The Role of Wastewater Treatment Plants in Dissemination of Antibiotic Resistance: Source, Measurement, Removal and Risk Assessment.

Antibiotic Resistance Genes (ARGs) are contaminants of emerging concern with marked potential to impact public and environmental health. This review focusses on factors that influence the presence, abundance, and dissemination of ARGs within Wastewater Treatment Plants (WWTPs) and associated effluents. Antibiotic-Resistant Bacteria (ARB) and ARGs have been detected in the influent and the effluent of WWTPs worldwide. Different levels of wastewater treatment (primary, secondary, and tertiary) show different degrees of removal efficiency of ARGs, with further differences being observed when ARGs are captured as intracellular or extracellular forms. Furthermore, routinely used molecular methodologies such as quantitative polymerase chain reaction or whole genome sequencing may also vary in resistome identification and in quantifying ARG removal efficiencies from WWTP effluents. Additionally, we provide an overview of the One Health risk assessment framework, as well as future strategies on how WWTPs can be assessed for environmental and public health impact.

Differential expressions of FASN, SCD, and FABP4 genes in the ribeye muscle of omega-3 oil-supplemented Tattykeel Australian White lambs.

BACKGROUND: The concept of the functional nutritional value of health-beneficial omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) is becoming a phenomenon among red meat consumers globally. This study examined the expressions of three lipogenic genes (fatty acid binding protein 4, FABP4, fatty acid synthase, FASN; and stearoyl-CoA desaturase, SCD) in the ribeye (Longissimus thoracis et lumborum) muscle of Tattykeel Australian White (TAW) lambs fed fortified omega-3 diets and correlations with fatty acids. To answer the research question, "are there differences in the expression of lipogenic genes between control, MSM whole grain and omega-3 supplemented lambs?", we tested the hypothesis that fortification of lamb diets with omega-3 will lead to a down-regulation of lipogenic genes. Seventy-five six-month old TAW lambs were randomly allocated to the (1) omega-3 oil-fortified grain pellets, (2) unfortified grain pellets (control) or (3) unfortified MSM whole grain pellets diet supplements to generate three treatments of 25 lambs each. The feeding trial lasted 47 days. RESULTS: From the Kruskal-Wallis test, the results showed a striking disparity in lipogenic gene expression between the three dietary treatments in which the FABP4 gene was significantly up-regulated by 3-folds in the muscles of lambs fed MSM Milling (MSM) whole grain diet compared to the omega-3 and control diets. A negative correlation was observed between FASN gene expression and intramuscular fat (IMF), eicosapentaenoic acid (EPA), total polyunsaturated fatty acids (PUFA), omega-6 polyunsaturated fatty acids (n-6 PUFA) and monounsaturated fatty acids (MUFA). The FABP4 gene expression was positively correlated (P < 0.05) with EPA and docosahexaenoic acid (DHA). CONCLUSION: Taken together, this study's results suggest that FABP4 and FASN genes perform an important role in the biosynthesis of fatty acids in the ribeye muscle of TAW lambs, and supplementary diet composition is an important factor influencing their expressions.

Prevalence and distribution of antimicrobial resistance in effluent wastewater from animal slaughter facilities: A systematic review.

The extensive use of antibiotics in food animal production and disposal of untreated wastewater from food animal slaughter facilities may create a shift in microbiomes of different ecosystems by generating reservoirs of antimicrobial resistance along the human-animal-environmental interface. This epidemiological problem has been studied, but its magnitude and impact on a global scale is poorly characterised. A systematic review was done to determine global prevalence and distribution patterns of antimicrobial resistance in effluent wastewater from animal slaughter facilities. Extracted data were stratified into rational groups for secondary analyses and presented as percentages. Culture and sensitivity testing was the predominant method; Escherichia spp., Enterococcus spp., and Staphylococcus aureus were the most targeted isolates. Variable incidences of resistance were detected against all major antimicrobial classes including reserved drugs such as ceftazidime, piperacillin, gentamicin, ciprofloxacin, and chloramphenicol; the median frequency and range in resistant Gram-negative isolates were: 11 (0-100), 62 (0-100), 8 (0-100), 14 (0-93) and 12 (0-62) respectively. Ciprofloxacin was the most tested drug with the highest incidences of resistance in livestock slaughterhouses in Iran (93%), Nigeria (50%) and China (20%), and poultry slaughterhouses in Germany (21-81%) and Spain (56%). Spatial global distribution patterns for antimicrobial resistance were associated with previously reported magnitude of antibiotic use in livestock or poultry farming and, the implicit existence of jurisdictional policies to regulate antibiotic use. These data indicate that anthropogenic activities in farming systems are a major contributor to the cause and dissemination of antimicrobial resistance into the environment via slaughterhouse effluents.

