Fecal excretion of Campylobacter jejuni by young dairy calves and the relationship with neonatal immunity and personality traits.

AIMS: Zoonotic pathogens in bovine herds are major concerns for human and animal health, but their monitoring in animals can be challenging in the absence of clinical signs. Our objective was to determine the association between fecal excretion of Campylobacter jejuni, neonatal immunity, and personality traits of calves. METHODS AND RESULTS: Forty-eight dairy calves were reared in three indoor pens from birth to 4 weeks of life. Microbial analyses of the fecal samples collected weekly revealed that the proportion of calves naturally contaminated with C. jejuni in each pen reached 70% after 3 weeks of life. High (>16 g l-1) levels of IgG levels in the serum of neonatal calves were negatively (P = .04) associated with fecal detection of C. jejuni over the trial period. Calves that spent more time interacting with a novel object tended to be positive (P = .058) for C. jejuni. CONCLUSIONS: Overall, the findings indicate that the immunity of neonatal dairy animals and possibly the animal's behavior may contribute to the fecal shedding of C. jejuni.

Evaluation of alternatives to cautery disbudding of dairy goat kids using physiological measures of immediate and longer-term pain.

We evaluated alternatives to cautery disbudding of goat kids using physiological measures of immediate and longer-term pain. Fifty Saanen doe kids were randomly assigned to 1 of 5 treatments (n = 10/treatment): (1) cautery disbudding (CAUT), (2) caustic paste disbudding (CASP), (3) liquid nitrogen disbudding (CRYO), (4) clove oil injected into the horn bud (CLOV), or (5) sham disbudding (SHAM). Serum cortisol and haptoglobin concentrations were measured from blood samples collected immediately before treatment (baseline) and at 15, 30, 60, and 120 min and then again at 6 and 24 h post-treatment. An infrared thermography camera was used to take images of the horn buds 24 h pre- and 24, 48, and 72 h post-treatment to measure skin temperature. Body weight was measured daily for 1 wk to assess weight change post-treatment. Images of the horn buds were taken at d 1, 2, and 7 and at 6 wk post-treatment to assess tissue damage and wound healing. Mean cortisol concentrations were elevated in CASP kids 1 h post-treatment relative to CAUT kids. Cortisol concentrations of CRYO kids were higher than those of CAUT kids 30 min post-treatment; concentrations for CLOV kids were similar to CAUT kids post-treatment. Mean haptoglobin concentrations were similar across treatments over time; however, CLOV kids had higher concentrations at 24 h post-treatment than all other treatments. Skin temperatures of CASP and CLOV kids were elevated relative to CAUT kids at all time points post-treatment, and all disbudded kids had skin temperatures above those of SHAM kids at 72 h post-treatment. Treatment did not influence weight gain. The CAUT kids had large, open wounds exposing bone; small scabs were still evident 6 wk post-treatment. The CASP kids had red and open, raw wounds that generated large eschars, apparent for up to 6 wk. The CRYO kids had closed, dry wounds initially, but over time lesions appeared that caused open wounds; small scabs were present 6 wk post-treatment. The CLOV kids had closed, dry wounds with blackened skin; healed skin and minimal scabs were present 6 wk post-treatment. Caustic paste and cryosurgical disbudding appeared to cause more pain compared with cautery disbudding; thus, these methods may not provide good alternatives to cautery disbudding. Clove oil appeared to cause a similar pain response as cautery disbudding and smaller wounds with earlier tissue repair; this method shows promise as an alternative to cautery disbudding.

Effect of isoflurane alone or in combination with meloxicam on the behavior and physiology of goat kids following cautery disbudding.

