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Background Numerous laboratory and fewer field-based studies have found that ixodid ticks develop more quickly and survive better at temperatures between 18 °C and 26 °C and relative humidity (RH) between 75 and 94%. Ixodes anatis Chilton, 1904, is an endophilic, nidicolous species endemic to North Island brown kiwi (Apteryx mantelli) (NIBK) and the tokoeka (Apteryx australis), and little is known about the environmental conditions required for its development. The aims of this study were to determine and compare the conditions of temperature and RH that ensure the best survival of the kiwi tick and the shortest interstadial periods, in laboratory conditions and outdoors inside artificial kiwi burrows. Methods Free-walking engorged ticks were collected off wild kiwi hosts and placed in the laboratory under various fixed temperature and humidity regimes. In addition, sets of the collected ticks at different developmental stages were placed in artificial kiwi burrows. In both settings, we recorded the times taken for the ticks to moult to the next stage. Results Larvae and nymphs both showed optimum development at between 10 °C and 20 °C, which is lower than the optimum temperature for development in many other species of ixodid ticks. However, larvae moulted quicker and survived better when saturation deficits were 94%); in comparison, the optimum saturation deficits for nymph development were 1–10 mmHg. Conclusions Our results suggest that the kiwi tick has adapted to the stable, but relatively cool and humid conditions in kiwi burrows, reflecting the evolutionary consequences of its association with the kiwi. Graphical Abstract

Coccidia (Eimeria spp.) in brown kiwi (Apteryx mantelli) cause significant morbidity and mortality in captive rearing facilities. Monitoring the abundance of this parasite in individual birds is crucial for successful management of kiwi. This research compares the abilities of centrifugal faecal flotations (CFF) and a modified Mini-FLOTAC protocol to detect oocysts. We hypothesised that the Mini-FLOTAC would detect higher oocyst counts. Kiwi dropping samples (n = 10) were homogenized in MgSO4 (SG 1.28) and oocyst counts made with CFFs and Mini-FLOTAC counting chambers, with three replicates for each method. For CFF, 0.5 g of droppings were examined using standard methods. Mini-FLOTAC counts were made using a modified sample preparation compared with the manufacturer’s protocol but still used a 1:20 dilution of droppings. Oocysts were quantified using light microscopy at ×100–300 magnification. A linear mixed-effects model by REML showed that oocyst per gram estimates via the Mini-FLOTAC method were 3.2 times higher (95% CI 2.4–4.5, p

Although currently exotic to New Zealand, the potential geographic distribution ofAmblyomma americanum(L.), the lone star tick, was modelled using maximum entropy (MaxEnt). The MaxEnt model was calibrated across the native range ofA. americanumin North America using present-day climatic conditions and occurrence data from museum collections. The resulting model was then projected onto New Zealand using both present-day and future climates modelled under two greenhouse gas emission scenarios, representative concentration pathways (RCP) 4.5 (low) and RCP 8.5 (high). Three sets of WorldClim bioclimatic variables were chosen using the jackknife method and tested in MaxEnt using different combinations of model feature class functions and regularization multiplier values. The preferred model was selected based on partial receiver operating characteristic tests, the omission rate and the lowest Akaike information criterion. The final model had four bioclimatic variables, Annual Mean Temperature (BIO1), Annual Precipitation (BIO12), Precipitation Seasonality (BIO15) and Precipitation of Driest Quarter (BIO17), and the projected New Zealand distribution was broadly similar to that ofHaemaphysalis longicornisNeumann, New Zealand’s only livestock tick, but with a more extensive predicted suitability. The climate change predictions for the year 2050 under both low and high RCP scenarios projected only moderate increases in habitat suitability along the mountain valleys in the South Island. In conclusion, this analysis shows that given the opportunity and licenseA. americanumcould and would successfully establish in New Zealand and could provide another vector for theileriosis organisms.

This article sets out to document and summarise the New Zealand epidemic and the epidemiological research conducted on the epizootic of bovine anaemia associated with Theileria orientalis Ikeda type infection, which began in New Zealand in August 2012. As New Zealand has no other pathogenic tick-borne cattle haemoparasites, the effects of the T. orientalis Ikeda type infection observed in affected herds and individual animals were not confounded by other concurrent haemoparasite infections, as was possibly the case in other countries. This has resulted in an unbiased perspective of a new disease. In addition, as both New Zealand’s beef and dairy cattle systems are seasonally based, this has led to a different epidemiological presentation than that reported by almost all other affected countries. Having verified the establishment of a new disease and identified the associated pathogen, the remaining key requirements of an epidemiological investigation, for a disease affecting production animals, are to describe how the disease spreads, describe the likely impacts of that disease at the individual and herd level and explore methods of disease control or mitigation.

Flytraps can be used on farms to monitor the populations of primary strike flies (Lucilia cuprina and Lucilia sericata) and, hence, offer a view regarding the incidence of flystrike on sheep. This study aimed to contrast the specificity and effectiveness of the LuciTrap with its combination of three chemical lures (Lucilures) and the Western Australian Trap with three bait types (LuciLure, Sheep liver with 30% sodium sulphide and squid). A mean model and rate model were fitted to the data. The mean model showed no difference (p &gt, 0.05) in the mean weekly catch for L. cuprina between the Western Australian Trap with LuciLures and the Western Australian Trap baited with sheep liver with 30% sodium sulphide (p &lt, 0.05). Whereas, for L. sericata, no difference (p &gt, 0.05) was found between the Western Australian Trap with LuciLures, the Western Australian Trap baited with sheep liver with 30% sodium sulphide and the LuciTrap. The rate model illustrated that the Western Australian Trap with sheep liver with 30% sodium sulphide and LuciTrap did not differ (p &gt, 0.05) for L. cuprina and L. sericata. Combined, these results indicate that New Zealand farmers can use either the LuciTrap or the Western Australian Trap with sheep liver with 30% sodium sulphide to monitor these target species.

This study aimed to investigate whether the infection intensity of Theileria orientalis Ikeda type organisms within Haemaphysalis longicornis larvae and nymph stages fluctuated over 6 mo after feeding as larvae on infected calves in the field. Naive larvae, hatched from eggs, were fed on infected calves for 5 days while contained within cotton socks glued over the calves' ears. Larvae were first sampled immediately post-feeding and then sampled every 3 wk for 23 wk in total, after molting to nymphs. All larvae and nymphs were tested for T. orientalis Ikeda organisms using quantitative PCR. The qPCR results showed that the infection intensity of Haemaphysalis longicornis larvae and nymphs was not constant over the sampling period, and after initially dropping after molting to nymphs, it then rose with fasting to a maximum at 17 and 23 wk post-feeding. The significant rise in T. orientalis Ikeda organisms observed at 23 wk postfeeding may explain why more severe clinical cases of bovine theileriosis in New Zealand are seen in the spring when nymphs are the predominant instar questing.

Monitoring activity patterns of animals offers the opportunity to assess individual health and welfare in support of precision livestock farming. The purpose of this study was to use a triaxial accelerometer sensor to determine the diel activity of sheep on pasture. Six Perendale ewe lambs, each fitted with a neck collar mounting a triaxial accelerometer, were filmed during targeted periods of sheep activities: grazing, lying, walking, and standing. The corresponding acceleration data were fitted using a Random Forest algorithm to classify activity (=classifier). This classifier was then applied to accelerometer data from an additional 10 ewe lambs to determine their activity budgets. Each of these was fitted with a neck collar mounting an accelerometer as well as two additional accelerometers placed on a head halter and a body harness over the shoulders of the animal. These were monitored continuously for three days. A classification accuracy of 89.6% was achieved for the grazing, walking and resting activities (i.e., a new class combining lying and standing activity). Triaxial accelerometer data showed that sheep spent 64% (95% CI 55% to 74%) of daylight time grazing, with grazing at night reduced to 14% (95% CI 8% to 20%). Similar activity budgets were achieved from the halter mounted sensors, but not those on a body harness. These results are consistent with previous studies directly observing daily activity of pasture-based sheep and can be applied in a variety of contexts to investigate animal health and welfare metrics e.g., to better understand the impact that young sheep can suffer when carrying even modest burdens of parasitic nematodes.

AIMS: The principle aim of this study was to examine the association between infection with Theileria orientalis Ikeda type and growth rates of suckled beef calves on four beef farms. In addition, associations between calf sex, sampling time, and individual farm and T. orientalis Ikeda type infection intensity and haematocrit (HCT) were investigated. METHODS: The study was a prospective longitudinal study in which 240 calves from four purposively selected beef farms in the North Island of New Zealand were blood sampled and weighed in late spring, mid-summer and early autumn. Two farms were from high-risk (A and B) and two from low-risk (C and D) tick areas. Blood samples were analysed to determine HCT, and the number of T. orientalis Ikeda type organisms/µL of blood (infection intensity) using a quantitative PCR assay. A calf was defined as infected if >415 organisms/µL were detected in a blood sample. Linear mixed models were used to examine associations between infection intensity, mean daily liveweight gain (MDG), HCT, calf sex and time of sampling on the four farms. RESULTS: On Farms A and B nearly all calves were infected at each sampling time, on Farm C T. orientalis Ikeda type infection in suckled beef calves.

