Host parasite relationships in Trypanosoma (duttonella) vivax with special reference to the influence of antigenic variation

A mouse model system was used to study various aspects of host and parasite relationships in Trypanosoma vivax infections. These included the phenomenon of antigenic variation, the variable parasite antigens responsible for this phenomenon, parasite-host adaption, host immune responses and the role of genes in the major histocompatibility complex in the control of infection. While the mouse model system has allowed investigation of these aspects of host parasite relationships, it is clear that the system is much more limited than those generally used in T. brucei spp and T. congolense infections. This is indicated by the discovery that not all VATs of T. vivax were equally infective for mice, though in some cases infectivity could be improved by bovine serum supplementation and/or immunosuppression of the mouse host. In the case of rats, infection was even restricted to a smaller number of the VATs studied. It was, however, possible to biochemically characterize the variable surface antigen carried by T. vivax and show its similarity to those carried by T. brucei and T. congolense. The H-2 complex was found not to influence acquired resistance of inbred strains. Cyclic transmissions of T. vivax infections to goats combined with chemotherapy were carried out in an attempt to induce protection to subsequent infection as has been shown in T. brucei and T. congolense infections. Such protection could, however, not be obtained, The failure of the metacyclic VATs to induce immunity, was perhaps due to rapid decrease in antibody titres to bloodstream VATs found after treatment and prior to rechallenge. The usefulness of the mouse model system in elucidating the mechanisms responsible for the non-H-2 linked differences in susceptibility to T. vivax infections should be further explored and its relevance to mechanisms of trypanotolerance in domestic ruminants defined.     

Key Factors Affecting Reproductive Success of Thoroughbred Mares and Stallions on a Commercial Stud Farm

To evaluate factors contributing to fertility of thoroughbred mares, data from 3743 oestrous periods of 2385 mares were collected on a large thoroughbred farm in Ireland. Fourteen stallions (mean age 8.3 years; range 4–15 years) had bred 2385 mares (mean age 9.4 years; range 3–24 years). Maiden mares accounted for 12%, mares with a foal at foot for 64%, and barren, slipped or rested mares for 24% of the total. The mean pregnancy rate per cycle was 67.8% (68.6% in year 1 and 66.9% in year 2). Backward stepwise multivariable logistic regression analysis was utilized to develop two models to evaluate mare factors, including mare age, reproductive status, month of foaling, dystocia, month of cover, foal heat, cycle number, treatments, walk‐in status and stallion factors including stallion identity, stallion age, shuttle status, time elapsed between covers and high stallion usage on the per cycle pregnancy rate and pregnancy loss. Old age (p < 0.001) and cover within 20 days post‐partum (p < 0.003) were associated with lowered pregnancy rates. High mare age (p < 0.05) and barren, slipped or rested reproductive status (p = 0.05) increased the likelihood of pregnancy loss. Uterine inflammation or infection, if appropriately treated, did not affect fertility. Only high usage of stallions (used more than 21 times in previous week) was associated with lowered (p = 0.009) pregnancy rates. However, shuttle stallions were more likely to have increased (p = 0.035) pregnancy survival, perhaps reflecting a bias in stallion selection. In conclusion, mare age exerted the greatest influence on fertility; nonetheless, thoroughbreds can be effectively managed to achieve high reproductive performance in a commercial setting.     

The effects of vaccination of Merino ewes with an attenuated Australian bluetongue virus serotype 23 at different stages of gestation

SUMMARY A cell culture attenuated Australian bluetongue virus serotype 23 (BLU23) prototype vaccine was assessed for its effects on pregnant Merino sheep. Seventy-six ewes were vaccinated at 5 different stages of gestation, and the failure to lamb at term was as follows: 35 to 43 days of gestation, 20/36 (56%); 57 to 64 days of gestation, 3/10 (30%); 81 to 88 days of gestation, 3/10 (30%); 109 to 116 days of gestation, 0/10 (0%); 130 to 137 days of gestation, 0/10 (0%). Of 30 ewes vaccinated with a cell culture supernatant fluid control between 35 and 43 days of gestation, 6.7% (2/30) failed to lamb at term. Two ewes vaccinated with BLU23 vaccine between 35 and 43 days of gestation had lambs with hydranencephaly. All other lambs born were clinically normal. Three ewes vaccinated with BLU23 aborted. Two of these were vaccinated between 35 and 43 days of gestation, the 3rd between 81 and 88 days of gestation. Five lambs were born with BLU group antibody. Four of these were from ewes vaccinated between 35 and 43 days of gestation, and 2 of these had hydranencephaly. The fifth was from a ewe vaccinated between 57 and 64 days of gestation. The vaccine did not produce disease in adult sheep, but was a potent cause of early foetal death and to a much lesser extent foetal malformation.     

Macquarie island and the cause of oceanic linear magnetic anomalies

Macquarie Islands is formed of probably Pliocene oceanic crust. Intruded into pillow lavas is a belt of harzburgite and layered gabbro mnasses cut by dike swarms. Similar belt-like structures may cause the linear magnetic anomalies of the ocean.     