Cautery disbudding of goat kids is painful, but may be alleviated with pain mitigation. We therefore evaluated the effect of administering general anesthesia (isoflurane) or a nonsteroidal anti-inflammatory drug (meloxicam) on goat kid behavior and physiology following cautery disbudding. Trial 1 (n = 12/treatment) evaluated behavioral responses in 72 female Saanen dairy goat kids (mean ± standard error of the mean; 3.9 ± 0.15 d old) and trial 2 (n = 10/treatment) evaluated physiological responses in 60 female Saanen dairy goat kids (4.3 ± 0.14 d old). Goat kids were randomly assigned to 1 of 6 treatment groups that were either (1) sham-handled only (simulated disbudding; SHAM) or disbudded with (2) no pain relief (CAUT), (3) isoflurane gas (ISO), (4) isoflurane and s.c. meloxicam combined (ISO+MEL), (5) meloxicam s.c. (0.5 mg/kg of body weight; I-MEL), or (6) oral meloxicam (0.2 mg/kg of body weight; O-MEL). Head shaking, head scratching, self-grooming, feeding, and body shaking were continuously video recorded for 24 h pre- and post-treatment. Lying behavior was recorded continuously for 24 h pre- and post-treatment using accelerometers. Plasma cortisol, glucose, and lactate concentrations were measured from blood samples collected immediately before treatment (baseline) and at 15, 60, and 120 min post-treatment. Body temperature was measured immediately after blood sampling at all blood sampling time points. Head shaking and body shaking frequencies were 50% higher in CAUT than SHAM kids 5 min post-treatment; ISO+MEL and ISO kids performed 25% less body shakes than CAUT kids. Head scratching durations 1 h post-treatment were higher in CAUT than SHAM kids, whereas O-MEL were similar to SHAM kids from 2 h post-treatment. Self-grooming, feeding, and lying did not differ between groups. Cortisol concentrations were higher in CAUT than SHAM kids (156.4 ± 26.41 and 104.1 ± 26.41 nmol/L, respectively), whereas ISO+MEL and ISO kids (88.3 ± 26.41 and 113.2 ± 26.41 nmol/L, respectively) had lower cortisol concentrations than CAUT kids over the 2-h sampling period. Moreover, O-MEL and I-MEL kids (163.0 ± 26.41 and 130.9 ± 26.41 nmol/L, respectively) had similar cortisol concentrations to CAUT kids. We found no evidence that plasma glucose and lactate concentrations or body temperature were affected by treatment. The administration of isoflurane, with or without meloxicam, appeared to reduce pain associated with cautery disbudding.

Assessing DNA methylation levels in animals: choosing the right tool for the job.

Selection of agricultural animals for improved performance based on genetics has seen significant progress made over the past few decades. Further improvements are likely by combining genetic selection with epigenetic selection or manipulation. However, before this can be undertaken, an understanding of epigenetic mechanisms is required, and this can be obtained only by precise and accurate analysis of epigenetic patterns. Even when one only considers a single epigenetic modification such as DNA methylation, the last 10 years have seen a wide array of technologies developed. For scientists whose primary training is in a field other than epigenetics, the choices can be confusing, and it can be challenging to determine which technology is best for the task at hand. There are many factors to take into consideration before beginning analysis of DNA methylation in animals. It is crucial that the most appropriate tools are selected to ensure that the best possible results are achieved. This review provides an overview of the most common methods of analysing DNA methylation in animals, when they are appropriate, what resolution of information they can provide and what their limitations are.

Medium-term storage of calf beddings affects bacterial community and effectiveness to inactivate zoonotic bacteria.

