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Ovine Johne's disease is a chronic debilitating disease of sheep caused by Mycobacterium avium subsp. paratuberculosis (Mptb) which results in diarrhoea, emaciation and mortalities in infected animals. Vaccination with Gudair® has been a key strategy for controlling the disease in Australia since its approval in 2002. Previous research conducted in Australia has demonstrated that the vaccine is quite effective in reducing sheep mortalities. While some farms have also been successful in reducing the prevalence of the disease in their flocks to undetectable levels, sheep in other flocks continue to shed Mptb in faeces even after an ongoing vaccination program . This study was conducted to investigate management, husbandry and biosecurity factors associated with paratuberculosis infection in Gudair® vaccinated sheep flocks in Australia. We enrolled 64 sheep farmers and interviewed them to obtain information about their management and biosecurity practices. Pooled faecal samples were collected from sheep at each farm and cultured to create two outcome variables: Mptb positive (yes/no) and disease prevalence level (nil, < 1 %, ≥ 1 %). Binary and ordinal logistic regression analyses were conducted to evaluate the association of management, husbandry and biosecurity factors with these outcome variables. Farms were more likely to have Mptb positive sheep and a higher disease prevalence in their flocks if they: (a) provided supplementary feed on the ground (instead of in a trough); (b) had a greater number of neighbours with sheep; and (c) had introduced rams from a greater number of sources. The results suggest the effectiveness of Gudair® vaccination to control OJD can be improved if sheep producers maintain other risk management strategies and biosecurity practices. Extension agencies should advise farmers not to relax their biosecurity practices and to purchase rams from only low-risk sources, even if they are continuing to vaccinate their flocks.

The current study evaluated the cardioprotective effect of the aqueous extract of seeds ofDatura metel(AESDM) on the acute cardiotoxicity induced with doxorubicin in the Wistar rats. Indeed, 30 rats have been randomized then, divided into 6 groups of 5 animals each: 3 control groups (normal, negative and positive) and 3 test groups (AESDM 100, 200 and 300 mg/kg). These animals received orally, six consecutive days, 1 mL/200 g distilled water (normal and negative control), 100 mg/kg Vitamin-E (positive control), AESDM at 100, 200 and 300 mg/kg (test groups). On the seventh day of treatment, animals in the different groups excepted the normal control received a single intraperitoneal injection of 15 mg/kg doxorubicin followed respective treatments up to the tenth day of the trial period. Serum parameters (ASAT/ALAT), biochemical markers of oxidative stress (MDA, SOD, CAT, GSH) and lipid parameters (total cholesterol, triglycerides, HDL and LDL) were evaluated. Histological sections of animal hearts were prepared. The results of this study showed that treatment with AESDM resulted in a significant decrease (P ˂ 0.001) in MDA, triglycerides and LDL-cholesterol levels, ASAT/ALAT activity and a significant increase (P ˂ 0.001) in HDL-cholesterol concentration and SOD, CAT and GSH activities, compared to the negative control. Histological observation revealed that AESDM protected the heart from doxorubicin-induced damage. In conclusion, AESDM would have a cardioprotective effect that could be attributed to its antioxidant potential.&nbsp

Background: An investigation aimed to confirm whether immunisation with the “benign” buffeli genotype of Theileria orientalis could reduce the parasitosis of the virulent ikeda genotype. Methods: Calves were inoculated intravenously or subcutaneously with bovine blood containing merozoites of T.orientalis buffeli. When recipients became positive, they and control animals were challenged with unfed nymphs of Haemaphysalis longicornis ticks infested as larvae with T.orientalis ikeda. Results: All calves became positive for the challenge within 12 days after tick application. In the immunised calves, the first wave of parasitosis with T.orientalis ikeda from 4-6 weeks was reduced significantly by >80% before the infestation declined into the carrier state by 9 weeks. Conclusions: The results confirm the field experience with theileriosis in endemic zones where the carrier state appears to prevent clinical disease despite repeated, seasonal tick infestations with virulent genotypes of the parasite. This method offers a means to reduce the severity of the first wave of theilerial parasitosis after tick challenge and recover associated production losses.

Background The tick vector Rhipicephalus microplus which transmits Babesia spp. and rickettsial pathogens has not been reported in Kenya since 1998. More recently, the pathogenic Babesia bovis has been detected in cattle blood DNA. The status of R. microplus in Kenya remains unknown. This study employed morphological and molecular tools to characterize R. microplus originating from Kenya and assess the genetic relationships between Kenyan and other African R. microplus genotypes. Methods Ticks were collected in south-eastern Kenya (Kwale County) from cattle and characterized to investigate the existence of R. microplus. Genetic and phylogenetic relationships between the Kenyan and other annotated R. microplus reference sequences was investigated by analysis of the cytochrome c oxidase subunit 1 (cox1) gene. To further characterize Kenyan ticks, we generated low coverage whole genome sequences of two R. microplus, one R. decoloratus and R. appendiculatus. A B. bovis specific TaqMan probe qPCR assay was used to detect B. bovis in gDNA from R. microplus ticks. Results Occurrence of R. microplus was confirmed in Kwale County, Kenya. The Kenyan R. microplus cox1 sequences showed very high pairwise identities (> 99%) and clustered very closely with reference African R. microplus sequences. We found a low genetic variation and lack of geographical sub-structuring among the African cox1 sequences of R. microplus. Four complete mitochondrial (mt) genomes for two R. microplus, one R. decoloratus and one R. appendiculatus were assembled from next generation sequence data. The mitochondrial genome sequences of the two Kenyan R. microplus ticks clustered closely with reference genome sequences from Brazil, USA, Cambodia and India forming R. microplus Clade A. No B. bovis was detected in the Kwale R. microplus DNA. Conclusions These findings confirm the presence of R. microplus in Kenya and suggest that R. microplus Clade A is prevalent in cattle in sub-Saharan Africa. These and other recent findings of widespread occurrence of R. microplus in Africa provide a strong justification for urgent surveillance to determine and monitor the spread of R. microplus and vector competence of Boophilus ticks for B. bovis in Africa, with the ultimate goal of strategic control.

This review addresses the research landscape regarding vaccines against scour worms, particularly Trichostrongylus spp. and Teladorsagia circumcincta. The inability of past research to deliver scour-worm vaccines with reliable and reproducible efficacy has been due in part to gaps in knowledge concerning: (i) host-parasite interactions leading to development of type-2 immunity, (ii) definition of an optimal suite of parasite antigens, and (iii) rational formulation and administration to induce protective immunity against gastrointestinal nematodes (GIN) at the site of infestation. Recent 'omics' developments enable more systematic analyses. GIN genomes are reaching completion, facilitating "reverse vaccinology" approaches that have been used successfully for the Rhipicephalus australis vaccine for cattle tick, while methods for gene silencing and editing in GIN enable identification and validation of potential vaccine antigens. We envisage that any efficacious scour worm vaccine(s) would be adopted similarly to "Barbervax™" within integrated parasite management schemes. Vaccines would therefore effectively parallel the use of resistant animals, and reduce the frequency of drenching and pasture contamination. These aspects of integration, efficacy and operation require updated models and validation in the field. The conclusion of this review outlines an approach to facilitate an integrated research program.