Since 2012, New Zealand has suffered from an epidemic of infectious bovine anaemia associated with T. orientalis (Ikeda), an obligate intracellular protozoan parasite of cattle. Despite widespread agreement that T. orientalis (Ikeda) infection has impacted beef and dairy farming in New Zealand there is very little quantitative data to support this conclusion. A randomised controlled experimental study of the effect of T. orientalis (Ikeda) infection on the live weight, haematocrit (HCT), temperature and activity of 2-year-old Friesian bulls was conducted at a Massey University Research farm, Palmerston North. Ten out of seventeen 2-year-old Friesian bulls were injected intravenously with 30 mL whole blood from 2 clinical cases of Theileria-associated bovine anaemia and then followed over a period of 20 weeks. The bulls were blood sampled, had rectal temperature recorded and weighed 3 times weekly for 13 weeks and then once weekly thereafter until the end of the trial. Infection intensity was monitored using qPCR. All 10 inoculated bulls were successfully infected with T. orientalis (Ikeda). The results showed that the live weight response to infection was varied and the bulls could be divided into two groups based on this response. Four infected bulls showed a significant weight reduction of 41.5 kg (p 0.0001), a financial loss of around NZ$112 per bull, compared with the other 6 bulls in the infected group, which were not different to the 7 uninfected controls. The live weight of the 4 poor growing bulls was significantly lower than the other 6 infected bulls from Day 71 post infection (p 0.05). All ten infected bulls showed a similar decrease in HCT, with the lowest HCT reached around Day 60 to 80 post-infection, however the four infected bulls that grew poorly did have a significantly elevated HCT for the first 1 to 3 weeks post infection (p 0.05). The 4 infected bulls which grew poorly also had a significantly higher infection intensity than the other infected bulls from Day 27 to Day 60 post-infection (p 0.05). There was no pyrexia recorded in the infected group or control groups, instead there was a tendency for the infected group to have a lower rectal temperature from Day 5 to 70 post infection. The infected bulls walked on average 239 steps per day less than the control bulls, although this difference was not significant (p = 0.35). Overall the study clearly showed, by controlling infection date and infectious dose, that a proportion of cattle infected with T. orientalis (Ikeda) have significantly decreased live weight gains.

The establishment rate of Cooperia oncophora related to host age and previous infection was investigated in young calves. Calves of similar age were kept on a feed pad and allocated into multiple groups, based on their age and weight. Two groups (each n = 16) received trickle infections with an ivermectin-susceptible C. oncophora isolate of 2000 or 10,000 infective stage larvae per week while another group (n = 16) was kept as an uninfected control. At intervals over a period of 11 months, two animals from each group were challenged with 15,000 infective stage larvae of an ivermectin-resistant isolate, 25 days later orally treated with ivermectin and 5 days after that slaughtered for worm counts. On three occasions additional calves (n = 2), subjected to the high trickle infection rate, received an ivermectin treatment to remove the existing worm burden, prior to challenge as above. Further calves (n = 4) of similar age were introduced at the beginning and the end of the experiment to determine the effect of larval age on establishment rate. The establishment in the two trickle infection groups declined to10% within the first three months, which was significantly different from the control group. In the animals receiving the high trickle infection, but an anthelmintic treatment before challenge the establishment rate was not significantly different from the controls. Over the duration of the experiment establishment in the control group declined from 53% to20%, which was similar to the decrease recorded at the beginning and the end of the experiment in the animals to determine the effect of larval age. The findings indicate that an existing C. oncophora burden had a strong effect on the establishment of incoming larvae in the trickle infected groups, but this was not observed if the existing burden was removed before the final challenge. The decline in establishment rate in the control group was attributed to the age of the larvae and not the age of the calves per se.

Theileria orientalis is a tick‒borne intracellular parasite of red blood cells that causes severe and mild infections in various ruminants worldwide. To date there have been 11 types identified within this species, of which 4 types are presently found in New Zealand cattle. Since 2012, New Zealand has suffered a substantial epidemic of infectious bovine anaemia in both dairy and beef cattle associated with the Ikeda type. The speed at which the disease spread through the North Island suggested that other species could have been involved in transmission. The aim of a series of related experiments was to test the null hypothesis that sheep cannot maintain T. orientalis Ikeda type infection or infect ticks that feed on them. Several studies were conducted over 2 years to address this hypothesis which together showed that sheep can have detectable levels of T. orientalis Ikeda type infection in both the acute and chronic phase and that Haemaphysalis longicornis larvae can become infected when feeding on sheep. No anaemia, weight loss or clinical disease was recorded in the sheep in the acute phase of infection. The levels of infection recorded in the sheep were much lower than those found in cattle, consistent with the sheep being asymptomatic carriers of T. orientalis Ikeda type infection.

Recent reports indicate that gastrointestinal nematodes (GIN) are contributing to significant losses in deer productivity and that anthelmintic resistance has become an issue of concern for deer farmers in New Zealand. The aim of this study was to evaluate cross-grazing of deer with sheep or cattle as an aid for control of gastrointestinal and pulmonary nematode parasites of farmed red deer (Cervus elaphus) in New Zealand. This was a field study replicated over two years (2012 and 2013) for 16 weeks each year at two locations (Massey University, Palmerston North and Invermay AgResearch Centre, Mosgiel). Each farm replicate included four groups (19-20 deer) at each location: red deer cross-grazing with cattle (Deer/Cattle); red deer cross-grazing with sheep (Deer/Sheep); red deer grazing on their own (DeerOwn); and red deer grazing on their own and treated with anthelmintics every two weeks to suppress worm burdens, as a positive control (DeerSup). The key outcome was the number of anthelmintic treatments (AT) given to deer. The decision to treat individual resident deer in Deer/Cattle, Deer/Sheep and DeerOwn groups was based on "trigger" criteria including faecal egg count (FEC)≥250 eggs/g or Dictyocaulus faecal larval count (FLC)≥100 larvae/g or when growth rate was less than 80 % of the mean of the DeerSup group in the previous two weeks. In addition, to quantify the species of parasites cycling in each group, sets of three "tracer" deer were introduced to graze with each group at the mid-point and again at the end of each 16 week period in both years at both locations. Least squares means (LSM) of the number of AT given per animal for Deer/Sheep (3.4) and DeerOwn (3.3) groups were significantly higher than for the Deer/Cattle (2.7) group (p0.001). In tracer animals, the LSM of abomasal Trichostrongylus spp. were significantly fewer in the DeerOwn (17), Deer/Cattle (37) and DeerSup (54) groups than in the Deer/Sheep (952; p0.001) group. The LSM of the nematodes in the subfamily Ostertagiinae (=Ostertagia-type) were significantly more in the DeerOwn (1950) than in Deer/Sheep (370; p = 0.003) and DeerSup (238; p0.001) groups, but the number in the Deer/Cattle group (689) was not different to DeerOwn (p0.05). The LSM of lungworm were fewer in Deer/Sheep (3), Deer/Cattle (4) and DeerSup (3; p0.001) groups than in DeerOwn (40) group. The Deer/Cattle and DeerSup groups had significantly higher LSM of liveweight gain over the 16 weeks (p0.001) than the other two groups. This study demonstrated that cross-grazing with either sheep or cattle aided control of lungworm and gastrointestinal nematodes in young deer during autumn. However, the advantages varied between the use of sheep or cattle and in the ability to control different species of parasites.

The aim of this study was to test the hypothesis that toltrazuril administered at 4 weeks post-turnout reduces the infection intensity of Theileria orientalis Ikeda type in dairy calves and so prevents serious clinical disease in these animals at 2–3 months of age. Two groups of 40 dairy calves on two separate dairy farms in the Waikato were followed for 16 weeks post-turnout onto pasture. On each farm, 20 calves were randomly selected and orally treated with toltrazuril (15 mg/kg) at 4 weeks post-turnout, whilst the remaining 20 calves were left untreated. All 40 calves were blood sampled and weighed at 2, 4, 6, 8, 12, and 16 weeks post-turnout i.e. 6 samplings per calf. A random subset of 10 calves from each treatment group on each farm were faecal sampled at each visit. The blood samples were used to estimate the T. orientalis Ikeda type infection intensity and haematocrit for each calf and the faecal samples were used to estimate the number of coccidia oocysts per gram of faeces. Three linear mixed effects models, to evaluate the effect of toltrazuril treatment on infection intensity, haematocrit (HCT) and weight respectively were fitted to the data. No calves on either farm developed clinical theileriosis or coccidiosis and the three mixed effects linear models, controlling for the effect of farm and days from turnout, showed that there was no effect of treatment on infection intensity (p = 0.81), on HCT (p = 0.99) and on weight gain (p = 0.79). In conclusion, this study showed no evidence supporting the use of toltrazuril to control T. orientalis Ikeda type infection levels and prevent disease.

