Ascaris suum draft genome.

Parasitic diseases have a devastating, long-term impact on human health, welfare and food production worldwide. More than two billion people are infected with geohelminths, including the roundworms Ascaris (common roundworm), Necator and Ancylostoma (hookworms), and Trichuris (whipworm), mainly in developing or impoverished nations of Asia, Africa and Latin America. In humans, the diseases caused by these parasites result in about 135,000 deaths annually, with a global burden comparable with that of malaria or tuberculosis in disability-adjusted life years. Ascaris alone infects around 1.2 billion people and, in children, causes nutritional deficiency, impaired physical and cognitive development and, in severe cases, death. Ascaris also causes major production losses in pigs owing to reduced growth, failure to thrive and mortality. The Ascaris-swine model makes it possible to study the parasite, its relationship with the host, and ascariasis at the molecular level. To enable such molecular studies, we report the 273 megabase draft genome of Ascaris suum and compare it with other nematode genomes. This genome has low repeat content (4.4%) and encodes about 18,500 protein-coding genes. Notably, the A. suum secretome (about 750 molecules) is rich in peptidases linked to the penetration and degradation of host tissues, and an assemblage of molecules likely to modulate or evade host immune responses. This genome provides a comprehensive resource to the scientific community and underpins the development of new and urgently needed interventions (drugs, vaccines and diagnostic tests) against ascariasis and other nematodiases.

The effects of direct and indirect road transport consignment in combination with feed withdrawal in young dairy calves.

Male dairy calves may be transported from their farm of origin at a young age. This process may involve an extended period off feed and indirect consignment through an intermediate facility, prompting potential welfare concerns. To assess the impact of transport, 59 male Holstein-Friesian dairy calves (5-9 d old) were either (1) held in situ on farm (control); (2) transported for 6 h; (3) transported for 12 h; or (4) transported for 1 h to a holding facility where they were kept for 6 h and then transported for 5 h. All treatments included a 30-h period of feed (milk) withdrawal, and calf responses were measured over time from before their last feed until the completion of the study after the transport and feed withdrawal periods. Apart from increases in serum creatine kinase in calves transported for 12 h, transported calves generally did not differ in blood concentrations of glucose, beta-hydroxybutyrate, lactate, total protein or in packed cell volume, compared with controls (P>0.05). Calf responses to the indirect consignment treatment did not differ from those of other transported calves. Withdrawal of feed for 30 h caused calves to lose 6% of body weight; blood glucose varied from 3.96 mmol/l immediately before daily feeding to 5.46 mmol/l at 3 h post feeding, and then declined to 3.43 mmol/l at 30 h. Calves lay down for 22-32% of the time during transport, and did not show a rebound effect in lying behaviour post arrival in comparison with controls. Best practice transport of 6-12 h duration, including indirect consignment via a holding facility, did not significantly affect calf blood biochemistry and metabolism in comparison with untransported animals. However, extending the time off feed beyond the daily feeding interval resulted in reduced blood glucose concentrations, suggesting that time off feed needs to be carefully managed in young transported dairy calves.

Immunohistochemical analysis of laryngeal muscle of horses clinically affected with recurrent laryngeal neuropathy.