Integrated parasite management (IPM) for pests, pathogens and parasites involves reducing or breaking transmission to reduce the impact of infection or infestation. For Theileria orientalis, the critical impact of infection is the first wave of parasitaemia from the virulent genotypes, Ikeda and Chitose, associated with the sequelae from the development of anaemia. Therefore, current control measures for T. orientalis advocate excluding the movement of naïve stock from non-endemic regions into infected areas and controlling the tick Haemaphysalislongicornis, the final host. In Australia, treatment of established infection is limited to supportive therapy. To update and expand these options, this review examines progress towards prevention and therapy for T. orientalis, which are key elements for inclusion in IPM measures to control this parasite.

Background In regions with endemic Theileria orientalis, recovered cattle are carriers, leaving newborn calves and introduced stock as the most susceptible groups to develop clinical disease after tick infestation with parasite transmission. Methods To gain information on the kinetics of infection and development and effects of theileriosis caused by virulent ikeda and chitose genotypes and the “benign” buffeli genotype of T.orientalis, this study sampled a total of 134 calves from 3 farms in Dorrigo, Australia, a region with multiple genotypes of the parasite. In addition, 30 introduced beef weaners were bled and weighed for 6 months after arrival. Results In both cohorts, parasitaemia with the ikeda and chitose (genotypes 1 & 2) of T.orientalis was detectable by PCR within 3–4 weeks in >95 % of the groups, with maximal gene copies generated around 5–7 weeks after birth or introduction, before declining. In contrast, parasitaemias of T.orientalis buffeli (genotype 3), increased slowly, with around 80 % of the population testing positive by PCR after 4 months. The parasitaemias of the buffeli genotype did not exceed 40,000 gene copies /ul and were similar to those exhibited by ikeda and chitose genotypes following the “first wave of parasitaemia” as cattle entered the carrier state. In the 30 introduced weaners, the early infection with T.orientalis ikeda and chitose also caused significant reductions in packed cell volume (PCV) and incurred an estimated 20Kg loss in weight gain; the latter had not been recovered by 6 months after introduction. Conclusions The results support previous findings in other endemic regions of theileriosis, and imply that amelioration of the early high levels of parasitosis by the virulent genotypes could reduce the initial impact of theileriosis on production, which appears much less affected once animals enter the carrier state.

At present diagnosis of true resistance and determination of drug efficacy in Fasciola hepatica infection rely solely on terminal experiments. The coproantigen ELISA (cELISA) has been reported previously as a sensitive and specific tool appropriate to detect treatment failure, and potentially drug resistance. Two studies were conducted to determine whether the cELISA was appropriate for on-farm efficacy and resistance testing in Australian Merino sheep. In Study 1 sheep were infected orally with 50 F. hepatica metacercariae on three occasions, twelve, six and two weeks prior to a single flukicide treatment with triclabendazole, closantel or albendazole. Sheep were sampled weekly for a further seven weeks prior to necropsy. Following effective treatment, no faecal antigen was detected from 1 week. When immature stages (≤6 weeks) survived treatment, coproantigen reappeared from 6 weeks post-treatment. Therefore, cELISA conducted 1-4 weeks after treatment will demonstrate obvious treatment failure against adult F. hepatica, but is not sufficiently sensitive to detect survival of immature fluke until these reach maturity. In study 2, fluke burdens of sheep necropsied 13 weeks post single infection were compared to fecal worm egg counts (FWEC) and cELISA at necropsy. Regression analysis demonstrated that cELISA correlated strongly with fluke burden, whilst FWEC correlated weakly with cELISA. The correlation between FWEC and fluke burden was also weak, although stronger than that of FWEC with cELISA. The cELISA is an appropriate tool for monitoring effectiveness of treatments against Fasciola hepatica if an adult infection is present, however when immature stages of the parasite are present it is not as reliable. Where immature parasites are present it is recommended that initial cELISA be followed with a secondary cELISA at least 6 weeks after treatment to ensure resistance to immature stages is detected. Further testing is justified for monitoring the effectiveness of control programs by detecting adult populations that have survived a treatment regime.

Aim. The effects of the water extract of the leaves of Euphorbia Poissonii (Euphorbiacea) on frog heart contraction and its possible mechanism action were investigated. Method. By using the Langerdhorff model of the isolated frog heart preparation, the strength and frequency of heart contractions as well as cardiac output were measured under perfusion with test drugs. Results and conclusion. When assessed with digoxin and adrenaline as positive controls, the water extract (3 mg/mL) exhibited significant positive inotropic (70.00% improvement compared to baseline) and chronotropic (17.79% improvement) effects. Additionally, heart contractility was inhibited in the presence of propranolol and amlodipine, suggesting that the testing extract is having adrenergic properties. Reversible heart arrest was noticed at the dose of 6 mg/mL.

The diagnosis, monitoring and flukicide efficacy testing of fasciolosis on-farm is reliant on non-terminal methods. The coproantigen ELISA (cELISA) has been recommended for diagnosis of fasciolosis and associated flukicide efficacy testing as an alternative to fluke egg counts for monitoring parasitism. Recently experimental multi-age infections have suggested that the reliability of efficacy results can be improved by a second cELISA testing at 6 weeks post-treatment (wpt) in addition to the generally accepted 1 wpt. A field study was conducted to determine the suitability of faecal fluke egg counts (FFEC) and cELISA as diagnostic, drug efficacy testing and epidemiological tools on Australian sheep and cattle farms. Faecal samples from sheep and/or cattle on three endemic farms were taken at monthly intervals for 12 months and examined by both methods. Normal farm management was maintained during the study period and opportunistic efficacy testing, in line with each farm’s normal flukicide management was undertaken. Additionally, the suitability of the Ollerenshaw Index as a predictive model for fasciolosis under Australian conditions was examined. While both diagnostics demonstrated their value in the farm environment, the current data demonstrate a distinct and significant increase in diagnostic sensitivity for epidemiological studies by using the two tests in parallel. The agreement between the two diagnostics was found to be higher in cattle, despite the poor sensitivity of FFEC in this species. Similar levels of agreement between the two tests were demonstrated at both sheep properties, regardless of the marked difference in the intensity of F. hepatica challenge. Linear regression models demonstrated the results of the two diagnostics utilized in parallel were explained substantially (R2 = 0.91) as were series data (R2 = 0.88) when the respective models were fitted. In contrast, the fitted models for FFEC (R2 = 0.54) and cELISA (R2 = 0.58) were poor explanations for test outcomes. The outcomes of these models support previous findings that suggest that the two diagnostic tests are best utilized together, particularly in parallel. The application of the Ollerenshaw Index to Australian conditions requires further investigation. Keywords: Coproantigen, ELISA, Efficacy, Fasciola hepatica, Triclabendazole, Prevalence

Background: Vitellaria paradoxa is a traditional medicinal plant of Cameroon. Several studies on this plant have focused on the cosmetic profile of its fruits. The present study focuses on the anti-inflammatory potency of stem barks extract of this plant. Objective: The objective was to evaluate the effect of methanolic extract of V. paradoxa (VPME) stem barks on inflammatory response in rats. Materials and Methods: Anti-inflammatory effects of VPME were evaluated in acute and chronic (28 days) inflammation induced in Wistar albino rats. The effects on hyperalgesia and locomotors activity were also quantified. The relative weight of lymphoid organs was obtained as well as some hematological parameters. Results: In the carrageenan-induced inflammation, VPME (75 mg/kg) exhibited a significant (66.67%) inhibition after 1 h. On the complete Freund's adjuvant-induced rheumatoid arthritis, VPME showed a significant protective effect with 8.12% inflammation against 25.00% for the control group after 2 days of the treatment. The extract (75 and 150 mg/kg) significantly reduced the score of arthritis with a maximum obtained on day 19 th of the experimentation. There was a significant increase in the reaction time of rats on the hot plate as well as the exploratory activities of the animals in the open field. This extract significantly prevented weight, hemoglobin and red blood cells losses, and spleen hypertrophy. A protective action against skin destruction and cartilage erosion was evident. Liquid chromatography-mass spectrometry analysis of the extract revealed the presence of catechins. Conclusions: These findings suggested that V. paradoxa may contribute to the reduction of the inflammatory response.