To assess the efficacy of toltrazuril against the Eimeria spp. affecting brown kiwi (Apteryx mantelli).Droppings were collected from three brown kiwi, aged6 months old, at a captive rearing facility in the North Island of New Zealand, between 22 February and 20 April 2017, on 14 sampling dates. Only droppings (n=30) that were excreted between 03:00 and 07:00, as determined using video surveillance, were included for analysis, reflecting the peak time for shedding of coccidial oocysts for brown kiwi. Oocysts were quantified in each sample and Eimeria species identified on the basis of oocyst morphology. All samples were collected between 2 and 10 days after the birds had been treated with 25 mg/kg toltrazuril.Eimeria spp. oocysts were identified in 28/30 individual samples and on 14/14 sampling dates. Oocyst counts varied from 0 to 328,080 oocysts per gram (opg), and at least one oocyst count10,000 opg was measured on 12/14 sampling dates. Three species of Eimeria were observed, with Eimeria apteryxii and E. kiwii most commonly encountered, whereas only one sample contained E. paraurii.In the three birds monitored at this research site, there was a high abundance of E. apteryxii and E. kiwii oocysts in droppings despite recent administration of toltrazuril. These results suggest that the populations of Eimeria spp. affecting brown kiwi at this location appear to possess an ability to survive exposure to toltrazuril. Toltrazuril is widely used at captive rearing facilities to limit the effects of coccidiosis in juvenile kiwi. If a lack of efficacy is confirmed, it will be necessary to investigate alternative treatment regimens alongside broader environmental management strategies.

The longhorned tick, Haemaphysalis longicornis, feeds upon a wide range of bird and mammalian hosts. Mammalian hosts include cattle, deer, sheep, goats, humans, and horses. This tick is known to transmit a number of pathogens causing tick-borne diseases, and was the vector of a recent serious outbreak of oriental theileriosis in New Zealand. A New Zealand-USA consortium was established to sequence, assemble, and annotate the genome of this tick, using ticks obtained from New Zealand's North Island. In New Zealand, the tick is considered exclusively parthenogenetic and this trait was deemed useful for genome assembly. Very high molecular weight genomic DNA was sequenced on the Illumina HiSeq4000 and the long-read Pac Bio Sequel platforms. Twenty-eight SMRT cells produced a total of 21.3 million reads which were assembled with Canu on a reserved supercomputer node with access to 12 TB of RAM, running continuously for over 24 days. The final assembly dataset consisted of 34,211 contigs with an average contig length of 215,205 bp. The quality of the annotated genome was assessed by BUSCO analysis, an approach that provides quantitative measures for the quality of an assembled genome. Over 95% of the BUSCO gene set was found in the assembled genome. Only 48 of the 1066 BUSCO genes were missing and only 9 were present in a fragmented condition. The raw sequencing reads and the assembled contigs/scaffolds are archived at the National Center for Biotechnology Information. Keywords: Tick genome, Pac Bio de novo assembly, Genome annotation, Cattle tick

Since 1982 there have been two epidemics of bovine anaemia associated with Theileria orientalis infection (TABA) in New Zealand. The latest and more serious epidemic started in 2012 and is associated with the T. orientalis (Ikeda) type. In 2013 the Ministry for Primary Industries (MPI) carried out several epidemiological investigations on the TABA epidemic, which included a series of on-farm herd prevalence studies. The aims of this paper are to present and summarise the results of these herd prevalence studies. The data were collected from March 2013 to September 2013 and comprised 26 farms. For 21/26 of these farms, a T. orientalis (Ikeda) positive index case of TABA precipitated the follow-up herd prevalence study. Each herd prevalence study involved a random sample of 10 cows for haematocrit, for piroplasm counts (number of Theileria infected RBCs per 1000 RBCs) and for PCR molecular diagnosis. Animals were diagnosed anaemic if the haematocrit ≤0.24. The results showed that for the index cases, the average haematocrit = 0.10 L/L and the average piroplasm count = 10.9 Theileria infected RBCs per 1000 RBCs, and for the herd prevalence studies the average haematocrit = 0.26 and the average piroplasm count = 6.9. The average herd prevalence of T. orientalis (Ikeda) PCR positive cattle and of anaemic cattle was 87% and 26% respectively. For dairy herds (n = 7 herds) it took, on average, 80 days for the within herd prevalence of anaemia to return to zero after the initial diagnosis. In conclusion the prevalence studies showed that infection rates and anaemia rates were already high in herds when the index case of TABA was observed. At the time, these prevalence studies significantly added to our understanding of the epidemiology of the evolving T. orientalis (Ikeda) epidemic and furthermore met the New Zealand MPI objective of being rapid and applied, fit for purpose and completed at a relatively low cost.

Previous studies and anecdotal reports have generally suggested that goats and deer have higher metabolic rates of anthelmintics as compared to sheep. If this is correct then these animals are effectively under dosed if treated with the sheep dose rate. Underdosing is an accepted risk factor for developing anthelmintic resistance as it allows resistant, and even some partly resistant parasites to survive. Despite this risk, there has been no study to date, which has quantified the pharmacokinetic profile of abamectin in sheep, goats and deer rates under similar conditions. The objective of this research was to compare the pharmacokinetic parameters of interest esp. Cmax, Tmax and AUC in sheep, goats and deer given 0.2 mg/kg abamectin per os and then manipulate the dose rate for any variation to match that of sheep. The blood samples obtained from these animals were analysed by HPLC-Fluorescence detector using a validated method of analysis. Goats at the same dose rate had significantly lower Cmax (11.5 ± 1.6 ng/mL) as compared to sheep (20.1 ± 7.3 ng/mL) and deer (26.4 ± 7.0 ng/mL) with the latter two not being different to each other. Goats dosed with a double oral dose of abamectin (0.4 mg/kg) achieved a Cmax (20.2 ± 5.69 ng/mL) which was similar sheep and deer orally dosed with 0.2 mg/kg abamectin. The results of this study suggest that goats are being potentially underdosed at the sheep dose rates and efficacy studies of abamectin should be conducted in goats at both the dosing regimens. In contrast, these results do not support the hypothesis that deer are metabolising abamectin more rapidly than sheep and hence this dose rate would seem to be appropriate for deer but should be confirmed by further study.

Captive rearing of wild brown kiwi (Apteryx mantelli) is widely carried out to assist in the recovery of this declining species. As a consequence, high densities of immunologically naïve kiwi are commonly housed in semi-captive conditions, with the potential to result in substantial morbidity and mortality from coccidiosis caused by multiple species of Eimeria. Previous research has described circadian variation in oocyst shedding across multiple avian host species. The aim of this research was to describe any circadian variation in oocyst shedding in brown kiwi. Droppings were collected from brown kiwi (n = 4) at a single captive rearing facility using video surveillance to determine the time of excretion, and oocyst counts were undertaken. Results show that two of the Eimeria spp. affecting brown kiwi exhibit a peak in oocyst shedding between 03.00 and 07.00 with few or no oocysts shed between 08.00 and midnight. These results are not able to be explained by the current hypotheses theorising the evolutionary forces behind the development of this adaptive trait. Our findings increase the current understanding of the biology of the Eimeria spp. affecting brown kiwi and have important implications for the management of captive-reared kiwi, in particular for the accurate interpretation of faecal oocyst counts.

