Olanzapine's effects on hypothalamic transcriptomics and kinase activity.

Olanzapine is a second-generation antipsychotic that disrupts metabolism and is associated with an increased risk of type 2 diabetes. The hypothalamus is a key region in the control of whole-body metabolic homeostasis. The objective of the current study was to determine how acute peripheral olanzapine administration affects transcription and serine/threonine kinase activity in the hypothalamus. Hypothalamus samples from rats were collected following the pancreatic euglycemic clamp, thereby allowing us to study endpoints under steady state conditions for plasma glucose and insulin. Olanzapine stimulated pathways associated with inflammation, but diminished pathways associated with the capacity to combat endoplasmic reticulum stress and G protein-coupled receptor activity. These pathways represent potential targets to reduce the incidence of type 2 diabetes in patients taking antipsychotics.

Retinoic Acid-Mediated Inhibition of Mouse Coronavirus Replication Is Dependent on IRF3 and CaMKK.

The ongoing COVID-19 pandemic has revealed the shortfalls in our understanding of how to treat coronavirus infections. With almost 7 million case fatalities of COVID-19 globally, the catalog of FDA-approved antiviral therapeutics is limited compared to other medications, such as antibiotics. All-trans retinoic acid (RA), or activated vitamin A, has been studied as a potential therapeutic against coronavirus infection because of its antiviral properties. Due to its impact on different signaling pathways, RA's mechanism of action during coronavirus infection has not been thoroughly described. To determine RA's mechanism of action, we examined its effect against a mouse coronavirus, mouse hepatitis virus strain A59 (MHV). We demonstrated that RA significantly decreased viral titers in infected mouse L929 fibroblasts and RAW 264.7 macrophages. The reduced viral titers were associated with a corresponding decrease in MHV nucleocapsid protein expression. Using interferon regulatory factor 3 (IRF3) knockout RAW 264.7 cells, we demonstrated that RA-induced suppression of MHV required IRF3 activity. RNA-seq analysis of wildtype and IRF3 knockout RAW cells showed that RA upregulated calcium/calmodulin (CaM) signaling proteins, such as CaM kinase kinase 1 (CaMKK1). When treated with a CaMKK inhibitor, RA was unable to upregulate IRF activation during MHV infection. In conclusion, our results demonstrate that RA-induced protection against coronavirus infection depends on IRF3 and CaMKK.

Developmental pyrethroid exposure disrupts molecular pathways for MAP kinase and circadian rhythms in mouse brain.

Neurodevelopmental disorders (NDDs) are a category of pervasive disorders of the developing nervous system with few or no recognized biomarkers. A significant portion of the risk for NDDs, including attention deficit hyperactivity disorder (ADHD), is contributed by the environment, and exposure to pyrethroid pesticides during pregnancy has been identified as a potential risk factor for NDD in the unborn child. We recently showed that low-dose developmental exposure to the pyrethroid pesticide deltamethrin in mice causes male-biased changes to ADHD- and NDD-relevant behaviors as well as the striatal dopamine system. Here, we used an integrated multiomics approach to determine the broadest possible set of biological changes in the mouse brain caused by developmental pyrethroid exposure (DPE). Using a litter-based, split-sample design, we exposed mouse dams during pregnancy and lactation to deltamethrin (3 mg/kg or vehicle every 3 days) at a concentration well below the EPA-determined benchmark dose used for regulatory guidance. We raised male offspring to adulthood, euthanized them, and pulverized and divided whole brain samples for split-sample transcriptomics, kinomics and multiomics integration. Transcriptome analysis revealed alterations to multiple canonical clock genes, and kinome analysis revealed changes in the activity of multiple kinases involved in synaptic plasticity, including the mitogen-activated protein (MAP) kinase ERK. Multiomics integration revealed a dysregulated protein-protein interaction network containing primary clusters for MAP kinase cascades, regulation of apoptosis, and synaptic function. These results demonstrate that DPE causes a multi-modal biophenotype in the brain relevant to ADHD and identifies new potential mechanisms of action.

Developmental pyrethroid exposure disrupts folate metabolism in mouse brain.

