Australian surveys on incidence and control of blowfly strike in sheep between 2003 and 2019 reveal increased use of breeding for resistance, treatment with preventative chemicals and pain relief around mulesing.

Blowfly strike or cutaneous myiasis, caused principally by the Australian sheep blowfly Lucilia cuprina is endemic to Australian sheep producing areas and is a major cause of suffering and economic loss. This article reports incidence and control practices as determined by three cross-sectional surveys of Australian sheep farmers covering the years 2003 (n = 1365), 2011 (n = 575) and 2018 (n = 354) with more questions relating to blowfly strike in the latter two surveys. Breech strike was reported at a higher rate than body strike in all survey years. Reported annual incidence of breech strike ranged from 2.2 to 4.7% of sheep nationally with minor differences between ewes and lambs. The incidence of body strike ranged from 1.0 to 7.1% with higher incidence in younger sheep in all years. The use of fixed time routine preventative chemical treatments increased during the survey period (43%, 46% and 66%, in 2003, 2011 and 2018 respectively) with dicyclanil (54%-58%) and cyromazine (23-36%) the main insecticides used for control since 2011. The use of the Mules operation (mulesing) to remove skin wrinkles from the breech area as a preventative modification did not change between 2011 and 2018 (47% in both), however, the use of pain relief at mulesing of replacement ewe lambs increased substantially between 2011 (59%) and 2018 (87%). The most commonly used methods to assist with flystrike control were timing of crutching (clipping of wool from susceptible areas) and shearing, with few respondents destroying maggots from treated sheep (21-26%) and very low use of fly traps (5-8%). Phenotypic selection of ewes for reduced susceptibility to flystrike based on visual traits was practiced by 61% of respondents in 2011 and 56% in 2018. Selection of rams for reduced susceptibility using estimated breeding values increased from 10% in 2011 to 17% in 2018. Breeding for resistance was the most commonly nominated (21%) most important change made to flystrike management in 2018. The results indicate that concern for sheep welfare has increased with almost universal use of pain relief at mulesing. Increased use of selection for blowfly resistance indicates farmer commitment to planning for a future without mulesing, although the practice remains prevalent. An increase in the reliance on preventative chemical treatments increases the risk of insecticide resistance.

Prevalence of sheep lice and trends in control practices across Australia - Australian sheep parasite control surveys from 2003 to 2019.

The sheep body louse (Bovicola ovis) is a parasite of major economic concern in Australia. This article reports lice incidence and control practices on Australian sheep farms as determined by three cross-sectional surveys of the years 2003, 2011 and 2018. The incidence of live lice detected within a flock was similar across the survey years with a slight decline in 2018 (2003: 20%, 2011: 18.6% and 2018: 13.9%). In an average year, most farmers did not see any direct or indirect evidence of lice infestation (2011: 59.3% and 2018: 69.5%), however, over a five-year period 38.9% (2003), 66.3% (in 2011) and 55.8% (in 2018) reported seeing evidence of lice infestation at least once with significant variation between sheep producing regions in 2011. In an average year, nearly three quarters of respondents treated for lice (2011:73.5% and 2018: 73.3%) with the majority treating immediately following shearing and very few treating sheep with long wool. Even higher percentages treated for lice at least once in the preceding five-year period (2011: 86.2% and 2018: 87.5%). Backliner was the most popular method of chemical application for lice control in sheep off-shears or with short wool (2003: 77%, 2011: 73% and 2018: 74.3%). For long wool treatments, hand jetting declined in popularity from 2003 (64%) to 2011 (54%) and 2018 (8.6%) as backliners became more popular (2003: 36%, 2011: 51% and 2018: 60%). The use of benzoylphenyl urea insect growth regulators (IGR) for off-shears/short wool treatment declined from 2003 (92.8%) to 2011 (51%) and 2018 (2.9%) and were largely replaced by neonicotinoids and spinosad for backliner/spray-on products. The use of organophosphates declined for plunge dipping (2003: 83.8%, 2011: 83% and 2018: 7.7%). Spinosad use for plunge dipping off-shears/short wool increased over the survey years (2003, 0%, 2011: 9% and 2018:46.2%). The use of IGRs declined for backliner application on long wool and were mainly replaced by spinosad in 2011 and 2018. Fewer respondents reported suspected resistance to lice control products in 2018 (8%) compared with 2003 (26%) and 2011 (13%) with most reporting suspected resistance to IGR and synthetic pyrethroids and emerging suspicions of resistance to neonicotinoids in 2018. Resistance to lice control products also reduced in importance as a reason for recurring lice infestations between 2011 (ranked 2nd) and 2018 (ranked 6th). Biosecurity was important to sheep producers with the highest ranked reason for recurring lice infestations being from stray or purchased sheep.