Commencing in 2012, an epidemic of infectious bovine anaemia associated with Theileria orientalis Ikeda type has been present in New Zealand. The aims of this study were to analyse the temporal and spatial effects of T. orientalis Ikeda type infection on the sample submission rates and haematocrits of infected cattle over the first two years of the New Zealand epidemic. The data were collected from 30/08/2012 to 28/11/2014 and included all samples that met the case definition for Theileria associated bovine anaemia (TABA) and tested positive for T. orientalis Ikeda type by PCR. The sample submission rates by month and farm type were highly seasonal with dairy farm submissions peaking in September a month before beef farm submissions peaked. A second lesser peak of dairy farm submissions in April was absent for beef farms. A mixed effects model was fitted to the data and showed a significant interaction between farm production type (dairy or beef) and month of sampling (p=0.006) and between latitude and month of sampling (p=0.024). The estimated haematocrit, adjusted for month and latitude, for dairy cattle=0.125 (95%CI 0.121-0.129) and for beef cattle=0.151 (95% CI 0.138-0.165), p

AIM To determine the most commonly used words in the clinical histories of animals naturally infected with Theileria orientalis Ikeda type; whether these words differed between cases categorised by age, farm type or haematocrit (HCT), and if there was any clustering of the common words in relation to these categories. METHODS Clinical histories were transcribed for 605 cases of bovine anaemia associated with T. orientalis (TABA), that were submitted to laboratories with blood samples which tested positive for T. orientalis Ikeda type infection by PCR analysis, between October 2012 and November 2014. χ2 tests were used to determine whether the proportion of submissions for each word was similar across the categories of HCT (normal, moderate anaemia or severe anaemia), farm type (dairy or beef) and age (young or old). Correspondence analysis (CA) was carried out on a contingency table of the frequency of the 28 most commonly used history words, cross-tabulated by age categories (young, old or unknown). Agglomerative hierarchical clustering, using Ward's method, was then performed on the coordinates from the correspondence analysis. RESULTS The six most commonly used history words were jaundice (204/605), lethargic (162/605), pale mucous membranes (161/605), cow (151/605), anaemia (147/605), and off milk (115/605). The proportion of cases with some history words differed between categories of age, farm type and HCT. The cluster analysis indicated that the recorded history words were grouped in two main clusters. The first included the words weight loss, tachycardia, pale mucous membranes, anaemia, lethargic and thin, and was associated with adult (p

Haemaphysalis longicornis is the only species of tick present in New Zealand which infests livestock and is also the only competent vector for Theileria orientalis. Since 2012, New Zealand has suffered from an epidemic of infectious bovine anaemia associated with T. orientalis, an obligate intracellular protozoan parasite of cattle and buffaloes. The aim of this study was to predict the spatial distribution of habitat suitability of New Zealand for the tick H. longicornis using a simple rule-based climate envelope model, to validate the model against published data and use the validated model to project an expansion in habitat suitability for H. longicornis under two alternative climate change scenarios for the periods 2046-2065 and 2081-2100, relative to the climate of 1981-2010. A rule-based climate envelope model was developed based on the environmental requirements for off-host tick survival. The resulting model was validated against a maximum entropy environmental niche model of environmental suitability for T. orientalis transmission and against a H. longicornis occurrence map. Validation was completed using the I-similarity statistic and by linear regression. The H. longicornis climate envelope model predicted that 75% of cattle farms in the North Island, 3% of cattle farms in the South Island and 54% of cattle farms in New Zealand overall have habitats potentially suitable for the establishment of H. longicornis. The validation methods showed an acceptable level of agreement between the envelope model and published data. Both of the climate change scenarios, for each of the time periods, projected only slight to moderate increases in the average farm habitat suitability scores for all the South Island regions. However, only for the West Coast, Marlborough, Tasman, and Nelson regions did these increases in environmental suitability translate into an increased proportion of cattle farms with low or high H. longicornis habitat suitability. These results will have important implications for the geographical progression of Theileria-associated bovine anaemia (TABA) in New Zealand and will also be of interest to Haemaphysalis longicornis researchers in Australia, Japan, Korea and New Zealand.

Red deer can be infected with some gastrointestinal nematodes (GIN) of cattle but it is unknown to what extent. An indoor study was conducted to determine the establishment rate of cattle GIN in young deer. Five young calves and 5 young red deer were used. They were effectively treated with anthelmintics when housed and then infected 2 weeks later. After four weeks they were killed for total worm counts. Establishment rates were assessed comparing worm counts to the infective dose which were identified morphologically, and to the relative establishment rate of different species. The establishment rates (%) in cattle and deer respectively were H. contortus (8.0, 18.7, p=0.18), Ostertagia ostertagi (30.8, 0.7, p0.001), Cooperia spp. (72.3, 2.3, p0.001) and Trichostrongylus spp. (19.0, 25.3, p=0.12). The majority (98%) of Trichostrongylus spp. were Trichostrongylus axei in both hosts and there were no differences between hosts for this species (p=0.11). In cattle98% of Cooperia were Cooperia oncophora and the mean burden was much higher than in deer (p0.01) where there were similar proportions of Cooperia oncophora, Cooperia punctata and C. curticei. Small numbers of Oesophagostomum venulosum were also present with 3X as many found in deer as in cattle (p0.05). This study has shown that some cattle-origin GIN can establish in red deer. In particular, the establishment of H. contortus and T. axei could allow sufficient burdens to build up to be clinically significant. Importantly, almost no cattle Ostertagia species or small intestinal species established in deer.

This study used morphological techniques to describe and name four new species of coccidia from the brown kiwi (Apteryx mantelli). Four distinct eimerian oocyst species were recovered that we describe as new species. The largest of these, Eimeria paraurii n. sp. measured 32.2 × 19.8 μm and is morphologically similar to gametocytes previously described histologically in colorectal polyps (Morgan et al. in Parasitol Res 111(4):1689-1699, 2012). Eimeria apteryxii n. sp. measured 23.9 × 14.9 μm and is similar to renal oocysts described histologically in brown, rowi (A. rowii) and Haast tokoeka kiwi (A. australis "Haast") (Morgan et al. in Avian Pathol 42(2):137-146, 2013). Eimeria kiwii n. sp. measured 14.8 × 13.9 μm and resembled gametocytes described previously in kiwi intestinal epithelial cells in brown kiwi (Morgan et al. in Parasitol Res 111(4):1689-1699, 2012). Eimeria mantellii n. sp. measured 17.8 × 10.7 μm and did not appear similar to any coccidia previously described in histological studies in kiwi. These are the first species of Eimeria to be described and named from brown kiwi. Because the morphological descriptions in the present study were determined from a limited number of kiwi droppings from two geographical locations, it is likely that these represent only a portion of Eimeria species present in other populations of both brown kiwi and other Apteryx species from around New Zealand.

Animals suffering from parasitism typically display altered grazing behaviour and a voluntary reduction in feed intake. These changes are potentially important as indicators of disease. Recent advances in sensor technologies provide the opportunity to objectively measure animal activity while on pasture. Tri-axial accelerometers measure body movement in terms of acceleration, which can then be used to estimate physical activity over time. This study investigated if tri-axial measures of overall activity can be used to assess the impact of gastrointestinal nematode (GIN) infection in young sheep. To address this, the overall activity, faecal nematode egg count (FEC) and body weight of two treatment groups of Romney X Suffolk ram lambs were compared. Animals were monitored for four days using tri-axial accelerometer sensors mounted on a ram mating harness after 42-days grazing on contaminated pasture. On Day 0, all lambs were given anthelmintics. Subsequently, a Suppressive Treatment Group (n = 12) was treated with anthelmintics every two weeks. An Untreated Group (n = 12) did not receive further anthelmintics. Overall activity levels were monitored from Day 42 – 46. Activity level was calculated as vectorial dynamic body acceleration (VeDBA). Anthelmintic treatment had a significant effect on FEC but there was no evidence found for a treatment effect on body weight growth over the 42-day period. An effect of treatment and lamb starting weight on overall activity was found (beta = –0.74, 95 % CI –1.17 to –0.30, p = 0.002), identifying a negative impact of parasitism on activity in heavier animals. These results highlight the usefulness of this approach in assessing the effect of GIN parasitism on sheep monitored remotely. If a threshold value of activity could be determined, it could provide a useful tool for farmers and managers that serves as an early indicator of parasitism in sheep.

The objective of this research was to evaluate comparative pharmacokinetics of doramectin in alpacas, after subcutaneous administration of 0.2 mg/kg dose. Six healthy adult alpacas, mean age of 5 years ± 1, (three female and three gelded males) of mean bodyweight of 62 kg ± 16 kg with an average body condition scored 2.8 ± 1 out of five, were used in this study. Serial blood samples were collected from the jugular vein before the administration until day 21 afterwards to establish the pharmacokinetics of doramectin after its subcutaneous administration at 0.2 mg/kg dose. The blood samples were analysed using high-performance liquid chromatography (HPLC), fluorescence detection method with precolumn derivatisation, validated for alpacas. The pharmacokinetic parameters were calculated using a noncompartmental model, and results showed Cmax (6.05 ± 5.34 ng/ml), Tmax (3.83 ± 2.48 days), AUC (62.12 ± 18.86 ng/ml × d), terminal half-life (6.2 ± 4.9 days) and MRT (11.56 ± 4.43 days). The results of this study showed that the Cmax and AUC were much lower than in cattle and sheep at the same dosage. Tmax remained similar to cattle and sheep. This study presents valuable information about pharmacokinetics of doramectin in alpacas, which can be utilised in its future efficacy studies.

