Caring for Canines: A Survey of Dog Ownership and Parasite Control Practices in Ireland.

Dogs are an important part of life for many people. Dog ownership can confer various health benefits to their owners, but it also presents some risks. In order to establish if dog owners were aware of these risks, an online survey of dog ownership practices was carried out. The survey was open during the month of January 2022 and received 662 responses regarding 850 individual dogs. Overall, 52% of people reported deworming their dog between zero and twice a year, which is unlikely to reduce the risk of infection to humans. The majority of dog owners (71%) reported disposing of faeces correctly; however, when comparing urban and rural environments, 33% of those in rural environments did not dispose of their dogs' faeces at all, compared with 3% of people in urban locations. People who obtained their dog during the pandemic brought their dog to the vet and dewormed them more frequently than those who obtained their dog before the pandemic. There were no differences in how faeces was disposed of between these groups. These results indicate that we, as researchers and veterinarians, have much work to do in terms of educating dog owners about the ways in which we can reduce the risk of infection to ourselves and our communities.

Efficient Aflatoxin B1 Sequestration by Yeast Cell Wall Extract and Hydrated Sodium Calcium Aluminosilicate Evaluated Using a Multimodal In-Vitro and Ex-Vivo Methodology.

In this work, adsorption of the carcinogenic mycotoxin aflatoxin B1 (AFB1) by two sequestrants-a yeast cell wall-based adsorbent (YCW) and a hydrated sodium calcium aluminosilicate (HSCAS)-was studied across four laboratory models: (1) an in vitro model from a reference method was employed to quantify the sorption capabilities of both sequestrants under buffer conditions at two pH values using liquid chromatography with fluorescence detection (LC-FLD); (2) in a second in vitro model, the influence of the upper gastrointestinal environment on the mycotoxin sorption capacity of the same two sequestrants was studied using a chronic AFB1 level commonly encountered in the field (10 µg/L and in the presence of feed); (3) the third model used a novel ex vivo approach to measure the absorption of 3H-labelled AFB1 in the intestinal tissue and the ability of the sequestrants to offset this process; and (4) a second previously developed ex vivo model readapted to AFB1 was used to measure the transfer of 3H-labelled AFB1 through live intestinal tissue, and the influence of sequestrants on its bioavailability by means of an Ussing chamber system. Despite some sorption effects caused by the feed itself studied in the second model, both in vitro models established that the adsorption capacity of both YCW and HSCAS is promoted at a low acidic pH. Ex vivo Models 3 and 4 showed that the same tested material formed a protective barrier on the epithelial mucosa and that they significantly reduced the transfer of AFB1 through live intestinal tissue. The results indicate that, by reducing the transmembrane transfer rate and reducing over 60% of the concentration of free AFB1, both products are able to significantly limit the bioavailability of AFB1. Moreover, there were limited differences between YCW and HSCAS in their sorption capacities. The inclusion of YCW in the dietary ration could have a positive influence in reducing AFB1's physiological bioavailability.

The Analysis of Docosahexaenoic Acid (DHA) in Dried Dog Food Enriched with an Aurantiochytrium limacinum Biomass: Matrix Extension Validation and Verification of AOAC Method 996.06.

BACKGROUND: Docosahexaenoic acid (DHA) plays an important role in brain and retinal development in dogs. However, supranutritional dietary supplementation can result in health issues, including gastrointestinal bleeding, making the accurate analysis of DHA in dog food important for nutritional and welfare regulatory compliance. OBJECTIVE: The aim of this study was to conduct a validation and verification of the AOAC 996.06 method, and hence establish its fitness for purpose, for the analysis of DHA in dried dog food supplemented with a heterotrophically grown unextracted DHA-rich Aurantiochytrium limacinum biomass. METHODS: The AOAC 996.06 method, which involves the use of gas chromatography coupled to flame ionization detection (GC-FID), was used to conduct a validation of the analysis of DHA in dried dog food and the results were verified in a second laboratory. RESULTS: The method was found to be linear over the ranges analyzed and results were found to be within the acceptance criteria for precision and accuracy, verifying the applicability for this matrix. The selectivity and sensitivity of the method were also determined. CONCLUSIONS: The AOAC 996.06 method is fit for purpose for the analysis of DHA in dry dog food kibble. HIGHLIGHTS: The method can be applied to various dog food samples, supplemented with an unextracted Aurantiochytrium limacinum biomass, using alternative manufacturing methods, i.e. pelleted and extruded with no significant matrix effects being observed.

Whole-Life or Fattening Period Only Broiler Feeding Strategies Achieve Similar Levels of Omega-3 Fatty Acid Enrichment Using the DHA-Rich Protist, Aurantiochytrium limacinum.

