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Biomedical subjects

D M West

Publications and source records attributed to D M West.

At least 37 records · Page 2Linked to original sources

Pulsed-field gel electrophoresis typing of Campylobacter fetus subsp. fetus from sheep abortions in the Hawke's Bay region of New Zealand.

AIM: To type Campylobacter isolates from sheep abortions from the Hawke's Bay region of New Zealand. METHODS: Campylobacter isolates were collected from aborted sheep fetuses from commercial farms in the Hawke's Bay region. Information on the Campylobacter vaccination status of flocks in the study was collected. Isolates were identified to species level using standard phenotypic tests, then typed using pulsed-field gel electrophoresis (PFGE). RESULTS: Eighty-one C. fetus subsp. fetus isolates were cultured from aborted sheep fetuses from 25 farms and four C. jejuni isolates were cultured from fetuses from three farms. The C. fetus subsp. fetus isolates were classified into six PFGE groups. A single pulsed-field type predominated amongst isolates from 19 of the 25 farms. The C. jejuni isolates comprised two types. CONCLUSIONS: A range of C. fetus subsp. fetus PFGE types was identified, and one type, B1, was found most frequently. Campylobacter fetus subsp. fetus was only isolated from samples from sheep that had not been vaccinated with C. fetus subsp. fetus vaccine that season.

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Chondrodysplasia of Texel sheep - a new disease of suspected genetic aetiology (abstract).

A chondrodysplasia characterised by dwarfism and varus deformity of the forelimbs has been recognised over four seasons in Texel and Texel-cross lambs on three related properties. Affected lambs appear normal at birth, but by 2 weeks of age have reduced growth rate, shortened neck and legs, varus forelimb deformities and a progressive reluctance to walk. Most die within the first 4 months of life. Some affected lambs have had normal co-twins. Post mortem examination of two severely affected 3-month-old lambs revealed extensive loss of articular cartilage and exposure of subchondral bone on the weight-bearing surfaces of the proximal humeri and femurs. Articular and physeal cartilages were thicker than normal, suggesting delayed or impaired endochondral ossification. Histopathologically, there was disorganisation of chondrocytes in articular and physeal cartilage of all bones examined. In addition, there were multiple foci of chondrolysis, which coalesced in some areas to form large clefts. The chondroid matrix surrounding chondrocytes had an abnormal fibrillar appearance. One lamb that exhibited early signs of the disease died of an unrelated cause at 3 weeks of age, and although there was no erosion of articular cartilage, characteristic microscopic lesions were present in articular and physeal cartilage. Although most of the affected lambs were born on a commercial property with a mix of different sheep breeds, all had Texel genes on both sire and dam sides of the pedigree. The farmer had been purchasing his Texel rams from the same flock for several seasons. Circumstantial evidence favours a genetic aetiology, characterised by either autosomal recessive inheritance or over dominance. Studies on the genetic and biochemical basis of the disease are continuing.

Journal Article↗

Microphthalmia in Texel lambs.

AIM: To describe an outbreak of congenital microphthalmia in Texel lambs in New Zealand. METHODS: Affected eyes were fixed in 10% formol saline, processed routinely for histopathology and stained with haematoxylin and eosin, Holme's silver and haematoxylin with and without luxol fast blue, and luxol fast blue alone. RESULTS: Macroscopically, the globes and optic nerves were hypoplastic. Histologically, all embryonic components of the eye were present, but the lens, ciliary body, iris and retina were dysplastic. Axons of the hypoplastic optic nerve were poorly myelinated. CONCLUSIONS: Congenital microphthalmia occurs in Texel sheep in New Zealand, and is likely inherited as an autosomal recessive trait.

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A multiplex allele specific polymerase chain reaction (MAS-PCR) on the dihydrofolate reductase gene for the detection of Cryptosporidium parvum genotypes 1 and 2.