Information on the susceptibility status of Fasciola hepatica isolates is lacking in the literature, even for those isolates considered to be laboratory reference strains. Four controlled efficacy studies were conducted on two Fasciola hepatica isolates from Australia, viz. 'Oberon' and 'Sunny Corner' with treatment at either 2, 6 or 10 weeks post-infection (wpi) as defined in each study. Fluke burdens and examination of livers occurred at necropsy in weeks 12 (Study 1) or 13 (Studies 2, 3 and 4) post-infection. The triclabendazole (TCBZ) resistance status of the Oberon isolate was confirmed in 6 and 10-week old F. hepatica, utilizing the drug alone (Fasinex; 71.5% and 31.1%, respectively) and in combination with oxfendazole (Flukazole C; 79.9% and 0%, respectively). The susceptibility of this isolate to albendazole, as well as salicylanilide and sulphonamide drugs was confirmed. The Sunny Corner isolate was confirmed as susceptible to TCBZ (>99% all stages) and closantel (>90% at ≥6 wpi).

Datura metel leaves are used by athletes to enhance their performances. The water extract of the leaves and seeds was tested for their cardiotonic effect. Langherdoff’s model for the measurement of the mechanical activity of isolated amphibian heart in ventricular perfusion was used to evaluate some parameters of cardiac contractility. The effect of the extracts was also compared to sympathetic agonist and antagonist. The leaves and seed extracts induced significant (p

Objective. The objective of the present study was to evaluate the in vivo wound healing effect of water extract of Tetrapleura tetraptera in stem-bark. Method: The healing activity was studied in 40 male rats using excision and incision wounds on normal and dexamethasone-suppressed wound healing. For each model, rats were divided in 4 groups as follows: control, dexamethasone, T. tetraptera and dexamethasone combined with T. tetraptera. Results: Data recorded exhibited a significant effect by the extract in the epithelialization time within 14 and 18 days of the normal and dexamethasone-induced healing delay rats respectively (p

Background Heartworm (Dirofilaria immitis) in dogs is considered endemic in Australia, but the clinical heartworm disease caused by the heartworm is rare and prevalence is low. The mainstream prevention of the heartworm is based on macrocyclic lactone (ML) administration. The aim of this study was to confirm endemism of the heartworm under current Australian conditions using a cohort of recent microfilaria-positive dogs which were on variable heartworm prevention. Methods A hotspot of canine heartworm antigen-positive and microfilaria-positive dogs has been detected recently in Queensland, Australia. Blood samples from 39 dogs from Queensland and two dogs from New South Wales were investigated for canine filarioids. Rapid antigen diagnostic tests capable of detection of D. immitis and real-time PCR for quantification and differentiation between D. immitis from Acanthocheilonema reconditum with quantification of microfilariae in canine blood samples, together with D. immitis specific real-time PCR assay, were applied to microfilaria-positive dogs. The P-glycoprotein genotype was determined to test whether Australian-sourced heartworm shared the same genetic markers as those suspected of ML-resistance in North America. Results Only D. immitis was detected in the samples from Queensland and New South Wales, Australia. Using high resolution melt real-time PCR and D. immitis specific real-time PCR, the calculated microfilaria concentration ranged from 1 to 44,957 microfilariae/ml and from 7 to 60,526 microfilariae/ml, respectively. DNA sequencing of the PCR products confirmed D. immitis. Fifteen of the examined dogs were on putative, rigorous ML prevention. For the remaining dogs, compliance with heartworm prevention was unknown or reported as inconsistent. Wild-type genotype AA-GG of the P-glycoprotein locus of D. immitis sequence has been obtained for three blood samples. Due to the incomplete history, any suggestion of a loss of efficacy of MLs must be treated as ‘remotely possible’. In the immediate future, records of preventative administration and annual antigen testing would be required to determine any problems with the efficacy of preventatives. Conclusions The prevalence of canine heartworm in Australia remains poorly understood. It is generally assumed to be low by veterinary practitioners. The localised increase in the study area confirms endemism of canine heartworm and a requirement for ongoing vigilance through annual heartworm testing to better understand the changing distribution of canine heartworm, client compliance, as well as to detect any change in ML-susceptibility. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1821-x) contains supplementary material, which is available to authorized users.

Ovine Johne's disease (OJD) is a degenerative wasting condition of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Similar to other pathogenic mycobacterial infections it is a slow-progressing chronic disease and subclinically infected individuals can spread the disease while appearing healthy. MAP infects macrophages and the host responds by mounting a cell-mediated immune response. Disease progression is associated with immune dysfunction but the reasons are unknown. Increasing our current knowledge of the immunological mechanisms involved in disease progression, including apoptotic responses, may allow advancements in the area of early diagnosis, identification of resistant animals and disease control. We describe lymphocyte apoptosis in peripheral blood mononuclear cells (PBMC) and lymph node cells from sheep exposed to MAP as well as from healthy non-exposed sheep. Apoptosis in intestinal lymph node cells from MAP-exposed infected sheep, but not in MAP-exposed uninfected sheep, increased in response to MAP antigen. In this first longitudinal study of lymphocyte apoptosis using an experimental infection model of MAP infection, we found that there was a transient increase of ex vivo PBMC apoptosis in MAP-exposed sheep soon after exposure to MAP (4 months post inoculation). MAP antigen-specific apoptosis occurred later, at 12 months post inoculation. The cells involved were mainly γδ and CD4+ T cells. Antigen-mediated lymphocyte apoptosis during mycobacterial disease progression could contribute to the immune dysfunction in Johne's disease.

Although a live attenuated vaccine has been used extensively to provide immunity against porcine proliferative enteropathy (PE) caused by Lawsonia intracellularis, the nature of the protective response is an area of considerable interest for the control of PE. Two trials investigated immune responses in pigs after oral and intramuscular (IM) vaccination followed by virulent L. intracellularis challenge. After an oral vaccination with 10(5.9) TCID50 organisms, significantly increased serum and mucosal secretions of IgM, IgG and higher mucosal TNF-α and TGF-β1 were detected by day 17, together with a trend towards higher levels of IFN-γ and IL-6. Pigs vaccinated IM produced elevated serum antibody titres but mucosal immune responses were not detected. After challenge with virulent L. intracellularis, non-vaccinated control pigs had higher PE lesion scores and excreted significantly higher numbers of L. intracellularis in faeces than the vaccinated pigs. Reduced intestinal pathology and faecal L. intracellularis shedding were evident in the vaccinated groups. The results indicated that protection was associated with mucosal cytokine and specific IgG and IgA responses after vaccination and that systemic antibody responses were boosted following challenge. However in the search for an immune correlate with protection, a causal association was not evident from a kinetic analysis of immune parameters in serum, ileal pathology and faecal shedding.