The fatty acid composition of broiler chicken tissues can be increased by adding omega-3 rich ingredients to their diets. The purpose of this study was to compare the levels of tissue enrichment observed following the supplementation of broilers with the docosahexaenoic acid (DHA)-rich protist, Aurantiochytrium limacinum (AURA) for their whole life (42 days) or for the final 21-day fattening period. Day-old chicks (n = 350) were distributed among 35 pens (10 birds per pen) with each pen randomly assigned to one of five treatments: Control; 0.5% AURA from day 0-42; 1% AURA from day 0-42; 0.5% AURA from day 21-42; 1% AURA from day 21-42. Production parameters were recorded over the course of the study and the fatty acid profile of the breast, thigh, liver, kidney and skin with adhering fat was quantified at the end of the feeding period. The level of supplementation had a significant impact on the degree of omega-3 tissue enrichment, however, no differences were observed when the same dose was provided for 21 or 42 days. These results indicate that supplementation with AURA for a period of 21 days does not negatively affect broiler productivity and is the most efficient strategy to increase the nutritional value of broiler products.

Tolerance of Broilers to Dietary Supplementation with High Levels of the DHA-Rich Microalga, Aurantiochytrium Limacinum: Effects on Health and Productivity.

It is well established that the docosahexaenoic acid (DHA) content of commonly consumed meats, such as chicken, can be increased through dietary supplementation with DHA-rich ingredients. The purpose of this study was to investigate the tolerance of broilers to dietary supplementation with the unextracted biomass of a DHA-rich microalgae Aurantiochytrium limacinum, so as to ensure its safety, since it is accumulated in broiler meat. Healthy day-old male Ross 308 chicks (n = 1120) were evenly distributed to 32 pens (35 chicks per pen), with pens randomly allocated to one of four dietary treatments, each having eight replicates. The dietary groups included one untreated control and three treatments corresponding to three inclusion levels (0.5, 2.5, and 5.0%) of All-G-Rich®, with the birds receiving the experimental diets ad libitum during the study (day 0⁻42). Bird survival, blood parameters, productivity, and breast and thigh DHA content were determined after 42 days of feeding. Supplementation at up to 10 times the intended use level had no negative effects on the mortality, blood parameters or productivity of the birds, while significant increases in the meat DHA content were observed. These results indicate that supplementation with Aurantiochytrium limacinum is a safe and effective way to increase broiler tissue DHA content.

Live weight as a basis for targeted selective treatment of lambs post-weaning.

Targeted selective treatment (TST) has been proposed as a sustainable method of gastrointestinal nematode control that reduces the number of anthelmintic treatments administered, thereby preserving a susceptible nematode population in refugia. In order to minimise the impact of withholding treatment on animal performance, animals that would benefit most should be selected for treatment. However, the most suitable criteria for selecting which animals to treat remain a subject of research. The impact of implementing a TST strategy based on lamb live weight was investigated, and whether heavy lambs were more resilient than light lambs if left untreated. The study was conducted using weaned lambs on 3 sheep farms, and over 2 years. On each farm lambs were weighed and divided into heavy, medium and light weight classes. Within the heavy (n = 225) and light (n = 218) weight classes lambs were randomly allocated to two treatments; anthelmintic treatment or no anthelmintic treatment. All lambs in the medium weight class were treated. Animal performance and parasitological parameters were assessed over a 28-day period. Anthelmintic treatment had a significant effect (P < 0.01) on faecal egg count, average daily weight gain, body condition score and dag score; there was no treatment by weight class interaction. Anthelmintic treatment had no effect on plasma pepsinogen concentration or on blood leukocyte numbers. Withholding anthelmintic treatment from lambs therefore had a negative effect on worm egg count and animal performance with no evidence that heavy lambs were more resilient than light lambs when left untreated.

The validation & verification of an LC/MS method for the determination of total docosahexaenoic acid concentrations in canine blood serum.

Docosahexaenoic acid (DHA), is an omega 3 fatty acid (n-3 FA) that has been shown to play a role in canine growth and physiological integrity and improvements in skin and coat condition. However, potential adverse effects of n-3 FA specifically, impaired cellular immunity has been observed in dogs fed diets with elevated levels of n-3 FA. As such, a safe upper limit (SUL) for total n-3 FAs (DHA and EPA) in dogs has been established. Considering this SUL, sensitive methods detecting DHA in blood serum as a biomarker when conducting n-3 FA supplementation trials involving dogs are required. In this study, an LC-ESI-MS/MS method of DHA detection in dog serum was validated and verified. Recovery of DHA was optimized and parallelism tests were conducted with spiked samples demonstrating that the serum matrix did not interfere with quantitation. The stability of DHA in serum was also investigated, with -80 °C considered suitable when storing samples for up to six months. The method was linear over a calibration range of 1-500 µg/mL and precision and accuracy were found to meet the requirements for validation. This method was verified in an alternative laboratory using a different analytical system and operator, with the results meeting the criteria for verification.