There is a desire to increase cattle production on Sumbawa Island but anecdotal reports from farmers indicate that calf mortality can be high. The aim of this paper is to report the occurrence and common causes of calf mortality, as well as to determine if colostral antibody transfer is sufficient in new-born calves. Personal interviews were conducted with 27 farmers. Immunoglobulins were measured in neonatal calves (n=18) using the FASTest IgG Bovine kit. The presence of various pathogens was investigated in fecal samples from calves (n=12) with signs of diarrhea between 1-2 months of age using the FASTest D4T bovine kit and the presence of gastrointestinal parasites was investigated using fecal floatation from pre-weaned calves (3 weeks-3 months of age; n=62). From the questionnaire calf losses ranged from 10–27%, with most losses occurring in older calves. Only one of the 18 calves demonstrated an insufficient concentration of IgG in serum, indicating the possibility of failure of passive transfer from the dam. Of the samples tested from scouring calves, 7 out of 12 samples tested positive for one of the microorganisms causing general diarrhoea in calves. A range of different gastrointestinal nematode parasites were found although very few coccidial oocysts were seen. The results of these studies indicate that calf mortality is high and will be a limitation on the production of beef cattle for slaughter in the Dompu region of Sumbawa Island. Further investigation is required to determine which pathogens are the cause of this calf loss.

The eggs of some species of the parasitic nematode Nematodirus require a period of chilling before they can hatch; N. filicollis is one such species. This study investigated this requirement for chilling in a New Zealand strain of this species. Eggs of N. filicollis were extracted from lamb's faeces and incubated at 20°C to allow development to the third stage larvae within the egg. These eggs were then placed into tissue culture plates and incubated at: 2.7°C (±0.99), 3.6°C (±0.90), 4.7°C (±0.35), 6.4°C (±0.37), 8.0°C (±1.54) or 9.9°C (±0.14) for up to 224 days. At 14day intervals until day 84, then every 28 days, one plate was removed from each temperature and placed at 13.1°C (±0.44) for 14 days. Eggs were then assessed for hatching. From this data, chill units were calculated by subtracting the culture temperature from a constant threshold of 11°C and multiplying by the number of days for which the sample was cultured; then the Gompertz model fitted. Even though hatching overall was low, a greater proportion of eggs hatched with chill accumulation. Maximum hatching of eggs required 800-1000 chill units. Consequently in the field, more than one season of chilling would be required before hatching. As such a generation time could take more than one year to complete. This is different to the hatching dynamics of N. spathiger, the other main species found in New Zealand sheep, which does not display this requirement for chilling and hatches immediately once the third stage larvae are developed.

Gastrointestinal nematodes are recognised as an animal health issue for farmed red deer. The aim of this study was to explore the range of species infecting farmed deer herds and their farm-level prevalence in New Zealand. Faecal samples were collected from 12–24-month-old deer (n = 6–26; mean 19) on 59 farms located in the North (n = 25) and South (n = 34) Islands. Sub-samples of faeces were pooled by farm and cultured to recover third stage larvae. Twenty four larvae were randomly selected and identified to species using a multiplex PCR (total = 1217 larvae). At farm-level the most prevalent nematodes were Oesophagostomum venulosum 83% (n = 49) and the deer-specific nematodes in the subfamily Ostertagiinae (=Ostertagia-type) including, Spiculoptera asymmetrica 73% (n = 43), Ostertagia leptospicularis 47% (n = 28), Spiculoptera spiculoptera 47% (n = 28). The recently identified Trichostrongylus askivali was present on 32% (n = 19) of the farms and Oesophagostomum sikae on 17% (n = 10). In the analysis of the total number of larvae identified, the proportion was in similar order, 45% (n = 548) were O. venulosum, 14% (n = 173) S. asymmetrica, 10% (n = 124) S. spiculoptera, 9% (n = 114) O. leptospicularis, T. askivali, 3% (n = 40) and only 2% were O. sikae (n = 20). This study is the first to show the farm-level prevalence of nematode species in deer in New Zealand and the first to use PCR as a diagnostic tool. It provides data consistent with cross-infection from sheep/cattle to deer, and provided tentative insights into the proportions of the main GIN species across the deer population including O. sikae and T. askivali which have only recently been identified in New Zealand.

There is an epidemic in New Zealand of infectious bovine anaemia associated with Theileria orientalis Ikeda type, an obligate intracellular protozoan parasite. To establish whether T. orientalis Ikeda type infection adversely affects fertility of bulls used for natural mating, a randomised controlled experimental study was conducted. Ten of 17 2-year-old Friesian bulls that had not been previously infected with T. orientalis were infected with T. orientalis Ikeda type and then evaluations occurred during a 20-week period. There were semen and libido evaluations every 2 weeks, starting 4 weeks before the date of infection. In addition, there were blood collections, for haematocrit and infection intensity evaluations, rectal temperatures recorded, and bulls weighed three times weekly for 13 weeks after infection and then once weekly until completion of the study. Physical activity meters were also attached from Days 9-60 and 65-124 post-infection. The ten bulls were successfully infected with T. orientalis Ikeda type and this resulted in a decrease in HCT to about 0.25 by 70 days post-infection. There were no effects of infection on semen quality; however, during the acute phase of infection, when the infection intensity was rapidly increasing, the infected bulls took a longer time period for repeated mounting of females, and were less dominant in the herd social heiracrchy. In conclusion, although the transitory effects on libido could reduce conception rates, the overall effects of T. orientalis Ikeda type infection on bull fertility will probably be little.

The aims of this study were to monitor the change in Theileria orientalis Ikeda type infection intensity, haematocrit, milk production and reproduction on three New Zealand spring calving dairy herds, over the 2014-2015 milking season. Three spring calving dairy farms, A, B and C, from high risk (endemically stable), low risk (endemically unstable), and zero risk (disease-free) tick areas respectively were followed through the 2014-2015 milking season. On Farms, A and B, 100 cows were randomly selected at the first visit, and the same cows blood sampled every month thereafter, whilst on Farm C, the whole herd was blood sampled bimonthly (140 cows). Blood samples were tested for haematocrit, by centrifugation, and Ikeda infection intensity, using qPCR. Animals that were Ikeda type PCR positive at the first sampling were described as prevalence cases and cows that were negative at the first sampling and became PCR positive during the sampling period were described as incidence cases. Production and reproduction data were accessed through LIC MINDA® and milk production data was standardised to energy corrected milk (ECM). In addition, the effect of buparvaquone (BPQ) treatment on milk production was estimated on Farm B. The prevalence of infection at the first sampling was 100 % on Farm A, 57 % on Farm B and 26 % on Farm C. The incidence risk of infection over the sampling period on Farms B and C was 25 % and 2 % and the incident rate was 0.026 and 0.002 cases per cow-month respectively. The average infection intensity for prevalence cases on all farms was low throughout the milking season,7000 Ikeda organisms/μL however, cases of anaemia still occurred. There was no direct effect of infection intensity on milk production or from being a prevalence case compared to an uninfected cow on milk production, across all farms. However, on Farm B there was a loss of 266 kg (95 % CI 82 ̶ 450) ECM (∼20 kg milk solids) for incidence cases and a loss of 458 kg (95 % CI 211 ̶ 710) of ECM for buparvaquone treated cows, compared to uninfected cows. No significant effect of Ikeda infection on reproduction could be shown for Farms B and C, reproductive data for Farm A was not available. The effect of T. orientalis Ikeda type infection on production and reproduction appears to be minimal once animals have passed through the acute phase of infection and reached the chronic, asymptomatic carrier phase of infection.

Ixodes anatis is a species of endophilic (nidicolous) tick species parasitizing brown kiwi (Apteryx mantelli). Even though they are endemic to New Zealand like their host, very little is known about these ticks or their population dynamics and relationships with their hosts. We conducted a study from May 2013 to June 2014 to evaluate the effect of shelter location (one of three gullies), habitat (forest, scrub and pasture) and type (tree, soil and surface) on the abundance of the different life stages of I. anatis. In total, 12,172 ticks were collected from 63 shelters, which were sampled monthly for 11 months over the 14 month period. Un-engorged larvae predominated over other stages accounting for 87.2% of the samples collected. We found that location, habitat in which the shelters were located, and the type of shelter were significant predictors of I. anatis abundance. Tree shelters in forests had significantly higher tick abundance than those in scrub and pasture. Tree and soil shelters in general had significantly more ticks than surface shelters. Shelters located in Kauri Bush a drier site, had higher abundances than those in wetter sites. While some of these changes can be explained with the movement of the host, we believe more research needs to be done on the effect of shelters' microclimate on I. anatis' life cycle.