Colchicine Does Not Reduce Abdominal Aortic Aneurysm Growth in a Mouse Model.

Background and Aims: The nacht domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome is upregulated in human abdominal aortic aneurysm (AAA), but its pathogenic role is unclear. The aims of this study were firstly to examine whether the inflammasome was upregulated in a mouse model of AAA and secondly to test whether the inflammasome inhibitor colchicine limited AAA growth. Methods: AAA was induced in eight-week-old male C57BL6/J mice with topical application of elastase to the infrarenal aorta and oral 3-aminopropionitrile (E-BAPN). For aim one, inflammasome activation, abdominal aortic diameter, and rupture were compared between mice with AAA and sham controls. For aim two, 3 weeks after AAA induction, mice were randomly allocated to receive colchicine (n = 28, 0.2 mg/kg/d) or vehicle control (n = 29). The primary outcome was the rate of maximum aortic diameter increase measured by ultrasound over 13 weeks. Results: There was upregulation of NLRP3 markers interleukin- (IL-) 1ß (median, IQR; 15.67, 7.11-22.60 pg/mg protein versus 6.87, 4.54-11.60 pg/mg protein, p = .048) and caspase-1 (109, 83-155 relative luminosity units (RLU) versus 45, 38-65 RLU, p < .001) in AAA samples compared to controls. Aortic diameter increase over 80 days (mean difference, MD, 4.3 mm, 95% CI 3.3, 5.3, p < .001) was significantly greater in mice in which aneurysms were induced compared to sham controls. Colchicine did not significantly limit aortic diameter increase over 80 days (MD -0.1 mm, 95% CI -1.1, 0.86, p = .922). Conclusions: The inflammasome was activated in this mouse model of AAA; however, daily oral administration of colchicine did not limit AAA growth.

Feedlot growth performance and carcass characteristics of steers backgrounded on buffel grass or buffel-Desmanthus mixed pastures.

Feedlot performance and carcass characteristics of tropical beef steers backgrounded on buffel grass (Cenchrus ciliaris) only or buffel grass oversown with desmanthus (Desmanthus spp. ; 11.5% initial sward botanical composition) were evaluated. It was hypothesized that tropical beef cattle steers backgrounded on buffel grass only or buffel grass oversown with desmanthus with similar backgrounding growth performance will not differ in feedlot growth performance and carcass quality. Three hundred and twelve Bos indicus × Bos taurus tropical composite steers, 20-23 months old and weighing 413 ± 24 kg, previously backgrounded on buffel grass only or buffel-desmanthus mixed pastures for 147 days were finished on a concentrate diet in the feedlot for 110 days before slaughter. Buffel-desmanthus backgrounded steers had a slightly higher average daily gain (ADG; 1.8 kg/day) than the buffel grass backgrounded steers that had 1.7 kg/day ADG (p < 0.01). However, the final live weight and dry matter intake were not different (p ≥ 0.59). All the carcass traits measured were not different (p ≥ 0.18). Only 4% buffel grass and 8% buffel-desmanthus backgrounded steers fell short of the Meat Standards Australia (MSA) index, a level that is within the 4-9% reported for cattle produced in Queensland and slaughtered between July 2019 and June 2020. These findings indicate that desmanthus can be used to background beef cattle in northern Australia vertosol soil regions, where there is a paucity of adapted pasture legumes, with no negative impact on feedlot performance and carcass quality. The hypothesis that tropical beef cattle steers backgrounded on buffel grass only pastures or buffel grass oversown with desmanthus with similar backgrounding growth performance will have similar feedlot growth performance and carcass quality was accepted.

Fortification of diets with omega-3 long-chain polyunsaturated fatty acids enhances feedlot performance, intramuscular fat content, fat melting point, and carcass characteristics of Tattykeel Australian White MARGRA lambs.