Dietary supplementation of finishing pigs with the docosahexaenoic acid-rich microalgae, Aurantiochytrium limacinum: effects on performance, carcass characteristics and tissue fatty acid profile.

OBJECTIVE: The aim of this experiment was to evaluate the effect of dietary supplementation with the docosahexaenoic acid (DHA)-rich microalgae, Aurantiochytrium limacinum (AURA) on pig performance, carcass traits, and the fatty acid composition of pork Longissimus lumborum (LL) and backfat. METHODS: A total of 144 Pig Improvement Company (PIC)×Goland finishing pigs (72 females and 72 castrated males) of mean weight 117.1 (±13.1) kg were blocked by sex and body weight and provided with 0% or 1% AURA in isonutritive and isocaloric diets. A total of 24 pens provided 12 replicates per treatment. Animals were weighed on day 0 and 28 with feed and water intake recorded per pen. After 31 days supplementation (28 days of study and 3 days until the slaughtering date) three animals per pen (n = 72) were slaughtered and the LL and backfat thickness, lean meat content and dressing percentage were recorded for the carcasses. The fatty acid (FA) profile of the LL and backfat was established by direct FA methyl ester synthesis. RESULTS: No differences were observed for any performance parameters or carcass traits. Supplementation with AURA resulted in significant changes to the FA profiles of both the LL and backfat with male and female pigs responding differently to supplementation in terms of particular FAs. Overall, pork LL samples had significantly higher eicosapentaenoic acid (p<0.001) and DHA concentrations (p<0.001), and higher omega-3 (n-3) FAs (p<0.001), as well as an increased omega3:omega6 (n-3:n-6) ratio (p = 0.001). For backfat, supplementation resulted in significantly higher amounts of DHA (p<0.001) and n-3 FAs (p<0.001). CONCLUSION: These results indicate that dietary supplementation with 1% AURA over a 31 day period can increase the FA composition of pork LL and backfat, specifically the DHA, with no major impact on growth performance and carcass traits.

Genetic basis of benzimidazole resistance in Teladorsagia circumcincta in Ireland.

Resistance to benzimidazole (BZ) anthelmintics is common in ovine nematodes of economic importance. Single nucleotide polymorphisms (SNP) at three positions in the isotype 1 ß- tubulin gene have been associated with BZ resistance and molecular tests for the detection of BZ resistance have been developed. In order to determine if such tests are practicable in Ireland the polymorphisms associated with BZ resistance must be identified. To this end, BZ-resistant nematodes were recovered from four farms in Ireland. Resistant Teladorsagia circumcincta, Cooperia curticei and Trichostrongylus colubriformis were recovered, with resistant T. circumcincta the most common and the only species studied further. Sequencing of the isotype 1 ß-tubulin gene from resistant T. circumcincta identified a T - A transition, resulting in an F200Y substitution known to be responsible for BZ-resistance, on three of the farms. However, on the fourth farm the frequency of the resistant A allele was only 0.33 indicating another BZ resistance mechanism may be present on this farm. An additional polymorphism resulting in a substitution of glutamate for leucine (E198L) was also found on this farm at low frequency (0.17). No polymorphisms at position 167 were identified on any farm. Therefore, molecular tests to detect BZ resistance in T. circumcincta in Ireland could prove useful; however, they may result in some instances of resistance remaining undetected.

A nationwide survey of anthelmintic treatment failure on sheep farms in Ireland.

