention from their dam.

The effect of different types of stressors during mid- and late pregnancy on lamb weight and body size at birth.

Mid-pregnancy shearing has consistently been shown to increase lamb birth weight, which can lead to an increase in lamb survival rates. However, shearing ewes during the winter months and under outdoor pastoral farming conditions can expose the recently shorn ewe to a greater risk of hypothermia. The aim of this study was to determine if exposure of ewes to repeated stressors, in mid- and late pregnancy, would result in an increase in lamb birth weight. This information may assist in the elucidation of the mechanism for the birth weight response to mid-pregnancy shearing, which in turn could assist in the design of management options to increase lamb birth weight without placing the ewe at risk. One hundred and forty-four twin-bearing Romney ewes were allocated to one of six mid-pregnancy treatments: control, isolation on 2 or 10 occasions, sham-shearing on 10 occasions, intramuscular cortisol injection on 10 occasions or shearing. Isolation, sham-shearing and cortisol treatments were conducted twice a week beginning, on average, day 74 of pregnancy and shearing occurred on day 76. During pregnancy, ewe treatment had no effect on ewe live weight. However, average ewe body condition scores were higher in the shorn group than in the sham-shorn or cortisol groups (P < 0.05). Intramuscular injections of cortisol had a greater effect on ewe plasma cortisol concentrations than all other treatments (P < 0.05). Shearing produced a greater plasma cortisol response than isolation × 10 and sham-shearing (P < 0.05). Ewe plasma cortisol responses decreased during the 5 weeks of isolation and sham-shearing but cortisol injections produced a greater response during the fifth treatment than the first or ninth treatments (P < 0.05). Lambs born to shorn ewes were heavier and had a longer crown rump, forelimb and hind limb lengths than all other lambs (P < 0.05). In addition, lambs born to ewes in the cortisol treatment were lighter than lambs born to control, isolation × 2, isolation × 10 and shorn ewes (P < 0.05). The plasma cortisol concentrations observed for ewes injected with cortisol were far greater than those observed in all other groups, which is likely to explain the low birth weights of lambs born to ewes in that group. These results indicate that the mechanism by which mid-pregnancy shearing increases lamb birth weight is unlikely to be repeated stressors.