Background Heartworm (Dirofilaria immitis) in dogs is considered endemic in Australia, but the clinical heartworm disease caused by the heartworm is rare and prevalence is low. The mainstream prevention of the heartworm is based on macrocyclic lactone (ML) administration. The aim of this study was to confirm endemism of the heartworm under current Australian conditions using a cohort of recent microfilaria-positive dogs which were on variable heartworm prevention. Methods A hotspot of canine heartworm antigen-positive and microfilaria-positive dogs has been detected recently in Queensland, Australia. Blood samples from 39 dogs from Queensland and two dogs from New South Wales were investigated for canine filarioids. Rapid antigen diagnostic tests capable of detection of D. immitis and real-time PCR for quantification and differentiation between D. immitis from Acanthocheilonema reconditum with quantification of microfilariae in canine blood samples, together with D. immitis specific real-time PCR assay, were applied to microfilaria-positive dogs. The P-glycoprotein genotype was determined to test whether Australian-sourced heartworm shared the same genetic markers as those suspected of ML-resistance in North America. Results Only D. immitis was detected in the samples from Queensland and New South Wales, Australia. Using high resolution melt real-time PCR and D. immitis specific real-time PCR, the calculated microfilaria concentration ranged from 1 to 44,957 microfilariae/ml and from 7 to 60,526 microfilariae/ml, respectively. DNA sequencing of the PCR products confirmed D. immitis. Fifteen of the examined dogs were on putative, rigorous ML prevention. For the remaining dogs, compliance with heartworm prevention was unknown or reported as inconsistent. Wild-type genotype AA-GG of the P-glycoprotein locus of D. immitis sequence has been obtained for three blood samples. Due to the incomplete history, any suggestion of a loss of efficacy of MLs must be treated as ‘remotely possible’. In the immediate future, records of preventative administration and annual antigen testing would be required to determine any problems with the efficacy of preventatives. Conclusions The prevalence of canine heartworm in Australia remains poorly understood. It is generally assumed to be low by veterinary practitioners. The localised increase in the study area confirms endemism of canine heartworm and a requirement for ongoing vigilance through annual heartworm testing to better understand the changing distribution of canine heartworm, client compliance, as well as to detect any change in ML-susceptibility.

The present study examined the anxiolytic and antidepressant effects of the aqueous extract ofAlafia multiflora Stapf(AM) stem barks (150 and 300 mg/kg, 7 days administration) on rats and mice, using experimental paradigms of anxiety and depression. In the open field, the aqueous extract increased significantly the number of center square crossed and the time spent at the center of the field as well as the rearing time, while the grooming time was reduced significantly. In the elevated plus maze, the aqueous extract increased the time spent and the number of entries in the open arms. All these effects were also completely reversed by flumazenil, an antagonist of benzodiazepine receptors and pindolol aβ-adrenoceptors blocker/5-HT 1A/1B receptor antagonist. The time spent in the light compartment, the latency time, and the number of the light-dark transitions increased significantly in the light/dark exploration test after the treatment with AM. The extract was able to reduce significantly the immobility time and increase swimming as well as climbing duration. Taken together, the present work evidenced anxiolytic effects of the aqueous extract of AM that might involve an action on benzodiazepine-type receptors and an antidepressant effect where noradrenergic mechanisms will probably play a role.

The capacity of a commercially available gamma interferon (IFNγ) assay to detect infected sheep early in the pathogenesis of Johne's disease enables the removal of such animals from the flock before bacterial shedding and pasture contamination. However, nonspecific IFNγ responses in the assay have meant that to achieve high-test specificity, there has been a reduction in sensitivity. Although the optimal conditions for the use of the assay in cattle have been well documented, there have been few studies optimizing the assay for use in sheep. The current study details the effect of anticoagulant, duration of incubation, cell concentration, blood storage temperature, time of stimulation of cells with antigen relative to time of sample collection, and temperatures during transit on IFNγ synthesis. Maximal IFNγ synthesis occurred with incubation periods of 48 hr in blood collected into heparinized tubes. Decreasing the leukocyte population by diluting the total peripheral blood leukocyte concentration was associated with a decreasing IFNγ response. Conversely, concentrating the peripheral blood concentration 2-fold resulted in an increase in the IFNγ production. In field studies, immediate incubation of blood samples with antigen at 37°C resulted in larger IFNγ responses; however, significantly lower IFNγ values were obtained if the samples were transported at ambient temperature. The results of this study indicate that optimization of the IFNγ assay may enable increased synthesis of IFNγ during the stimulation phase of the assay and that future work may determine whether this translates to increased sensitivity of the assay in detecting early infections in sheep.

The objective of the present study was to evaluate the in vivo wound healing effect of water extract of Stereospernum kunthianum -bark. The plant is mainly used by the local people of Mayo-Danay, Cameron as wound healing agent. The healing activity was evaluatedin rats using excision and incision wound on normal dexamethasone-delayed wound healing. Control animals received normal saline topically while other groups received dexamethasone (i.m), water extract orally or dexamethasone with the plant extract. The extract treated wounds were found to epithelize faster than those of controls dexamethasone-treated rats. Significant increase in skin tensile strength was observed in earlier than in later groups. Histological examination of incision wounds of extract-treated group showed dense collagen cross-linking and numerous blood vessels. The present study concludes that the stem bark of Stereospernum kunthianum possesses excellent wound healing potential that may substantiate its use in the traditional medicine.

Reoffending by prisoners returning to the community is a key driver in crime and prison statistics. The Community Chaplaincy project targeted at the three‐month bridging period has produced promising results in engaging clients, supporting resettlement and promoting rehabilitation. The Spider Assessment System has proved invaluable both as an assessment tool and as a therapeutic instrument, as it is easy to apply, interactive and provides clients a form of self‐expression.

The control of ovine Johne's disease (OJD) is important for domestic trade, the viability of farming units and possibly also for public health. Current strategies in Australia have included quarantine and pasture spelling to decrease prevalence and shedding rates and reduce numbers of Mycobacterium paratuberculosis (Mptb) ingested by susceptible sheep. However, alternative procedures are needed and vaccination with Gudair® has recently commenced. This review examines prospects for the control of OJD by chemotherapy, vaccination and mycophages. Current chemotherapeutic regimes for treatment of M. paratuberculosis in ruminants are prohibitively expensive and of dubious efficacy, and apart from environmental concerns, mycophage therapy lacks a track record of success against intracellular bacteria. There is substantial evidence that live and killed mycobacterial vaccines reduce the incidence of clinical disease and shedding rates in OJD. An appraisal of recent experimental results suggests that neonatal vaccination with a defined dose of M. paratuberculosis offers the best prospects for the induction of protective Th1-type immunity.