BACKGROUND: Between 2013 and 2015 the Department of Agriculture, Food and the Marine (DAFM) administered a sheep technology adoption programme (STAP), with the aim of increasing profitability on Irish sheep farms by encouraging the adoption of best management practices. One of the options available to STAP participants was to test the efficacy of the anthelmintic treatment (benzimadazole, levamisole or macrocyclic lactone) used in their flocks by means of a drench test, which is a modification of the faecal egg count reduction test; individual faecal samples were collected from the same group of lambs before and after anthelmintic treatment, the number of eggs present pre and post treatment was subsequently determined from a pooled sample. RESULTS: In total, 4211 drench tests were undertaken by farmers during the 3 years of the programme. Information on the anthelmintic product used was available for 3771 of these tests; anthelmintics from the classes benzimidazole (BZ), levamisole (LV) and macrocyclic lactone (ML) (avermectins (AVM) plus moxidectin (MOX)) were used in 42.0%, 23.4% and 32.5% of tests, respectively. The remaining 2.1% of tests involved an inappropriate product. The efficacy of treatment against 'other trichostrongyles' (excluding Nematodirus spp and Strongyloides papillosus.) could be established for 1446 tests, and 51% of these tests were considered effective (i.e. a reduction of faecal egg count (FEC) ≥ 95%). There was a significant difference among the drug groups in efficacy; 31.5%, 51.9%, 62.5% and 84% of treatments were considered effective for BZ, LV, AVM, MOX, respectively. The efficacy of treatment against Nematodirus spp. could be established for 338 tests and the overall efficacy was 96%. CONCLUSIONS: Due to the significant difference among the anthelmintic classes for efficacy against 'other trichostrongyles' along with the high level of efficacy against Nematodirus spp., a genus for which anthelmintic resistance is rarely reported, it is concluded that anthelmintic resistance was responsible for the majority of the anthelmintic treatment failures observed.

Characterisation of ivermectin and multi-drug resistance in two field isolates of Teladorsagia circumcincta from Irish sheep flocks.

Ivermectin resistant Teladorsagia circumcincta were isolated from two Irish sheep farms. Parasite naïve lambs were artificially challenged with the isolates in order to test their sensitivity to benzimidazole (BZ), levamisole (LEV) and ivermectin (IVM) by means of a faecal egg count reduction test (FECRT). Both isolates were found to be resistant to all three anthelmintics with reductions of 47% (C.I. 10-68), 92% (C.I. 74-97) and 50% (C.I. 24-68) for the Farm A isolate and of 85% (C.I. 59-94), 89% (C.I. 69-96) and 73% (C.I. 49-86) for the Farm B isolate for BZ, LEV and IVM, respectively. A controlled efficacy test (CET) was undertaken to confirm resistance to IVM and to determine the efficacy of moxidectin (MOX). In agreement with the FECRT, IVM resistance was confirmed for both isolates, with reductions in worm burden of 78% (C.I. 67-85) and 80% (C.I. 45-93) for the Farm A and B isolates respectively. Both isolates were found to be MOX susceptible, with reductions in worm burden of 97% (C.I. 91-99) and 100% for the Farm A and B isolates respectively. Additionally, in vitro tests confirmed the results of the FECRT; results from the larval development assay (LDA) confirmed that both isolates were resistant to BZ and LEV with development observed at the discriminating dose for each drug, while the larval migration inhibition assay (LMIA) confirmed IVM resistance and indicated MOX susceptibility in both isolates. These results represent the first confirmed cases of IVM resistance on Irish sheep farms as well as the first report of multiple anthelmintic resistant gastrointestinal nematodes in Ireland.

High level of treatment failure with commonly used anthelmintics on Irish sheep farms.

BACKGROUND: In 2013 a Technology Adoption Program for sheep farmers was established to encourage the implementation of best management practices on sheep farms in Ireland. There were 4,500 participants in this programme in 2013. As part of this programme, farmers had the option to carry out a drench test to establish the efficacy of their anthelmintic treatment. RESULTS: Flock faecal samples were collected before and after treatment administration and gastrointestinal nematode eggs enumerated. In total there were 1,893 participants in the task, however only 1,585 included both a pre- and post-treatment faecal sample. Of those, 1,308 provided information on the anthelmintic product that they used with 46%, 23% and 28% using a benzimidazole (BZ), levamisole (LEV) and macrocyclic lactone (ML) product respectively. The remaining farmers used a product inapplicable for inclusion in the task such as a flukicide or BZ/LEV combination product. Samples were included for analysis of drench efficacy if the pre-treatment flock egg count was ≥200 eggs per gram and the interval post-sampling was 10-14 days for BZ products, 4-7 days for LEV products and 14-18 days for ML products. These criteria reduced the number of valid tests to 369, 19.5% of all tests conducted. If the reduction post-treatment was ≥95% the treatment was considered effective. Only 51% of treatments were considered effective using this criterion. There was a significant difference in efficacy between the anthelmintic drug classes with BZ effective in only 30% of treatments, LEV effective in 52% of cases and ML effective in 76% of cases. CONCLUSIONS: Gastrointestinal nematode anthelmintic treatments, as practiced on Irish farms, have a high failure rate. There was a significant difference between the efficacies of the anthelmintic classes with BZ the least effective and ML the most effective.