A cost-benefit analysis of foot and mouth disease control program for smallholder cattle farmers in Cambodia.

The Cambodian government is attempting to mobilise government, donor and private sector funding to implement a coordinated FMD vaccination program (FMDVP). A necessary first step is to convince the farmers of the benefits of participating in and potentially financially supporting this program. Information was collected from 300 farmers in order to estimate the on-farm benefits and costs of their participation in an FMDVP. Implementing a successful vaccination program is difficult, and farmers understand from previous experience that there may be institutional, social, technical and financial constraints which limit its success. A benefit-cost analysis needs to take into account that outbreaks do not occur every year, not all cattle will be successfully vaccinated, not all sick animals successfully treated and sometimes sick animals simply sold. This study sensitises these variables in order to give a realistic estimation of the farmer participation benefits in an FMDVP. A general result is that it is worthwhile for farmers to participate in the FMDVP if there are average annual outbreaks, or at least two major outbreaks, in the ensuing 5 years. However, the results are influenced by the interaction of vaccination success and treatment success and coverage. Ineffective coverage and poor treatment of sick animals reduce the benefits of an FMDVP. It is also important that farmers do not sell sick stock and, if they do, that they are able to breed replacements rather than purchase replacements. There are many factors in the smallholder cattle farming system that will influence the success of an FMDVP; farmers will only choose to participate if they can be convinced of the short and long-term economic benefits.

Investigation of the combined efficacy of two Haemonchus contortus vaccines in weaner Merino sheep.

This study was conducted to investigate whether co-administration of Barbervax® (Bvax) with Haemonchus contortus surface larval antigen (HcsL3) would increase the protective efficacy and duration of protection against H. contortus infection in weaner Merino sheep. A total of 132 10-month-old weaned Merino ewe lambs were randomly allocated into six treatment groups (n = 22). Sheep were vaccinated four times with either Barbervax® (Bvax), H. contortus L3 surface larval antigen (HcsL3), combined vaccination (Bvax + HcsL3), Bvax + AlOH, HcsL3 + Saponin or remained as unvaccinated controls. Aluminium hydroxide (AlOH) and saponin adjuvants were included in HcsL3 and Bvax vaccines respectively. The first three vaccinations were given at 4 week intervals and the fourth vaccination provided as booster, 9 weeks later. All animals were treated with Zolvix™ (monepantel 25 mg/mL, Elanco) at the third vaccination and commencing two weeks later, artificially trickle infected with H. contortus L3. Worm egg count (WEC), packed cell volume (PCV), antibody titre and bodyweight were measured throughout the study as was specific antibody directed against each antigen using ELISA. The administration of Bvax and HcsL3, alone or in combination, induced an antibody response against HcsL3 but only the Bvax and the combined treatment elicited an antibody response to the Bvax antigen. The targeting of HcsL3 by each vaccine was confirmed by immunofluorescence staining of H. contortus L3. However, only the booster vaccination in the Bvax treatments reduced WEC to levels below untreated controls. The HcsL3 vaccine did not reduce WEC in this experiment and co-administration with Bvax did not improve the efficacy and duration of protection against H. contortus infection.

Benchmarking Australian sheep parasite control: Changes in gastrointestinal nematode control practices reported from surveys between 2003 and 2019.

