Investigation of pre-natal Chlamydophila abortus (Chlamydia psittaci) exposure of female lambs and the outcome of their first pregnancy.
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Abortion, Veterinary↗
Biomedical subjects
M J Clarkson
Publications and source records attributed to M J Clarkson.
The specificity of antibody responses in cattle naturally exposed to Fasciola hepatica.
Fasciola hepatica causes significant morbidity and mortality in dairy cattle in the Andean region of Cajamarca, Peru, where prevalence of infection of up to 78% has been reported. ELISA and Western blot analyses were used to characterise antibody responses in dairy cattle to adult F. hepatica to excretory-secretory (E/S), somatic (SO) and surface (SU) antigens. Three groups of dairy cattle - calves, heifers and adult cows - naturally exposed to F. hepatica in this region, were monitored every 2 months over a 2-year period. Calves, heifers and adult cows all had antibodies which recognised a 28kDa protein in the SO preparation, whereas only adult cows had antibodies that recognised a 28kDa protein in E/S products. All three groups of cattle responded to a 60-66kDa group of proteins in E/S and SU preparations and a 17kDa antigen in SO products was recognised by antibodies from cows and heifers but not calves. The total antibody response to E/S antigens measured by ELISA, increased over time in calves and remained constantly high over the 2-year period in all three groups of cattle. Slight fluctuations in the antibody response occurred in the group of heifers and cows coinciding with seasonal changes in the level of challenge.
Animals↗
Distinct importin recognition properties of histones and chromatin assembly factors.
Synthesis of the protein components of nuclear chromatin occurs in the cytoplasm, necessitating specific import into the nucleus. Here, we report the binding affinities of the nuclear localisation sequence (NLS)-binding importin subunits for a range of histones and chromatin assembly factors. The results suggest that import of histones to the nucleus may be mediated predominantly by importin beta1, whereas the import of the other components probably relies on the conventional alpha/beta1 import pathway. Differences in recognition by importin beta1 were observed between histone H2A and the variant H2AZ, as well as between histone H3/4 with or without acetylation. The results imply that different histone variants may possess distinct nuclear import properties, with acetylation possibly playing an inhibitory role through NLS masking.
Acetylation↗
Crystal structure of a nucleosome core particle containing the variant histone H2A.Z.
Activation of transcription within chromatin has been correlated with the incorporation of the essential histone variant H2A.Z into nucleosomes. H2A.Z and other histone variants may establish structurally distinct chromosomal domains; however, the molecular mechanism by which they function is largely unknown. Here we report the 2.6 A crystal structure of a nucleosome core particle containing the histone variant H2A.Z. The overall structure is similar to that of the previously reported 2.8 A nucleosome structure containing major histone proteins. However, distinct localized changes result in the subtle destabilization of the interaction between the (H2A.Z-H2B) dimer and the (H3-H4)(2) tetramer. Moreover, H2A.Z nucleosomes have an altered surface that includes a metal ion. This altered surface may lead to changes in higher order structure, and/or could result in the association of specific nuclear proteins with H2A.Z. Finally, incorporation of H2A.Z and H2A within the same nucleosome is unlikely, due to significant changes in the interface between the two H2A.Z-H2B dimers.
Amino Acid Sequence↗
Regions of variant histone His2AvD required for Drosophila development.
One way in which a distinct chromosomal domain could be established to carry out a specialized function is by the localized incorporation of specific histone variants into nucleosomes. H2AZ, one such variant of the histone protein H2A, is required for the survival of Drosophila melanogaster, Tetrahymena thermophila and mice (R. Faast et al., in preparation). To search for the unique features of Drosophila H2AZ (His2AvD, also referred to as H2AvD) that are required for its essential function, we have performed amino-acid swap experiments in which residues unique to Drosophila His2AvD were replaced with equivalently positioned Drosophila H2A.1 residues. Mutated His2AvD genes encoding modified versions of this histone were transformed into Drosophila and tested for their ability to rescue null-mutant lethality. We show that the unique feature of His2AvD does not reside in its histone fold but in its carboxy-terminal domain. This C-terminal region maps to a short alpha-helix in H2A that is buried deep inside the nucleosome core.
Amino Acid Sequence↗
The effect of cyclic temperatures on the growth of Fasciola hepatica and Lymnaea viatrix.
