Chronic inflammation decreases arcuate kisspeptin expression in male sheep.

Lipopolysaccharide (LPS) from Gram-negative bacteria induces an immune response and impairs reproduction through suppression of gonadotropin releasing hormone (GnRH), subsequently luteinizing hormone (LH) secretion. While there is evidence that acute inflammation inhibits kisspeptin, little is known about the impact of chronic inflammation on this key reproductive neuropeptide in livestock species. Thus, we sought to examine a central mechanism whereby LPS suppresses LH secretion in sheep. Twenty wethers were randomly assigned to one of five treatment groups: control (CON; n=4), single acute IV LPS dose (SAD; n=4), daily acute IV LPS dose (DAD; n=4), daily increasing IV LPS dose (DID; n=4), and chronic subcutaneous LPS dose (CSD; n=4). On Days 1 and 7, blood samples were collected every 12 minutes for 360 minutes using jugular venipuncture. Following blood collection on Day 7, all animals were euthanized, brain tissue was perfused with 4% paraformaldehyde, and hypothalamic blocks were removed and processed for immunohistochemistry. On Day 1, LH pulse frequency was significantly lower (p=0.02) in SAD (0.25 ± 0.1 pulses/hour), DAD (0.25 ± 0.1 pulses/hour), DID (0.35 ± 0.1 pulses/hour), and CSD (0.40 ± 0.1 pulses/hour) compared to CON (0.70 ±0.1 pulses/hour). On Day 7, only DID animals (0.35 ± 0.1 pulses/hour) had significantly lower (p=0.049) LH pulse frequency compared to controls (0.85 ± 0.1 pulse/hour). Furthermore, only DID animals (33.3 ± 10.9 cells/section/animal) had significantly fewer (p=0.001) kisspeptin-immunopositive cells compared to controls (82.6 ± 13.6 cells/section/animal). Taken together, we suggest that daily increasing doses of LPS is a powerful inhibitor of kisspeptin neurons in young male sheep and a physiologically relevant model to examine the impact of chronic inflammation on the reproductive axis in livestock.

Surgical management of superficial digital flexor tendon luxation in dogs: 48 cases (2005-2020).

OBJECTIVES: To report the outcome, frequency of complications and potential prognostic factors associated with surgical repair of superficial digital flexor tendon (SDFT) luxation in dogs. MATERIALS AND METHODS: Medical records from 10 referral hospitals were reviewed retrospectively for cases of SDFT luxation in dogs that underwent surgical stabilisation. Signalment, clinical presentation, diagnostic imaging, surgical method, type and length of post-operative limb immobilisation, nature of and length of exercise restriction, presence of post-operative complications and outcomes were recorded. Data were summarised descriptively and prognostic risk factors assessed for association with surgical outcome using risk ratios. RESULTS: Forty-eight cases were included. A successful surgical outcome was recorded in 35 of 48 (73%) cases. Re-luxation of the SDFT occurred in seven of 48 (15%). Six out of 48 (13%) had a persistent lameness despite a stable non-luxating SDFT. A high frequency of post-operative complications occurred (71%), with the majority resolved medically. The risk of surgical failure was 60% higher (risk ratio 1.6, 95% confidence interval 1.1 to 2.4) where absorbable suture material was used compared to non-absorbable suture material. Surgical failure was more common in cases managed with non-rigid immobilisation post-operatively (57% failure) compared to cases managed with rigid immobilisation (19% failure), although this result was not statistically significant. Limb immobilisation of 6 weeks or longer did not significantly affect surgical outcome, compared to shorter periods of exercise restriction or limb immobilisation. CLINICAL SIGNIFICANCE: A good outcome can be expected following surgical stabilisation of SDFT luxation. The use of non-absorbable suture was associated with a more successful surgical outcome.

Analysis of adoption trends of in-parlor technologies over a 10-year period for labor saving and data capture on pasture-based dairy farms.