Cross-sectional surveys of sheep parasite control practices in Australia were conducted in 2004, 2012 and 2019 to document parasite problems, control practices and measure change over time. This article reports the results pertaining to gastrointestinal nematode infection; comparisons between years are mostly descriptive and not based on statistical inference. There was a general increase in the use of grazing management to prepare clean pastures for sheep to control gastrointestinal nematode infections with the largest increases in the use of: cropping, long acting anthelmintics to prepare clean pastures, feeding strategies, selecting rams for resistance to nematodes, and leaving some sheep untreated. The proportion of respondents using faecal worm egg count monitoring (WEC) and the number of WEC monitors per year were similar in 2003 (weaners: 3.0 WEC/year, adult ewes: 2.6 WEC/year) and 2018 (lambs and weaners and adult ewes both 3.1 WEC/year) but lower in 2011(lambs and weaners: 2.0 WEC/year, adult ewes: 2.9 WEC/year), whilst there was a higher number of anthelmintic treatments given in 2011 than 2003 and 2018 which were similar. There was an increase in the proportion of respondents carrying out WEC monitors themselves from 2011 (21%) to 2018 (30%) and a corresponding reduction in the use of private laboratories, government laboratories and veterinarians with regional differences in who carried out WECs. The top three anthelmintic groups used did not differ between the three survey years with macrocyclic lactones the most used followed by benzimidazoles (BZ) and levamisole (LEV) although the percentage of MLs used in 2011 and 2018 was lower than in 2003 with higher proportions of respondents using BZ and LEV groups in the latter two survey years. Moxidectin was in the most commonly used active in all survey years. There was an increase in the use of combination of anthelmintics from different groups, especially for a combination of three anthelmintics (2003: 4.4%, 2011: 19.1%, 2018: 21.5%), with lower use of single anthelmintics in 2011 (57.0%) and 2018 (55.4%) compared with 2003 (74.5%). The use of testing for anthelmintic resistance was generally low across the survey years with a lower proportion of respondents using tests in 2011 and 2018 than in 2003 (2003:48%, 2011: 29%, 2018: 37%). Time of year, results from WEC and seasonal weather condition were the most important factors when deciding when to apply anthelmintic treatment.

Protective efficacy of Barbervax® in Merino weaner sheep trickle infected with five doses of Haemonchus contortus infective larvae.

Barbervax® protects sheep against H. contortus infection; however, the level of protection afforded by the vaccine at different levels of larval challenge under field conditions has not been reported. Therefore, this study was designed to investigate the protective efficacy of Barbervax® at increasing doses of trickle infection with H. contortus third stage larvae (L3). Merino lambs (220) were randomly allocated to challenge treatment groups and received Barbervax® vaccinations at approximately 8, 11, 15 and 22 weeks of age or not vaccinated (control group). Animals within each treatment group were subjected to one of five levels of H. contortus L3 trickle infection (0, 300, 600, 1200 and 2400 L3/week in two split doses). Trickle infections started two weeks after the third vaccination (week 0). Worm egg count (WEC), packed cell volume (PCV), antibody titre and bodyweight were measured at week 2 (start of trickle infection), week 5 and then every two weeks for 11 weeks. Vaccinated weaners had a significantly (P < 0.0001) lower faecal WEC than unvaccinated control animals. Vaccination induced significant specific antibody responses that were not influenced by level of trickle infection. Vaccination significantly reduced the number of weaners requiring salvage anthelmintic treatment (9.4% vaccinated weaners versus 33.6% unvaccinated). Vaccine protective index based on WEC was similar at all challenge levels (overall mean> 60%) and at the higher challenge levels did not reduce H. contortus infection to levels that would not require anthelmintic treatment. Therefore, it is suggested that under conditions of high larval challenge the use of Barbervax® should be integrated with other control methods.

Viability and development of Ascaridia galli eggs recovered in artificial media followed by storage under different conditions.