A multiplex allele specific polymerase chain reaction (MAS-PCR) based on the Cryptosporidium parvum dihydrofolate reductase (dhfr) gene sequence differentiates genotype 1 ('Human') from 2 ('Cattle') in a 1-step reaction. The MAS-PCR was validated on a panel of 34 microscopically positive C. parvum faecal samples of human and animal origin in comparison with 2 published PCR-restriction fragment length polymorphism (RFLP) methods targeting dhfr and the oocyst wall protein (cowp) genes. A validation panel of 37 negative faecal samples of human and animal origin was also tested in comparison with the cowp PCR-RFLP. MAS-PCR was found to be as sensitive for species detection as the most sensitive of the other tests, and detected more mixed genotype infections than the two other tests combined. In addition the MAS-PCR showed equivalent detection sensitivity in comparison with a published nested RFLP targeting the SSU rRNA gene, on a panel of prepared mixed genotype samples. The 1-step reaction is simpler and less expensive to perform than the RFLP methods, while the C. parvum specific amplicons and those for genotypes 1 and 2 (575, 357 and 190 bp respectively) can be easily distinguished on agarose gel.

Alleles↗

Effects of Brucella ovis infection on semen characteristics of 16-month-old red deer stags.

AIM: To determine the effects of Brucella ovis infection on semen characteristics of 16-month-old red deer stags (Cervus elaphus). METHODS: At monthly intervals during March, April and May, semen was collected using electro-ejaculation from 9 yearling red deer stags that had been artificially infected with B. ovis 3 months previously. In March, semen was also collected from 6 non-infected stags from the same peer group for comparison. Semen was evaluated for gross appearance, percentage of sperm showing forward motility, sperm morphology and sperm density/ml of semen, and the presence of white blood cells was determined. In addition, at the time of semen collection, the epididymes of each stag were palpated and lesions recorded. RESULTS: Grossly visible purulent material was present in semen from 6/9 infected stags and the percentage of sperm showing forward motility did not exceed 20% in any of these samples. Increased numbers of white blood cells and cellular debris were evident in semen from 8/9 infected stags. Compared with non-infected stags, sperm motility in semen from infected stags was significantly reduced (p<0.05). There were no differences in percentage of morphologically normal sperm between groups, but infected stags had more detached sperm heads present in semen than non-infected stags (p<0.05). Only 2 of the infected stags had lesions of epididymitis evident on scrotal palpation. CONCLUSIONS: B. ovis infection is likely to have a detrimental effect on stag semen quality. While there was variation between individual animals, overall sperm motility was decreased and semen from infected stags had increased numbers of white blood cells and detached sperm heads. CLINICAL RELEVANCE: The fertility of breeding stags may be reduced if they are infected with B. ovis and this possibility should be considered when investigating reproductive problems of deer.

Journal Article↗

Effects of vaginal Brucella ovis infection of red deer hinds on reproductive performance, and venereal transmission to stags.