Land-spreading of animal faecal wastes -such as animal beddings- can introduce zoonotic enteropathogens into the food system environment. The study evaluated the effectiveness of animal beddings naturally contaminated by calf manure to reduce E. coli O157:H7 or Salmonella enterica. The two pathogens were introduced separately as a four strains-cocktail and at high (>6.5 Log10 g-1) concentration into bedding materials, and their inactivation over a 10 weeks-period was monitored by using a Most Probable Number (MPN) enumeration method. Inactivation of E. coli O157:H7 was more effective in the bedding inoculated immediately after collection from calf pens than in the beddings inoculated after a 2 months-pre-storage period: E. coli O157:H7 levels were reduced by 6.6 Log10 g-1 in unstored bedding (0.5 Log10 g-1 recovered; 95%CI: 0.0-1.2), and by 4.9 Log10 g-1 in pre-stored bedding (2.2 Log10 g-1 recovered; 95%CI: 1.5-2.8) with a significant (p<0.05) difference between unstored and pre-stored. S. enterica was inactivated less effectively as counts were reduced by one order of magnitude, with no significant difference in inactivation between unstored and pre-stored beddings. Low levels of naturally occurring E. coli O157 and Salmonella spp. were detected in the non-inoculated beddings, as well as in the straw prior to use in the animal facility. To better understand the possible biological processes involved, the bacterial community present in the beddings was characterised by short-read 16S rRNA sequencing. Pre-storage of the bedding affected the composition but not the diversity of the bacterial community. Analyses of the key bacterial phyla suggested that the presence of a diverse and stable bacterial community might facilitate inactivation of the introduced pathogens, and a possible role of bacterial orders associated with lignocellulolytic resources. Overall, the study contributed to the understanding of the fate of zoonotic bacteria introduced in animal beddings during storage and identified bedding storage practices pre-and post-use in animal facilities that could be important to prevent the risk of zoonosis dissemination to the environment or to the dairy herds.

Worms and welfare: Behavioural and physiological changes associated with gastrointestinal nematode parasitism in lambs.

Parasitism with gastrointestinal nematodes (GIN) is a worldwide issue impacting negatively on animal production, health, and welfare. Therefore, early diagnostic signs of parasitism are required to allow for timely interventions. The objective of this study was to evaluate the behavioural and physiological changes in lambs associated with GIN infection. We used 30, 8-month-old Romney-cross wethers, that were administered anthelmintics until faecal egg counts (FEC) were zero and housed in an indoor facility. The study lasted 9 weeks, which comprised a 3-week pre-treatment, and a 6-week treatment phase. Lambs were randomly assigned to one of two treatments (n = 15/treatment) trickle-dosed with: 1) 1500 infective third stage larvae (L3) three days/week for 6 weeks (27,000 total L3; challenged), or 2) water 3 days/week for 6 weeks (control). Within each pen there were 5 pairs of lambs (balanced for liveweight), with each pair comprising a challenged and control lamb. Blood, faecal, and saliva samples were collected 1 week pre-treatment and weekly for 6 weeks of treatment. Behaviour was observed (e.g., feeding, lying, standing) from video-camera recordings using scan sampling every 5 min for 8 h, 1 day pre-treatment and on the day immediately prior to physiological sampling across the 6-week treatment phase (7 days in total). Accelerometers were attached to each lamb to continuously monitor behaviour from 3 weeks pre-treatment and for the remainder of the study. Liveweight, body condition, faecal soiling and faecal consistency scoring were performed weekly as was lipidomic analysis of plasma samples. From week 2 of treatment, challenged lambs spent less time feeding and more time lying than control lambs until week 5 of treatment (P ≤ 0.01). At week 3 of treatment, elevated lipids (mainly triglycerides and phospholipids), loose faeces and faecal soiling around the anus were observed in challenged lambs compared with controls (P ≤ 0.05). From week 4 of treatment, FEC were elevated in the challenged compared to control lambs (P ≤ 0.05). There was also lower liveweight gain at 4 and 5 weeks of treatment in the challenged lambs compared with control lambs (P ≤ 0.05). These results show a clear timeline of changes in behaviour (e.g., feeding and lying), lipids such as triglycerides, and digestive function (e.g., faecal soiling) suggestive of GIN subclinical disease, which show promise for use in future studies on early identification of subclinical GIN parasitism in lambs.

Sub-acute feeding study of a tall fescue endophyte in a perennial ryegrass host using mice.