Eggs oviposited by Ascaridia galli females in artificial media are commonly used as a source of infective material. We investigated the rate of egg production by cultured mature females (n = 223), and changes in egg viability under different storage and incubation conditions. Eggs recovered after 1, 2 or 3 days of culture were subjected to either (1) storage in water at 4°C (1, 4 or 8 weeks) followed by incubation in 0.1 N H2SO4 at 26°C (2, 4 or 6 weeks); or (2) prolonged storage at 4°C (up to 14 weeks). Egg development and viability was assessed by morphology coupled with a viability dye exclusion test of hatched larvae. Of the 6,044 eggs recovered per mature female 49.2, 38.5 and 12.3% were recovered on days 1, 2 and 3 of worm incubation respectively with similar initial viability (≥99%) between days. Eggs recovered on different days had only minor differences in viability after storage. The prolonged storage period at 4°C significantly affected both viability and embryonation ability resulting in decline in viability of 5.7-6.2% per week. A smaller but significant decline in egg (2.0%) and hatched larval (1.4%) viability per week of incubation at 26°C was also observed. We conclude that storage and incubation conditions, not the day of egg recovery, are the main factors affecting A. galli egg viability. Our findings indicate that under aerobic conditions storage at 26°C may be preferable to 4°C whereas other studies indicate that under anaerobic conditions storage at 4°C is preferable.

A simplified Barbervax® vaccination regimen in lambs to evoke immunological protection to Haemonchus contortus.

This study assessed the feasibility of altering the pre-weaning vaccination schedule of the commercially available Barbervax® vaccine directed against Haemonchus contortus, to avoid the 2nd priming vaccination which typically falls between lamb marking and weaning. Merino lambs (n = 175) born to maiden ewes, located in the Northern Tablelands, New South Wales, were randomly allocated to treatment groups (n = 35) and subjected to five different vaccination regimes. One group remained as unvaccinated controls and another had the full set of three priming doses. The other three groups were vaccinated only at marking and weaning receiving a double dose of vaccine at marking and/or weaning. The hypothesis tested was that reducing the interval between lamb marking and weaning to 6 weeks, and providing a double dose of vaccine at marking, weaning or both would remove the need for the second priming vaccination between lamb marking and weaning. This priming vaccination in the vaccination protocol necessitates an additional mustering of ewes with young lambs at foot and is a time consuming and costly exercise which increases the risk of mis-mothering. Blood and faecal samples were collected at frequent intervals for worm egg count (WEC), larval differentiation and H. contortus vaccine specific ELISA antibody analysis. Overall, the results supported the hypothesis, and it was found that antibody titres and WEC reductions equivalent to the registered vaccine regimen were achieved by the alternative regimens. This finding requires further investigation under a wider range of conditions. Deviation from the registered vaccination protocol would constitute off-label usage, and at this time and until further evaluations are done these deviations are not recommended.

Evaluation of Barbervax® vaccination for lambing Merino ewes.

The Barbervax® vaccine, directed against Haemonchus contortus, has been commercially available in Australia since 2014. However, to date the optimal timing for booster vaccination of lambing ewes has not been identified. Therefore the aim of this experiment was to compare the efficacy of Barbervax® when administered to ewes one week prior to the commencement of lambing versus at lamb marking 7 weeks later. A total of 400 single bearing Merino ewes, run on a commercial sheep property in the New England region of New South Wales, previously primed with Barbervax® were enrolled in the project. The experiment incorporated a cross-over design with two treatments imposed pre-lambing; Barbervax® booster vaccination with anthelmintic (n = 200) or anthelmintic alone (n = 200). At lamb marking, there were four treatment groups (n = 100) and anthelmintic was administered to all treatment groups. The four treatment combinations were Barbervax® booster vaccination at pre-lambing and lamb marking (V-V), booster vaccination only at pre-lambing (V-A), booster vaccination only at lamb marking (A-V) or anthelmintic only at both at pre-lambing and lamb marking (A-A). Pre-lambing Barbervax® booster vaccination of ewes induced an antibody response and, when used in conjunction with anthelmintic, significantly reduced worm egg counts (WEC) assessed at 7 weeks (lamb marking) and 13 weeks later, relative to treatment with anthelmintic alone. Booster vaccination plus anthelmintic treatment at lamb marking was not effective at reducing WEC compared to anthelmintic treatment alone. Body weight and packed cell volume did not differ between the treatment groups throughout the trial. In conclusion Barbervax® booster vaccination at pre-lambing provides additional protection for reproducing ewes given an effective short acting anthelmintic at this time.

Flock use of the range is associated with the use of different components of a multi-tier aviary system in commercial free-range laying hens.