BACKGROUND: As myosin heavy chain (MyHC) profile of muscle fibres is heavily influenced by neural input, changes in MyHC expression are expected in horses clinically affected with recurrent laryngeal neuropathy (RLN) yet, this has not been thoroughly investigated. OBJECTIVES: To describe the changes in MyHC and fibre diameter in left cricoarytenoideus dorsalis (L-CAD) muscle of horses with clinical signs of RLN. STUDY DESIGN: Observational cohort study. METHODS: Immunohistochemistry was used to assess the MyHC-based fibre-type proportion, size and grouping in the L-CAD of 10 Thoroughbred horses, five clinically affected with RLN and five unaffected controls based on resting endoscopic examination. The Mann-Whitney U test was used to compare the two groups. RESULTS: Compared to controls (of mean age 3.0 ± 1.7 years) which only expressed type I, IIA and IIX MyHC, the L-CAD of affected horses (of mean age 2.8 ± 0.8 years) had obvious fibre-type grouping, and despite apparent compensatory hypertrophy of a small number of fibres, a decrease in overall fibre diameter (median difference -35.2 µm, 95% CI -47.4 to -7.9, P = .02) and diameter of type IIA fibres (median difference -46.8 µm, 95% CI -52.1 to -5.0, P = .03) was observed. Anti-fast MyHC (MY32) cross-immunoreacted with embryonic-MyHC. Whereas MY32-positive fibres were identified as type IIX in controls, in affected horses these fibres were less than 50 µm diameter with internal nuclei and were MYH3-positive for embryonic myosin indicating depletion of type IIX fibres, yet active regeneration and fibre renewal. MAIN LIMITATIONS: Small sample size that did not include subclinical cases. Fibre size and appearance rather than staining colour were relied upon to differentiate embryonic from type IIX MyHC. CONCLUSIONS: Horses clinically affected with RLN have overall atrophy of fibres, loss of IIX fibres and expression of embryonic myosin indicating regenerative capacity. Despite hypertrophy of some remaining fibres, the overall decline in the bulk of fibres, including those most fatigue-resistant, may be the critical change that results in failure to maintain arytenoid abduction during exercise although direct comparison to subclinical cases is needed to confirm this.

Oncospheral penetration glands and secretory blebs are the sources of Taenia ovis vaccine antigens.

Taenia ovis is a cestode parasite infecting primarily sheep as intermediate hosts and dogs as definitive hosts. The first highly effective, recombinant vaccine against a parasitic organism was developed against T. ovis infection in sheep. Three separate host-protective antigens (To16, To18, and To45W) have been cloned from the oncosphere of the parasite. We localize these antigens in the oncosphere by using quantitative immunogold labeling and transmission electron microscopy. The three antigens were uniquely associated with penetration gland cells. The cytoplasm and secretory granules of both penetration gland type 1 and type 2 cells exhibited statistically significant levels of staining for each of the three antigens. The intensity of labeling of the penetration gland type 1 cell was approximately three to five times greater (P < 0.01) compared to the level of staining intensity seen in the penetration gland type 2 cell. In activated oncospheres, secretory blebs were found to contain granules with a structure similar to those observed in the penetration gland cells. The granules within the secretory blebs were shown to stain specifically for the presence of each of the three host-protective antigens. The absence of surface location of the T. ovis antigens suggests that the parasite may not be susceptible to vaccine-induced antibody- and complement-mediated attack until some postoncospheral development has occurred after infection of the intermediate host.

Use of stored equine colostrum for the treatment of foals perceived to be at risk for failure of transfer of passive immunity.

OBJECTIVE-To assess the use of stored equine colostrum for the treatment of foals perceived to be at risk for failure of transfer of passive immunity (FTPI). DESIGN-Cohort study. ANIMALS-232 Thoroughbred foals and 191 Thoroughbred mares (41 mares gave birth to 1 foal on 2 occasions). PROCEDURES-Postpartum, presuckle colostrum samples were collected from mares; samples with a colostral refractive index (cRI) > or = 23% were frozen (-20 degrees C [-4 degrees F]) and stored for > or = 7 days but < 2 years. Foals of dams that produced colostrum with a cRI value < 20% were treated with > or = 300 mL of stored colostrum that was thawed and administered via nasogastric tube on 1 to 4 occasions within 6 hours after parturition. Serum samples were obtained from colostrum-treated and nontreated foals 24 hours after treatment or suckling, respectively, for determination of serum IgG (sIgG) concentration. RESULTS-8 foals and their respective dams were excluded from the analyses. For the remaining 30 treated and 194 nontreated foals, mean +/- SD sIgG concentration was 1,597 +/- 574 mg/dL. Thirteen (5.8%) foals had sIgG concentrations < 800 mg/dL, of which 1 (0.4%) had an sIgG concentration < 400 mg/dL. Nine of these foals had suckled mares producing colostrum with a cRI value > or = 20%, and 2 foals had been treated with stored colostrum. CONCLUSIONS AND CLINICAL RELEVANCE-Treatment with stored colostrum appeared to be effective for prevention of FTPI in at-risk foals. However, foals were still at risk for FTPI despite suckling of or treatment with colostrum with adequate cRI values.