The use of precision technology is increasingly seen as an option to improve productivity, animal welfare, resource use efficiency, and workplace features on dairy farms. There is limited research related to longitudinal adoption patterns of precision dairy technologies and reasons for any patterns. The aim of this analysis was to investigate trends in technology adoption regarding both the amount (number of farms with a technology) and intensity (number of technologies per farm) of adoption. Surveys of parlor technology adoption were conducted on New Zealand dairy farms in 2008, 2013, and 2018, with 532, 500, and 500 respondents, respectively. Technologies were grouped into labor-saving (LS, such as automatic cluster removers) or data-capture (DC, such as in-line milk meters) categories. Trends were examined for farms that had only LS, only DC, or LS+DC technologies. Technology adoption increased over time; the likelihood of technology adoption in 2018 (and 2013 in parentheses) increased by 21 (22), 7 (68), and 378% (165) for LS, DC, and LS+DC technology groups, respectively, compared to 2008. Farms with LS+DC technologies also had a greater proportion of LS technologies compared to non-LS+DC farms, although this relationship declined over the 10-yr period. The use of a rotary versus herringbone parlor was estimated to be associated with 356 and 470% increase in the likelihood of adopting LS technologies and LS+DC, respectively, from 2008 to 2018. Regional differences in adoption were also found, with the likelihood of adopting DC and LS+DC technologies found to be 46 and 59% greater, respectively, in the South Island of New Zealand, compared to the base region of Waikato. The results highlight the importance of understanding spatial and temporal farm characteristics when considering future effect and adoption of precision dairy technologies. For example, the analysis indicates the occurrence of 2 trajectories to technology investment on farms, where larger farms are able to take advantage of technology opportunities, but smaller farms may be constrained by factors such as lack of economies of scale, limited capital to invest, and inability to retrofit technology into aging parlor infrastructure.

Orthogonal bone plate stabilization for limb-sparing surgery.

This report describes limb-sparing surgery in a 35 kg, six-year-old Hungarian Vizsla with a distal radial lytic bone lesion. Preoperative biopsy had suggested a bone cyst, however histopathology on the excised bone segment was indicative of an osteosarcoma. Following excision of the tumour, the bone defect was filled with a composite bone graft and stabilized with a custom-made dorsal 3.5/2.7 mm pancarpal arthrodesis plate and an orthogonally positioned medial 2.7 mm compression plate. This technique has not previously been described for limb-sparing procedures. No complications were encountered, and despite the owners declining adjunctive chemotherapy, the dog was alive 34 months postoperatively with near normal limb function.

Evaluation of the Threshold of Toxicological Concern (TTC)--challenges and approaches.

Thresholds of Toxicological Concern (TTCs) have been used in the risk assessment of chemicals to which humans are exposed at very low levels. TTC values were developed using data from rodent cancer bioassays and from oral chronic and sub-chronic toxicity studies for non-cancer effects. The workshop assessed the adequacy and fitness for purpose of the TTC approach and the potential for future modifications of critical aspects. The current TTC value for cancer was considered adequate and fit for purpose because it is derived by linear extrapolation from the lowest TD(50) for each compound in the largest available rodent carcinogenicity database. The database on non-cancer endpoints was considered adequate and fit for purpose because the chemical domain, the distributions of NOAELs and the calculated TTC values are comparable across different databases. Application of the TTC approach gives conclusions compatible with the risk assessment approaches currently used by international advisory committees. The workshop recognised the desirability of developing better tools to assess the comparability of the chemical domain covered in different toxicological databases, and the need to develop an internationally acceptable framework and databases for updating the aspects critical to application of the TTC approach.

Host-parasite interactions in a fragmented landscape.