AIMS: To investigate the effects of vaginal Brucella ovis infection on the reproductive performance of red deer (Cervus elaphus) hinds. To determine whether stags may become infected with B. ovis by venereal transmission from mating infected hinds. METHODS: Thirty mixed-age red deer hinds serologically negative for B. ovis antibodies were synchronised for oestrus on 22 March 2000. B. ovis was inoculated into the vagina of each hind at oestrus and again, 18 days later. At oestrus, hinds were randomly allocated to six groups, each joined with a 16 month-old red deer stag seronegative for B. ovis, for 55 days. Hinds were blood sampled and scanned for pregnancy using rectal ultrasonography at monthly intervals. Six pregnant and four non-pregnant hinds were slaughtered pre-calving and three hinds were slaughtered post-calving. Reproductive tracts and foetuses were examined grossly, histologically and microbiologically. Calves were identified and blood sampled within 3 days of birth. Hinds and calves were blood sampled in February and May 2001 and vaginal swabs were collected from hinds for B. ovis culture. Blood was collected from stags, 5 and 19 days after mating and semen was collected for B. ovis culture. The 17 remaining hinds were mated in 2001 to two mixed-age wapiti (Cervus canadensis) stags. Both stags were blood sampled after mating. Sera were tested in a B. ovis complement fixation test (CFT) and enzyme-linked immunosorbent assay (ELISA). RESULTS: All 30 hinds developed B. ovis antibody levels, measurable using either the CFT or ELISA, but these did not remain elevated. There was no evidence of infection, either by gross pathology, histopathology or microbiological culture in the ten hinds or six foetuses slaughtered pre-calving. All remaining 20 hinds produced normal calves, 15 of which survived until weaning. Three hinds experienced dystocia and gave birth to dead calves and two calves died within 4 days of birth. One hind which had dystocia was euthanased. Samples from this hind and from 3/5 dead calves showed no evidence of B. ovis infection. B. ovis was cultured from the vagina of 1/19 hinds 48 weeks after inoculation, at which time B. ovis CFT and ELISA results for this hind were negative. Most calves had B. ovis serum antibodies at 1-3 days of age but levels were negligible when sampled at 10-15 weeks of age. Foetuses and dead calves were all seronegative. Three of the five red deer stags used for mating became infected with B. ovis. The two wapiti stags used to mate the remaining 17 hinds the following year remained seronegative. CONCLUSIONS: B. ovis is unlikely to have significant detrimental effects on the reproductive performance of red deer hinds. Venereal transmission via the vagina of hinds is a possible route of transmission between stags. It is possible that survival of the organism in the vagina of some hinds could create difficulties in disease control programmes. CLINICAL RELEVANCE: B. ovis infection of hinds at the time of mating is unlikely to cause significant reproductive losses. Venereal transmission of B. ovis between stags via the hinds may occur when groups of hinds are joined with more than one stag.

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Consequences of anthelmintic resistance on liveweight gain of lambs on commercial sheep farms.

AIM: To assess the consequences of anthelmintic resistance on commercial sheep farms on liveweight (LW) gains and susceptibility to diarrhoea of growing lambs. METHODS: Five farms with a history of resistance to benzimidazole drenches were selected. On each farm, 150 ewe lambs were randomly allocated to 1 of 3 treatment groups (n=50). Group 1 was treated 5 times at 28-day intervals with oxfendazole (to which nematode resistance had been demonstrated); Group 2 was treated 5 times at 28-day intervals with levamisole or levamisole + oxfendazole (to which nematode resistance had not been demonstrated) and; Group 3 was treated with a combination of slow-release albendazole capsules and oral moxidectin (intended to totally suppress nematode infection). Differentiated faecal egg counts (FECs), FEC reduction tests (FECRT), LW gain and degree of faecal soiling around the breech (dag scores) were determined and compared between groups. RESULTS: Pre-treatment FECs in Group 1 were similar to or slightly lower than those of Group 2. Efficacy of oxfendazole in Group 1 varied between farms and over time within farms, the reduction in FECs after treatment varying from 0% to 95%. The genera identified as resistant on each farm also varied over time. Lambs from Group 3 had lower FECs, lower dag scores and higher growth rates than lambs from Group 2, which in turn had higher LW gains than lambs from Group 1. The overall LW gains during the 5 months of the study were 7.03+/-0.27 kg for Group 1, 8.33+/-0.27 kg for Group 2, and 9.91+/-0.31 kg for Group 3 (mean+/-SEM; means differed significantly from each other, p<0.01). CONCLUSIONS: The decrease in growth rates associated with the continued use of an incompletely effective anthelmintic were surprisingly small in situations where all lambs grazed together.

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The difference in efficacy of ivermectin oral, moxidectin oral and moxidectin injectable formulations against an ivermectin-resistant strain of Trichostrongylus colubriformis in sheep.