Comparison of bronchoalveolar lavage fluid examination and other diagnostic techniques with the Baermann technique for detection of naturally occurring Aelurostrongylus abstrusus infection in cats.

OBJECTIVE: To evaluate the diagnostic sensitivity and specificity of bronchoalveolar lavage (BAL) fluid examination and other diagnostic techniques, compared with the use of the Baermann technique performed on fecal samples as the reference standard, for detection of naturally occurring Aelurostrongylus abstrusus infection in a population of cats. DESIGN: Cross-sectional study. SAMPLE POPULATION: Cadavers of 80 semiferal domestic cats. PROCEDURES: BAL fluid collection and analysis, necropsy, examination of fecal samples and minced lung tissue via the Baermann technique, fecal sedimentation-flotation, and histologic examination of lung tissue were performed. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for detection of A abstrusus infection were calculated. RESULTS: On the basis of fecal Baermann test results, prevalence of infection was 13.8%. Sensitivity (NPV) of tests was as follows: Baermann technique on minced lung tissue, 81.8% (97.2%); fecal flotation-sedimentation, 63.6% (94.5%); stereomicroscopic examination of BAL fluid combined with cytologic examination of BAL fluid, 54.5% (93.2%); stereomicroscopic examination of BAL fluid alone, 45.4% (92.0%); cytologic examination of BAL fluid alone, 36.4% (90.8%); histologic examination of lung tissue, 45.4% (91.8%); and gross lung appearance, 36.4% (90.8%). Specificity and PPV of all tests were 100%, with the exception of histologic examination of lung tissue (specificity, 97.1%; PPV, 71.4%), which identified infected cats that had negative fecal Baermann test results. CONCLUSIONS AND CLINICAL RELEVANCE: The Baermann technique was the most sensitive test for detection of A abstrusus infection. On the basis of the prevalence of 13.8% in this study, A abstrusus infection should be considered in pet cats.

Vaccination with recombinant oncosphere antigens reduces the susceptibility of sheep to infection with Taenia multiceps.

Taenia multiceps is a cestode parasite, the larval stage of which encysts in the brain of sheep, goats and cattle causing an often fatal condition. The parasite also causes zoonotic infections in humans. Homologues of the recombinant oncosphere vaccine antigens from Taenia ovis and other Taenia species were identified in T. multiceps. Sequencing of the associated T. multiceps genes and cloning of the encoding mRNA has revealed conserved features in the genes and proteins. The T. multiceps oncosphere proteins, designated Tm16 and Tm18, contain a predicted secretory signal and fibronectin type III domain. The recombinant Tm16 and Tm18 proteins were successfully expressed in Escherichia coli as fusion proteins with GST. The antigens, formulated with Quil A adjuvant, were tested in a vaccine trial in sheep. The antigens stimulated immunity in sheep against challenge infection with T. multiceps eggs. Five of nine control sheep died due to a challenge infection with T. multiceps whereas none of 20 vaccinated animals died as a result of the parasite challenge (P=0.001). In addition, vaccination with the Tm16 protein, or Tm16 plus Tm18, induced significant protection against the number of parasites encysting in the brain as a result of the challenge infection (P=0.023, P=0.015, respectively). No clear relationship was apparent between the level of specific serum antibody in vaccinated animals and either the presence or absence of parasites or the number of parasites that occurred in some of the vaccinated animals. We believe this study is the first description of recombinant vaccine-related investigations for T. multiceps. The recombinant oncosphere antigens identified may allow development of effective vaccination strategies against T. multiceps infection in sheep. They raise the potential for the development of a combined vaccine with the Echinococcus granulosus EG95 antigen for prevention of T. multiceps as well as preventing the transmission of cystic hydatid disease.