The experiment aimed to measure the effect of constant and variable temperatures on the growth of Lymnaea viatrix snails, on the development of a Peruvian isolate of Fasciola hepatica eggs and on the development of F. hepatica in the snails. This was carried out by cultivating infected and uninfected snails and fluke eggs in artificial, temperature controlled chambers. L. viatrix snails were found to develop at a rate dependent on environmental temperature, but developed at least as well under conditions of varying temperature as at the same mean constant temperature. F. hepatica eggs held at constant or varying temperatures, developed at a rate comparable to other reports. However, eggs developing at varying temperatures appeared to have reduced hatchability. Parasite development within the snails was slow, though within the limits calculated from the literature, and varying temperature did not appear to reduce development compared to constant temperatures.
Animals↗
Strategic control of fasciolosis in the inter-Andean valley of Cajamarca, Peru.
Fasciolosis is recognised as a major problem in dairy cattle in Cajamarca, Peru. The infection has an annual cycle, with the major period of infection from January to March. A control programme, involving two doses of the fasciolicide triclabendazole aimed at reducing the passage of Fasciola hepatica eggs on to the pasture, together with the use of the molluscicide, niclosamide, was evaluated against traditional treatment programmes. The double treatment regimen did not significantly reduce the overall parasite burden, as measured by faecal egg counts, but did control the parasites to a level similar to that achieved by traditional programmes averaging 3-7 treatments per year. The cows' eosinophil counts and serum liver enzyme activities were significantly lower than in animals treated by the traditional programmes. The use of niclosamide in addition to the triclabendazole treatments gave no clear advantage, although there was a significant reduction in the number of intermediate host snails after the application of the molluscicide.
Animal Husbandry↗
Acute phase toxoplasma abortions in sheep.
Within 13 days of the experimental infection of 15 oestrus-synchronised ewes with 2000 sporulated oocysts of Toxoplasma gondii at 80 to 90 days of gestation 11 had aborted. The infection induced pyrexia and specific antibody in all the ewes. One ewe resorbed its fetus, 11 ewes aborted and three delivered, at full term, live congenitally infected lambs whose pre-colostral serum was antibody-positive. Tissues from the aborted fetuses and placentae from the live lambs were examined for toxoplasma infection by polymerase chain reaction (PCR) amplification of the B1 gene and by mouse inoculation. The live lambs were all shown to be infected by both methods, but there was no evidence of infection in any of the tissues from the acute phase abortions, suggesting that these fatalities occurred before the placenta or the fetus had been invaded by T gondii. Such toxoplasma-induced, acute-phase abortions are likely to be impossible to diagnose from fetal tissues. These results have implications not only for the diagnosis of naturally occurring ovine abortions but also for the understanding of the pathogenesis of toxoplasma-induced abortion.
Abortion, Veterinary↗
Diagnosis of toxoplasma abortion in ewes by polymerase chain reaction.
Eighteen oestrus-synchronised ewes were infected experimentally with 1500 sporulated oocysts of Toxoplasma gondii between 80 and 90 days of gestation. The infection induced pyrexia and specific antibody in all the ewes. One ewe resorbed its fetus, five ewes aborted and 12 delivered live, congenitally-infected lambs whose pre-colostral serum was antibody-positive. Tissues from the aborted fetuses and placentae from the live lambs were examined for toxoplasma infection by polymerase chain reaction (PCR) amplification of the B1 gene and by mouse inoculation. Using a simple protocol of tissue preparation without DNA extraction and a nested format, PCR was as sensitive as mouse inoculation. Placental cotyledon gave a higher sensitivity of detection than brain, lung or liver, and 16 of 19 placentae were positive by PCR compared with 13 of 18 by mouse inoculation. In mock-infected tissues, as few as 10 tachyzoites could be detected. The PCR could be applied to tissues unfit for mouse inoculation.
Abortion, Veterinary↗
Experimental infection of pregnant ewes with Chlamydia pecorum.