Monoclonal antibodies (MAbs) were developed against recombinant ovine interleukin-5 (IL-5) produced in the baculovirus expression vector system. One MAb, D11 (isotype IgG1), neutralised the activity of both recombinant and native sources of IL-5 in a biological assay (Baf cell assay) but was only weakly reactive in immunocytochemistry. Conversely, a second MAb, A8 (isotype IgA), successfully detected IL-5 in immunocytochemistry but did not display neutralising activity. The development of these MAbs will enable the assay of ovine IL-5 in vitro and permit studies into the role of hypersensitivity reactions in sheep by neutralisation of IL-5 in vivo.

In an on-farm study, 40 weaned piglets aged 3 weeks were vaccinated with Lawsonia intracellularis vaccine orally, IM or IP while a fourth group remained unvaccinated. All vaccinated animals showed increased serum levels of L. intracellularis-specific IgG antibodies, but significantly elevated concentrations of specific IgG, IgA and cytokines were generated in ileal mucosal secretions from the orally and IP vaccinated pigs when examined at 17 days after vaccination.

In order to analyse Th2-type immune responses in sheep by the assay of interleukin (IL)-5 in biological fluids, the ovine IL-5 gene was cloned and expressed in Spodoptera frugiperda Sf9 insect cells using the baculovirus expression vector system. The recombinant product was purified as BAC-OV-IL-5 from the supernatant fluid. The ovine IL-5 was biologically active in a bioassay using IL-5-dependent Baf cells, which have been used previously to specifically detect human IL-5. The specificity of Baf cells for ovine IL-5 was examined by two methods. First, Baf cells only proliferated in response to BAC-OV-IL-5 and did not respond to addition of recombinant ovine cytokines granulocyte-macrophage colony stimulating factor (GM-CSF), IL-1beta, IL-2, IL-3, IL-6, IL-8, stem cell factor (SCF) or IFN-gamma at doses from 0.01 to 1 microg/well. Second, the rat monoclonal antibody to murine IL-5, TRFK-5, neutralized murine, but not ovine, IL-5. However, rabbit antisera to BAC-OV-IL-5 neutralized murine and ovine recombinant IL-5 and abolished responses of Baf cells to IL-5 activity in supernatant fluids from mesenteric lymph node cells (MLNC) of parasitized sheep. The bioassay had a sensitivity to detect 8 ng in a 200 microL assay (40 ng/mL). Thus, the specificity of Baf cells to detect human IL-5 also extends to ovine IL-5 and therefore provides a method for monitoring the production of Th2 immune reactivity in sheep.

The ability of young Merino lambs to achieve protective immunity following vaccination via viable nematode infections was assessed. Lambs were infected from 1 month of age by repeated continuous low dose (trickle) administration of Trichostrongylus colubriformis or Haemonchus contortus infective larvae (L3), or by truncated infections with high doses of viable T. colubriformis L3. After 7 weeks all groups were drenched with anthelmintic and at 3 months of age they were re-infected with the homologous species. Protection was assessed by faecal egg counts at 3, 4, 5, 6 and 7 weeks after challenge, and worm count at 7 weeks after challenge. Young lambs were partially protected by 3 months of age against Trichostrongylus by trickle infection. This protection correlated with local mast cell and T-cell priming, increased numbers of local antigen-presenting cells and T-cells and increased worm-specific antibody titres in the intestine. However, there was no evidence that young lambs were capable of immunologically recognising H. contortus antigens following trickle infection, nor did trickle infection significantly protect young lambs against Haemonchus challenge.

We evaluated the anxiolytic and antidepressant effects of the aqueous extract of the bark ofTapinanthus dodoneifolius(TAE) (Danser) (25, 50, and 100 mg/kg), using open field, elevated plus maze, and forced swimming tests. Effect of TAE was compared to standard drugs diazepam (2 mg/kg) and imipramine (10 mg/kg). Additionally, the same doses of TAE were evaluated on rat's memory using Y-maze task. Results showed a significant (P<0.05; 100 mg/kg) increase in the percentage of open arm entry and the time spent in the open arms in the elevated plus maze, suggesting an anxiolytic activity of the extract. In a dose-dependant manner, TAE at 25 mg/kg significantly (P<0.05) decreased the number of lines crossed and the rearing behavior in the open field test, suggesting its possible sedative activity. In the forced swimming test, the immobility time of the animal was significantly reduced (P<0.05) by TAE (100 mg/kg), compared to control, and this effect was quite comparable to that of imipramine. In the Y-maze paradigm, TAE at 50 mg/kg caused a significant increase in the spontaneous alternations but with a significant decrease in exploratory behavioral pattern. Taking these results together, TAE improved the spatial memory and showed anxiolytic, antidepressant, and sedative activities. The present results support the anxiolytic and antidepressant activities of TAE and, to our knowledge, for the first time, demonstrate its enhancing effect on memory.

An ongoing study was made of 804 primary Stanmore total hip prostheses implanted in 839 patients between 1973 and 1991. The earliest surviving implants were brought back for radiologic and clinical review in 1995 at an average of 17 years after surgery. The remainder of the patients still living were sent a questionnaire to assess their current status. Survivorship was 95% at 10 years, 85% at 15 years, and 73% at 20 years. The average Merle d'Aubigne-Postel score was excellent up until 14 years. Patient satisfaction remained high until 22 years. Overall, 10% of the prostheses had failed. The results of this study suggest that the Stanmore prosthesis is capable of producing satisfactory long-term results that compare favorably with those of other cemented prostheses.

This study examined the role of CD8+ and WC1+ T-cells and of interferon (IFN)-gamma in the development of protective immunity against infection with the enteric nematode parasite Trichostrongylus colubriformis in sheep. Monoclonal antibodies (mAb) were administered during induction of the immune response to deplete or neutralise these components. Protection against the primary and challenge infections was assessed by faecal egg count and total worm count. Prolonged administration of mAb recognising IFN-gamma and CD8 resulted in significantly increased protection during the 6 week primary infection and following challenge. CD8+ cells were depleted from blood but not from intestinal mucosa. After injection of mAb (CC15) recognising the surface antigen WC1, WC1+ and Tcr gamma delta + cells were depleted from blood but not markedly from enteric mucosa, and protection against challenge, although variable, was increased by up to 88%. It appears that CD8+ and WC1+/gamma delta+ cells and IFN-gamma all retard the potential development of naturally acquired immunity against the parasite.

Soluble antigens that protected guinea pigs against experimental challenge with Trichostrongylus colubriformis were found to be less effective if injected as emulsions in Freund's adjuvants. This occurred despite the production of higher antibody titres in guinea pigs given emulsified antigen. Investigations of this phenomenon showed that an intraperitoneal injection of Freund's complete adjuvant (FCA) delayed rejection of primary infections and partially abrogated resistance to challenge infection. Administration of FCA/antigen emulsion to infected guinea pigs resulted in a prolonged blood eosinophilia which paralleled the increased longevity of the parasitosis. These findings suggest the need for caution in the selection of adjuvants for vaccination against gastrointestinal nematodes.