Prevalence and Clinical Significance of Herpesvirus Infection in Populations of Australian Marsupials.

Herpesviruses have been reported in several marsupial species, but molecular classification has been limited to four herpesviruses in macropodids, a gammaherpesvirus in two antechinus species (Antechinus flavipes and Antechinus agilis), a gammaherpesvirus in a potoroid, the eastern bettong (Bettongia gaimardi) and two gammaherpesviruses in koalas (Phascolarctos cinereus). In this study we examined a range of Australian marsupials for the presence of herpesviruses using molecular and serological techniques, and also assessed risk factors associated with herpesvirus infection. Our study population included 99 koalas (Phascolarctos cinereus), 96 eastern grey kangaroos (Macropus giganteus), 50 Tasmanian devils (Sarcophilus harrisii) and 33 common wombats (Vombatus ursinius). In total, six novel herpesviruses (one alphaherpesvirus and five gammaherpesviruses) were identified in various host species. The overall prevalence of detection of herpesvirus DNA in our study population was 27.2% (95% confidence interval (CI) of 22.6-32.2%), but this varied between species and reached as high as 45.4% (95% CI 28.1-63.7%) in common wombats. Serum antibodies to two closely related macropodid herpesviruses (macropodid herpesvirus 1 and 2) were detected in 44.3% (95% CI 33.1-55.9%) of animals tested. This also varied between species and was as high as 92% (95% CI 74.0-99.0%) in eastern grey kangaroos. A number of epidemiological variables were identified as positive predictors for the presence of herpesvirus DNA in the marsupial samples evaluated. The most striking association was observed in koalas, where the presence of Chlamydia pecorum DNA was strongly associated with the presence of herpesvirus DNA (Odds Ratio = 60, 95% CI 12.1-297.8). Our results demonstrate the common presence of herpesviruses in Australian marsupials and provide directions for future research.

The prevalence and clinical significance of Chlamydia infection in island and mainland populations of Victorian koalas (Phascolarctos cinereus).

Chlamydia infection is known to impact the health of koalas (Phascolarctos cinereus) in New South Wales (NSW) and Queensland, but the clinical significance of Chlamydia infections in Victorian koalas is not well described. We examined the prevalence of Chlamydia infection and assessed associated health parameters in two Victorian koala populations known to be Chlamydia positive. The same testing regimen was applied to a third Victorian population in which Chlamydia had not been detected. We examined 288 koalas and collected samples from the urogenital sinus and conjunctival sacs. Detection and differentiation of Chlamydia species utilized real-time PCR and high-resolution melting curve analysis. Chlamydia pecorum was detected in two populations (prevalences: 25% and 41%, respectively) but only from urogenital sinus swabs. Chlamydia was not detected in the third population. Chlamydia pneumoniae was not detected. Chlamydia pecorum infection was positively associated with wet bottom (indicating chronic urinary tract disease) in one Chlamydia-positive population and with abnormal urogenital ultrasound findings in the other Chlamydia-positive population. The prevalence of wet bottom was similar in all populations (including the Chlamydia-free population), suggesting there is another significant cause (or causes) of wet bottom in Victorian koalas. Ocular disease was not observed. This is the largest study of Chlamydia infection in Victorian koalas, and the results suggest the potential for epidemiologic differences related to Chlamydia infections between Victorian koalas and koalas in Queensland and NSW and also between geographically distinct Victorian populations. Further studies to investigate the genotypes of C. pecorum present in Victorian koalas and to identify additional causes of wet bottom in koalas are indicated.

Oriental theileriosis in dairy cows causes a significant milk production loss.