Pregnant ewes were infected in midpregnancy with three isolates of Chlamydia pecorum derived from the feces of healthy lambs from three different farms. Oral infection, alone or together with Fasciola hepatica, did not result in tissue invasion, since all placental and fecal samples were negative for chlamydiae. Intravenous infection resulted in placental infection in 16 of 18 ewes in that chlamydiae were cultured from placentas or vaginal swabs. Two ewes bore dead lambs after a shortened gestation time. The chlamydiae isolated were all C. pecorum. There were no significant differences between the weights of the lambs from the infected groups and those from uninfected control ewes. Most ewes showed no serological evidence of infection by the complement fixation test; therefore, it is unlikely that the enteric subtype of C. pecorum is responsible for the cross-reactions sometimes seen in flocks being tested for C. psittaci infection.
Animals↗
Isolation of faecal chlamydia from sheep in Britain and their characterization by cultural properties.
Faeces samples were taken per rectum from sheep on 26 farms in England and Wales and examined for the presence of chlamydia by culture in McCoy cell monolayers. Thirteen of the farms were known to have had abortion outbreaks associated with Chlamydia psittaci (enzootic abortion) and 13 were free of this infection. The chlamydia isolated were characterized by cultural techniques. Chlamydia were isolated from the faeces of lambs on all 26 farms and the prevalence of infection varied form 5-50% on individual farms. There was no significant difference between the proportion of infected lambs on farms where enzootic abortion was present or absent. Lambs first showed infection when they were 3 months old and the prevalence rate of infection increased up to 9 months old. No chlamydia were isolated from the faeces of 316 adult ewes. The chlamydia were identified as enteric rather than abortion type and thus were C. pecorum rather than C. psittaci.
Abortion, Veterinary↗
Epidemiology of lameness in dairy cattle: the influence of cubicles and indoor and outdoor walking surfaces.
A survey of cubicles and indoor and outdoor walking surfaces on 37 farms served by four veterinary practices in Somerset, Cheshire, Wirral and west. Wales was carried out in 1989 to 1991. A study of the space requirements of Friesian/Holstein cows at pasture showed that they required approximately 240 cm x 120 cm lying space and a further 60 cm lunging space for rising. By these standards, 87 per cent of the cubicles were too short and 50 per cent were too wide or too narrow. Over 1500 observations on cows lying down, rising and standing indicated that only 12 per cent of the cubicles permitted real freedom of movement; 91 per cent of top partition rails were judged to be too low and 70 per cent of bottom rails too low or too high. In addition, the kerb was very high in 76 per cent of the cubicles. As a result, 10 per cent of cows appeared moderately or severely restricted when lying down, 33 per cent when rising and 55 per cent when standing. Over 2000 cubicle beds were also studied; 75 per cent had a concrete base and of those, 63 per cent were judged to have too little bedding and 11 per cent next to none. Higher incidences and prevalences of lameness were associated with limited borrowing space (P < 0.01) low bottom rails (P < 0.05), high kerbs (P < 0.05) and inadequate bedding (P < 0.01). Of 3190 walking surfaces, only 25 per cent were classified as satisfactory in the first winter and 34 per cent in the second. In general, surfaces in silage bays were too rough and those in other sites were too smooth. The farms with the smoothest indoor walking surfaces had a significantly higher incidence of lameness (P < 0.01). Of 3335 outdoor walking surfaces only 25 per cent were classified as satisfactory, and 70 per cent were too rough. The incidence of lameness was not significantly related to these findings.
Animals↗
Epidemiology of lameness in dairy cattle: description and analysis of foot lesions.
Information from 37 dairy farms, in four regions of England and Wales provided data on 8991 lesions and the preventive trimming of 4837 cows' feet. Of the total of 13,828 forms returned, veterinary surgeons treated 32 per cent and farmers or stockmen 46 per cent. Of the 8645 lesions associated with episodes of lameness, lesions in the hindlimbs accounted for 92 per cent, of which 65 per cent were in the outer claw, 20 per cent in the skin and 14 per cent in the inner claw. Sole ulcers (40 per cent) and white line lesions (29 per cent) were the predominant diseases of horn, and digital dermatitis (40 per cent) was the most common disease of the skin. Subjective assessments showed that sandcrack, penetration of the sole by foreign bodies and interdigital necrobacillosis were associated with the most severe cases of lameness. There was a significant seasonal effect in the reporting of lesions.
Animals↗
Incidence and prevalence of lameness in dairy cattle.