Theory suggests that habitat fragmentation should reduce the risk of being parasitised due to reduced size and increased isolation of the host population. It is predicted that a threshold host population size exists, below which parasites will not be able to persist. Small mammals were trapped and their ecto-parasites removed in 14 field margins of varying widths over 2 years in a highly fragmented agro-ecosystem. No evidence to suggest the presence of a threshold in parasite prevalence was found, which may be due to the high rate of host movement and transiency within the system. Contrary to expectation, the probability of infestation decreased with host abundance and the abundance of alternative hosts, suggesting a dilution effect. The relatively long life cycle of small mammal specialist tick and flea species present under the prevailing environmental conditions may have left the parasites unable to keep up with the rate of reproduction and dispersal of the host. It is important to consider changes in the behaviour of the host and the presence of alternative hosts when predicting the effects of habitat fragmentation on disease spread.

Refining perception-based farmer typologies with the analysis of past census data.

Perception-based typologies have been used to explore the decision making process of farmers and to inform policy design. These typologies have been criticised, however, for not fully capturing true farmer behaviour, and are consequently limited for supporting policy formulation. We present a method that develops a typology, using a social survey approach based on how farmers perceive their environment (e.g. birds and agri-environmental schemes). We then apply time-series census data on past farm strategies (i.e. land use allocation, management style and participation into agri-environmental schemes) to refine these typologies. Consequently, this offers an approach to improving the profiling of farmer types, and strengthens the validity of input into future agricultural policies. While the social survey highlights a certain degree of awareness towards birds with respect to farmer types, the analysis of past farm strategies indicated that farmers did not entirely follow their stated objectives. External factors such as input and output price signals and subsidy levels had a stronger influence on their strategies rather than stated environmental and social issues. Consequently, the refining of farmer types using this approach would aid the design of policy instruments, which integrate ecological issues within planning.

In silico QTL mapping of maternal nurturing ability with the mouse diversity panel.

Significant variation exists for maternal nurturing ability in inbred mice. Although classical mapping approaches have identified quantitative trait loci (QTL) that may account for this variation, the underlying genes are unknown. In this study, lactation performance data among the mouse diversity panel were used to map genomic regions associated with this variation. Females from each of 32 inbred strains (n = 8-19 dams/strain) were studied during the first 8 days of lactation by allowing them to raise weight- and size-normalized cross-foster litters (10 pups/litter). Average daily weight gain (ADG) of litters served as the primary indicator of milk production. The number of pups successfully reared to 8 days (PNUM8) also served as a related indicator of maternal performance. Initial haplotype association analysis using a Bonferroni-corrected, genome-wide threshold revealed 10 and 15 associations encompassing 11 and 13 genes for ADG and PNUM8, respectively. The most significant of these associated haplotype blocks were found on MMU 8, 11, and 19 and contained the genes Nr3c2, Egfr, Sec61g, and Gnaq. Lastly, two haplotype blocks on MMU9 were detected in association with PNUM8. These overlapped with the previously described maternal performance QTL, Neogq1. These results suggest that the application of in silico QTL mapping is a useful tool in discovering the presence of novel candidate genes involved in determining lactation capacity in mice.

Treatment of lumbosacral discospondylitis by surgical stabilisation and application of a gentamicin-impregnated collagen sponge.

This report describes a case of lumbosacral discospondylitis in a two-year-old boxer dog. The dog had been presented with chronic hindlimb lameness and signs of lumbar spinal pain. The diagnosis was confirmed with a magnetic resonance imaging scan and positive blood culture. Following unsuccessful conservative management, the dog was treated with surgical stabilisation using screws and polymethylmethacrylate, and implantation of a gentamicin-impregnated collagen sponge into the L7-S1 disc space. This technique has not previously been described. The dog had a successful long-term outcome with complete resolution of clinical signs.

Preliminary experiences of the triple tibial osteotomy procedure: tibial morphology and complications.