BACKGROUND: Oriental theileriosis is a tick-borne, protozoan disease of cattle caused by members of the Theileria orientalis-complex. Recent outbreaks of this disease in eastern Australia have caused major concerns to the dairy and beef farming communities, but there are no published studies of the economic impact of this disease. On a farm in Victoria, Australia, we assessed whether oriental theileriosis has an impact on milk production and reproductive performance in dairy cows. METHODS: Blood samples collected from all 662 cows on the farm were tested using an established molecular test. For individual cows, milk production and reproductive performance data were collected. A clinical assessment of individual cows was performed. Based on clinical findings and molecular test results, the following groups of cows were classified: group 1, with cardinal clinical signs of oriental theileriosis and molecular test-positive for T. orientalis; group 2, with mild or suspected signs of theileriosis and test-positive; group 3, with no clinical signs and test-positive; and group 4, with no clinical signs and test-negative. Milk production and reproductive performance data for groups 1, 2 and 3 were each compared with those for group 4 using linear and logistic regression analyses, respectively. RESULTS: At 100 days of lactation, group 1 cows produced significantly less milk (288 l; P = 0.001), milk fat (16.8 kg; P < 0.001) and milk protein (12.6 kg; P < 0.001) compared with group 4. At this lactation point, group 2 also produced significantly less milk fat (13.6 kg; P = 0.002) and milk protein (8.6 kg; P = 0.005) than group 4. At 305 days of lactation, group 1 cows produced significantly less milk (624 l; P = 0.004), milk fat (42.9 kg; P < 0.001) and milk protein (26.0 kg; P < 0.001) compared with group 4 cows. Group 2 cows also produced significantly less milk fat (21.2 kg; P = 0.033) at this lactation point. No statistically significant difference in reproductive performance was found upon pairwise comparisons of groups 1-3 with group 4 cows. CONCLUSIONS: The present findings demonstrate that clinical oriental theileriosis can cause significant milk production losses in dairy cattle.

Compressive fatigue life of subchondral bone of the metacarpal condyle in thoroughbred racehorses.

In racehorses, fatigue related subchondral bone injury leads to overt fracture or articular surface collapse and subsequent articular cartilage degeneration. We hypothesised that the fatigue behaviour of equine subchondral bone in compression follows a power law function similar to that observed in cortical and trabecular bone. We determined the fatigue life of equine metacarpal subchondral bone in-vitro and investigated the factors influencing initial bone stiffness. Subchondral bone specimens were loaded cyclically in compression [54MPa (n=6), 66MPa (n=6), 78MPa (n=5), and 90MPa (n=6)] until failure. The fatigue life curve was determined by linear regression from log transformed number of cycles to failure and load. A general linear model was used to investigate the influence of the following variables on initial Young's Modulus: age (4-8years), specimen storage time (31-864days), time in training since most recent rest period (6-32weeks), limb, actual density (1.6873-1.8684g/cm(3)), subchondral bone injury grade (0-3), and cause of death (fatigue injury vs. other). Number of cycles to failure was (median, range) 223,603, 78,316-806,792 at 54MPa; 69,908, 146-149,855 at 66MPa; 13204, 614-16,425 at 78MPa (n=3); and 4001, 152-11,568 at 90MPa. The fatigue life curve was σ=112.2-9.6 log10Nf, (R(2)=0.52, P<0.001), where Nf is number of cycles to failure and σ is load. Removal of the three horses with the highest SCBI grade resulted in: σ=134.2-14.1 log10Nf, (R(2)=0.72, P<0.001). Initial Young's Modulus (mean±SD) was 2500±494MPa (n=22). Actual density (ρ) was the only variable retained in the model to describe initial Young's Modulus (E): E=-8196.7+5880.6ρ, (R(2)=0.34, P=0.0044). The fatigue behaviour of equine subchondral bone in compression is similar to that of cortical and trabecular bone. These data can be used to model the development of SCBI to optimize training regimes.

Description of technique and lower reference limit for magnetic resonance imaging of hippocampal volumetry in dogs.