Meat eating quality indices such as intramuscular fat content (IMF) and fat melting point (FMP) of the Longissimus thoracis et lumborum muscle and the feedlot performance, carcass traits, and commercial wholesale cuts of lot-fed Tattykeel Australian White (TAW) MARGRA lambs as a result of dietary fortification of the diet with omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) were evaluated. A total of 75 TAW MARGRA lambs at 6 months of age with an average liveweight of 30 ± 1.2 kg were used. The lambs were randomly allocated to the following three dietary treatments of 25 lambs each in a 47-day feeding trial using a completely randomized experimental design: (1) control diet of hay plus pellets without omega-3 oil, (2) hay plus commercial whole grain pellets (MSM) without omega-3 oil, and (3) hay plus pellets fortified with omega-3 oil. It was hypothesized that dietary supplementation with omega-3 fortified pellets will improve feedlot performance, meat-eating quality indices of IMF, FMP, and carcass characteristics. Lot-fed lambs on the MSM whole grain had the highest feed intake of 1.69 kg/day, followed by the control at 1.57 kg/day and the lowest in the omega-3 diet at 1.01 kg/day (p = 0.0001). However, the omega-3 diet had the highest average daily gain of 230 g/head/day (p = 0.0001), indicating the greatest feed efficiency since it had the best growth response with minimal feed intake. Post-slaughter evaluation of the Longissimus thoracis et lumborum muscle revealed significant treatment variations in IMF (p = 0.0001), FMP (p = 0.0001), pH (p = 0.0380), and wholesale French rack primal cut (p = 0.0001). Strong correlations (p < 0.05) between liveweight, temperature, pH, FMP, and IMF were observed. Similarly, significant correlations between carcass characteristics of total saleable meat yield, lean trim, fat trims, bones, and leg shank were evident (p < 0.05). However, there were no treatment differences in the final liveweight, GR fat depth, hot standard carcass weight, or dressing percentage. The findings indicate that feedlot performance, meat-eating quality traits such as IMF and FMP, and commercial wholesale French rack cuts can be further improved during feedlot finishing of TAW MARGRA lambs through dietary supplementation with omega-3 oils, and hence the tested hypothesis of improved meat quality attributes is partially confirmed.

Insight into the Enterovirus A71: A review.

Enterovirus A71 is a major causative pathogen of hand, foot and mouth disease. It has become a global public health threat, and is especially important for infants and young children in the Asian-Pacific countries. The enterovirus A71 is a non-enveloped virus of the Picornaviridae family having a single-stranded positive-sense RNA genome of about 7.4 kb which encodes the structural and nonstructural proteins. Currently there are no US FDA-approved vaccines or antiviral therapy available against enterovirus A71 infection. Although enterovirus A71 vaccines have been licenced in China, clinically approved vaccines for widespread vaccination programs are lacking. Substantial progress has recently been achieved on understanding the structure and function of enterovirus A71 proteins together with information on the viral genetic diversity and geographic distribution. The present review is intended to provide an overview on our current understanding of the molecular biology and epidemiology of enterovirus A71 which will aid the development of vaccines, therapeutics and other control strategies so as to bolster the preparedness for future enterovirus A71 outbreaks.

Effect of incremental proportions of Desmanthus spp. in isonitrogenous forage diets on growth performance, rumen fermentation and plasma metabolites of pen-fed growing Brahman, Charbray and Droughtmaster crossbred beef steers.

Desmanthus (Desmanthus spp.), a tropically adapted pasture legume, is highly productive and has the potential to reduce methane emissions in beef cattle. However, liveweight gain response to desmanthus supplementation has been inconclusive in ruminants. This study aimed to evaluate weight gain, rumen fermentation and plasma metabolites of Australian tropical beef cattle in response to supplementation with incremental levels of desmanthus forage legume in isonitrogenous diets. Forty-eight Brahman, Charbray and Droughtmaster crossbred beef steers were pen-housed and fed a basal diet of Rhodes grass (Chloris gayana) hay supplemented with 0, 15, 30 or 45% freshly chopped desmanthus forage on dry matter basis, for 140 days. Varying levels of lucerne (Medicago sativa) hay were added in the 0, 15 and 30% diets to ensure that all diets were isonitrogenous with the 45% desmanthus diet. Data were analyzed using the Mixed Model procedures of SAS software. Results showed that the proportion of desmanthus in the diet had no significant effect on steer liveweight, rumen volatile fatty acids molar proportions and plasma metabolites (P ≥ 0.067). Total bilirubin ranged between 3.0 and 3.6 µmol/L for all the diet treatments (P = 0.67). All plasma metabolites measured were within the expected normal range reported for beef cattle. Rumen ammonia nitrogen content was above the 10 mg/dl threshold required to maintain effective rumen microbial activity and maximize voluntary feed intake in cattle fed low-quality tropical forages. The average daily weight gains averaged 0.5 to 0.6 kg/day (P = 0.13) and were within the range required to meet the target slaughter weight for prime beef markets within 2.5 years of age. These results indicate that desmanthus alone or mixed with other high-quality legume forages can be used to supplement grass-based diets to improve tropical beef cattle production in northern Australia with no adverse effect on cattle health.