OBJECTIVE: To document the effect of variable tibial morphology on the planning and execution of the triple tibial osteotomy (TTO) procedure in dogs and complications encountered while becoming familiar with the technique. METHODS: The records of 21 consecutive cases that had a TTO performed were reviewed. Preoperative tibial morphology was assessed. Modifications in the described technique and complications were documented. Postoperative outcome was evaluated using a client-based questionnaire. RESULTS: The size of the tibial wedge ostectomy was modified because of variable tibial morphology in four dogs (19 per cent) to avoid over or under correction of the tibial plateau angle (TPA). Intraoperatively, fracture through the caudal tibial cortex occurred in nine cases (41 percent) and through the distal tibial crest cortex in four cases (18 percent). Major complications occurred in five (23 per cent) of cases with four (18 per cent) requiring further surgery. The client questionnaire results demonstrated significant improvements in all parameters and no significant difference from the preinjury status. CLINICAL SIGNIFICANCE: TPA measurement is recommended when planning a TTO to avoid over or under correction of the TPA. Despite frequent minor complications, it appears that the TTO is an effective procedure for management of cranial cruciate ligament rupture in the dog.

Risk assessment of micronutrients.

Risk assessment of micronutrients has to take into account two different intake-response relationships; the risk of deficiency, which decreases with increase in intake, and the risk of toxicity, which increases with increase in intake. The available databases on micronutrients tend to focus on benefits at low intakes, and there are usually few reliable data on hazard identification and characterisation at high intakes. Application of the usual default uncertainty factors for species differences, human variability and database inadequacy could result in "recommended" upper intake levels that would cause deficiency. There have been a number of comprehensive reviews that have used low, and largely arbitrary, uncertainty factors to establish tolerable upper intake levels for vitamins and minerals. A recent FAO/WHO Workshop developed a structured approach to the application of a single composite uncertainty factor. Risk-benefit approaches have been developed recently that balance the risk of toxicity against the risk of deficiency, and offer the potential for more scientifically based methods.

Risk assessment of dietary exposures to compounds that are genotoxic and carcinogenic--an overview.

The process of risk assessment of dietary exposures to genotoxic carcinogens is summarised. Exposures to six genotoxic carcinogens in food (acrylamide, aflatoxin B1, benzo(a)pyrene, dimethylnitrosamine, ethyl carbamate, PhIP) have been used to illustrate the process. The margin of exposure (MOE) approach is seen as a useful method to be used in the risk characterisation step of assessing exposures to genotoxic carcinogens. This approach combines information on animal potency and human exposure, and can be used to indicate levels of concern and also the ranking between various exposures to such agents. Both the T25 and the BMDL10 methods may be used as a reference point. Should a specific MOE value be developed as a cut-off between levels of concern and levels of low concern, the value using T25 data is proposed to be 2.5-times higher than using BMDL10 data. Linear low-dose extrapolation using either T25 or BMDL10, may also be applied. However, it should be understood that this approach should not be interpreted as giving a precise estimate of human risk. For exposures to mutagens in food lacking carcinogenicity data, it is proposed to apply the MOE approach to the lowest effective dose (LED) for in vivo genotoxicity.

The use of a sweetener substitution method to predict dietary exposures for the intense sweetener rebaudioside A.

There are more published dietary exposure data for intense sweeteners than for any other group of food additives. Data are available for countries with different patterns of sweetener approvals and also for population groups with high potential intakes, such as children and diabetic subjects. These data provide a secure basis for predicting the potential intakes of a novel intense sweetener by adjustment of the reported intakes of different sweeteners in mg/kg body weight by their relative sweetness intensities. This approach allows the possibility that a novel sweetener attains the same pattern and extent of use as the existing sweeteners. The intakes by high consumers of other sweeteners allows for possible brand loyalty to the novel sweetener. Using this method, the estimated dietary exposures for rebaudioside A in average and high consumers are predicted to be 1.3 and 3.4mg/kg body weight per day for the general population, 2.1 and 5.0mg/kg body weight per day for children and 3.4 and 4.5mg/kg body weight per day for children with diabetes. The temporary ADI defined by the JECFA for steviol glycosides [JECFA, 2005. Steviol glycosides. In: 63rd Meeting of the Joint FAO/WHO Expert Committee on Food Additives. World Health Organization (WHO), Geneva, Switzerland, WHO Technical Report Series 928, pp. 34-39] was set at 0-2mg/kg body weight (expressed as steviol equivalents); after correction for the difference in molecular weights, these estimated intakes of rebaudioside A are equivalent to daily steviol intakes of less than 2mg/kg. In consequence, this analysis shows that the intakes of rebaudioside A would not exceed the JECFA temporary ADI set for steviol glycosides.