Epichloë endophytes in grass associations express a myriad of secondary metabolites which can affect the health of grazing animals and reduce the impact of insect pests on pasture. The ideal endophyte-grass association must have a favourable chemical profile such that the impact on animal health is minimised while the beneficial, deterrent effect on insect pests is maximised. A number of endophyte-perennial ryegrass associations have been successfully commercialised but research is on-going to further improve production in farming systems. Secondary metabolites expressed by endophyte-infected tall fescue include lolines, an animal-safe class of compound which imparts a potent effect on insects. Since endophyte-infected perennial ryegrass does not express lolines, a tall fescue endophyte, AR501, was inoculated into perennial ryegrass in an attempt to improve the insect resistance of this pasture type. In addition to animal safety, it is imperative that consideration is given to the safety of humans consuming animal products derived from livestock grazing the novel pasture. Although pure loline alkaloids have previously been tested on mice it is essential that the entire AR501 endophyte-infected perennial ryegrass matrix is tested since this will result in the exposure of both known and unknown secondary metabolites to mice. Three treatment groups each containing 6 male and 6 female mice were fed diets containing AR501 endophyte-infected perennial ryegrass seed (30%), perennial ryegrass seed containing no endophyte (30%) or a diet without seed (control) for 3 weeks. Mice fed control diet ate more than either of the treatment groups fed a diet containing seed. Male mice fed diet containing Nil endophyte seed ate more than those eating AR501 endophyte-infected perennial ryegrass seed although there was no difference observed in the food intake of female mice. While a few statistically significant differences were observed in the haematology and serum biochemical data, in every instance the difference was restricted to only one gender so is considered unlikely to be of toxicological significance. Mice fed AR501 endophyte-infected perennial ryegrass seed remained healthy throughout the experimental period despite consuming 62,000 mg/kg lolines and 4600 mg/kg peramine per day as well as the wide array of other unknown secondary metabolites expressed by this endophyte. Although animal products may contain additional metabolites as a result of animal metabolism, this experiment raises no food safety concerns for AR501 endophyte-infected perennial ryegrass.

Welfare Assessment of 30 Dairy Goat Farms in the Midwestern United States.

Dairy goat animal welfare assessment protocols have been developed and conducted in Europe and the United Kingdom for dairy goats; however, there are no published reports of large-scale welfare assessment for dairy goats on farms in the Midwestern United States (US). Therefore, the objective of this study was to perform welfare assessment of lactating dairy goats and identify the most prevalent welfare issues on 30 farms across the Midwestern US. Thirty dairy goat farms (self-selected) were enrolled in the study if they shipped milk for human consumption (regardless of herd size). The number of lactating does on each farm ranged from 34 to 6,500 goats, with a median number of 158 lactating does (mean ± SD: 602 ± 1,708 lactating does). The protocol used was developed from available literature on goat welfare assessment but modified for use in the Midwestern US. Observations were made without handling the animals and included 22 animal-based indicators evaluated at the group- and individual-level. The observations were conducted during ~3-5 h during a milking session (either morning or afternoon) and time in the home pen. Principal components analysis (PCA) was carried out on the welfare assessment data from each farm. The first two dimensions of the PCA explained 34.8% of the variation. The PCA biplot indicated correlations between indicators. The most prevalent conditions observed across the 30 farms included any knee calluses (80.9%), any claw overgrowth (51.4%), poor hygiene (14.9%), skin lesions (8.9%), poor hair coat condition (8.3%) and any ear pathology (8.0%). These results are the first to provide the Midwestern US dairy goat industry with information to improve commercial dairy goat welfare.

Sub-Acute Feeding Study of Saxitoxin to Mice Confirms the Effectiveness of Current Regulatory Limits for Paralytic Shellfish Toxins.

Regulatory limits for shellfish toxins are required to protect human health. Often these limits are set using only acute toxicity data, which is significant, as in some communities, shellfish makes up a large proportion of their daily diet and can be contaminated with paralytic shellfish toxins (PSTs) for several months. In the current study, feeding protocols were developed to mimic human feeding behaviour and diets containing three dose rates of saxitoxin dihydrochloride (STX.2HCl) were fed to mice for 21 days. This yielded STX.2HCl dose rates of up to 730 µg/kg bw/day with no effects on food consumption, growth, blood pressure, heart rate, motor coordination, grip strength, blood chemistry, haematology, organ weights or tissue histology. Using the 100-fold safety factor to extrapolate from animals to humans yields a dose rate of 7.3 µg/kg bw/day, which is well above the current acute reference dose (ARfD) of 0.5 µg STX.2HCl eq/kg bw proposed by the European Food Safety Authority. Furthermore, to reach the dose rate of 7.3 µg/kg bw, a 60 or 70 kg human would have to consume 540 or 630 g of shellfish contaminated with PSTs at the current regulatory limit (800 µg/kg shellfish flesh), respectively. The current regulatory limit for PSTs therefore seems appropriate.