There is growing concern that given the high frequency with which anthelmintics are being administered to many horses, anthelmintic resistance amongst equine helminth populations will be an increasing problem, rendering many of the currently available products unusable with little prospect of new products becoming available, at least in the near future. Worldwide, much reliance has been placed on the macrocyclic lactone (ML) group of anthelmintics, but resistance has been reported to these products as well as to the two other major anthelmintic classes used in this species, the benzimidazoles (BZ) and the tetrahydropyrimidines (e.g. pyrantel). In New Zealand, resistance has been reported to the ML and BZ groups, but not yet to pyrantel. As an alternative to interval-based anthelmintic regimens, the highly overdispersed nature of parasite populations in horses can be utilised to decide whether treatment is required, based on whether or not animals exceed a predetermined level of shedding of parasite eggs. If well managed, such a targeted and selective approach can be utilised to eliminate the majority of egg output whilst still providing a refuge for susceptible parasites to persist. Such a system would require that an adequate standard of monitoring be in place and cognisance needs to be taken of parasites or their lifecycle stages that cannot be diagnosed by routine methods. At the same time, using anthelmintics with high levels of efficacy, avoiding practices such as under-dosing, as well as utilising non-chemical means of parasite control when possible, e.g. regular removal of faeces from pasture, should all be considered. Combinations of anthelmintics, specifically of anthelmintics that target the same or a similar spectrum of parasite species, should play an important role in parasite control in horses. As well as providing arguably the highest levels of efficacy, combinations may also slow the rate at which anthelmintic resistance develops.

Recent reports indicate N. caninum has a possible role in causing abortions in sheep in New Zealand. Knowledge about the mode of transmission of neosporosis in sheep in New Zealand is limited. This study aimed to determine the rate of vertical transmission that would occur in lambs born from experimentally inoculated ewes and to determine if previous inoculation would protect the lambs from N. caninum infection. A group of 50 ewes was divided into 2 groups with one group being inoculated with 5 × 106 N. caninum tachyzoites prior to pregnancy in Year 1. In Year 2, each of these groups was subdivided into 2 groups with one from each original group being inoculated with 1 × 107 N. caninum tachyzoites on Day 120 of gestation. Inoculation of N. caninum tachyzoites into ewes prior to mating resulted in no congenital transmission in lambs born in Year 1 but without further inoculation, 7 out of 11 lambs in Year 2 were positive for N. caninum infection. Ewes that were inoculated in both years resulted in all 12 lambs born in Year 2 being positive for N. caninum infection. This indicates that previous inoculation in Year 1 did not result in any vertical transmission in that year but did not provide any protection against vertical transmission in Year 2. These results suggest that vertical transmission occurs readily once the ewe is infected.

AIMS: To present the haematology and biochemistry profiles for cattle in New Zealand naturally infected with Theileria orientalis Ikeda type and investigate if the results differed between adult dairy cattle and calves aged T. orientalis Ikeda type by PCR, that were submitted to veterinary laboratories in New Zealand between October 2012 and November 2014. Data sets for haematology and biochemistry results were prepared for adult dairy cattle (n=62 and 28, respectively) and calves aged T. orientalis Ikeda type were consistent with extravascular haemolytic anaemia. Adult dairy cattle were more likely to be severely anaemic than calves. There were differences in haematology and biochemistry profiles between adult dairy cattle and calves, but most of these differences likely had a physiological rather than pathological basis. Overall, the haematological changes in calves aged

European hedgehogs (Erinaceus europaeus) were introduced into New Zealand from Britain during the period from 1869 to the early 1900s. The only mite found on New Zealand hedgehogs in early studies was Caparinia tripilis, with Sarcoptes scabiei first being reported in 1996. The aim of this study was to investigate the prevalence of Sarcoptes infestation on hedgehogs in New Zealand, the number of mites found and the degree of mange observed. Dead hedgehogs were collected from veterinary clinics, rescue centres, members of the public and from road-kill. Twenty-one (55.3%) of the animals examined had visible skin lesions. Both Caparinia and Sarcoptes mites were identified on microscopic examination with Sarcoptes the most common, being found on over 70% of animals examined (n = 38). The numbers of mites recovered after brushing the head and body ranged from 1 to 5659 (median = 341 mites) with only six animals (22.2%) having fewer than 10 Sarcoptes mites found. Caparinia mites were seen on fewer animals and generally in very low numbers. These findings indicate a change in the mite populations on hedgehogs in New Zealand and that infected animals develop the debilitating hyperkeratotic form of sarcoptic mange without an accompanying hypersensitivity response limiting numbers of mites. Analysis of the cox 1 gene of Sarcoptes from two hedgehogs showed close alignment to sequences derived from a pig with one and from a dog with the second. More work needs to be undertaken to identify the source(s) of the Sarcoptes found on hedgehogs in New Zealand and whether other mammalian hosts may be infected from contact with hedgehogs.

To compare the prevalence of Nematodirus spathiger and N. filicollis on a sample of farms in the lower South Island with the lower and central North Island of New Zealand by way of a presence/absence survey.A presence/absence non-random survey was conducted in which pooled faeces from lambs from 50 farms in the lower and central North Island (n=22) and lower South Island (n=28) were cultured, with and without a post-culture period of chilling, to produce third-stage parasitic nematode larvae (L3). After recovery using the Baermann technique, individual L3 were identified to Nematodirus spp. using a PCR assay. This identified the species that were present on each farm that were then used to compare the regions.Nematodirus filicollis was present in 38/50 (76%) samples from which Nematodirus spp. larvae were cultured, compared with 50/50 (100%) in the case of N. spathiger. No regional differences were observed in the prevalence of either species (p=0.74). Of the Nematodirus spp. L3 recovered from the unchilled samples, 415/428 (97%) were N. spathiger and 13/428 (3%) N. filicollis. After chilling 594/695 (85%) of the Nematodirus spp. L3 were N. spathiger and 101/695 (15%) were N. filicollis.Despite the non-random nature of the study, the evidence suggests that both these species are likely to occur sympatrically on most sheep farms throughout New Zealand. In general N. filicollis eggs did not hatch without a period of chilling and this has implications for the identification of these larvae using conventional culture methods.

SUMMARYRecent work has highlighted and enumerated the economic annual losses due to Neospora caninum abortions worldwide, which should provide strong motivation for the control of bovine neosporosis. However, with the recent withdrawal from sale of the only commercially available vaccine, control options for N. caninum have become more restricted. While researchers continue to work on developing alternative efficacious vaccines, what are the control options presently available for the cattle industries? At the practical level, recommendations for ‘Test-and-cull’, or ‘not breeding from seropositive dams’ stand diametrically opposed to alternative options put forward that suggest a primary producer is better advised to keep those cows in the herd that are already seropositive, i.e. assumed to be chronically infected, and indeed those that have already aborted once. Treatment with a coccidiostat has been recommended as the only economically viable option, yet no such treatment has gained official, regulatory approval. Dogs are central to the life cycle of N. caninum and have repeatedly been associated with infection and abortions in cattle by epidemiological studies. Knowledge and understanding of that pivotal role should be able to be put to use in control programmes. The present review canvasses the relevant literature for evidence for control options for N. caninum (some of them proven, many not) and assesses them in the light of the authors’ knowledge and experience with control of N. caninum.

To ascertain whether resistance was present in Parascaris equorum to the macrocyclic lactone anthelmintic ivermectin, using faecal egg count reduction tests.Thirty-nine foals aged between 11 and 28 weeks on three Thoroughbred stud farms (Farms A; n=20, B; n=5 and C; n=14) were treated with ivermectin (Day 0) and faecal egg counts (FEC) were monitored before and for 21 (Farms A and B) or 14 (Farm C) days after treatment. On Farms A and B, the foals were treated with a macrocyclic lactone/benzimidazole/praziquantel combination on Day 21 and FEC assessed on Day 35. The three farms were all in the Manawatu region in the southern half of New Zealand's North Island.Of the 39 foals, 15 were not shedding P. equorum eggs on the day they were treated with ivermectin, but all 15 did so post-treatment. The FEC on Farms A and B showed no evidence of a reduction at any time between Day 7 and 21; egg output increased steadily over this period. Following combination treatment FEC were reduced by 94% on farm A and 100% on Farm B. On Farm C, there was a 69% reduction in P. equorum FEC on Day 14. Twelve of the 14 foals on Farm C were shedding strongylid eggs on Day 0 and on Day 14 FEC were reduced by 84%.Without comparison to untreated control animals, these results do not allow a reliable estimation of the exact level of efficacy of ivermectin against P. equorum, but they nevertheless show that, within the farms studied, ivermectin no longer achieved a complete kill of egg-laying adults and that foals treated with this drug may continue to shed considerable numbers of eggs after treatment. The results also indicate that ivermectin had sub-optimal efficacy against strongylid nematodes on one of the farms.