OBJECTIVE: To evaluate the use of high-resolution MRI for hippocampal volumetry in dogs and to define a lower reference limit for hippocampal formation (HF) volume. ANIMALS: 20 dogs (with no history of seizures and no underlying structural brain disease) that underwent MRI of the brain. PROCEDURES: The MRI protocol included a high-resolution T1-weighted 3-D ultrafast gradient-echo sequence aligned in a dorsal plane perpendicular to the long axis of the HF. Images obtained with MRI were retrospectively analyzed by 2 observers (A and B). Intraobserver and interobserver agreement were calculated with the Lin concordance correlation coefficient. Volume measurements of the HF were adjusted for intracranial volume, and a lower 95% reference limit for adjusted HF volume was calculated. RESULTS: There was substantial intraobserver agreement (Lin concordance correlation coefficient, 0.97 [95% confidence interval {CI}, 0.94 to 0.99]) but poor interobserver agreement (Lin concordance correlation coefficient, 0.63 [95% CI, 0.37 to 0.79]). The lower 95% reference limit for adjusted HF volume was 0.56 cm(3) (90% CI, 0.52 to 0.60 cm(3)) for the right HF and 0.55 cm(3) (90% CI, 0.52 to 0.58 cm(3)) for the left HF. CONCLUSIONS AND CLINICAL RELEVANCE: HF volumes should be adjusted for intracranial volume to account for the large variation in canine skull size. The amount of time required to perform HF volumetry and low interobserver agreement may restrict this technique to research applications, such as the investigation of epileptic patients for hippocampal sclerosis or other cognitive disorders.

Epidemiological survey following oriental theileriosis outbreaks in Victoria, Australia, on selected cattle farms.

This study investigated Theileria orientalis following outbreaks of oriental theileriosis in cattle in the state of Victoria, Australia, from September 2010 to January 2012, using traditional and molecular methods of diagnosis. A questionnaire was used to collect epidemiological information from cattle farms. Blood samples (n=301), collected from individual symptomatic and asymptomatic cattle from 19 cattle farms, were examined for the presence of Theileria on stained blood smears and tested using a PCR-based approach, employing a region within the major piroplasm surface protein (MPSP) gene as a marker. The microscopic examination of stained blood smears detected stages consistent with Theileria piroplasms in 28.1% (79/281) of the samples. PCR products were amplified from 70.8% (213/301) of the samples. Mutation scanning analysis of all amplicons displayed seven distinct profiles. Following the direct sequencing of representative amplicons, the genotypes ikeda, chitose, buffeli and type 5 were detected in 91.1%, 32.9%, 2.4% and 1.4% of 213 blood samples, respectively. The distribution of these four genotypes varied among the 19 farms; genotype ikeda was detected on all farms, whereas genotypes chitose, buffeli and type 5 were detected on 14, 3 and 2 farms, respectively. Mix infections with genotypes ikeda and chitose were common (21.6%). Survey results revealed that oriental theileriosis affected mainly beef cows of more than two years of age, prior to calving, and disease was associated with abortion and cow deaths. Future investigations should focus on developing improved tools for investigating and managing oriental theileriosis.

The effects on claw health of supplement feeding grazing dairy cows on feed pads.

The effects of feeding and management systems on the health and welfare of grazing dairy cows were investigated by comparing the claw health of cows fed grain during milking and pasture silage in the paddock (Control), with cows fed a grain-based partial mixed ration (PMR) on a concrete feed pad. Cows were assessed on three occasions during lactation: (1) early lactation (20-81 days in milk [DIM]) before allocation to feeding treatments; (2) mid-lactation (97-158 DIM) immediately following an intensive feeding experiment, and (3) late lactation (173-243 DIM) several months after return to initial management groups. At the final examination, claw puncture resistance was measured. The results showed that for the most prevalent lesions (white line disease, paintbrush haemorrhage and traumatic bruising), there was no effect of feeding system or amount of supplement on the presence of the moderate to severe forms in early lactation, but cows were more likely to have a particular lesion at the second assessment if it was present in early lactation. Puncture resistance of the claw was not related to presence of a lesion for any of the most prevalent lesion types. It was concluded for this herd that for most indicators of claw health, there was no overall effect of different feeding systems (supplement fed during milking or on a feed pad) or amount of supplement.

Association between canine pancreatic-specific lipase and histologic exocrine pancreatic inflammation in dogs: assessing specificity.