AIM: To evaluate the efficacy of ivermectin oral, moxidectin oral and moxidectin injectable formulations against an ivermectin-resistant strain of Trichostrongylus colubriformis in sheep. METHODS: Twenty-four mixed breed lambs were infected with 15,000 infective third-stage larvae of an ivermectin-resistant strain of T. colubriformis which had originally been isolated from a goat farm in Northland in 1997. Twenty-six days post infection, the lambs were divided into 3 treatment groups and a control group (n=6 lambs/group). Treatment consisted of either ivermectin oral formulation (0.2 mg/kg), moxidectin oral formulation (0.2 mg/kg), or moxidectin injectable formulation (0.2 mg/kg). Faecal egg counts (FECs) were determined at 0, 3, 5, 7 and 10 days after treatment. All animals were necropsied 12 days after treatment and worm counts were performed. Larval development assays were conducted 24 days post infection. A further 3 lambs were infected with 15,000 infective third-stage larvae of a fully susceptible strain of T. colubriformis for comparative purposes in the larval development assay. The efficacy of the moxidectin injectable formulation was also confirmed in these 3 lambs. RESULTS: The FEC reduction test at day 10 after treatment revealed 62%, 100% and 0% reductions in arithmetic-mean FECs for ivermectin oral, moxidectin oral and moxidectin injectable groups, respectively. The ivermectin oral, moxidectin oral and moxidectin injectable formulations achieved 62%, 98% and 4% reductions in arithmetic-mean worm burdens, respectively. Larval development assays showed resistance ratios for ivermectin of 4:1, avermectin B2 of 2.7:1, ivermectin aglycone of 37:1, moxidectin of 1.4:1, thiabendazole of 14.6:1 and levamisole of 1.8:1. CONCLUSIONS: The moxidectin oral formulation provided a high degree of control against ivermectin-resistant T. colubriformis whereas the moxidectin injectable formulation had very low efficacy. Ivermectin aglycone was the analogue of choice for diagnosis of ivermectin resistance in T. colubriformis in the larval development assay.

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Oral mucosal Langerhans' cells as target, effector and vector in HIV infection.

The mechanism underlying a transition of the oral cavity mucosal epithelium towards susceptibility to opportunistic infections in HIV-seropositive patients was investigated. Phenotypic markers CD1a, HLA-DR, and CD86 of oral mucosal Langerhans' cells (LCs), p17 core protein of human immunodeficiency virus (HIV), and CD45RO of memory T cells were labeled on oral hairy leukoplakia lesional biopsies and clinically normal autologous tissue of HIV-infected patients. HIV p17 protein was detected in association with mucosal LCs, mainly within the lesional epithelium. There were significant correlations between the detection of HIV p17 and the depletion of LCs, and between the depletion of LCs and the presence of hairy leukoplakia lesions. Conjugates of activated LCs and memory T cells were also evident in the submucosal area of lesional biopsies. The findings from this study support the hypothesis that oral mucosal LCs are also the target of HIV infection. Cytopathic changes of LCs caused by productive HIV infection may contribute to selective depletion of LCs, which may impair the mucosal immunologic protection against colonization by microorganisms causing HIV-associated oral mucosal lesions.

AIDS-Related Opportunistic Infections↗

Transmission of Brucella ovis from rams to red deer stags.

AIM: To determine whether B. ovis will transmit from infected rams to non-infected red deer stags (Cervus elaphus) grazing together in the same paddock. METHODS: Six rams artificially infected with B. ovis were grazed with six non-infected 14-month-old red deer stags for a four and a half month period from March 4 to July 20, 1999. Stags were blood sampled at one- to six-weekly intervals to test for B. ovis antibodies using a complement fixation test. Stags that seroconverted were semen sampled to test for B. ovis infection by bacteriological culture. RESULTS: Between day 92 and day 124 of grazing together (June 4 and July 6), sera from five of the six stags became positive in the B. ovis complement fixation test. B. ovis was cultured from semen samples from four of the seropositive stags. CONCLUSIONS: Brucella ovis can be transmitted from infected rams to non-infected stags grazing in the same paddock, suggesting that B. ovis infection in farmed deer in New Zealand initially came from infected rams. Whether transmission occurs from direct contact between rams or stags, or indirectly by environmental contamination needs to be established.