The Use of Hides during and after Calving in New Zealand Dairy Cows.

Isolation during calving is a common dairy cow behavior, however it has not been examined in large outdoor group settings. The provision of "hides" was monitored for its impact on calving location and cow-calf behavior. Stocking density and bedding management were either controlled (Phase 1) or managed according to farm practice (Phase 2). Hides were used for calving by 18% (Phase 1) and 22% (Phase 2) of the cows; a further 59% and 44% of cows moved into the hides after calving (Phase 1 and 2, respectively). When hides were not available, cows calved near the edges of the calving area. In Phase 2, as stocking density increased, cows tended to use the hides less. Older cows were less likely to isolate regardless of management. Cow-calf interaction with other cows and calves was lower when hides were available. There was no evidence that hides reduced incorrect matching of cows and calves by staff, however cases of "mismothering" (i.e., calves being taken by other dams) were observed. Since the majority of cows used the hides at some stage before or after calving, we suggest opportunities for seclusion should be provided in large calving groups.

Can Isoflurane and Meloxicam Mitigate Pain Associated with Cautery Disbudding of 3-Week-Old Goat Kids?

: We evaluated the effect of pain mitigation strategies (isoflurane and meloxicam) on the behaviour and physiology of 3-week-old disbudded goat kids. Fifty Saanen does (mean ± SD, 21 ± 3 days old) were randomly allocated to one of five treatments: (1) cautery-disbudded (CAUT), (2) CAUT + isoflurane (ISO), (3) CAUT + isoflurane + meloxicam (ISO+MEL), (4) CAUT + meloxicam (MEL), and (5) handled without disbudding or pain relief (SHAM). Blood samples were taken immediately prior to treatment and at 15-, 60- and 120-min post-treatment to assess cortisol, glucose and lactate concentrations. Behaviour (head shaking and scratching, body shaking, feeding and self-grooming) was observed for 1 h pre- and post-treatment using video-cameras. ISO + MEL and ISO kids had lower cortisol concentrations than CAUT kids 15 min post-treatment (p ≤ 0.05). There was no effect of treatment or time for glucose and lactate concentrations (p ≥ 0.62). At 35 min post-treatment, CAUT, MEL and ISO kids performed more head shakes than SHAM kids (p ≤ 0.05). Isoflurane, with or without meloxicam, may reduce acute stress associated with disbudding of 3-week-old goat kids. More research is needed to assess whether isoflurane (with or without meloxicam) can provide sufficient pain relief for disbudding 3-week-old kids.

Evaluation of infrared thermography as a non-invasive method of measuring the autonomic nervous response in sheep.

Eye temperature measured using infrared thermography (IRT) can be used as a non-invasive measure of autonomic nervous system (ANS) activity in cattle. The objective of this study was to evaluate if changes in eye temperature (measured using IRT) can be used to non-invasively measure ANS activity in sheep. Twenty, 2 to 4-year-old, Romney ewes were randomly assigned to receive either epinephrine (EPI) or physiological saline (SAL) for 5 min administered via jugular catheter (n = 10 ewes/treatment). Eye temperature (°C) was recorded continuously using IRT for approximately 25 min before and 20 min after the start of infusion. Heart rate and heart rate variability, measured using the root mean square of successive differences (RMSSD) and the standard deviation of all inter-beat intervals (SDNN), were recorded for 5 min before and up to 10 min after the start of infusion. Blood samples were taken before and after the infusion period to measure plasma epinephrine, norepinephrine, cortisol and packed cell volume (PCV) concentrations. During the infusion period, maximum eye temperature was on average higher (P<0.05) in sheep that received epinephrine than those that received saline. On average, heart rate was higher (SAL: 87.5 beats/min, EPI: 123.2 beats/min, SED = 7.07 beats/min; P<0.05), and RMSSD (SAL: 55.3 ms, EPI: 17.3 ms, SED = 14.18 ms) and SDNN (SAL: 54.3 ms, EPI: 21.5 ms, SED = 10.00 ms) lower (P<0.05) in ewes during the 5 min post-infusion period compared with ewes that received saline. An infusion of epinephrine resulted in higher geometric mean epinephrine (P<0.05) and cortisol (P<0.05) but not norepinephrine (P>0.05) concentrations in ewes compared to an infusion of saline. PCV concentrations were higher (P<0.001) by 7 ± 1.0% (mean±SED) in ewes after an epinephrine infusion. These results suggest that heart rate variability is a sensitive, non-invasive method that can be used to measure ANS activity in sheep, whereas change in eye temperature measured using IRT is a less sensitive method.