Environmental and genetic risk factors, and their interactions, contribute significantly to the etiology of neurodevelopmental disorders (NDDs). Recent epidemiology studies have implicated pyrethroid pesticides as an environmental risk factor for autism and developmental delay. Our previous research showed that low-dose developmental exposure to the pyrethroid pesticide deltamethrin in mice caused male-biased changes in the brain and in NDD-relevant behaviors in adulthood. Here, we used a metabolomics approach to determine the broadest possible set of metabolic changes in the adult male mouse brain caused by low-dose pyrethroid exposure during development. Using a litter-based design, we exposed mouse dams during pregnancy and lactation to deltamethrin (3 mg/kg or vehicle every 3 days) at a concentration well below the EPA-determined benchmark dose used for regulatory guidance. We raised male offspring to adulthood and collected whole brain samples for untargeted high-resolution metabolomics analysis. Developmentally exposed mice had disruptions in 116 metabolites which clustered into pathways for folate biosynthesis, retinol metabolism, and tryptophan metabolism. As a cross-validation, we integrated metabolomics and transcriptomics data from the same samples, which confirmed previous findings of altered dopamine signaling. These results suggest that pyrethroid exposure during development leads to disruptions in folate metabolism in the adult brain, which may inform both prevention and therapeutic strategies.

In Silico Transcriptome-based Screens Identify Epidermal Growth Factor Receptor Inhibitors as Therapeutics for Noise-induced Hearing Loss.

Noise-Induced Hearing Loss (NIHL) represents a widespread disease for which no therapeutics have been approved by the Food and Drug Administration (FDA). Addressing the conspicuous void of efficacious in vitro or animal models for high throughput pharmacological screening, we utilized an in silico transcriptome-oriented drug screening strategy, unveiling 22 biological pathways and 64 promising small molecule candidates for NIHL protection. Afatinib and zorifertinib, both inhibitors of the Epidermal Growth Factor Receptor (EGFR), were validated for their protective efficacy against NIHL in experimental zebrafish and murine models. This protective effect was further confirmed with EGFR conditional knockout mice and EGF knockdown zebrafish, both demonstrating protection against NIHL. Molecular analysis using Western blot and kinome signaling arrays on adult mouse cochlear lysates unveiled the intricate involvement of several signaling pathways, with particular emphasis on EGFR and its downstream pathways being modulated by noise exposure and Zorifertinib treatment. Administered orally, Zorifertinib was successfully detected in the perilymph fluid of the inner ear in mice with favorable pharmacokinetic attributes. Zorifertinib, in conjunction with AZD5438 - a potent inhibitor of cyclin dependent kinase 2 - produced synergistic protection against NIHL in the zebrafish model. Collectively, our findings underscore the potential application of in silico transcriptome-based drug screening for diseases bereft of efficient screening models and posit EGFR inhibitors as promising therapeutic agents warranting clinical exploration for combatting NIHL. Highlights: In silico transcriptome-based drug screens identify pathways and drugs against NIHL.EGFR signaling is activated by noise but reduced by zorifertinib in mouse cochleae.Afatinib, zorifertinib and EGFR knockout protect against NIHL in mice and zebrafish.Orally delivered zorifertinib has inner ear PK and synergizes with a CDK2 inhibitor.

Antipsychotics impair regulation of glucose metabolism by central glucose.

Hypothalamic detection of elevated circulating glucose triggers suppression of endogenous glucose production (EGP) to maintain glucose homeostasis. Antipsychotics alleviate symptoms associated with schizophrenia but also increase the risk for impaired glucose metabolism. In the current study, we examined whether two acutely administered antipsychotics from different drug classes, haloperidol (first generation antipsychotic) and olanzapine (second generation antipsychotic), affect the ability of intracerebroventricular (ICV) glucose infusion approximating postprandial levels to suppress EGP. The experimental protocol consisted of a pancreatic euglycemic clamp, followed by kinomic and RNA-seq analyses of hypothalamic samples to determine changes in serine/threonine kinase activity and gene expression, respectively. Both antipsychotics inhibited ICV glucose-mediated increases in glucose infusion rate during the clamp, a measure of whole-body glucose metabolism. Similarly, olanzapine and haloperidol blocked central glucose-induced suppression of EGP. ICV glucose stimulated the vascular endothelial growth factor (VEGF) pathway, phosphatidylinositol 3-kinase (PI3K) pathway, and kinases capable of activating KATP channels in the hypothalamus. These effects were inhibited by both antipsychotics. In conclusion, olanzapine and haloperidol impair central glucose sensing. Although results of hypothalamic analyses in our study do not prove causality, they are novel and provide the basis for a multitude of future studies.