1. The objective of this study was to investigate the association of using a multi-tier aviary system and access to range on flock uniformity in free-range laying hens, and to determine whether the extent of range use or flock uniformity can be predicted from the use of different levels of the aviary system.2. A total of 13,716 Lohmann Brown hens from five commercial free-range flocks housed in identical houses on the same farm were individually weighed at 16 weeks of age and allocated to five replicate areas within each house. Hen movement in the multi-tier aviary system and on the range was individually monitored using radio frequency identification (RFID). All hens had access to the range from 18 to 22 weeks of age and were exposed to the same management conditions.3. Whilst only one flock significantly changed its flock uniformity with time, they differed from each other in uniformity and body weight (P = 0.001).4. Hens spent most of their available time on the lower aviary tier (7.29 ± 0.029 h/hen/day) and on the upper aviary tier (4.29 ± 0.024 h/hen/day) while the least amount of time was spent on the range and in the nest boxes (0.93 ± 0.005 h/hen/day and 1.48 ± 0.007 h, respectively, P = 0.001).5. Range use was negatively correlated (r = -0.30) to the time spent on the upper aviary tier and positively correlated (r = 0.46) to the time spent on the lower aviary tier (P = 0.001). Bivariate analysis revealed that range and upper aviary resp. lower aviary tier usage had a significant curvilinear association.6. In conclusion, the study showed that range use was associated to the time hens spent on the different tiers of the aviary system. Flock uniformity varied between flocks but was not associated with either range and aviary system usage.

Tissue distribution, shedding and environmental detection of infectious bursal disease virus genome following infection of meat chickens at two ages.

OBJECTIVE: To compare the effects of infectious bursal disease virus (IBDV) infection of commercial meat chickens at 0 and 16 days old (d.o.) and determine if IBDV vRNA is quantifiable in litter and dust samples. METHODS: Ross meat chickens (n = 60) were orally infected or not with IBDV at 0 or 16 d.o. Blood and faecal samples were collected longitudinally to 28 days post infection (dpi) from six chickens and tissues collected weekly from three euthanased chickens. Relative bursal weight was recorded postmortem. IBDV antibody titres in sera were measured using ELISA and VCN was determined in tissues, faeces, litter and dust using qRT-PCR. RESULTS: Chickens infected at 16 d.o. had earlier and more severe bursal atrophy, earlier and higher IBDV vRNA load in lymphoid organs and an earlier and greater antibody response to infection than those infected at 0 d.o. Faecal shedding of IBDV between 2 and 6 dpi was observed in both groups followed by cessation with the 0 d.o. group and re-initiation of shedding at 28 dpi. IBDV was readily detected and quantified in litter and dust samples. CONCLUSIONS: The presence of significant maternal antibody (MAb) titres in 0 d.o. chickens provided protection against IBDV replication and bursal atrophy at 7 and 14 days post infection. The reduced titres of MAb present at 16 d.o. did not prevent rapid IBDV replication and early marked bursal atrophy. The observed resistance of 0 d.o. chickens is likely to be a combination of MAb inhibition of IBDV and true age resistance of neonatal chicks. Measurement of IBDV in litter and dust may have research or diagnostic application.

Pathogenicity, tissue distribution, shedding and environmental detection of two strains of IBDV following infection of chickens at 0 and 14 days of age.

Infectious bursal disease virus (IBDV) is endemic to most poultry-producing countries worldwide. Immunosuppressive classical and variant IBDV strains endemic to Australia are genetically distinct from other international strains. We report the results of infection experiments with Australian classical strain 06/95 and variant strain 02/95 in SPF chickens. We tested the effects of strain and age of infection on bursal atrophy, viral RNA (vRNA) load in bursa of Fabricius (bursa), spleen, thymus, caecal tonsils, faeces, litter and exhaust dust as determined by real-time reverse transcriptase polymerase chain reaction. The two IBDV strains did not differ in the degree of bursal atrophy induced, lymphoid organ distribution and faecal shedding but variant strain 02/95 induced a greater antibody response to the infection than classical strain 06/95 which was associated with a more rapid decline in IBDV vRNA genome copy number (VCN) in lymphoid organs and faeces. Infection at 14 days of age induced greater bursal atrophy and higher vRNA copy number in lymphoid tissues than infection on the day of hatching, indicating true age susceptibility independent of maternal antibody (Mab) status. The direction of the association between rankings for IBDV vRNA load in bursa and relative bursal weight changed from positive at 3 and 6 days post-infection to negative at 28 days post-infection. Intra-tracheal administration of dust collected from chickens infected with IBDV resulted in successful transmission of IBDV. IBDV vRNA was detected successfully at high levels in the environmental litter and dust samples.

