Biomedical subjects
Publications and source records attributed to A J Trees.
Examination of ovaries and testes from adult Onchocerca ochengi, O. gutturosa, O. armillata and O. lienalis revealed five pairs of chromosomes, but in contrast O. volvulus and O. gibsoni had only four pairs.
Laboratory reared nulliparous female flies of six temperate species of Simuliidae were examined for their susceptibility to infection with Onchocerca ochengi by intrathoracic injection of cryopreserved skin microfilariae obtained from cattle in Mali. Three species (S. equinum, S. ornatum and S. erythrocephalum) supported development to the infective stage, one species (S. variegatum) allowed partial development and the remaining two species (S. reptans and S. aureum) were insusceptible to infection. The most suitable surrogate vectors were S. equinum and S. ornatum which had survival rates of 44% and 49%, proportions of microfilariae developing to third stage larvae of 6.4% and 3%, and infection rates with infective larvae of 13.5% and 14% respectively. O. volvulus infective larvae, produced by intrathoracic microfilarial injection in S. ornatum, were 586-760 microns (mean 687 microns) long and were significantly shorter (p less than 0.02) than the O. ochengi infective larvae (645-880 microns, mean 756 microns). No constant differences in the posterior or anterior morphology, or in the acid phosphatase staining patterns between O. ochengi and O. volvulus, were seen. These results raise the possibility that the presence of O. ochengi in a population of infective larvae from vector flies in endemic onchocerciasis zones might be identifiable on the basis of their length alone.
Previous studies of bovine Onchocerca spp. in cattle in Sierra Leone indicated that only O. gutturosa was transmitted in the forest zone at high intensity. To determine its vector(s) and the extent to which Onchocerca-like infections in Simulium damnosum were likely to be of bovine origin, three lines of investigation were pursued. Firstly, a study was made of the biting flies attacking an ox bait animal over a 14-month period at Njala University Campus, near Bo. Secondly, attempts were made to infect the dominant local forest cytospecies of S. damnosum s.l. with O. gutturosa by feeding them on an infected ox under a bed-net. Thirdly, S. damnosum s.l. were infected by intra-thoracic injection of O. gutturosa microfilariae (mff). In 113 collections made at dawn and dusk at weekly intervals from the ox bait, 624 simuliids and 7740 Culicoides spp. were collected. Almost all the simuliids were S. damnosum s.l. which, on the basis of iso-enzyme examination and knowledge of local breeding sites, were identified as S. soubrense 'B'. Although this cytospecies fed readily on the ox at ventral sites where O. gutturosa mff occurred and the bed-net experiments showed that 16.1% of engorged S. soubrense 'B' ingested an average of 3.3 O. gutturosa mff each, no development occurred. The refractoriness of S. damnosum s.l. to O. gutturosa was confirmed by the intra-thoracic injection experiments. The predominant Culicoides spp. attacking the ox bait were C. krameri, C. trifasciellus and C. fulvithorax, with smaller numbers of C. schultzei. In 5803 dissected Culicoides spp., natural infections of Onchocerca-like larvae, presumed to be O. gutturosa, were found in 0.3% of C. fulvithorax, 0.1% of C. trifasciellus and 0.06% of C. krameri. It was concluded that, in the forest zone of Sierra Leone, S. damnosum s.l. is not a vector of O. gutturosa and all Onchocerca-like larvae in S. damnosum are likely to be O. volvulus, while the natural vectors of O. gutturosa are the Culicoides species C. fulvithorax, C. trifasciellus and C. krameri.
The antibody response of 20 pregnant ewes to oocyst infection with Toxoplasma gondii was determined by the latex agglutination test (LAT) and compared with the indirect fluorescent antibody test (IFAT) and a commercially available indirect haemagglutination test (IHAT). The LAT and IFAT showed a similar rapid response with antibody first appearing by two to three weeks after infection and titres that correlated closely (r = 0.81, P less than 0.001). The IHAT response was slower and less consistent up to seven weeks after infection. The LAT response was biphasic in seven of the sheep. Sera were fractionated using a minicolumn gel filtration technique and specific IgM and IgG titres determined by LAT. IgM titres peaked three weeks after infection and IgG titres exceeded IgM titres at a mean time of 4.7 weeks after infection (range 3 to 7). Eleven sheep exhibited fetopathy with abortion/parturition 12 to 53 days after infection; in nine of them IgG titres exceeded IgM at that time. A non-specific anti-toxoplasma reaction associated with IgM antibody occurred at low titre in one sheep. The results indicate that used from a dilution of 1/64 the LAT is a sensitive, reliable and rapidly responsive serological test for toxoplasma infection in ewes and it may be utilised with sample fractionation techniques to determine IgM titres. It is suggested that the best time to examine ewe sera to assist diagnosis of toxoplasma abortion is one week after abortion. While the determination of specific IgM titres in ewe sera may assist epidemiological studies and, sometimes, diagnosis, in the majority of aborting ewe sera it is unlikely to aid diagnosis.