Detection of Giardia and helminths in Western Europe at local K9 (canine) sites (DOGWALKS Study).

BACKGROUND: Intestinal parasite contamination from infected dogs can place other dogs and humans at risk. A study was initiated to estimate the prevalence of canine intestinal parasitism by collecting fecal samples in cities across Western Europe. METHODS: Fresh fecal samples were collected from 2469 dogs visiting 164 parks in 33 cities across 12 countries. Each owner responded to a questionnaire focusing on their dog's signalment and recent anthelmintic treatment history. The collected samples were examined for hookworms, whipworms, ascarids and Giardia using a coproantigen diagnostic immunoassay and microscopy following centrifugal flotation. RESULTS: Nematodes or Giardia were detected in at least one sample from 100% of cities and in 93.3% of parks. Nematodes were detected in 57% of parks. Overall, 22.8% of dogs tested positive for an intestinal parasite, with Giardia being the most commonly identified parasites (17.3% of dogs, 83.5% of parks). For nematode infection, 7.6% of all dogs tested positive, with 9.9% of dogs aged < 1 year infected, 7.7% of those aged 1-3 years, 7.3% of those aged 4-6 years and 6.6% of those aged ≥ 7 years. Among the nematodes detected, ascarids were the most prevalent (3.6% of dogs, parks, 28.7% of parks), being most common in dogs aged < 1 year but also present in older dogs, including those aged ≥ 7 years. Hookworms and whipworms were detected in 3.2% and 2.3% of dogs of all ages, respectively, and in 37.2% and 17.7% of parks, respectively. A larger proportion of fecal samples tested positive with the coproantigen immunoassay than with centrifugal flotation. Positive test results for Giardia were sevenfold higher when both diagnostic tests were used than when centrifugal flotation alone was used, and there were 60% more positive test results for nematodes when both tests were used than when flotation alone was used. Overall, 77.2% of owners reported previous anthelmintic treatment, among whom at least 62.7% failed to follow recommended treatment frequency. Dogs receiving anthelmintic within the previous month had a lower percentage of nematode infection than those in which > 1 month had passed since the previous dose. CONCLUSIONS: The prevalence estimates of intestinal parasite infections in dogs reported here highlight the need for owner education concerning guidelines for regular testing and treatment, even in older dogs. Failure to adhere to guidelines can result in ongoing transmission of these infections, including those with zoonotic potential. Combining coproantigen immunoassay with centrifugal flotation for diagnostic testing and regular anthelmintic treatment are important measures for ensuring optimal intestinal parasite control.

Variability in non-core vaccination rates of dogs and cats in veterinary clinics across the United States.

Vaccination guidelines for dogs and cats indicate that core vaccines (for dogs, rabies, distemper, adenovirus, parvovirus; for cats, feline parvovirus, herpes virus-1, calicivirus) are essential to maintain health, and that non-core vaccines be administered according to a clinician's assessment of a pet's risk of exposure and susceptibility to infection. A reliance on individual risk assessment introduces the potential for between-practice inconsistencies in non-core vaccine recommendations. A study was initiated to determine non-core vaccination rates of dogs (Leptospira, Borrelia burgdorferi, Bordetella bronchiseptica, canine influenza virus) and cats (feline leukemia virus) in patients current for core vaccines in veterinary practices across the United States. Transactional data for 5,531,866 dogs (1,670 practices) and 1,914,373 cats (1,661 practices) were retrieved from practice management systems for the period November 1, 2016 through January 1, 2020, deidentified and normalized. Non-core vaccination status was evaluated in 2,798,875 dogs and 788,772 cats that were core-vaccine current. Nationally, median clinic vaccination rates for dogs were highest for leptospirosis (70.5%) and B. bronchiseptica (68.7%), and much lower for canine influenza (4.8%). In Lyme-endemic states, the median clinic borreliosis vaccination rate was 51.8%. Feline leukemia median clinic vaccination rates were low for adult cats (34.6%) and for kittens and 1-year old cats (36.8%). Individual clinic vaccination rates ranged from 0 to 100% for leptospirosis, B. bronchiseptica and feline leukemia, 0-96% for canine influenza, and 0-94% for borreliosis. Wide variation in non-core vaccination rates between clinics in similar geographies indicates that factors other than disease risk are driving the use of non-core vaccines in pet dogs and cats, highlighting a need for veterinary practices to address gaps in patient protection. Failure to implement effective non-core vaccination strategies leaves susceptible dogs and cats unprotected against vaccine-preventable diseases.