A survey was made of 37 dairy farms in Wirral, mid-Cheshire, mid-Somerset and Dyfed, Wales, to assess the incidence and prevalence of lameness in the cows between May 1989 and September 1991. The incidence was obtained from records made whenever a cow was examined for lameness or received preventive foot-trimming. The mean annual incidence was 54.6 new cases per 100 cows with a range from 10.7 to 170.1 and the mean values during summer and winter were 22.9 and 31.7, respectively. The prevalence of lameness was measured by regular visits at which locomotion was scored on a scale of 1 to 5, and the prevalence of lameness was calculated for each visit as the proportion of cows with scores of 3 or more. The mean annual prevalence over the whole period was 20.6 per cent with a range from 2.0 to 53.9 per cent for the 37 farms. The mean prevalences during summer and winter were 18.6 and 25.0 per cent, respectively. The prevalence measured at a single visit in midsummer or midwinter was significantly correlated with the mean prevalence over the whole corresponding period and may be useful as an assessment of the extent of lameness in a herd and the efficacy of control measures. There was evidence that training farmers to recognise early cases of lameness and request veterinary treatment resulted in a marked reduction in the duration of cases of lameness.
Animals↗
Analyses of the genomes of chlamydial isolates from ruminants and pigs support the adoption of the new species Chlamydia pecorum.
Analysis of the genomic DNAs of chlamydial isolates from sheep, cattle, and pigs was performed by Southern blot hybridization and by restriction endonuclease (RE) profiling of DNA amplified by PCR. The hybridization probes were derived from whole genomic DNA, the major outer membrane protein (MOMP) gene, the 16S rRNA gene, and an avian Chlamydia psittaci isolate plasmid. The PCR analysis used targets in the MOMP gene, the 16S rRNA gene, and the 60-kDa cysteine-rich protein gene. Together, the results showed that although there was considerable heterogeneity in the DNA sequence in the MOMP gene region, all the isolates had the same underlying total genomic RE profiles and yielded identical RE profiles for the rRNA and 60-kDa-protein gene regions. Most of the isolates were found to hybridize with the plasmid probe. Comparison of the MOMP sequence of one of the isolates (P787) with that of a known Chlamydia pecorum strain together with the results of the RE analyses allowed the conclusion that the isolates should all be classified within this new species.
Amino Acid Sequence↗
Spontaneous change from overt to covert infection of Chlamydia pecorum in cycloheximide-treated mouse McCoy cells.
Some isolates of Chlamydia pecorum from sheep feces failed to produce inclusions on passage in cycloheximide-treated monolayers, but chlamydiae could be recovered several weeks later. Chlamydia psittaci from sheep abortions did not show this phenomenon.
Animals↗
Responses of the post-term arterial duct to oxygen, prostaglandin E2, and the nitric oxide donor, 3-morpholinosydnonimine, in lambs and their clinical implications.
BACKGROUND: Nitric oxide is a potent dilator of the pulmonary vasculature. There have been no previous reports on the action of nitric oxide on the arterial duct. OBJECTIVES: To determine the responses of isolated post-term arterial duct rings from lambs to oxygen, prostaglandin E2 (PGE2) and the nitric oxide donor, 3-morpholinosydnonimine (SIN-1). SETTING: Experimental laboratory. SUBJECTS: Six neonatal lambs. METHODS: Lambs aged 1-5 days were killed and the arterial duct and aorta excised and cut into rings. These were mounted on tension gauges in organ baths containing Krebs-Henseleit solution. Rings were exposed to increasing concentrations of oxygen, PGE2 and after preconstriction with potassium (40 mmol/l) to SIN-1. Tension and relaxation responses were recorded. RESULTS: Increased oxygen tension resulted in increased tension in the ductal rings above 88.9 mm Hg as previously described. No response to PGE2 occurred before or after ductal rings were exposed to oxygen. SIN-1 caused relaxation of smooth muscle in the arterial duct to a similar degree as that in the aortic rings. CONCLUSIONS: As previously shown, oxygen is a potent constrictor of the arterial duct. The post-term arterial duct does not relax in response to PGE2 possibly as a result of inactivation by oxygen of the special sensitivity of the duct to PGE2. SIN-1 is a potent smooth muscle relaxing agent in the term arterial duct and may have a role in the initial management of neonates with duct dependent pulmonary circulation.
Animals↗