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Effect of an injectable microencapsulated Vitamin B12 on serum and liver Vitamin B12 concentrations in calves.

AIM: To evaluate the efficacy of a long-acting injectable microencapsulated formulation of Vitamin B12 in dairy calves. METHOD: Fifty calves, average liveweight 110kg, were randomly allocated to 5 groups of 10 animals and injected subcutaneously in the anterior neck with 0.12, 0.18, 0.24 and 0.3 mg Vitamin B12/kg liveweight using a formulation of microencapsulated Vitamin B12 in a lactide: glycolide copolymer. The untreated calves were injected with the same vehicle, without Vitamin B12. Subsequent changes in serum and liver Vitamin B12 concentrations were followed for 244 days. RESULTS: The microencapsulated Vitamin B12 significantly increased, then maintained serum and liver Vitamin B12 concentrations higher than those of untreated controls for at least 110 days. CONCLUSIONS: Injection of the microencapsulated Vitamin B12 at dose rates of 0.12 to 0.24 mg/kg liveweight will increase and maintain the Vitamin B12 status of calves for at least 110 days.

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Attempted transmission of Brucella ovis between red deer stags by successive grazing or adjacent-paddock grazing.

AIM: To determine whether Brucella ovis can be transmitted from stag to stag by successive grazing of infected and noninfected stags in the same paddock, or by grazing infected and non-infected stags in adjacent paddocks. METHODS: Six red deer stags (Cervus elaphus) were artificially infected with B. ovis and 5 were confirmed to be shedding the organism in their semen. Infected stags alternated paddocks, and therefore grazing and wallows (successive grazing), once or twice weekly with 6 non-infected stags from 3 March to 18 August, 1999. Direct contact between the 2 groups of animals was prevented. The 2 groups alternated paddocks 32 times. Six other non-infected stags were grazed in a paddock adjacent to the infected stags throughout this period, separated by a standard deer fence. Non-infected stags were blood sampled at 2to 6-week intervals to test for B. ovis antibodies using a complement fixation test and an enzyme linked immunosorbent assay. RESULTS: No stag from either non-infected group became infected with B. ovis. CONCLUSIONS: The risk of stags becoming infected with B. ovis by successive grazing of the same paddock as, or by grazing in paddocks adjacent to, infected stags appears to be low. We conclude from this result, and similar experimental evidence and experience of this disease in sheep, that transmission of B. ovis requires animals to be grazed or confined together in a way that allows direct contact between animals. CLINICAL RELEVANCE: It is likely that infected and non-infected stags can be managed on the same property without transmission occurring between the groups, provided that they do not come into direct contact with one another.

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Campylobacter fetus fetus abortions in vaccinated ewes.

AIMS: To investigate the cause of an outbreak of ovine abortion in 1996 in a flock of 300 two-tooth (rising 2-year-old) ewes vaccinated against Campylobacter fetus fetus infection and to subsequently characterise the strain of C. fetus fetus isolated from aborted foetuses. METHODS: Standard bacteriological methods were used to identify C. fetus fetus isolates which were then antigenically typed and subjected to pulsed-field gel electrophoresis (PFGE) and compared to the vaccine strain. RESULTS: C. fetus fetus was identified as the causal agent of the abortions despite the ewes having been vaccinated before ram introduction and at the time of ram removal. Four isolates cultured from aborted material were indistinguishable when compared using antigenic typing and PFGE, but all differed from the vaccine strain. CONCLUSIONS: Within the limitations of the available typing systems, it is proposed that PFGE may be a useful tool to establish the distribution and strain variation of C. fetus fetus. CLINICAL RELEVANCE: This field case indicates the need for further study of non-vaccine C. fetus fetus strains which cause abortion in vaccinated ewes, and of the importance of these strains to the New Zealand sheep industry.

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Comparison of three serological tests and an interferon-gamma assay for the diagnosis of paratuberculosis in experimentally infected sheep.