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Monensin was fed to 69 pregnant ewes from 81 to 84 days gestation until lambing, at an estimated rate of nil, 16.8 or 27.9 mg per head per day. Ten days after the start of this regime, groups of the ewes were dosed orally with nil, 2000 or 12,000 sporulated Toxoplasma gondii oocysts. Twenty ewes given T. gondii alone (T ewes) produced 29 lambs or aborted foetuses, 16 (55.2 per cent) of which were born dead. The 39 ewes given monensin and T. gondii (M + T ewes) produced 48 lambs or aborted foetuses, 8 (16.7 per cent) of which were born dead. The 10 ewes given monensin alone produced 12 live lambs. No difference of effect was apparent between the two doses of monensin given, nor between the two doses of Toxoplasma oocysts used. Monensin alone caused no discernible problems. Not only were proportionately more live lambs born to M + T ewes than to T ewes, but they were also heavier, possibly due to a lesser "weight" of infection within the gravid uterus. We conclude that monensin fed at about 16 mg per head per day to pregnant ewes can significantly reduce losses at lambing time due to experimentally administered T. gondii.
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Onchocerca gutturosa and O. lienalis infections in British cattle were studied by examination of cattle post-mortem originating from North Wales and Cheshire (north west England). In 463 adult animals, the microfilarial (mf) prevalence was 28.5%. In 95.3% of the mf infected animals, gravid worms could not be found at either the ligamentum nuchae or the gastro-splenic omentum. Dermal mf at the head were identified as O. gutturosa on the basis of their highly significant association with the presence of gravid O. gutturosa at the ligamentum nuchae, which were found in only 3.2% of cattle. Mfs were isolated from different skin sites and from adult worms and a minimum of 10 mfs from each isolate were examined for width and acid phosphatase (AP) staining pattern. The width of O. gutturosa dermal mf was less than 4 micron (4 isolations), narrower than that of putative O. lienalis mf isolated from umbilical skin of cattle without evidence of O. gutturosa, which were in 20/22 isolations greater than 4 micron wide. The dermal mf were also distinguished on the basis of different AP staining patterns which, for each species, correlated closely with that of hatched intrauterine mf from their respective adult female worms. Based on the criteria of morphology and AP staining patterns the mf species prevalences in the survey population were estimated as O. lienalis 24.1% and O. gutturosa 2.2%, with a further 2.2% of cattle infected with both species. The results indicate that the predilection site of adult O. lienalis is not the gastro-splenic omentum. In North Wales, the distribution of the two species was different; O. lienalis was widely distributed in all cattle rearing areas both lowland and upland, whereas O. gutturosa was largely restricted to valleys close to major rivers.
A problem of high levels of liver condemnations which had affected bought-in lambs fattened on a North Yorkshire farm for several years was investigated. Because the farmer spread pig slurry on the land, migrating Ascaris suum larvae were initially suspected as a cause but the investigation eliminated this possibility. Migrating Taenia hydatigena larvae were found in tissue sections and isolated from livers. A combination of parasitological and serological evidence using oncospheral antigen in an ELISA suggested that infection was occurring immediately before the arrival of lambs on the farm, although a source of infection on the farm was also identified. Peak liver condemnations occurred in December-January. Possible reasons for this are suggested and the relevance of these findings to national liver condemnation losses are discussed.
Using a three-layer immunofluorescent test, class-specific, parasite-specific circulating antibody responses to Eimeria tenella were investigated following oocyst infection, drug-truncated oocyst infection and the injection of non-living antigens. Following all three means of antigenic stimulation, IgG, IgM and IgA antibodies were detected. The response to drug-truncated infections was dose-dependent. The sequence of appearance of antibody was IgM, IgA, IgG, whilst the relative quantities were IgG greater than or equal to IgM greater than IgA.
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In splenectomised calves infected with Babesia divergens IFA titres appeared after peak parasitaemia and maximum titres were observed 35 days after infection: the IFA response of adult non-splenectomised cattle was similar. It was concluded that the absence of a spleen did not markedly inhibit the IFA response of cattle to B divergens infection.
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By microinjection of cryopreserved microfilariae (mf) into nulliparous flies, a comparison of the lengths of the infective larvae (L3) of Onchocerca volvulus and O. ochengi from the head of Simulium damnosum s.l. (presumed S. sirbanum) has been made. The suitability of S. sirbanum as a host was similar for both Onchocerca spp. The mean length +/- standard deviation of O. ochengi infective larvae measured in aqueous medium after storage of infected flies in liquid nitrogen was 762 +/- 63 microns (n = 39), significantly longer (P much less than 0.0001) than those of a savanna isolate of O. volvulus (676 +/- 56 microns, n = 26). Although the frequency distributions of the lengths of larvae of the 2 species overlapped, a critical value for discrimination of 719 microns applied to normally distributed populations with means and standard deviations of these samples would result in correct classification of 78% of true O. volvulus and 75% of true O. ochengi. A discriminant function analysis incorporating width measurements did not usefully improve the level of accuracy of discrimination. Larvae from flies stored in 70% ethanol and stained with acid haemalum were about 10% shorter, but O. ochengi infective larvae were still proportionately longer than those of O. volvulus (693 +/- 40 microns, n = 45 compared to 580 +/- 38 microns, n = 6, respectively). These data show that the infective L3 of O. volvulus and O. ochengi differ morphologically. Although the population length distributions overlap, by classifying larvae greater than 719 microns long as O. ochengi and those less than 719 microns long as O. volvulus a more accurate estimation of true O. volvulus infection rates in S. damnosum s.l. can be derived than is currently possible.