Responses of isolated mucosal mast cells (MMC) during infections with either Trichostrongylus colubriformis or Haemonchus contortus were examined by measuring the release of sheep mast cell protease (SMCP) in a degranulation assay. MMC from sheep immune to T. colubriformis released maximal amounts of SMCP and histamine within 0.5h of incubation with larval antigen whereas maximum secretion of leukotrienes occurred 3h after addition of antigen. It was only after 8 weeks of a primary T. colubriformis infection, that MMC released significantly elevated levels of SMCP (23%); this occurred when the worm burden was being rejected. In contrast, the SMCP release from MMC of immune sheep was significantly higher at 40%, and occurred within 1-4 days after challenge (DAC). The SMCP release peaked at 6-8 DAC at 51%, and declined after 56 DAC to < 25%. MMC isolated from the duodenum and mid-small intestine of immune sheep released 2-3 times higher proportion of SMCP than did cells recovered from the terminal ileum. Mast cell numbers were similar in the 3 regions but the quantity of globule leucocytes (GL) was 2.5 times higher in the duodenum. During infections with H. contortus in the abomasum, MMC isolated from the small intestine released greater levels of SMCP when incubated with larval antigens than did abomasal MMC. There was no increased release during the first 12 weeks of a primary infection although the SMCP release (23%) from immune MMC at 7-10 DAC was significantly enhanced. Once again the release from MMC isolated from the three intestinal regions of sheep immune to H. contortus was lowest in the terminal ileum.(ABSTRACT TRUNCATED AT 250 WORDS)

A mAb (IFN-9) which neutralizes biologically active ovine and bovine IFN-gamma was used to deplete levels of the cytokine in vivo in sheep and examine the consequences for immune responses to adjuvanted antigen and skin reactivity to Bacillus-Calmette-Guerin (BCG). Groups of sheep were immunized with ovalbumin in the adjuvants, Quil A or dextran sulphate (MW 500,000; DXS), both of which elicit production of IFN-gamma. MAb anti-IFN-gamma or an isotype control mAb (anti-carbonic anhydrase positive particles) were inoculated i.v. during primary and/or secondary responses. Reactions monitored in efferent prefemoral lymph indicated that anti-IFN-gamma effectively depleted levels of IFN-gamma in lymph but had no effects on the magnitude and kinetics of lymphocyte and lymphoblast traffic, or total or isotypic titres of specific Ig. When incubated in vitro with ovalbumin, antigen-reactive cells from anti-IFN-gamma treated sheep did not produce IFN-gamma, suggesting an on-going modification to cytokine production. In contrast, skin reactions to purified-protein derivative in sheep immunized with BCG were reduced by < 40% by anti-IFN-gamma. The results indicate that IFN-gamma production may not be obligatory for delayed-type hypersensitivity reactions or for the adjuvant action of Quil A or DXS, and that specific mAb can alter the profile of cytokines produced by antigen-reactive cells in sheep.

The effects of protein supplementation and infection with Trichostrongylus colubriformis on production responses and immune function in young sheep and on nematode population dynamics were assessed. Eighty-four 3-month-old Merino wether sheep were housed in individual pens and fed ad libitum chopped oaten hay containing 0.5% urea, together with 50 g day-1 lucerne meal. Fish meal (FM) was given as a source of protected protein at 0, 50 or 100 g day-1 (FM0, FM50, FM100; from Days --28 to 140). From Days 1 to 140, 0 or 1000 T. colubriformis infective larvae were given on Mondays, Wednesdays and Fridays. Infected sheep were slaughtered after 35, 70, 105, or 140 days of infection. Live-weight gain was reduced significantly by infection with T. colubriformis in sheep given FM0, but not in sheep given FM50 or FM100. Greasy wool production and fibre diameter were increased by FM, whereas the effects of infection with T. colubriformis on wool measurements depended on the level of FM given. Worm egg concentrations in faeces were significantly lower for sheep given FM100 than for those given FM0 or FM50 during the last 28 days of infection. Similarly, the apparent rate of worm expulsion was considerably higher in sheep given FM than in those not given FM. The rate of expulsion of T. colubriformis correlated with levels of circulating eosinophils as well as with the concentration of intestinal sheep mast cell proteases. Levels of parasite-specific and non-specific circulating antibodies were either unaffected or reduced as a result of supplementation with FM, although lymphocyte stimulation in vitro in response to T. colubriformis third stage larval antigen was enhanced significantly in infected animals given FM100. It was concluded that supplementary feeding with FM substantially reduced the production losses attributable to infection with T. colubriformis and was associated with enhanced expulsion of the parasite burden.

Helminthologically naive merino sheep were given either a single infection of 30,000 or a trickle infection of 6000 Trichostrongylus colubriformis infective larvae (TcL3) per week. Faecal egg counts started to fall after 8 weeks in the single infection and after 11 weeks in the trickle infection. Small intestinal contents were collected from indwelling intestinal fistulae over the next 14 weeks. Concentrations of sheep mast cell protease (SMCP) in these contents increased to highest levels 9–11 weeks and 6–10 weeks after the single infection and from the start of the trickle infection, respectively. Similarly, peptidyl leukotriene (PLT) concentrations were highest at 6 weeks and at 6–9 and 13 weeks, respectively. Histamine concentrations increased slightly after both infections to peak values at 7 weeks and 9 weeks, respectively. Inhibition of migration of larvae in vitro was increased in contents sampled at 8 weeks after the single infection and after 6–10 weeks of the trickle infection. Another 2 groups of sheep were immunised by repeated infections with TcL3. Gut contents from 1 group sampled immediately before and after challenge with 30,000 TcL3 at 0 and 18 days had increased levels of larval migration inhibitory (LMI) activity throughout the 35 day period, especially 7–14 days after challenge (DAC). The mediators SMCP increased significantly 5–7 DAC while PLT increased 7–14 DAC. In the second group of immunised sheep, levels of SMCP and PLT increased rapidly within 1 DAC and further increased 3–14 DAC. Increased concentrations of platelet activating factor PAF occurred only at 14 DAC, while LMI activity was greatest in mucus collected 5–8 DAC. Since increased concentrations of these mediators occurred around the time when egg counts started to decline in primary infections and were increased after challenge of immune sheep, the results suggest roles for the mediators and/or their cellular source in the expression of immunity.

The genetic relationship amongst Tritrichomonas foetus isolated from domestic cats and cattle was investigated by DNA sequencing of the internal transcribed region of the ribosomal DNA unit and the TR7/TR8 variable-length repeat. The results reject the hypothesis that T. foetus from domestic cats is genetically identical to T. foetus in cattle. We suggest recognition of a 'cat genotype' and a 'cattle genotype' of T. foetus. Review of public nucleotide repositories revealed that the 'cat genotype' has not been isolated from cattle nor the 'cattle genotype' recovered from cats. However, at least one cat isolate has been shown to induce disease in experimentally infected cattle. We conclude that these genotypes fall within the single species T. foetus.