Development of a Yersinia entomophaga bait for control of larvae of the porina moth (Wiseana spp.), a pest of New Zealand improved grassland systems.

BACKGROUND: Porina is the common name for moths and larvae of the genus Wiseana (Lepidoptera: Hepialidae), some of which are significant pasture pests in New Zealand. Because of environmental concerns and the non-target effects of insecticide control measures, biological alternatives for the control of insect pests such as porina are required. RESULTS: Using a food preference assay and time-lapse photography, a range of low-cost food ingredients were assessed for their palatability to porina larvae. Lead candidates were combined into extruded bait variants, allowing assessment of their palatability to porina larvae. A composite bait consisting of palatable ingredients was developed, into which the porina-active entomopathogen Yersinia entomophaga was incorporated. A 7 day minimum median lethal dose of approximately 6.0 × ±1 × 106 Y. entomophaga cells per 0.02 g of bait was defined. Field trials showed that the mean change in larval density over time differed between treatments, with Y. entomophaga bait applied at 87 kg ha-1 resulting in a mean 65% reduction in larval density relative to the control plots, and diflubenzuron treatment resulting in a mean 77% reduction relative to the control plots. The mean dry matter yields over the course of the trial were highest for diflubenzuron (5029 kg ha-1 ), followed by the Y. entomophaga (4783 kg ha-1 ) and control (4673 kg ha-1 ) treatments. CONCLUSIONS: The bacterium Y. entomophaga applied as a composite bait offers an environmentally sustainable approach for porina pest control. © 2019 Society of Chemical Industry.

Cow Lying Behaviour and Bedding Quality Changes during Five Weeks on a Stand-Off Pad.

Bedding quality and cow lying time were measured during five weeks in a normal farm practice (NFP) off-paddock system with no bedding refreshment. Two groups of 100 non-lactating dairy cows were compared to groups of 8 cows with fresh bedding (FB). The cows were on a woodchip pad for 18 h/d at a space allowance of 5.4 m2/cow, with 6 h/d on pasture for 5 weeks. Lying times were recorded continuously for 60 cows per group using accelerometers. Bedding moisture content was measured weekly. Data for each NFP group were analysed and compared with those of their respective FB group using repeated measures. The lying time declined over five weeks from 11.6 h/day during the first week to 5.6 h/day during the fifth week (SED = 0.3; F1,25 = 351.56; p < 0.001). The moisture content of the bedding increased over the five weeks and was significantly higher for both NFP groups (NFP Group 1: F5,59 = 8.33; p < 0.001; NFP Group 2: F5,61 = 5.54; p < 0.001) than those of the respective FB groups. The percentage of total time lying when in the paddock increased for the NFP groups, reaching 15% in the last week of the trial. During five weeks on a stand-off pad, bedding quality deteriorated, and cows lay down less, to such an extent that welfare was compromised.

The effectiveness of clove oil and two different cautery disbudding methods on preventing horn growth in dairy goat kids.