Early onset of pre-lethal effects of lotilaner (Credelio®) on Amblyomma americanum ticks on experimentally infested dogs.

BACKGROUND: The speed with which acaricides paralyze and kill ticks is relevant to impeding pathogen transmission. The objective of this study was to assess early-onset lotilaner effects on the motility and weights of Amblyomma americanum ticks collected from treated dogs. METHODS: Twelve healthy dogs were randomized between two groups to receive either lotilaner (Credelio®) on Day 0 or to be sham treated. On Day 7, 25 male and 25 female A. americanum were placed under bandages, two on each flank of each dog. After 30 or 45 min, all unattached ticks were removed and T = 0 was set. At T = 2, 4, 8 and 24 h post attachment, 5 attached ticks removed from each bandage on each dog were weighed, assessed by blinded observers for righting ability and movement recorded. RESULTS: After the infestation period significantly fewer treated than control dogs had 20 ticks attached (50.0% versus 91.7%, P = 0.0015). At 24 h post attachment, mean weights of ticks from treated dogs (males 1.69 mg; females 2.72) were significantly less than ticks from controls (males 2.66 mg; females 4.67) (Pmale = 0.0002; Pfemale < 0.0001). Mean tick weights from the treated group were significantly lower at 24 h than at earlier time points (Pmale < 0.0307; Pfemale = 0.0021). At 4 and 8 h, significantly fewer ticks from treated (14.3%, 0.0%, respectively) than from control dogs could right (73.3%, 70.0%) (P4h < 0.0001; P8h = 0.0024) (at 24 h, all ticks from treated dogs were dead), and distance moved was significantly less at all time points (P2h = 0.0413; P4h, P8h < 0.0001). Mean and maximum velocity of ticks from treated dogs were significantly lower, relative to controls, at 4 and 8 h (P ≤ 0.0001). Within the treated group, collected ticks had significantly lower mean and maximum velocities at 4 and 8 h compared to 2 h (Pmean < 0.0042; Pmax < 0.0194). CONCLUSION: The observed changes indicate that lotilaner may disrupt tick attachment. In ticks that attached, a progressive impairment of neuromuscular processes began within 2 h. Those irreversible changes could substantially reduce the risk of pathogen transmission from tick to host.

Correction to: Detection of gastrointestinal parasitism at recreational canine sites in the USA: the DOGPARCS study.

An amendment to this paper has been published and can be accessed via the original article.

Detection of gastrointestinal parasitism at recreational canine sites in the USA: the DOGPARCS study.

BACKGROUND: The rapid growth in off-leash dog parks provides opportunity for canine socialization activities but carries risk of exposure to intestinal parasites. This study assessed the prevalence of these infections in dogs visiting off-leash dog parks. METHODS: Fresh defecations were collected from dogs visiting parks in 30 metropolitan areas across the USA. Samples were analyzed by coproantigen immunoassay (CAI) (Fecal Dx® and Giardia Test, IDEXX Laboratories, Inc.) and zinc sulfate centrifugal flotation (CF). Owners responded to a questionnaire on their dog's signalment and use of heartworm/intestinal parasite control medications (HWCM). RESULTS: Samples were examined from 3006 dogs, 87.9% aged at least 12 months, visiting 288 parks. At least one intestinal parasite was detected in 622 (20.7%) samples, nematodes in 263 (8.8%), with hookworms, whipworms and ascarids in 7.1, 1.9 and 0.6% of samples, respectively. A sample positive for one or more intestinal parasites was found in 245 (85.1%) parks, with nematodes found in 143 (49.7%). Combined, CAI and CF detected 78.4% more intestinal nematode infections than CF alone. Hookworm and whipworm infections were detected in all age groups, but ascarids were only detected in dogs less than 4 years-old. Approximately 42% of dogs aged less than 1 year were positive for nematodes or Giardia. Based on owner reports, HWCM was current for 68.8% of dogs, dogs previously diagnosed with intestinal parasitism were more likely to be receiving a HWCM than those without such history, and a significantly lower (P = 0.0003) proportion of dogs receiving a HWCM were positive for intestinal nematodes compared with those not on such medication. CONCLUSIONS: Intestinal parasites, the most common of which were Giardia, Ancylostoma caninum and Trichuris vulpis, were found in 20% of dogs and 85% of dog parks across the USA. Enhanced detection of canine intestinal parasitism was achieved by combining CF and CAI. Canine intestinal parasites are common across the USA and dog health can be improved by regular testing of fecal samples and routine administration of medications effective against the most common infections.