Lipid Metabolism, Carcass Characteristics and Longissimus dorsi Muscle Fatty Acid Composition of Tropical Crossbred Beef Cattle in Response to Desmanthus spp. Forage Backgrounding.

Lipid metabolism, carcass characteristics and fatty acid (FA) composition of the Longissimus dorsi (loin eye) muscle were evaluated in tropical crossbred steers backgrounded on Desmanthus spp. (desmanthus) with or without feedlot finishing. It was hypothesized that steers backgrounded on isonitrogenous diets augmented with incremental proportions of desmanthus will produce carcasses with similar characteristics and FA composition. Forty-eight Brahman, Charbray and Droughtmaster crossbred beef steers were backgrounded for 140 days on Rhodes grass (Chloris gayana) hay augmented with 0, 15, 30 or 45 percent desmanthus on dry matter basis. Lucerne (Medicago sativa) hay was added to the 0, 15 and 30 percent desmanthus diets to ensure that they were isonitrogenous with the 45 percent desmanthus diet. After backgrounding, the two heaviest steers in each pen were slaughtered and the rest were finished in the feedlot for 95 days before slaughter. Muscle biopsy samples were taken at the beginning and end of the backgrounding phase. Carcasses were sampled at slaughter for intramuscular fat (IMF) content, fat melting point (FMP) and FA composition analyses. Increasing the proportion of desmanthus in the diet led to a linear increase in docosanoic acid (p = 0.04) and omega-6/omega-3 polyunsaturated FA ratio (n-6/n-3 PUFA; p = 0.01), while docosahexaenoic acid decreased linearly (p = 0.01). Feedlot finishing increased hot carcass weight, subcutaneous fat depth at the P8 site and dressing percentage (p ≤ 0.04). The n-6/n-3 PUFA ratio was within the recommended < 5 for human diets. IMF was within the consumer-preferred ≥3% level for palatability. The hypothesis that steers backgrounded on isonitrogenous diets augmented with incremental proportions of desmanthus will produce similar carcass characteristics and FA composition was accepted. These findings indicate that a combination of tropical beef cattle backgrounding on desmanthus augmented forage and short-term feedlot finishing produces healthy and highly palatable meat.

Next Generation Sequencing of Single Nucleotide Polymorphic DNA-Markers in Selecting for Intramuscular Fat, Fat Melting Point, Omega-3 Long-Chain Polyunsaturated Fatty Acids and Meat Eating Quality in Tattykeel Australian White MARGRA Lamb.

Meat quality data can only be obtained after slaughter when selection decisions about the live animal are already too late. Carcass estimated breeding values present major precision problems due to low accuracy, and by the time an informed decision on the genetic merit for meat quality is made, the animal is already dead. We report for the first time, a targeted next-generation sequencing (NGS) of single nucleotide polymorphisms (SNP) of lipid metabolism genes in Tattykeel Australian White (TAW) sheep of the MARGRA lamb brand, utilizing an innovative and minimally invasive muscle biopsy sampling technique for directly quantifying the genetic worth of live lambs for health-beneficial omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), intramuscular fat (IMF), and fat melting point (FMP). NGS of stearoyl-CoA desaturase (SCD), fatty acid binding protein-4 (FABP4), and fatty acid synthase (FASN) genes identified functional SNP with unique DNA marker signatures for TAW genetics. The SCD g.23881050T>C locus was significantly associated with IMF, C22:6n-3, and C22:5n-3; FASN g.12323864A>G locus with FMP, C18:3n-3, C18:1n-9, C18:0, C16:0, MUFA, and FABP4 g.62829478A>T locus with IMF. These add new knowledge, precision, and reliability in directly making early and informed decisions on live sheep selection and breeding for health-beneficial n-3 LC-PUFA, FMP, IMF and superior meat-eating quality at the farmgate level. The findings provide evidence that significant associations exist between SNP of lipid metabolism genes and n-3 LC-PUFA, IMF, and FMP, thus underpinning potential marker-assisted selection for meat-eating quality traits in TAW lambs.