The effectiveness of clove oil and cautery disbudding on horn growth was evaluated in goat kids. The study used 243 Saanen doe kids (4±1 days old; mean±SD) on two goat farms that were disbudded with either (i) clove oil injection (CLOVE), (ii) a cautery iron and bud removed (BUDOFF), or (iii) a cautery iron with bud left intact (BUDON). Each kid received a different treatment per bud, which were balanced between buds (left/right) and randomly allocated. A trained observer monitored bud growth following treatment for 3 months recording either: N: no growth, H: normal horn, S: abnormal horn (scur), or SC: soft, fibrous lump (scorn). After the final observation, buds were assessed for the probability of detecting (i) success (no growth), (ii) scurs, (iii) horns or (iv) scorns [with 95% CI]. The probability of success for BUDOFF (0.77 [0.63, 0.87]) was higher than for BUDON (0.20 [0.11, 0.34]) and CLOVE (0.09 [0.04, 0.18]; P ≤ 0.05). Furthermore, the probability of success for BUDON was higher than for CLOVE (P ≤ 0.05). The probability of scurs was higher for CLOVE (0.72 [0.63, 0.80]) than BUDOFF (0.25 [0.17, 0.34]) and BUDON (0.30 [0.21, 0.39]; P ≤ 0.05). There was no difference in the probability of scurs for BUDOFF and BUDON (P > 0.05). The probability of horns was higher for CLOVE (0.21 [0.15, 0.29]) than BUDON (0.02 [0.01, 0.06]; P ≤ 0.05); horns were not observed for BUDOFF. The probability of scorns for BUDON, the only treatment that led to scorns, was 0.41 (0.25, 0.60). These results suggest that BUDOFF was more effective at preventing growth than CLOVE and BUDON and appears the most effective method, of the methods tested, for disbudding kids. Future research should explore other alternatives to cautery disbudding that may be both efficacious and cause less pain.

Does White Clover (Trifolium repens) Abundance in Temperate Pastures Determine Sitona obsoletus (Coleoptera: Curculionidae) Larval Populations?

To determine if host plant abundance determined the size of clover root weevil (CRW) Sitona obsoletus larval populations, a study was conducted over 4 years in plots sown in ryegrass (Lolium perenne) (cv. Nui) sown at either 6 or 30 kg/ha and white clover (Trifolium repens) sown at a uniform rate of 8 kg/ha. This provided a range of % white clover content to investigate CRW population establishment and impacts on white clover survival. Larval sampling was carried out in spring (October) when larval densities are near their spring peak at Lincoln (Canterbury, New Zealand) with % clover measured in autumn (April) and spring (September) of each year. Overall, mean larval densities measured in spring 2012-2015 were 310, 38, 59, and 31 larvae m-2, respectively. There was a significant decline in larval populations between 2012 and 2013, but spring populations were relatively uniform thereafter. The mean % white clover measured in autumns of 2012 to 2015 was 17, 10, 3, and 11%, respectively. In comparison, mean spring % white clover from 2012 to 2015, averaged c. 5% each year. Analysis relating spring (October) larval populations to % white clover measured in each plot in autumn (April) found the 2012 larval population to be statistically significantly larger in the ryegrass 6 kg/ha plots than 30 kg/ha plots. Thereafter, sowing rate had no significant effect on larval populations. From 2013 to 2015, spring larval populations had a negative relationship with the previous autumn % white clover with the relationship highly significant for the 2014 data. When CRW larval populations in spring 2013 to 2015 were predicted from the 2013 to 2015 autumn % white clover, respectively, based on their positive relationship in 2012, the predicted densities were substantially larger than those observed. Conversely, when 2015 spring larval data and % clover was regressed against 2012-2014 larval populations, observed densities tended to be higher than predicted, but the numbers came closer to predicted for the 2013 and 2014 populations. These differences are attributed to a CRW population decline that was not accounted by % white clover changes, the CRW decline most likely due to biological control by the Braconid endoparasitoid Microctonus aethiopoides, which showed incremental increases in parasitism between 2012 and 2015, which in 2015 averaged 93%.

Temperature and Plant Genotype Alter Alkaloid Concentrations in Ryegrass Infected with an Epichloë Endophyte and This Affects an Insect Herbivore.