Field studies of the detection, persistence and spread of the Rispens CVI988 vaccine virus and the extent of co-infection with Marek's disease virus.

OBJECTIVE: To use specific real-time qPCR to determine (1) the vaccination success of Rispens CVI988 vaccine in feathers and dust; (2) persistence of Rispens infection in vaccinated layer chickens; (3) extent of co-infection with wild-type Marek's disease virus (MDV) in vaccinated layers; and (4) presence of Rispens virus in unvaccinated broiler flocks. METHODS: Feather, dust and serum samples were collected from birds aged 3 days to 91 weeks from three layer farms. qPCR was used to detect MDV and Rispens in DNA extracted from dust and feathers. Previously tested MDV-positive dust samples from 100 broiler flocks were tested for the presence of Rispens using qPCR, while serum samples were used to detect anti-MDV antibody using ELISA. RESULTS: Overall, 66% and 93% of feather and dust samples, respectively, from Rispens-vaccinated layers were Rispens-positive. Viral load in these samples varied between farms during early life, reaching readily detectable levels at 2-3 weeks of age. Vaccinated chickens maintained a high Rispens load in feathers and dust and high MDV antibody levels until 91 weeks of age. MDV infection was detected in 6.7% of feather samples from vaccinated chickens. Rispens virus was detected in 7% of samples from unvaccinated broiler flocks. CONCLUSION: Vaccine take can be measured effectively by Rispens-specific qPCR of feathers or dust from approximately 3 weeks post vaccination. Infection with Rispens is persistent, with lifelong shedding and serological response. The detectable infection rate of vaccinated chickens with MDV is low and there is preliminary evidence of escape of Rispens virus to unvaccinated flocks.

Immune-mediated responses account for the majority of production loss for grazing meat-breed lambs during Trichostrongylus colubriformis infection.

The hypothesis tested in this experiment was that Trichostrongylus colubriformis infection would reduce growth rates of grazing meat-breed lambs; however production loss would be reduced by suppression of the host immune response. The experiment had a 3×2 factorial design using 6-7 month old meat-breed lambs which remained uninfected or infected (IFY) with 2000 or 4000 T. colubriformis L3/week for 12 weeks and were immunosuppressed (SUPY) using methylprednisolone acetate once weekly or remained non-immunosuppressed (SUPN). Immunosuppression increased worm egg counts (WEC) of infected lambs (SUPY 2421 eggs per gram (epg), SUPN 1154 epg on day 84, p<0.05) and T. colubriformis burdens (p<0.05-0.10) and reduced circulating eosinophils (p<0.05 on days 11, 42, 56 and 84) and intestinal total antibody titres (p<0.02). There was a significant (p<0.05) interaction between the main effects of infection and immunosuppression with infection having a larger negative effect on the liveweight of non-immunosuppressed lambs. The immunological response of the host to T. colubriformis infection accounted for 75% of the overall cost of infection (3.1kg) with the majority of this cost occurring during the first 35 days of infection. In contrast, most of the cost associated with the direct effect of infection occurred after day 35. These results confirm in grazing meat-breed lambs that the host's immunological response to T. colubriformis infection is the major component of production loss.

In vivo characterisation of two Australian isolates of Marek's disease virus including pathology, viral load and neuropathotyping based on clinical signs.