Are Interactions between Epicardial Adipose Tissue, Cardiac Fibroblasts and Cardiac Myocytes Instrumental in Atrial Fibrosis and Atrial Fibrillation?

Atrial fibrillation is very common among the elderly and/or obese. While myocardial fibrosis is associated with atrial fibrillation, the exact mechanisms within atrial myocytes and surrounding non-myocytes are not fully understood. This review considers the potential roles of myocardial fibroblasts and myofibroblasts in fibrosis and modulating myocyte electrophysiology through electrotonic interactions. Coupling with (myo)fibroblasts in vitro and in silico prolonged myocyte action potential duration and caused resting depolarization; an optogenetic study has verified in vivo that fibroblasts depolarized when coupled myocytes produced action potentials. This review also introduces another non-myocyte which may modulate both myocardial (myo)fibroblasts and myocytes: epicardial adipose tissue. Epicardial adipocytes are in intimate contact with myocytes and (myo)fibroblasts and may infiltrate the myocardium. Adipocytes secrete numerous adipokines which modulate (myo)fibroblast and myocyte physiology. These adipokines are protective in healthy hearts, preventing inflammation and fibrosis. However, adipokines secreted from adipocytes may switch to pro-inflammatory and pro-fibrotic, associated with reactive oxygen species generation. Pro-fibrotic adipokines stimulate myofibroblast differentiation, causing pronounced fibrosis in the epicardial adipose tissue and the myocardium. Adipose tissue also influences myocyte electrophysiology, via the adipokines and/or through electrotonic interactions. Deeper understanding of the interactions between myocytes and non-myocytes is important to understand and manage atrial fibrillation.

Nutritional Enhancement of Health Beneficial Omega-3 Long-Chain Polyunsaturated Fatty Acids in the Muscle, Liver, Kidney, and Heart of Tattykeel Australian White MARGRA Lambs Fed Pellets Fortified with Omega-3 Oil in a Feedlot System.

The aim of this research was to evaluate the nutritional enhancement of omega-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA) composition of edible lamb Longissimus thoracis et lumborum muscle, heart, kidney, and liver in response to dietary supplementation of lot-fed lambs with or without omega-3 oil fortified pellets. The hypothesis tested was that fortifying feedlot pellets with omega-3 oil will enhance the human health beneficial n-3 LC-PUFA composition of edible lamb muscle tissue and organs. Seventy-five Tattykeel Australian White lambs exclusive to the MARGRA brand, with an average body weight of 30 kg at six months of age, were randomly assigned to the following three dietary treatments of 25 lambs each, and lot-fed as a cohort for 47 days in a completely randomized experimental design: (1) Control grain pellets without oil plus hay; (2) Omega-3 oil fortified grain pellets plus hay; and (3) Commercial whole grain pellets plus hay. All lambs had ad libitum access to the basal hay diet and water. Post-slaughter fatty acid composition of the Longissimus thoracis et lumborum muscle, liver, kidney, and heart were determined using thee gas chromatography-mass spectrophotometry technique. Results indicated significant variations (p < 0.05) in fatty acid profiles between tissues and organs. Omega-3 oil fortified pellets significantly (p < 0.05) increased ≥C20 n-3 LC-PUFA (C20:5n-3 eicosapentaenoate, EPA + C22:5n3 docosapentaenoate, DPA + C22:6n3 docosahexanoate DHA); C18:3n-3 alpha-linolenate, ALA; C18:2 conjugated linoleic acid, CLA; total monounsaturated fatty acids, MUFA; polyunsaturated fatty acids, PUFA contents; and reduced the ratio of omega-6 to omega-3 fatty acids in all lamb organs and tissues without impacting shelf-life. The findings demonstrate that the inclusion of omega-3 oil in feedlot diets of lambs enhances the human health beneficial omega-3 long-chain polyunsaturated fatty acid profiles of edible muscle tissue and organs without compromising meat quality.

Growth Performance and Plasma Metabolites of Grazing Beef Cattle Backgrounded on Buffel or Buffel-Desmanthus Mixed Pastures.