Microbial hydrolysis of steviol glycosides.

A review of the role of gut microbiota in the metabolism of the steviol glycosides, stevioside and rebaudioside A, indicates that they are not absorbed intact but undergo hydrolysis by the intestinal microflora to steviol. Steviol is not metabolized by the intestinal flora and is absorbed from the intestine. The rate of hydrolysis for stevioside is greater than for rebaudioside A. Recent studies using mass spectrometry have shown that steviol-16,17-epoxide is not a microbial metabolite of steviol glycosides. Bacteroides species are primarily responsible for hydrolysis via their beta-glucosidase activity. Fecal incubation studies with both human and animal mixed flora provide similar results, and this indicates that the rat is an appropriate model for studies on steviol glycosides. Given the similarity in the microbial metabolism of stevioside and rebaudioside A with the formation of steviol as the single hydrolysis product that is absorbed from the intestinal tract, the toxicological data on stevioside are relevant to the risk assessment of rebaudioside A.

The Threshold of Toxicological Concern (TTC) in risk assessment.

The Threshold of Toxicological Concern (TTC) is a level of human intake or exposure that is considered to be of negligible risk, despite the absence of chemical-specific toxicity data. The TTC approach is a form of risk characterisation in which uncertainties arising from the use of data on other compounds are balanced against the low level of exposure. The approach was initially developed by the FDA for packaging migrants, and used a single threshold value of 1.5 microg/day (called the threshold of regulation). Subsequent analyses of chronic toxicity data resulted in the development of TTC values for three structural classes with different potentials for toxicity (1,800, 540 and 90 microg/day). These TTC values have been incorporated into the procedure that is used internationally for the evaluation of flavouring substances. Further developments included additional TTC values for certain structural alerts for genotoxicity (0.15 microg/day), and for the presence of an organophosphate group (18 microg/day). All of these TTC values were incorporated into an extended decision tree for chemicals, such as contaminants, which might be present in human foods. The TTC approach has been shown to have potential applications to risk assessments of cosmetic ingredients, household products and impurities in therapeutic drugs.

Comparative toxicokinetics and metabolism of rebaudioside A, stevioside, and steviol in rats.

The toxicokinetics and metabolism of rebaudioside A, stevioside, and steviol were examined in rats for comparative purposes to determine whether toxicological studies conducted previously with stevioside would be applicable to the structurally-related glycoside, rebaudioside A. Single, oral doses of the radiolabelled compounds were extensively and rapidly absorbed with plasma concentration-time profiles following similar patterns for stevioside and rebaudioside A. Elimination of radioactivity from plasma was essentially complete within 72h. All plasma samples had similar metabolite profiles; the predominant radioactive component in all samples was steviol, with lower amounts of steviol glucuronide(s) and low levels of one or two other metabolites. Rebaudioside A, stevioside, and steviol were metabolized and excreted rapidly, with the majority of the radioactivity eliminated in the feces within 48h. Urinary excretion accounted for less than 2% of the administered dose for all compounds in both intact and bile duct-cannulated rats, and the majority of the absorbed dose was excreted via the bile. After administration of the compounds to intact and bile duct-cannulated rats, radioactivity in the feces was present primarily as steviol. The predominant radioactive compound detected in the bile of all cannulated rats was steviol glucuronide(s), indicating de-conjugation in the lower intestine. Overall, the data on toxicokinetics and metabolism indicate that rebaudioside A and stevioside are handled in an almost identical manner. These studies support the use of toxicological safety studies conducted with stevioside for the safety assessment of rebaudioside A.