The current study aimed to determine the specificity, and to a lesser extent the sensitivity, of canine pancreatic-specific lipase (cPL) concentration in dogs with various disease conditions. Dogs were presented for postmortem examination and had serum collected for cPL concentration within 6 hr preceding death or immediately postmortem. Pancreatic tissue was collected postmortem, and sections from the left lobe, right lobe, and body of the pancreas were examined histologically. Inflammation and fibrosis in each section were assessed to determine a total pancreatic inflammatory score and pancreatic fibrosis score in each dog. Correlations between these scores and the cPL concentration were made, as well as determination of specificity. A total of 32 dogs were included in the analysis, 20 of whom had no to minimal pancreatic inflammation. The specificity of cPL with a cutoff value of 200 µg/l was 80% (95% confidence interval [CI]: 56-94%), while with a cutoff of 400 µg/l, the specificity was 90% (95% CI: 68-99%). There was a significant but rather low correlation between cPL concentration and the pancreatic inflammation score, but not with the fibrosis score. Canine pancreatic-specific lipase concentration has good specificity overall in dogs without pancreatitis. This test is less useful in dogs with milder pancreatitis, and both false-positive and false-negative results occur. Results indicated that dogs with clinical signs suggestive of pancreatitis would require abdominal imaging in addition to serum cPL testing to evaluate the cause (or causes) of clinical signs.

Clinicopathologic evidence of myocardial injury in horses with acute abdominal disease.

OBJECTIVE: To determine whether there is evidence of myocardial injury in horses with acute abdominal disease. DESIGN: Prospective case series. ANIMALS: 18 healthy horses and 69 horses with acute abdominal disease. PROCEDURES: 18 healthy horses had been admitted to the hospital for investigation and were assigned to group 1. Horses examined for acute abdominal disease were assigned to 3 groups: strangulating obstruction, nonstrangulating obstruction, or inflammatory disease (groups 2, 3, and 4, respectively). Heart rate, Hct, and blood lactate and cardiac troponin I (cTnI) concentrations were measured at initial examination. Myocardial function was assessed by echocardiographic measurement of fractional shortening and left ventricular ejection time (LVET). Heart rhythm was evaluated via ECG. RESULTS: The proportion of horses with high (> 0.03 ng/mL) cTnI concentration was significantly greater among horses with strangulating (9/25 [36%]) or inflammatory (9/19 [47%]) lesions, compared with healthy horses (0/18). The proportion of horses with high cTnI concentration was significantly greater among nonsurvivors (12/24 [50%]) than among survivors (10/45 [22%]). Serum cTnI concentration was positively correlated with Hct, heart rate, and blood lactate concentration and negatively correlated with LVET. CONCLUSIONS AND CLINICAL RELEVANCE: Evidence of myocardial injury was observed in horses with acute abdominal disease, and this injury was associated with severity of illness. Recognition of myocardial injury could improve treatment of acute abdominal disease in horses.

Vaccination of bovines against Echinococcus granulosus (cystic echinococcosis).

Hydatid disease is an important human zoonosis. Humans become infected from carnivores that are infected with the Echinococcus granulosus tapeworm. Carnivores become infected after consuming hydatid cysts from grazing animals, which are generally sheep, goats and cattle. A vaccine, known as EG95, can protect sheep and goats against cystic echinococcosis. This paper describes the adaptation of the EG95 vaccine for use in cattle. The monitoring of results used serology and also infection with E. granulosus eggs, followed by necropsy. Immunisation with living E. granulosus oncospheres showed that cattle could be immunised against E. granulosus. Immunisation of cattle with EG95 plus QuilA was also successful. A dose-response and adjuvant trial showed best results were achieved with 250 µg of antigen and 5mg of the adjuvant QuilA, which was 5 times the recommended sheep dose. After two vaccinations given one month apart, 90% protection was maintained for 12 months. At 12 months a third vaccination boosted protection to 99% which was maintained for a further 11 months.

Establishment of a robotic, high-throughput platform for the specific diagnosis of gastrointestinal nematode infections in sheep.