Dietary crude protein and dry matter digestibility are among the major factors limiting feed intake and weight gain of cattle grazing native and improved pastures in the subtropics of Northern Australia during the dry season. Incorporating a suitable legume into grasses improves pasture quality and cattle weight gain, but only a limited number of legume pastures can establish and persist in cracking clay soils. This study aimed to evaluate the effect of Desmanthus inclusion in buffel grass (Cenchrus ciliaris) pastures on the plasma metabolite profile and growth performance of grazing beef cattle during the dry season. We hypothesised that backgrounding steers on buffel grass-Desmanthus mixed pastures would elicit significant changes in plasma glucose, bilirubin, creatinine, non-esterified fatty acids and ß-hydroxybutyrate, resulting in higher liveweight gains than in steers on buffel grass only pastures. Four hundred tropical composite steers were assigned to buffel grass only (n = 200) or buffel grass oversown with Desmanthus (11.5% initial sward dry matter) pastures (n = 200) and grazed for 147 days during the dry season. Desmanthus accounted for 6.2% sward dry matter at the end of grazing period. Plasma metabolites results showed that changes in ß-hydroxybutyrate, creatinine, bilirubin, glucose and non-esterified fatty acids were within the expected normal range for all the steers, indicating that with or without Desmanthus inclusion in the diet of grazing steers, animal health status was not compromised. It was also evident that Desmanthus inclusion in buffel grass pastures had no impact on the plasma metabolite profile, liveweight and daily weight gain of grazing steers. Therefore, our tested hypothesis of higher changes in plasma metabolite profile and higher liveweight gains due to backgrounding on low-level buffel grass-Desmanthus mixed pastures does not hold.

Testudines as Sentinels for Monitoring the Dissemination of Antibiotic Resistance in Marine Environments: An Integrative Review.

Dissemination of antibiotic resistance (AR) in marine environments is a global concern with a propensity to affect public health and many ecosystems worldwide. We evaluated the use of sea turtles as sentinel species for monitoring AR in marine environments. In this field, antibiotic-resistant bacteria have been commonly identified by using standard culture and sensitivity tests, leading to an overrepresentation of specific, culturable bacterial classes in the available literature. AR was detected against all major antibiotic classes, but the highest cumulative global frequency of resistance in all represented geographical sites was against the beta-lactam class by a two-fold difference compared to all other antibiotics. Wastewater facilities and turtle rehabilitation centres were associated with higher incidences of multidrug-resistant bacteria (MDRB) accounting for an average of 58% and 49% of resistant isolates, respectively. Furthermore, a relatively similar prevalence of MDRB was seen in all studied locations. These data suggest that anthropogenically driven selection pressures for the development of AR in sea turtles and marine environments are relatively similar worldwide. There is a need, however, to establish direct demonstrable associations between AR in sea turtles in their respective marine environments with wastewater facilities and other anthropogenic activities worldwide.

A systematic review of experimental evidence for antiviral effects of ivermectin and an in silico analysis of ivermectin's possible mode of action against SARS-CoV-2.

Viral infections remain a major cause of economic loss with an unmet need for novel therapeutic agents. Ivermectin is a putative antiviral compound; the proposed mechanism is the inhibition of nuclear translocation of viral proteins, facilitated by mammalian host importins, a necessary process for propagation of infections. We systematically reviewed the evidence for the applicability of ivermectin against viral infections including SARS-CoV-2 regarding efficacy, mechanisms and selective toxicity. The SARS-CoV-2 genome was mined to determine potential nuclear location signals for ivermectin and meta-analyses for in vivo studies included all comparators over time, dose range and viral replication in multiple organs. Ivermectin inhibited the replication of many viruses including those in Flaviviridae, Circoviridae and Coronaviridae families in vitro. Real and mock nuclear location signals were identified in SARS-CoV-2, a potential target for ivermectin and predicting a sequestration bait for importin ß, stopping infected cells from reaching a virus-resistant state. While pharmacokinetic evaluations indicate that ivermectin could be toxic if applied based on in vitro studies, inhibition of viral replication in vivo was shown for Porcine circovirus in piglets and Suid herpesvirus in mice. Overall standardized mean differences and 95% confidence intervals for ivermectin versus controls were -4.43 (-5.81, -3.04), p < 0.00001. Based on current results, the potential for repurposing ivermectin as an antiviral agent is promising. However, further work is needed to reconcile in vitro studies with clinical efficacy. Developing ivermectin as an additional antiviral agent should be pursued with an emphasis on pre-clinical trials in validated models of infection.

MARGRA Lamb Eating Quality and Human Health-Promoting Omega-3 Long-Chain Polyunsaturated Fatty Acid Profiles of Tattykeel Australian White Sheep: Linebreeding and Gender Effects.