OBJECTIVE: To evaluate the pathogenicity of Australian Marek's disease virus (MDV) isolate MPF23 (1985) against the reference strain MPF57 based on pathology, viral load and neuropathotyping on the basis of clinical signs. PROCEDURE: Two MDV challenge isolates (MPF57 or MPF23) were administered to unvaccinated specific-pathogen free (SPF) layer chicks on day 5 after hatch at three challenge doses (500, 2000 or 8000 plaque-forming units (pfu)/chick). Mortality, body weight, immune organ weights, MDV load in peripheral blood lymphocytes (PBL) and clinical signs were measured to 56 days post challenge (dpc). RESULTS: MPF23 was the more pathogenic of the two viruses, inducing higher mortality (81% vs 62%) and incidence of MD lesions (100% vs 76%). MPF23 induced earlier, more sustained and more severe neurological signs in the period 26-56 dpc. However, there were few differences during the 0-23 dpc used in the neuropathotyping classification under test. The observed pattern during this earlier period classified both viruses as neuropathotype B, consistent with a very virulent pathotype. MDV load in PBL at 7 and 44 dpc did not differ between virus isolates, but the load at 7 dpc was significantly and negatively associated with time to euthanasia or death. CONCLUSION: MPF23 appears to be as, or more, virulent than the MDV strains isolated over the subsequent two decades. The neuropathotyping system developed in the USA did not clearly differentiate between the two isolates under test; however, extension of the period of assessment of clinical signs beyond 26 dpc did reveal clear differences.

Use of developmental temperature and gastrointestinal tract location to isolate pure Trichostrongylus vitrinus from mixed, naturally acquired trichostrongylid infections in sheep.

OBJECTIVE: To isolate Trichostrongylus vitrinus from naturally-acquired mixed trichostrongylid infections (predominantly Trichostrongylus spp. and Teladorsagia circumcincta) in sheep using differential larval development temperatures and the anatomical location of adults. METHODS: Faeces were collected from ewes with mixed, naturally acquired trichostrongylid infection, and incubated at low temperatures (4°C or 8°C) for 20-68 days depending on temperature. Harvested infective larvae were passaged through worm-free lambs, from which the first 5 m of small intestine was collected and adult worms recovered. Purity of infection with T. vitrinus was assessed by examination of spicules on 100 adult male worms. Eggs of recovered adult female worms were cultured at 25°C and harvested larvae were re-passaged through worm-free lambs. RESULTS: Low temperature incubation (≤8°C) successfully exploited variation in the developmental temperature required by pre-parasitic stages of small intestinal nematodes, allowing T. vitrinus to develop while others did not. Differential organ harvest from infected sheep allowed isolation of T. vitrinus from other gastrointestinal nematodes, with amplification of pure T. vitrinus achieved by passage through a further generation of lambs. CONCLUSION: A successful method for the isolation of pure T. vitrinus from mixed infections is described. The resulting pure strain of T. vitrinus has been amplified and is held by two institutions for use in research (CSIRO reference SARDI 2011 strain).

Effects of chronic infection with Trichostrongylus vitrinus and immune suppression with corticosteroid on parasitological, immune and performance variables in crossbred meat lambs.

The effects of, and interactions between chronic Trichostrongylus vitrinus infection and immune suppression with methylprednisolone were investigated for a period of 112 days in a grazed flock of 176 crossbred meat lambs. Worm egg count of non-immune-suppressed lambs increased rapidly from days 21 to 42 post-initial infection, and then steadily declined. Infection was associated with significantly decreased fat depth, eye muscle area and cold carcase weight, and increased circulating anti-T. vitrinus IgG and IgA. Immune suppression led to sustained increases in WEC, and significantly greater worm count, liver weight, fat depth and carcase dressing percentage, and significantly reduced IgG and IgA anti-T. vitrinus titres, lymphocyte counts, adrenal weight, eye muscle area and cold carcase weight. Both infection and immune suppression were associated with significant body weight reductions. Only 39% of reduced growth rate due to infection was attributable to the host immune response to T. vitrinus.

Risk factors for Merino ewe mortality on the Northern Tablelands of New South Wales, Australia.