In-home assessment of flea control and dermatologic lesions in dogs provided by lotilaner (Credelio®) and spinosad (Comfortis®) in west central Florida.

Post-launch field investigations of recently-approved flea control products establish an efficacy baseline and in subsequent years can detect any efficacy decline suggestive of emerging resistance. As part of a continuing program of yearly assessment of flea control products in west central Florida, this study, using client-owned dogs, investigated the efficacy of lotilaner and spinosad in controlling fleas and in alleviating dermatologic signs likely associated with flea infestations. Forty-four qualifying households were randomized to either a lotilaner (Credelio®) (minimum dose rate 20 mg/kg) or a spinosad (Comfortis®) (30 mg/kg) group, with 33 and 36 dogs in each group, respectively. On Days 0 and 28 (±2) all dogs in each household were treated with the allocated product according to label directions, and all household cats received spinetoram (Cheristin®). On Day 0 and at weekly intervals through Day 56 (±2), on-animal and premises flea burdens were enumerated, a veterinary dermatologist scored integumental changes using canine atopic dermatitis extent and severity index (CADESI)-4 and flea allergy dermatitis (FAD) scales, and owners scored pruritus using the validated canine pruritus severity scale (CPSS). At study entry geometric mean flea counts were 33.2 and 29.9 in the lotilaner and spinosad groups, respectively. For both groups, reductions in flea counts were > 99% at the first post-treatment assessment (Week 1), and 100% from Week 6 through the final assessment (Week 8) when all study dogs were flea-free. For both groups, at each timepoint, flea counts on dogs and in traps were significantly reduced compared to the initial assessment (p < 0.001), as were improvements in median CADESI-4, FAD and CPSS scores (p ≤ 0.001). At Week 4, the geometric mean flea count on dogs in the lotilaner group (0.1) was significantly lower than that of dogs in the spinosad group (0.6) (p = 0.027), significantly fewer dogs in the lotilaner group were found to have fleas (p = 0.034), and mean owner-rated pruritus scores were significantly lower (p = 0.025). Under field conditions favoring heavy flea challenge, two consecutive monthly treatments of dogs with either lotilaner or spinosad produced a 100% reduction in canine flea infestations and dramatic improvements in dermatologic lesions and pruritus, based on scoring by a veterinary dermatologist and by dog owners. Household flea burdens were driven to extinction in all but one home in each treatment group.

In-home assessment of flea control and dermatologic lesions in dogs provided by lotilaner (Credelio®) and spinosad (Comfortis®) in west central Florida.

Post-launch field investigations of recently-approved flea control products establish an efficacy baseline and in subsequent years can detect any efficacy decline suggestive of emerging resistance. As part of a continuing program of yearly assessment of flea control products in west central Florida, this study, using client-owned dogs, investigated the efficacy of lotilaner and spinosad in controlling fleas and in alleviating dermatologic signs likely associated with flea infestations. Forty-four qualifying households were randomized to either a lotilaner (Credelio®) (minimum dose rate 20 mg/kg) or a spinosad (Comfortis®) (30 mg/kg) group, with 33 and 36 dogs in each group, respectively. On Days 0 and 28 (±2) all dogs in each household were treated with the allocated product according to label directions, and all household cats received spinetoram (Cheristin®). On Day 0 and at weekly intervals through Day 56 (±2), on-animal and premises flea burdens were enumerated, a veterinary dermatologist scored integumental changes using canine atopic dermatitis extent and severity index (CADESI)-4 and flea allergy dermatitis (FAD) scales, and owners scored pruritus using the validated canine pruritus severity scale (CPSS). At study entry geometric mean flea counts were 33.2 and 29.9 in the lotilaner and spinosad groups, respectively. For both groups, reductions in flea counts were > 99% at the first post-treatment assessment (Week 1), and 100% from Week 6 through the final assessment (Week 8) when all study dogs were flea-free. For both groups, at each timepoint, flea counts on dogs and in traps were significantly reduced compared to the initial assessment (p < 0.001), as were improvements in median CADESI-4, FAD and CPSS scores (p ≤ 0.001). At Week 4, the geometric mean flea count on dogs in the lotilaner group (0.1) was significantly lower than that of dogs in the spinosad group (0.6) (p = 0.027), significantly fewer dogs in the lotilaner group were found to have fleas (p = 0.034), and mean owner-rated pruritus scores were significantly lower (p = 0.025). Under field conditions favoring heavy flea challenge, two consecutive monthly treatments of dogs with either lotilaner or spinosad produced a 100% reduction in canine flea infestations and dramatic improvements in dermatologic lesions and pruritus, based on scoring by a veterinary dermatologist and by dog owners. Household flea burdens were driven to extinction in all but one home in each treatment group.