Despite significant conservation intervention, the kiwi (Apteryx spp.) is in serious population decline. To increase survival in the wild, conservation management includes rearing of young birds in captivity, safe from introduced mammalian predators. However, an increase in density of immunologically naïve kiwi increases the risk of exposure to disease, including coccidia. Intestinal coccidiosis has recently been described in the kiwi, and although extra-intestinal coccidiosis was first recognized in kiwi in 1978, very little is known about this disease entity. This study used archived histological tissues and reports from routine necropsies to describe the pathology of naturally occurring extra-intestinal coccidiosis. At least 4.5% of all kiwi necropsied during 1991 to 2011 (n=558) were affected by extra-intestinal coccidiosis, and it is estimated that it caused death in 0.9 to 1.2% of kiwi in the study group. Four forms were recognized: renal, hepatic, and, less commonly, splenic and pulmonary. At necropsy, renal coccidiosis was associated with miliary white streaks and foci through the kidneys, renomegaly, and renal pallor or congestion. Renal meronts and gametocytes were confined to the distal convoluted tubules and collecting ducts, and were associated with renal tubular necrosis and tubular obstruction. Hepatic miliary pinpoint foci were present throughout the hepatic parenchyma associated microscopically with macromeronts measuring 304×227 µm. In two cases, clusters of splenic meronts were identified, and a similar lesion was identified in the pulmonary interstitium of another case. Juvenile, captive kiwi were most often affected with extra-intestinal coccidiosis, illustrating an increased expression of disease with population manipulation for conservation purposes.

International audience; Resistance against macrocyclic lactones such as ivermectin is widespread among parasitic gastrointestinal nematodes of small ruminants and is rapidly increasing in cattle parasites. ABC transporters of the subfamily B, the so-called P-glycoproteins (Pgps) have been frequently implicated in ivermectin resistance and are a major cause of multi-drug resistance in protozoa and helminths. The Pgp inhibitor verapamil (VPL) dramatically enhanced susceptibility of the cattle parasitic nematode Cooperia oncophora to ivermectin in vitro as measured in a larval developmental assay and a larval migration inhibition assay using third stage larvae. Moreover, VPL completely restored susceptibility to ivermectin in a resistant isolate resulting in virtually identical dose-response curves of susceptible and resistant isolates in the presence of VPL. Further characterisation of the molecular mechanisms resulting in Pgp-mediated ivermectin resistance is still hampered by the lack of molecular and biochemical information for Pgps of parasitic nematodes. Using PCR with degenerate primers, fragments of four different C. oncophora Pgps could be amplified and the Conpgp-2, previously implicated in macrocyclic lactone resistance in Haemonchus contortus, and Conpgp-3 full-length cDNAs were obtained by RACE PCR. The pgp sequences presented here contribute important data required to systematically screen resistant C. oncophora isolates for up- or down-regulation of Pgps and for the detection of single nucleotide polymorphisms in Pgps to detect selection of specific Pgp alleles by anthelmintics as early as possible.

To determine the prevalence of benzimidazole resistance in Nematodirus spathiger and N. filicollis from a sample of New Zealand farms.The efficacy of albendazole (ABZ) against Nematodirus spp. was assessed by faecal nematode egg count reduction (FECR) tests undertaken in lambs aged 3-8 months old on 27 sheep farms throughout New Zealand. On each farm, groups of 10-16 lambs were either treated with ABZ (4.75 mg/kg) or remained as untreated controls. Faecal samples were collected from all animals at the time of treatment and 7-10 days later. Faecal nematode egg counts (FEC) were performed using a modified McMaster technique. Larvae were cultured from pooled faecal samples, collected 7-10 days after treatment from each group, by incubation at 20°C for 6 weeks, 4°C for 26 weeks then 13°C for 2 weeks. The resulting third stage larvae were identified to species using a multiplex PCR assay, that identified species-specific sequences in the second internal transcribed spacer region of ribosomal DNA. The efficacy of ABZ for N. spathiger and N. filicollis was calculated from the proportion of the two species in culture and the group mean FEC before and after treatment. Only farms with a mean of 10 epg for each species in untreated samples were included for analysis. Resistance was defined as an efficacy95%.On farms that met the threshold of 10 epg in faecal samples, benzimidazole resistance was found on 20/21 (95%) farms for N. spathiger compared with 4/10 (40%) farms for N. filicollis (p0.05). In samples collected following treatment, a mean of 83 (min 46, max 100)% of Nematodirus spp. larvae recovered from the untreated groups were N. spathiger, compared with 94 (min 45, max 100)% in the ABZ treated groups (p=0.03). This change in percentage was not influenced by the overall efficacy of treatment based on the FECR test (p=0.324).The results confirm the high level of resistance in N. spathiger in New Zealand and that benzimidazole resistance was more common in N. spathiger than N. filicollis. While resistance to benzimidazole anthelmintics has been reported previously in New Zealand, this is the first report of N. filicollis being resistant to benzimidazole anthelmintics.

Infections of ruminants with Fasciola hepatica are considered to be of regional importance within New Zealand but there is very little recent information on its prevalence or severity other than anecdotal reports. Generally they are considered to be of secondary importance compared to gastrointestinal nematode infections. Utilizing data from Virtual Climate Stations (n = 11491) distributed on a 5 km grid around New Zealand a growing degree-day model was used to describe the risk of infection with liver fluke from 1972 to 2012 and then to apply the predictions to estimate the risk of fluke infections within New Zealand for the years 2040 and 2090. The growing degree-day model was validated against the most recent survey of infection within New Zealand in 1984. A strong positive linear relationship for 1984 between F. hepatica prevalence in lambs and infection risk (p 2 = 0.71) was found indicating the model was effective for New Zealand. A linear regression for risk values from 14 regions in New Zealand for 1972–2012 did not show any discernible change in risk of infection over this time period (p > 0.05). Post-hoc comparisons indicate the risk in Westland was found to be substantially higher (p F. hepatica infection risk in 2040 and 2090 were detected although they did vary between different climate change scenarios. The highest average percentage changes in infection risk were found in regions with low initial risk values such as Canterbury and Otago; in these regions 2090 infection risk is expected to rise by an average of 186% and 184%, respectively. Despite the already high levels of infection risk in Westland, values are expected to rise by a further 76% by 2090. The model does show some areas with little change with Taranaki predicted to experience only very minor increases in infection risk with average 2040 and 2090 predicted changes of 0% and 29%, respectively. Overall, these results suggest the significance of F. hepatica in New Zealand farming systems is probably underestimated and that this risk will generally increase with global warming following climate change.

Increasing threats from anthelmintic resistant nematode populations warrant and motivate a reappraisal of chemotherapeutic strategies for nematode control in ruminant livestock and horses. The objective of this paper is to present a guideline for the evaluation of products containing two or more constituent anthelmintic actives in a single dosage form for the treatment of nematode infections in these animals. At present, regulatory policies on the approval of such products vary across jurisdictions, and this World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) guideline should enable the harmonization of the requirements. This guideline makes clear recommendations on the minimal standards needed, but stresses that registration dossiers for combination anthelmintic products submitted for approval must conform to the standards and practices already established in existing guidelines for anthelmintics.

Enteric coccidiosis may cause significant morbidity and mortality in juvenile brown kiwi (Apteryx mantelli). Morphology of sporulated oocysts indicates that at least two Eimeria species are able to infect the brown kiwi. A histological study of the endogenous stages of coccidia was undertaken in the intestinal tracts of ten naturally infected young kiwi. Sequential sectioning of the entire intestinal tract allowed identification and recording of the distribution of the various coccidial life stages. Macromeronts measuring 268 × 162 μm when mature were found mainly within the lamina propria of the proximal one third of the small intestine. A smaller form of lamina propria meront was also identified (8.7 × 6.4 μm) with a similar distribution to the macromeronts. Small meronts (4.4 × 3.8 μm) were also identified in mucosal epithelial cells, with the overall peak in distribution within the intestinal tract being distal to the lamina propria meronts. Three morphologically distinctive gametocytes were identified. Type A gametocytes contained within epithelial cells shared the same distribution as the epithelial meronts. Polyps containing large numbers of type B gametocytes within the distal intestinal tract were found in two cases, and type C gametocytes were identified throughout the entire intestinal tract in one case only. The observational nature of this study precludes complete knowledge of the parasite life cycles using histology alone. However, it is likely that each of the three morphologically distinct gametocytes represents a separate species of enteric coccidia.