Health-conscious consumers increasingly demand healthier, tastier, and more nutritious meat, hence the continuous need to meet market specifications and demand for high-quality lamb. We evaluated the longissimus dorsi muscle of 147 Tattykeel Australian White (TAW) sheep fed on antioxidant-rich ryegrass pastures exclusive to MAGRA lamb brand for meat eating quality parameters of intramuscular fat (IMF) content, fat melting point (FMP) and omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA). The aim was to assess the impact of linebreeding and gender on pasture-fed lamb eating quality and to test the hypothesis that variation in healthy lamb eating quality is a function of lamb gender and not its antioxidant status or inbreeding coefficient (IC). After solid-phase extraction and purification, phenolics and antioxidant enzyme activities were analysed by high-performance liquid chromatography and mass spectrometry. IMF and fatty acid composition were determined using solvent extraction and gas chromatography, respectively. IC was classified into low (0-5%), medium (6-10%) and high (>10%) and ranged from 0-15.6%. FMP and IMF ranged from 28 to 39 °C and 3.4% to 8.2%, with overall means of 34.6 ± 2.3 °C and 4.4 ± 0.2%, respectively, and n-3 LC-PUFA ranged from "source" to "good source" levels of 33-69 mg/100 g. Ewes had significantly (P ˂ 0.0001) higher IMF, C22:5n-3 (DPA), C22:6n-3 (DHA), C18:3n-6, C20:3, C22:4n-6, C22:5n-6, total monounsaturated (MUFA), PUFA and Σn-3 fatty acids and lower total saturated fatty acids (SFA) and FMP, than rams. As IC increased, there were no differences in FMP and IMF. Folin-Ciocalteu total phenolics, ferric reducing antioxidant power and antioxidant activities of glutathione peroxidase, catalase and superoxide dismutase enzymes did not differ by either gender or IC. This study provides evidence that IC is inconsequential in affecting antioxidant status, IMF, FMP and n-3 LC-PUFA in linebred and pasture-fed TAW sheep because the observed variation in individual fatty acids was mainly driven by gender differences between ewes and rams, hence the need to accept the tested hypothesis. This finding reinforces the consistent healthy eating quality of MARGRA lamb brand from TAW sheep regardless of its linebred origin.

Genetics of Omega-3 Long-Chain Polyunsaturated Fatty Acid Metabolism and Meat Eating Quality in Tattykeel Australian White Lambs.

Meat eating quality with a healthy composition hinges on intramuscular fat (IMF), fat melting point (FMP), tenderness, juiciness, flavour and omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) content. These health-beneficial n-3 LC-PUFA play significant roles in optimal cardiovascular, retinal, maternal and childhood brain functions, and include alpha linolenic (ALA), eicosapentaenoic (EPA), docosahexaenoic (DHA) and docosapentaenoic (DPA) acids. The primary objective of this review was to access, retrieve, synthesise and critically appraise the published literature on the synthesis, metabolism and genetics of n-3 LC-PUFA and meat eating quality. Studies on IMF content, FMP and fatty acid composition were reviewed to identify knowledge gaps that can inform future research with Tattykeel Australian White (TAW) lambs. The TAW is a new sheep breed exclusive to MARGRA brand of lamb with an outstanding low fat melting point (28-39°C), high n-3 LC-PUFA EPA+DHA content (33-69mg/100g), marbling (3.4-8.2%), tenderness (20.0-38.5N) and overall consumer liking (7.9-8.5). However, correlations between n-3 LC-PUFA profile, stearoyl-CoA desaturase (SCD), fatty acid binding protein 4 (FABP4), fatty acid synthase (FASN), other lipogenic genes and meat quality traits present major knowledge gaps. The review also identified research opportunities in nutrition-genetics interactions aimed at a greater understanding of the genetics of n-3 LC-PUFA, feedlot finishing performance, carcass traits and eating quality in the TAW sheep. It was concluded that studies on IMF, FMP and n-3 LC-PUFA profiles in parental and progeny generations of TAW sheep will be foundational for the genetic selection of healthy lamb eating qualities and provide useful insights into their correlations with SCD, FASN and FABP4 genes.

Diet and Genetics Influence Beef Cattle Performance and Meat Quality Characteristics.