Determination of the digestible energy intake and apparent absorption of macroelements in pasture-fed lactating Thoroughbred mares

AIM: To measure the nutritive value of pasture in terms of digestible energy (DE) intake (DEI) and dry matter (DM) digestibility, and to determine the apparent absorption of macroelements in lactating Thoroughbred mares grazed on pasture. METHODS: DM intake (DMI) and DEI were determined from daily faecal DM output measured in grazing mares, divided by the DM indigestible fraction (1-digestible DM), measured in a digestibility trial using pasture-fed mares. Eight lactating mares and their foals, that had a mean age of 40 days, were grazed separately on 50x100 m areas of pasture and daily faecal DM outputs were recorded for 8 days. Five mares and their foals were then placed in individual bare 20x20 m corrals containing custom-made feeding stations for 14 days to determine the indigestible DM fraction. DM, gross energy content, crude protein (CP), soluble carbohydrate, acid detergent fibre (ADF), neutral detergent fibre (NDF), lipid, and macroelement composition of the pasture offered and faeces were determined and their digestibility and/or apparent absorption calculated. RESULTS: DM digestibility of the pasture was 0.6 and the DMI and DEI of a grazing 560 kg mare in early lactation nursing a foal growing at 1.34 kg/day was 13.6 (SE 0.8) kg/day and 146.9 (SE 8.4) MJ DE/day, respectively. Apparent absorptions of the macroelements measured were: Ca 0.75, P 0.43, Mg 0.63, Na 0.78, and K 0.72. CONCLUSIONS: Good quality ryegrass-white clover pasture that had a DE content of 10.8 MJ/kg DM, and a macroelement composition (g/kg) of Ca 3.33, P 3.0, Mg 1.67, Na 1.67, and K 24.2, will provide adequate DMI, DEI, and macroelement intake to lactating Thoroughbred mares.     

Development and evaluation of an enzyme-linked immunosorbent assay for the serodiagnosis of pythiosis in dogs

Pythiosis (caused by the aquatic oomycete Pythium insidiosum) is a devastating and often fatal cause of either severe transmural gastroenteritis or locally invasive subcutaneous disease in dogs living in the southeastern United States. Although early diagnosis is essential for successful treatment, tools available for this task are limited. Therefore, we developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-P insidiosum antibodies in canine serum. A soluble mycelial extract of P insidiosum was utilized as antigen in the ELISA, which was used to evaluate serum from 43 dogs with pythiosis, 8 dogs with lagenidiosis (another canine oomycosis), 16 dogs with nonoomycotic fungal or algal infections, 22 dogs with nonfungal gastrointestinal or skin disease, and 55 healthy dogs. Results were expressed as percent positivity (PP) relative to a strong positive control serum run on each plate. Medians and ranges for each of the 5 groups were as follows: pythiosis (81.7%, 50.6-98.5%), lagenidiosis (17.3%, 11.3-29.2%), other fungal or algal infections (8.2%, 4.7-15.4%), nonfungal gastrointestinal or skin disease (6.2%, 3.9-20.7%), and healthy dogs (6.7%, 3.0-15.2%). When using a cutoff value of 40% PP, the sensitivity and specificity of the ELISA both were 100%. In addition, ELISA values measured after successful surgical therapy in 2 dogs showed a decrease of anti-P insidiosum antibody concentrations into the normal range as early as 2 months after treatment. We conclude that the ELISA is a sensitive and specific test for the diagnosis of canine pythiosis, and may be a useful tool for monitoring response to medical or surgical therapy.     

Producing progeny from endangered birds of prey: treatment of urine-contaminated semen and a novel intramagnal insemination approach.

Wild raptors brought into an ex situ environment often have poor semen quality that is further compromised by urine contamination. Generally, it is believed that in birds, artificial insemination into the cloaca or caudal vagina of females requires large doses of high-quality spermatozoa to maximize fertility. In an effort to define and overcome some of the challenges associated with reproduction in wild raptors, the objectives of this study were to 1) evaluate the frequency, impact, and remediation of urine contamination in fresh ejaculates for the purpose of maintaining sperm motility and viability in vitro, and 2) develop a deep insemination method that allows low numbers of washed sperm to be placed directly into the magnum to increase the probability of producing fertilized eggs. The species evaluated include golden eagle (Aquila chrysoetos), imperial eagle (A. adalberti), Bonelli's eagle (Hiernaetus fasciatus), and peregrine falcon (Falco peregrinus). Semen samples were collected and pooled by species, and a minimum of 25 pooled ejaculates per species were evaluated for urine contamination, pH, sperm viability, and sperm motility; the samples were either unwashed or washed in neutral (pH 7.0) or alkaline (pH 8.0) modified Lake's diluent. Female golden eagles and peregrine falcons were inseminated via transjunctional, intramagnal insemination with washed spermatozoa from urine-contaminated samples. Urine contamination occurred in 36.8 +/- 12.8% (mean +/- SEM) golden eagle, 43.1 +/- 9.1% imperial eagle. 28.7 +/- 16.1% Bonelli's eagle, and 48.2 +/- 17.3% peregrine falcon ejaculates. The pH in urine-contaminated semen samples ranged from 6.48 +/- 0.3 to 6.86 +/- 0.2, and in noncontaminated samples it ranged from from 7.17 +/- 0.1 to 7.56 +/- 0.1. Sperm viability and motility were reduced (P < 0.05) in all species for unwashed vs. washed sperm after 30 min incubation at room temperature. Two peregrine falcon chicks and one golden eagle chick hatched after intramagnal insemination. This study demonstrates that urine contamination, a common and lethal acidifier in manually collected raptor ejaculates, can be circumvented by immediate, gentle seminal washing. Furthermore, these processed sperm, when deposited by transjunctional intramagnal insemination, can produce live young.     

Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

Imaging of intermittency in ripple-wave turbulence

The dynamics of a fluid surface filled with high-amplitude ripples were studied with a technique (diffusing light photography) that resolves the height at all locations instantaneously. Even when nonlinearities are strong enough to generate a (Kolmogorov) cascade from long wavelength (where energy is input) to shorter wavelength, the resulting turbulent state contains large coherent spatial structures. The appearance of these structures in a thermal equilibrium state (with the same average energy) would be highly improbable.