Immune-mediated scouring due to ingested parasite larvae is a major concern for sheep producers in Mediterranean climates. We investigated immune-mediated scouring in parasite-resistant Merino sheep in Australia. Forty-adult, parasite-resistant Merino rams were judged to be either susceptible or non-susceptible to immune-mediated scouring on the basis of dag scores taken under field conditions. We hypothesised that the susceptible rams would have lower faecal dry matter during larval challenge than non-susceptible rams and that, at post-mortem examination, inflammatory mediators and granulocytes would be negatively correlated with both faecal dry matter and worm numbers. In pens, the rams received a dose of 500 Teladorsagia circumcincta L3 and 500 Trichostrongylus colubriformis L3 each day for 6 weeks before euthanasia. Ten rams acted as unchallenged controls. Challenging sheep with larvae reduced faecal dry matter at 2, 3 and 4 weeks after challenge began and the greatest reductions were with the sheep susceptible to scouring. The sheep showed good resistance to the parasite challenge as evidenced by low faecal worm egg counts and low total worm counts at post-mortem, with the numbers of T. colubriformis particularly low. Sheep with low faecal dry matter had significantly higher numbers of eosinophils in small intestine tissue. Sheep with low total worm counts had significantly higher levels of bradykinin in abomasum mucus. Sheep with more granulocytes in tissue and inflammatory mediators in mucus tended to have fewer numbers of T. circumcincta but there was little relationship with numbers of T. colubriformis. Our results show that dag scores are correlated to a reduction in faecal dry matter, which can be attributed to the challenge with infective parasite larvae. Inflammation during worm infection is associated with rejection of the worm challenge and may result in more fluid faeces and consequently diarrhoea. Therefore, sheep breeders should focus on breeding for both low worm egg counts and also low dag scores.

Introduction: The management of unstable slipped upper femoral epiphysis (SUFE) is controversial, with a high risk of developing avascular necrosis (AVN). We meta-analysed two areas of concern: reduction of the slip and the timing of treatment. Methods: A search of Medline, CINAHL and Embase identified only retrospectively relevant studies: four regarding the role of reduction and five regarding the timing of treatment. The incidence of AVN was compared between reduced and unreduced SUFEs, and between those treated within 24 h of symptom onset and those treated thereafter. Areas of agreement: Analysis of the pooled data gave an odds ratio of 2.20 (P ¼ 0.290) in favour of the unreduced group, who had a lower risk of developing AVN. The odds ratio was 0.50 in favour of the group treated within 24 h from symptom onset (P ¼ 0.441). However, though clinically important, these effects were not statistically significant. Areas of controversy: The timing of treatment is somewhat inconsistent: two studies favour management more than 24 h after the onset of symptoms, while for three unstable SUFEs are best managed within 24 h. Growing points: Despite the non-significant results from the meta-analysis, it can be suggested that, if reduction is to be performed, it should be undertaken cautiously, as it may be associated with increased AVN. The ideal time for management of unstable slip is probably within 24 h of symptom onset.

Johne's disease is a severe chronic enteritis of ruminants caused by Mycobacterium avium subspecies paratuberculosis (Map). Repeatable infections of known duration are required for validation of new diagnostic tests, evaluation of pathogenesis and development of improved vaccines. In the first study of its type, a standardised experimental model for Johne's disease was developed based on a lyophilised, low passage, pure culture, seedstock of Map. Experimental inoculations of sheep with accurately enumerated doses of Map resulted in infection outcomes across multiple trials that were modulated by the interval between inoculation and examination. Compared to an inoculum consisting of an intestinal mucosal homogenate from a naturally affected sheep, clinical signs from the pure culture of Map were manifested later, but other measures of infection were similar. Immunological assays showed that most of the inoculated animals were IFN-gamma positive in the early stages of the infection. Over time, an increasing number of sheep became Map-specific antibody positive, developed typical histopathological lesions and shed Map in their faeces. The repeatability and utility of this experimental infection model will enable study of many aspects of Johne's disease. It is the first study to show that models for Johne's disease can be standardised in relevant species using traditional microbiological approaches to production and storage of seedstock. It is recommended that an international bank of master seedstock be established, containing low passage isolates that are representative of the major strains of Map, S and C.

Proliferative enteropathy (PE) is an enteric disease of pigs that results in diarrhoea, reduced growth rate, reduced feed conversion efficiency and sometimes death. A survey of 13 pig veterinary practitioners in Australia was conducted to determine: (1) PE control strategies (antibiotics and vaccination), (2) how the efficacies of these strategies are evaluated and (3) how reliance on antibiotics could be reduced by increasing vaccine adoption. Antibiotics were routinely prescribed in the diets of weaner, grower and finisher pigs by 9/13, 10/13 and 8/13 veterinarians, respectively, if vaccination was not implemented. Water-soluble antibiotics were prescribed less frequently than in-feed antibiotics. Efficacy of control strategies was assessed most often through reduced clinical signs (diarrhoea, ‘tail-ender’ pigs, death) and reduced lesion incidence at abattoir postmortem inspection. Twelve practitioners had recommended PE vaccination to their clients in the previous 6 months. Barriers to continued vaccine implementation included perceived lack of efficacy for pigs housed in bedded systems, high cost relative to medication and difficulties in vaccinating pigs post-weaning.

Lymphatic cannulation and serial biopsy were used to examine changes in cell traffic and lymphocyte populations after injection of adjuvant in order to explain the mechanisms of the subversive effect that Freund's adjuvants (FA) have on protective immunity against nematode parasites in sheep. Within 4 days of intradermal injection of complete or incomplete FA into sheep there was a selective depletion of the T19+ TCY gamma delta+ T lymphocyte subpopulation from efferent lymph draining the local lymph node. Transient depletion of CD8+, T19+ and CD5+ cells from jugular blood was evident at day 2. The adjuvant-induced granuloma was a connective tissue/macrophage/T lymphocyte phenomenon, with no B cell follicles, and the T cell content continued to increase for the 21 days. The early granuloma (days 2-4) contained a disproportionate number of T19+ gamma delta + lymphocytes. It appeared that the adjuvants induced selective sequestration of this T cell subset during the initial period of granuloma formation.

Merino sheep were immunized against the intestinal nematode, T. colubriformis, by repeated infections, and proliferative responses of their peripheral blood lymphocytes (PBL) against parasite extracts and excretory-secretory (ES) antigens were monitored over 130 days. Maximal responses occurred 7–14 days after challenge. The ability of soluble proteins and parasite antigens to induce proliferation was compared with that of antigen-bearing particles obtained after antigen was adsorbed onto nitrocellulose. Blank particles increased c.p.m. two- to three-fold above that obtained in medium alone, and to elicit proliferative responses of comparable magnitude between 10 and 100 times more antigen was required when antigen-bearing particles were used instead of soluble extracts or denned proteins. Blood leucocytes as well as T-cell lines established by stimulation with parasite antigens in vitro reacted to moieties of from 5000 to 38,000 mol. wt in ES antigens on nitrocellulose particles. Direct comparisons of T-lymphocyte responses with antibody responses as assessed by immunoblots revealed different profiles of immunogenicity among ES proteins within individual sheep, but the 10,000, 30,000 and 75,000–90,000 mol. wt proteins were immunodominant. These proteins were also those consistently recognized by T-lymphocytes and sera from sheep immunized with ES proteins in adjuvant. Thus, this technique can be applied to identify parasite material which is immunogenic for T-lymphocytes, but the sensitivity of the procedure in sheep is less than reported in human studies.