OBJECTIVE: To compare the diagnostic performance of a complement fixation test, an agar gel immunodiffusion test, an enzyme-linked immunosorbent assay, and a whole-blood interferon-gamma assay for paratuberculosis in 14 sheep experimentally infected with Mycobacterium avium subsp paratuberculosis. EXPERIMENTAL DESIGN: Longitudinal study. RESULTS: The IFN-gamma assay detected more experimentally infected sheep, and earlier, than any of the serological tests. None of the antibody assays was able to detect all sheep with histologically confirmed paratuberculosis. CONCLUSIONS: The superior performance of the IFN-gamma assay in detecting infected sheep in this small experimental population warrants its further evaluation in a larger population of sheep naturally exposed to M a paratuberculosis.

Animals↗

Vaccination against paratuberculosis of lambs already infected experimentally with Mycobacterium avium subspecies paratuberculosis.

OBJECTIVE: To assess the protective value of a live-attenuated vaccine in sheep already exposed to Mycobacterium avium subsp paratuberculosis and to investigate the progression of a systemic immune response in experimentally infected sheep. STUDY DESIGN: Twenty-eight lambs, aged 1 to 1.5 months, were dosed via stomach tube with approximately 4.4 x 10(8) M a paratuberculosis organisms. Two weeks later, 14 of these 28 animals received subcutaneous injections of 1 mL of a live-attenuated vaccine. Thirteen additional lambs were neither dosed nor vaccinated (negative controls). Antigen-induced production of IFN-gamma in blood, and antibody concentrations in serum were sequentially monitored in vaccinated, unvaccinated and control animals for 1 year. Each sheep was examined for infection by an IS900-based PCR test on samples of ileum and ileocaecal lymph node and histological examination at the time of necropsy. RESULTS: Seven of 14 unvaccinated and two of 14 vaccinated sheep developed clinical paratuberculosis that was later confirmed by histological examination and/or the IS900-based PCR test. The granulomatous inflammation in the jejunal and ileal mucosa was less severe in vaccinated than in unvaccinated sheep. Acid-fast organisms were detected only in the unvaccinated group. The PCR assay on ileal samples gave positive reactions in two vaccinated and eight unvaccinated sheep. Both the antibody response and IFN-gamma response were detected earlier and were more substantial in vaccinated than in unvaccinated sheep. Furthermore, in experimentally infected but unvaccinated sheep, the IFN-gamma concentrations were higher in those animals without acid-fast organisms than in those with them. CONCLUSIONS: Vaccination of lambs with live-attenuated vaccine 2 weeks after oral inoculation with M a paratuberculosis stimulated the host response against the organism and led to a reduced mycobacterial burden. The diminished IFN-gamma responses in experimentally infected sheep with acid-fast organisms suggest a positive relationship between the magnitude of the systemic cell-mediated immune response and an animal's ability to control infection.

Animals↗

Use of the polymerase chain reaction assay for the detection of Mycobacterium avium subspecies paratuberculosis in blood and liver biopsies from experimentally infected sheep.

OBJECTIVE: To evaluate a polymerase chain reaction-based assay for the detection of Mycobacterium avium subsp paratuberculosis in blood and liver biopsies from subclinically infected sheep. STUDY DESIGN: A direct PCR assay for the detection of M a paratuberculosis was applied to liver biopsy specimens and to samples of blood that were sequentially collected over 53 weeks from 14 sheep infected experimentally with the organism. RESULTS: Of 117 blood samples from the 14 experimentally infected sheep, two tested positive in the PCR assay. Both positive results were obtained in two subclinically infected sheep that had paratuberculosis later confirmed by histological examination at necropsy. However, the assay failed to detect the target DNA in samples of blood from five other sheep with histologically confirmed paratuberculosis. Similarly, the PCR assay on liver biopsy specimens collected 32 weeks after administration of M a paratuberculosis gave only two positive results, both of which were obtained in sheep with histological evidence of paratuberculosis. CONCLUSIONS: The PCR assay on blood and liver biopsies does not provide a useful method for the diagnosis of M a paratuberculosis infection in subclinically infected sheep.

Animals↗