OBJECTIVE: To measure the association of worm egg count (WEC), body weight (BW) and fat score (FS) with mortality in Merino ewes. METHODS: On six farms, Merino ewes (n = 1440) were yarded at approximately 3-month intervals for 2 years for a range of experimental measurements. Mortality was defined by failure to present at two consecutive measurement days and at next shearing. Survival analysis was conducted with phenotypic traits measured both as continuous variables and categorised appropriately for industry guidelines. RESULTS: Overall mortality was 10%, with most deaths occurring in autumn followed by winter. Age class (2006 drop vs mature-age ewes) did not affect mortality. High WEC, low BW and low FS significantly increased the risk of mortality. WEC > 1200 epg significantly increased risk of mortality in summer (risk ratio 3.76) and autumn (risk ratio 3.61) compared with the reference group. Risk of mortality increased 1.1-fold for each kilogram unit decrease in BW and 7-fold for each unit decrease in FS. CONCLUSIONS: Mortality of Merino ewes can be significantly reduced by management methods that reduce WEC, maintain BW and increase the FS. For Merino ewes on the Northern Tablelands of NSW, improved animal welfare and economic benefits may be achieved by maintaining sheep with WEC < 1200 epg, BW > 35 kg and FS > 2.5.

Translation of H. contortus and T. colubriformis from egg to establishment in grazing sheep is unaffected by rainfall timing, rainfall amount and herbage height under conditions of high soil moisture in the Northern Tablelands of NSW.

A field experiment was conducted at Armidale in the Northern Tablelands of NSW, Australia to determine the effects of simulated rainfall amount (0, 12 and 24 mm), rainfall timing (days -1, 0 and 3 relative to plot contamination) and herbage height (4 and 12 cm), on translation of Haemonchus contortus and Trichostrongylus colubriformis from egg to established stages in grazing sheep under conditions of high soil moisture (22-23%). The experiment was conducted in summer when temperature was not anticipated to be a limiting factor for development success. Development success was assessed using tracer sheep and expressed as percentage recovery of parasitic stages relative to egg output on pasture (translation%). For both species, translation (0.11% H. contortus; 0.55% T. colubriformis) was observed in the absence of simulated rainfall and was unaffected by treatment effects of rainfall amount and timing, and herbage height. We suggest that soil moisture (>20%) alone was sufficient to support development and translation (from eggs to parasitic stages in the gut of tracer animals) of these species which contrasts with expectations for development success on dry soils. These findings identify the importance of taking soil moisture into account when predicting the likely effects of rainfall and herbage height on development to L3 and ultimately in predictive epidemiological models of ovine gastrointestinal nematodiasis.

Pathotyping of Australian isolates of Marek's disease virus in commercial broiler chickens vaccinated with herpesvirus of turkeys (HVT) or bivalent (HVT/SB1) vaccine and association with viral load in the spleen and feather dander.

OBJECTIVE: To pathotype Australian isolates of Marek's disease virus (MDV) in commercial broiler chickens using standard methods and to evaluate early markers of pathotype. METHODS: A complete 3 × 4 factorial experiment with two replicates was conducted using 648 Cobb broiler chickens in 24 isolators. The experimental factors were vaccination (unvaccinated, herpesvirus of turkeys (HVT), bivalent (HVT + SB1 strain of serotype 2 MDV)) and MDV challenge (unchallenged or 500 plaque-forming units of isolates MFP57, 02LAR or FT158). Mortality, body weight, immune-organ weights and viral load were measured to 56 days post challenge (dpc). Vaccinal protective index (PI) and virulence rank (VR) were calculated based on gross Marek's disease (MD) pathology. RESULTS: The PIs provided by the HVT and bivalent vaccines against challenge with MPF57, 02LAR, and FT158 were 84.6% 56%, 61.4% and 82.2%, 60.8%, 57.7%, respectively, leading to putative pathotypes of virulent MDV for MPF57 and very virulent MDV for 02LAR and FT158. Significantly more of the unvaccinated chickens (85.7%) had MD lesions than chickens vaccinated with either the HVT (26.8%) or bivalent vaccine (27.6%). Strong linear relationships were observed between the incidence of MD at 56 dpc and MDV load in the spleen at 7 dpc (R(2) = 0.71) and MDV load in the isolator exhaust dust at 14 dpc (R(2) = 0.57) and 21 dpc (R(2) = 0.51). Immune organ weights had a weaker association with subsequent MD incidence. CONCLUSION: Pathotyping results in broiler chickens with maternal antibody broadly agreed with those in specific-pathogen-free chickens in other studies, with some important differences. MDV load in the spleen at 7 dpc and in isolator dust at both 14 and 21 dpc was a powerful early predictor of subsequent MD incidence.