A comprehensive review of the impact of tropical pasture grazing, nutritional supplementation during feedlot finishing and fat metabolism-related genes on beef cattle performance and meat-eating traits is presented. Grazing beef cattle on low quality tropical forages with less than 5.6% crude protein, 10% soluble starches and 55% digestibility experience liveweight loss. However, backgrounding beef cattle on high quality leguminous forages and feedlot finishing on high-energy diets increase meat flavour, tenderness and juiciness due to improved intramuscular fat deposition and enhanced mono- and polyunsaturated fatty acids. This paper also reviews the roles of stearoyl-CoA desaturase, fatty acid binding protein 4 and fatty acid synthase genes and correlations with meat traits. The review argues that backgrounding of beef cattle on Desmanthus, an environmentally well-adapted and vigorous tropical legume that can persistently survive under harsh tropical and subtropical conditions, has the potential to improve animal performance. It also identifies existing knowledge gaps and research opportunities in nutrition-genetics interactions aimed at a greater understanding of grazing nutrition, feedlot finishing performance, and carcass traits of northern Australian tropical beef cattle to enable red meat industry players to work on marbling, juiciness, tenderness and overall meat-eating characteristics.

A volume-pressure tail cuff method for hemodynamic parameters: Comparison of restraint and light isoflurane anesthesia in normotensive male Lewis rats.

INTRODUCTION: A volume-pressure sensor and tail-cuff method for monitoring blood pressure is non-invasive and inexpensive. This method requires animals to be restrained or subjected to anesthesia, but comparative effects of these manipulations on hemodynamic parameters have not been documented. METHODS: Using a volume-pressure sensor and tail-cuff, we serially measured blood pressure and heart rate in normotensive adult male Lewis rats after light isoflurane-induced anesthesia (5% induction, 1% maintenance) and, following untrained restraint. Blood pressure was recorded until the acquisition of three complete measurements without the range of replicate mean arterial pressures exceeding 15 mmHg (steady-state). RESULTS: Averages for the entire series of consecutive measurements indicated that restraint yielded significantly higher systolic and diastolic blood pressure than anesthesia (P < .05), but heart rate was not affected. Following stabilization at steady-state, there were no significant differences in intra- or inter-day hemodynamic values between the restraint and isoflurane groups. The inter-day coefficient of variation for systolic pressure was 13-23% for isoflurane and 9-14% for restraint. Bland-Altman analysis showed wide limits of agreement (±59 mmHg systolic; ±49 mmHg diastolic pressure) between restraint and isoflurane measurements. DISCUSSION: Isoflurane caused more variability but there was agreement in BP evaluation by the isoflurane and restraint methods. Using the VPR system, light isoflurane-induced anesthesia and restraint could effectively be used to screen and quantify overt changes in hemodynamic parameters for cardiovascular research utilizing laboratory rodents.

Bacteriophage versus antibiotic therapy on gut bacterial communities of juvenile green turtle, Chelonia mydas.

Green turtles are endangered marine herbivorous hindgut fermenters that contribute to a variety of marine ecosystems. Debilitated turtles are often rehabilitated in turtle hospitals. Since accurate diagnosis of disease is difficult, broad-spectrum antibiotics are routinely used as a general treatment, potentially causing collateral damage to the gut microbiome of the patient. Here, we evaluated the concept of the application of bacteriophage (phages) to eliminate targeted intestinal bacteria as an alternative to a broad-spectrum antibiotic (enrofloxacin) in clinically healthy, captive green turtles. Additionally, the impact of a broad-spectrum antibiotic (enrofloxacin) and phage therapy on the gut bacterial communities of green turtles was evaluated. Gut bacterial communities in faecal samples were analysed by sequencing the V1-V3 regions of the bacterial 16S rRNA. Bacteria-specific phage cocktails significantly (P < 0.05) reduced targeted Acinetobacter in phage-treated turtles during the therapy. Compared to control, no significant difference was observed in the bacterial diversity and compositions in phage-treated turtles. In contrast, bacterial diversity was significantly (P < 0.05) reduced in antibiotic-treated turtles at day 15 and throughout the trial. The alteration in the bacterial microbiota of antibiotic-treated turtles was largely due to an increase in abundance of Gram-positive Firmicutes and a concurrent decrease in Gram-negative Bacteroidetes, Proteobacteria and Verrucomicrobia. Additionally, we observed the relative abundance of several bacteria at lower taxonomic level was much less affected by phages than by antibiotics. These data offer the proof of concept of phage therapy to manipulate transient as well as indigenous bacterial flora in gut-related dysbiosis of turtles.