Neospora caninum is an obligate intracellular parasite and is recognised as the leading cause of bovine abortion worldwide. Natural infection with N. caninum has been described in sheep but it has generally not been regarded as a significant cause of abortion. Recently, there have been several New Zealand cases of foetal abortions where N. caninum was detected which strongly suggested the involvement of Neospora in these abortions. However, there is minimal information about the prevalence of N. caninum infection naturally occurring in New Zealand sheep flocks and particularly its impact on reproduction success. Thus, this present study provides preliminary data on the role that Neospora is playing in ovine reproductive failure by establishing the prevalence of N. caninum antibodies and DNA in ewe blood and foetal material present in 21 New Zealand sheep farms with ongoing unexplained abortion problems and 10 farms with consistently high fertility levels. The results of this study demonstrated an overall seroprevalence of 1.4% which varied between Aborting/non-pregnant (1.8%), age-matched pregnant controls (0.6%) and high fertility (2.1%) ewes. However, despite the variation observed, there was no statistical difference between the three groups. In addition, Neospora DNA was detected by PCR in 13% of submitted foetal brains and in ewe blood from aborting/non-pregnant (6.9%), age-matched pregnant controls (3.6%) and high fertility pregnant (2.1%) ewes. When the PCR results were considered with the IFAT and IDEXX ELISA results, there was no correlation between serology positive and PCR positive blood samples. Taken together, these results reveal that reliance on ELISA-based serology or PCR alone may underestimate the involvement of Neospora. Furthermore, determining the involvement of Neospora appears to require a multi-facetted approach where diagnostic methods and serological cut-off values may need to be adjusted as further information about the effect of natural infections with N. caninum in the ovine host is elucidated.

Two indoor feeding experiments with young sheep were conducted to compare effects of feeding condensed tannin (CT) containing willow ( Salix spp.) or chaffed lucerne hay ( Medicago sativa ) upon death of established parasites and parasite fecundity. In Experiment 1, 24 parasite-free weaned male lambs were individually penned, fed chaffed lucerne hay ad libitum for 5 weeks and then fed either lucerne ( n = 12) or willow ( n = 12) diets for a further 5 weeks. Twelve days prior to willow feeding, all lambs were infected with a mixed population of 22,300 gastrointestinal nematodes comprising predominantly Teladorsagia spp. (0.93), with the remainder being Trichostrongylus , Cooperia and Haemonchus species. Voluntary feed intake, apparent digestibility, blood composition, faecal nematode egg counts, production of eggs and hatched larvae, female worm fecundity and worm burden at slaughter were measured. In Experiment 2, Teladorsagia eggs were added to the faeces of parasite-free sheep fed either lucerne or willow ( n = 9) to measure proportional recovery. Recovery of added Teladorsagia eggs was 0.85 in lucerne-fed lambs and 0.53 willow-fed lambs (P versus 0.83 of eggs added). In Experiment 1, total CT concentration in willow was 27 g/kg DM, with only traces in lucerne, while organic matter digestibility was 0.648 versus 0.599 (P Haemonchus contortus (P Teladorsagia circumcinta (P Haemonchus and Teladorsagia spp. (P i.e. , eggs produced/worm/day) for H. contortus (P in utero as a measure of fecundity in both T. circumcincta and Trichostrongylus spp. (P 3 larvae for both H. contortus and T. circumcincta (P Feeding willow to parasitised young sheep reduced nematode worm burdens and female worm fecundity, with abomasal dwelling species being most affected and conventional methods of measuring FEC underestimated nematode eggs in the faeces of willow-fed sheep.

SUMMARYThe occurrence of a gastrointestinal illness among a class of 96 undergraduate veterinary students in New Zealand prompted laboratory and questionnaire-based investigations.Cryptosporidium parvumwas the only enteropathogen identified in 4/7 faecal specimens analysed. TheC. parvumisolates carried a rareIIaGP60 allele, indicating a point-source outbreak. The infection source could not be microbiologically traced, but the investigation suggested contact with calves during a practical class as the most likely exposure. A total of 25/80 respondents to a questionnaire were defined as cases using a clinical case definition (31% attack rate). The inferred median incubation period was 5 days (range 0–11 days), and the median illness duration was 5–6 days (range 2–23 days), corroborating previous observations in experimental cryptosporidiosis. Disease was self-limiting, characterized by abdominal discomfort, diarrhoea, and in some cases, vomiting. Originating from a rural area and having had previously handled ruminants were associated with a significant risk reduction in males. All the three students who reported chronic use of steroid inhalers for treatment of asthma were cases. This case highlighted, once again, the potential hazard for explosive outbreaks of cryptosporidiosis.

Summary Reasons for performing study: Cyathostomins are the principal pathogenic nematode of equidae worldwide. In other species mast cell (MC) proteinases, in particular chymases, appear to have protective roles. Knowledge of the equine intestinal immune response to cyathostomins is limited. Objective: To investigate MC numbers and proteinase expression in equine cyathostomin-infected large intestine. Hypothesis: MC populations in the large intestine are positively associated with cyathostomin burden and predominantly express chymase. Methods: The caecal cyathostomin burden of naturally infected horses (n = 25) was determined by luminal counts and pepsin digest (mural count). MC were identified and enumerated in caecal tissue using toluidine blue (TB). Immunofluorescent labelling with polyclonal rabbit antibodies was used to demonstrate expression of equine tryptase and the chymase equine mast cell proteinase-1 (eqMCP-1) in Carnoy's fixed caecal sections. Results: Significant positive linear relationships were found between TB-stained mucosal and submucosal MC counts and total cyathostomin burden (P 36%), and both luminal (P 25%) and mural (P 36%) larval counts. Similar relationships were found with mucosal and submucosal chymase and tryptase-labelled MC counts (total: P 29%; luminal: P 30%; and mural: P 19%). With all three MC labels, mean MC counts were higher in the submucosa compared to the mucosa (P

The FLOTAC flotation technique has been introduced as a new diagnostic tool to detect parasitic elements from faeces. Samples from naturally infected young deer were used for counting Dictyocaulus larvae and strongylid eggs. The FLOTAC technique, using 11 different flotation solutions with specific gravities (sg) between 1.20 and 1.45, was compared with the Baermann technique and the saturated sodium chloride (sg 1.20)-based McMaster method. In addition, a comparison was made between the FLOTAC technique with magnesium sulphate (sg 1.28) and the Baermann technique for larval recovery from faeces that were examined on the day of collection or after 7 days storage at 4 degrees C. On the whole egg counts between the FLOTAC using different flotation solutions and the McMaster were unremarkable. In contrast, variations of larval counts were detected between different flotation solutions as well as with the Baermann technique. Most flotation solutions with a specific gravity of 1.20 floated significantly fewer lungworm larvae (p0.05) compared to flotation solutions with a higher specific gravity. Magnesium sulphate (sg 1.28) consistently produced the highest mean larval counts in all conducted experiments. Larval counts using magnesium sulphate (sg 1.28) were higher than with the Baermann technique both on the day of collection and after 7 days. Overall, the use of magnesium sulphate (sg 1.28) with FLOTAC for larval counts resulted in higher counts than the Baermann recovery technique and was the better choice of those flotation solutions examined. Furthermore, magnesium sulphate (sg 1.28) was also reliable for strongylid egg detection with the FLOTAC apparatus.

A 131-day rotational grazing experiment was conducted in the summer autumn of 2007/2008 to compare effects of feeding condensed tannin (CT)-containing willow (Salix spp.) fodder blocks (i.e., silvopastoral system) or perennial ryegrass (Lolium perenne)/white clover (Trifolium repens) control pasture upon the immune response to gastrointestinal nematode parasite infection in Romney weaned lambs. Groups of lambs (n = 40) were allocated to either willow fodder blocks or control pasture; half of each group (n = 20) were regularly-drenched with anthelmintic at approximately 21 day intervals, whilst the remaining 20 weaned lambs were not drenched unless pre-determined faecal nematode egg counts (FEC) were reached, when all weaned lambs in that group were drenched with anthelmintic (i.e., trigger-drenching). Metabolizable energy and CT concentrations were higher in willow fodder versus pasture herbages. Weaned lambs grazing willow fodder blocks had lower live weight gain (92 g/day) and carcass weight (14.4 kg) than those grazing control pasture (134 g; 15.3 kg), with no effect of anthelmintic drenching. Regular anthelmintic treatment maintained similar and low FEC up to day 82, which then increased, whilst trigger-drenched lambs grazing willow fodder blocks had higher FEC than lambs grazing control pasture on three out of eight occasions. As judged by faecal larval cultures, grazing willow fodder blocks reduced the relative proportions of abomasal-dwelling parasites (Haemonchus contortus and Teladorsagia spp.). Trigger-drenched willow fodder block-fed sheep had higher platelet, eosinophil, total white blood cell and lymphocyte counts, greater CD21+ and greater γδ (Gamma Delta) TCR+ (T cell receptor) lymphocyte subsets than control pasture-fed sheep, and higher plasma levels of Immunoglobulin A (IgA) specific for carbohydrate larval antigen (CarLa) on day 105 (P