Two experiments were conducted to elucidate the timing and nature of the sheep immune response to Haemonchus contortus (Barber's pole worm). The first experiment examined the establishment of H. contortus populations and the immune response by comparing a bolus infection of third-stage larvae in naive sheep with a group previously primed by a trickle infection. The second experiment used staggered doses of ivermectin-resistant larvae to compare the development of adult worms during different durations of trickle infection with ivermectin-sensitive larvae. Infections successfully generated pathological signs of haemonchosis such as anaemia. Image analysis software was used to measure the area and perimeter of worms collected at post-mortem, and the number of eggs present in individual adult females (fecundity) was significantly correlated with worm size. A significant inverse correlation was found between blood lymphocyte counts and worm fecundity. The absence of correlation between worm fecundity and other leukocyte and erythrocyte counts highlighted the specificity of the lymphocyte response. This is the first report of a link between haematology profiles and worm fecundity in haemonchosis. The correlation observed between adult worm size and egg content leads to the hypothesis that egg production in H. contortus is limited by immune regulation of worm size and presumably growth. Mean worm size and fecundity declined as sheep received more prolonged trickle infections before necropsy, confirming previous reports that immune responses to adult worms are enhanced by ongoing larval challenge. Immunohistochemical results showed trends consistent with a Th2 (humoral) immune response which has been implicated in reducing nematode burdens in several species.

The J558L cell line, previously transfected with the ovine Cepsilon gene, was induced to secrete a chimeric IgE protein composed of the ovine heavy chain and a mouse light chain with MW of approximately 80 and 26 kDa, respectively. After purification, the chimeric protein was used to immunise BALB-c mice and monoclonal antibodies (mAbs) were generated. The mAb 2F1, which had greatest anti-IgE activity in preliminary screens, was chosen for further characterisation and an examination of systemic and local IgE responses to the intestinal nematode, Trichostrongylus colubriformis. The chimeric IgE protein was not recognised in enzyme linked immunosorbent assay (ELISA) by mAbs raised against ovine IgG1, IgG2, IgA or IgM. However, 2F1 was highly specific to the chimeric IgE protein, and did not cross-react with ovine IgG1, IgG2 or IgA. Western blot analysis also showed that 2F1 and secretory IgA (sIgA) did not cross-react, and that 2F1 and the anti-IgA mAb identified different MW bands from colostrum (approximately 200 and 400 kDa, respectively). 2F1 bound to mucosal mast cells (MMC) isolated from the intestines of lambs infected with T. colubriformis, but cultured bone marrow-derived mast cells (BMMC) required prior incubation with the chimeric IgE protein for this binding to occur. Distinctive staining of plasma cells and putative mast cells were observed using 2F1 on immunohistological sections of mesenteric lymph node and jejunum.ELISA incorporating 2F1 was able to detect >0.4 ng chimeric protein. Total IgE in ovine colostrum and intestinal homogenates was quantified using a capture ELISA, with known amounts of chimeric protein used to produce a standard curve. Colostrum from outbred Merino ewes had 0.55-11.05 ng ml(-1) total IgE, and their lambs, at necropsy after infection with a total of 18,000 T. colubriformis infective larvae over a 9-week period, had 45-620 ng g(-1) total IgE in intestinal tissue. Compared to genetically susceptible lambs, antigen-specific levels of IgE were significantly higher in genetically resistant lambs after infection with 4500 T. colubriformis infective larvae (TcL3) per week for 9 weeks (161.4 versus 44.8 geometric mean titres; P=0.043). In western blots, distinctive bands (19-21 and 27 kDa) from T. colubriformis larval antigen were differentially recognised by IgE, as identified by 2F1, in intestinal homogenates from genetically resistant animals. These results have demonstrated the value of 2F1 for quantification of IgE responses in samples derived from ovine fluids and tissues using ELISA, western blots and immunohistology. In this respect, it recognises native ovine IgE and does not require pre-treatment of the sample with denaturing agents or ammonium sulphate.

The objective of the present study was to evaluate the antioxidant potential and the wound healing effect of the ethanolic extract of the bark of Calotropis procera. The antioxidant study was evaluated in vitro, using DPPH and deoxyribose degradation assays. Wound healing was studied using excision and incision wound on normal and dexamethasone-suppressed wound healing rodent models. Alkaloids, flavonoids, proteins and phenols where screened in the extract used whereas saponins and true tannins where absent. The extract contains only 12.5 gallic acid equivalent and 399.54 rutin equivalent. It was found to inhibit DPPH and deoxyribose oxidation (IC50= 24.24 and 5.40 respectively). In vivo study demonstrated a significant reduction in the epithelialization time (P

Aim: The objective of the present study was to evaluate the wound healing effect of a methanol extract of Allium cepa Linn. in vivo. Materials and Methods: The healing activity was studied using incision wound on diabetes suppressed wound healing rodent model. Results: Data recorded exhibited a significant improvement of the blood glucose level and scare breaking strength in diabetic mice at 300 and 400 mg/kg onion extract. Histological examination of the general morphology of wounds of extract treated group has shown a normal histological pattern of healthy skin at the same doses. Conclusion: This study illustrated an excellent potential of the onion consumption on dermal wound healing in diabetic patients, with a tentative mechanism of action related to tensile strength improvement and anti-hyperglycemic effect.

Following challenge with Trichosirongylus colubrifonizis, increased numbers of T-cells and immunoglobulin responses are seen in the intestine of sheep immunised by repeated infection with live worms. IL-6 mRNA expression in the small intestine from T. colubriformis-immunised and naive sheep was determined by in situ hybridisation, whereas CD4(+), IgA(+), IgG(+) cells in the gut were evaluated by immunohistochemistry. There was constitutive expression of IL-6 mRNA by cells in the naive gut, and the number of these cells was increased by parasite challenge. There were corresponding increases in numbers of CD4(+) and TCR gamma/delta(+) T-cells and IgG(+) B-cells. Our data are consistent with a role for IL-6, perhaps produced by CD4(+) and/or TCR gamma/delta(+) T-cells or B-cells, in B-cell terminal differentiation. Infiltration of B-cells, particularly IgG(+) B-cells, may reflect parasite immunity in the host.

This study examined whether infective larvae (L 3 ) of Trichostrongylus colubriformis could establish throughout the small intestine and were not restricted to the anterior duodenum in susceptible and resistant sheep. The location of worms was similar in susceptible animals given doses of T. colubriformis between 10,000 and 80,000 T. colubriformis larvae, with 90% of worms located in the proximal 3 m of the small intestine. Those worms recovered from resistant sheep were also found in the first 9 m of the intestine. However, worms recovered from immune sheep were significantly ( P=0.0074 ) relocated posteriorly from the first 3 m into the next 6 m of the intestine. By the surgical introduction of worms, it was found that T. colubriformis could establish at any site in the small intestine and to some extent in the caecum. Immunity was generated principally in the site of predilection in the anterior 3 m of the small intestine and effectively expelled challenges given at distal sites and the caecum, indicating dissemination of immunity throughout the gastrointestinal tract. Moreover, the rejection episode had removed worms from the entire small intestine within 2 h of introduction through the pylorus.