Asexual Epichloë endophytes colonize agricultural forage grasses in a relationship which is mutually beneficial and provides the host plant with protection against herbivorous insects. The endophyte strain AR37 (Epichloë festucae var. lolii) produces epoxy-janthitrem alkaloids and is the only endophyte known to provide ryegrass with resistance against porina larvae (Wiseana cervinata (Walker)), a major pasture pest in cooler areas of New Zealand. This study examined the effect of temperature on concentrations of epoxy-janthitrems in AR37-infected ryegrass and determined how the resulting variations in concentration affected consumption, growth and survival of porina larvae. Twenty replicate pairs of perennial (Lolium perenne L.) and Italian ryegrass (L. multiflorum Lam.) plants with and without endophyte were prepared by cloning, with one of each pair grown at either high (20°C) or low (7°C) temperature. After 10 weeks, herbage on each plant was harvested, divided into leaf and pseudostem, then freeze dried and ground. Leaf and pseudostem material was then incorporated separately into semi-synthetic diets which were fed to porina larvae in a bioassay over 3 weeks. Epoxy-janthitrem concentrations within the plant materials and the semi-synthetic diets were analyzed by high performance liquid chromatography. AR37-infected ryegrass grown at high temperature contained high in planta concentrations of epoxy-janthitrem (30.6 µg/g in leaves and 83.9 µg/g in pseudostems) that had a strong anti-feedant effect on porina larvae when incorporated into their diets, reducing their survival by 25-42% on pseudostems. In comparison, in planta epoxy-janthitrem concentrations in AR37-infected ryegrass grown at low temperature were very low (0.67 µg/g in leaves and 7.4 µg/g in pseudostems) resulting in a small anti-feedant effect in perennial but not in Italian ryegrass. Although alkaloid concentrations were greatly reduced by low temperature this reduction did not occur until after 4 weeks of exposure. Alkaloid concentrations were slightly lower in Italian than in perennial ryegrass and concentrations were higher in the pseudostems when compared with the leaves. In conclusion, epoxy-janthitrems expressed by the AR37 endophyte show strong activity against porina larvae. However, when ryegrass plants are grown at a constant low temperature for an extended period of time in planta epoxy-janthitrem concentrations are greatly reduced and are less effective against this pasture pest.

Detection of Invertebrate Suppressive Soils, and Identification of a Possible Biological Control Agent for Meloidogyne Nematodes Using High Resolution Rhizosphere Microbial Community Analysis.

White clover (Trifolium repens) is the key legume component of New Zealand pastoral agriculture due to the high quality feed and nitrogen inputs it provides. Invertebrate pests constrain white clover growth and this study investigated rhizosphere-associated fungal controls for two of these pests and attempts to disentangle the underpinning mechanisms. The degree of suppressiveness of 10 soils, in a latitudinal gradient down New Zealand, to added Meloidogyne hapla and Costelytra zealandica scarab larvae was measured in untreated soil. Most of the soils showed no suppressive activity against these pests but two showed activity against M. hapla and two against C. zealandica. Rhizosphere fungi responsible for pest suppressive responses were elucidated via next-generation sequencing. In the M. hapla-suppressive soils nematode-trapping Orbiliomycetes fungi were present in significantly greater abundance than non-suppressive soils and their abundance increased further with addition of M. hapla. A comparison of plant growth and the rhizosphere fungal community between untreated and irradiated soil was carried out on 5 of the 10 soils using Pyronota as the scarab larvae. Soil irradiation either: reduced (by 60-70%); increased (16×) or made no difference to white clover growth across the five soils tested, illustrating the range of microbial impacts on plant production. In one of the M. hapla suppressive soils irradiation resulted in a significant increase in nematode galling suggesting that Orbiliomycetes fungi were indeed responsible for the suppressive effect. Lack of consistent changes in soil macronutrients and pH post-irradiation suggest these were not responsible for plant or invertebrate responses. The use of next generation sequencing in controlled pot trials has allowed identification of a potential biological control organism and bioindicator for M. hapla suppression.