The accurate diagnosis of strongylid nematode infections is central to investigating their epidemiology and for parasite control. To overcome major limitations in sensitivity or specificity of traditional methods, including faecal egg count (FEC) and/or larval culture (LC), we evaluated and established a semi-automated, high throughput multiplexed-tandem PCR (MT-PCR) platform for the diagnosis of gastrointestinal strongylid nematode infections in sheep, and established its diagnostic sensitivity (100%) and specificity (87.5%) based on the testing of 100 faecal DNA samples from helminth-free sheep and 30 samples from sheep with infections confirmed by necropsy. Subsequently, the platform was employed to test 219 faecal samples from sheep with naturally acquired infections from various geographical localities within Australia and the results compared with those from conventional LC using 139 of the 219 samples. The results obtained using both MT-PCR and LC correlated significantly for most nematodes examined, but revealed that Oesophagostomum venulosum and Chabertia ovina (parasites of the large intestine) were significantly under-represented in the LC results. The results showed that Trichostrongylus spp. (87%), Teladorsagia circumcincta (80%) and Haemonchus contortus (67%) had the highest prevalences, followed by O. venulosum (51%) and C. ovina (12%). The molecular-diagnostic platform established can be used for species- or genus-specific diagnosis of patent nematode infections within 24h (compared with 7-10 days for LC), and is a sensitive and cost effective tool for routine application in research and service laboratories.

A molecular diagnostic tool to replace larval culture in conventional faecal egg count reduction testing in sheep.

The accurate diagnosis of parasitic nematode infections in livestock (including sheep and goats) is central to their effective control and the detection of the anthelmintic resistance. Traditionally, the faecal egg count reduction test (FECRT), combined with the technique of larval culture (LC), has been used widely to assess drug-susceptibility/resistance in strongylid nematodes. However, this approach suffers from a lack of specificity, sensitivity and reliability, and is time-consuming and costly to conduct. Here, we critically assessed a specific PCR assay to support FECRT, in a well-controlled experiment on sheep with naturally acquired strongylid infections known to be resistant to benzimidazoles. We showed that the PCR results were in close agreement with those of total worm count (TWC), but not of LC. Importantly, albendazole resistance detected by PCR-coupled FECRT was unequivocally linked to Teladorsagia circumcincta and, to lesser extent, Trichostrongylus colubriformis, a result that was not achievable by LC. The key findings from this study demonstrate that our PCR-coupled FECRT approach has major merit for supporting anthelmintic resistance in nematode populations. The findings also show clearly that our PCR assay can be used as an alternative to LC, and is more time-efficient and less laborious, which has important practical implications for the effective management and control strongylid nematodes of sheep.

Evaluation and application of a molecular method to assess the composition of strongylid nematode populations in sheep with naturally acquired infections.

We evaluated the performance of a PCR method for the diagnosis of naturally acquired strongylid nematode infections in sheep (n = 470; in a temperate climatic zone of south-eastern Australia), using a panel of 100 'negative control' samples from sheep known not to harbour parasitic helminths. We compared the diagnostic sensitivity (98%) and specificity (100%) of this assay against a conventional faecal flotation method and also established a system to rank the contribution of particular strongylid nematodes to the faecal egg counts (FECs) from 'mixed infections' in individual sheep. The testing of faecal samples herein revealed that Teladorsagia circumcincta (80%) and Trichostrongylus spp. (66%) were most prevalent, followed by Chabertia ovina (33%), Oesophagostomum venulosum (28%) and Haemonchus contortus (1%). For the majority of sheep in this study, T. circumcincta and Trichostrongylus spp. represented the largest proportion of strongylid eggs in faecal samples from individual sheep. This is the first large-scale prevalence survey of gastrointestinal nematodes in live sheep using a molecular tool. The ability to rapidly rank strongylid nematodes according to their contribution to mixed infections represents a major advantage over routine coprological methods. This PCR tool has the potential to replace the conventional technique of larval culture. Future efforts will focus on enhancing and adapting this molecular method for high throughput application in routine, diagnostic settings.