Effectiveness and residual speed of flea kill of a novel spot on formulation of spinetoram (Cheristin®) for cats.

BACKGROUND: A spot-on spinetoram formulation (Cheristin®) was developed to eliminate fleas from infested cats. This paper describes three spinetoram studies: two for registration (Studies 1 and 2), and one comparing residual speed of kill (SOK) with topically applied fipronil/(S)-methoprene (FSM) and imidacloprid (Study 3). METHODS: Cats were randomized to treatment based on flea counts from infestations placed within 2 weeks prior to treatment. In Studies 1 and 2, groups were untreated control and spinetoram; in Study 3, groups were untreated control, spinetoram, FSM and imidacloprid, all applied per label on Day 0. Cats were infested the day before treatment. In Studies 1 and 2, counts were completed 48 h post-treatment and after weekly challenges through 5 weeks. In Study 3, infestations were completed weekly through Day 28, with counts 1, 4, 8 and 12 h after treatment or post-infestation (PI). Efficacy was determined on geometric mean flea count reductions compared with controls, and in Study 3 mean flea counts in spinetoram-groups were compared with those in FSM and imidacloprid groups. RESULTS: In Studies 1 and 2, spinetoram effectiveness was 100% against existing infestations, and at least 96% through Day 37. In Study 3 mean counts were not significantly different from controls in any group until 8 h post-treatment when imidacloprid counts were significantly lower than spinetoram counts, which were in turn significantly lower than FSM counts (P < 0.05). At 1 h PI spinetoram-group counts were significantly lower (P < 0.05) than counts in: controls, all days; imidacloprid, Days 7, 14, and 28; FSM, Days 14 and 28. At 4 h PI, spinetoram mean counts were significantly lower (P < 0.05) relative to: controls, all days; imidacloprid, Days 7, 14 and 21; FSM, Days 7, 14, 21 and 28 (P < 0.05). On multiple occasions, at 8 and 12 h PI, mean counts were significantly lower (P < 0.05) for spinetoram than for imidacloprid and FSM; at no point were FSM or imidacloprid significantly more effective than spinetoram against new infestations. All treatments were well tolerated. CONCLUSIONS: Spinetoram was highly effective for at least 1 month post-treatment and provided more rapid month-long residual SOK than FSM or imidacloprid.

Management of Head Louse Infestations in the United States-A Literature Review.

UNLABELLED: Head lice are a source of scalp irritation, social disruption, and loss of school time. Health care providers need authoritative information to help avoid the costs and risks of ineffective treatment. A review was completed to provide relevant information on infestation treatments available in the United States. Three major biomedical databases were searched from 1985, when current products were first available, to 2014, focusing on U.S. REPORTS: A total of 579 references remained after duplicates were removed. A search of the U.S. Food and Drug Administration website and labels of approved products were reviewed. A marked decline in the effectiveness of permethrin and synergized pyrethrins was found, probably because of resistance arising from widespread and indiscriminate use, and the emergence of knockdown resistance mutations. The potential toxicity of lindane in the setting of readily available, safer, and more effective alternatives, should limit its use. Prescription products shown to be safe and effective with a single application, without nit combing, are topical ivermectin, malathion, and spinosad, whereas benzyl alcohol requires two applications. Home remedies such as mayonnaise, and essential oils, have not been demonstrated to be safe or effective, and may carry potential for severe adverse events. The high risk of failure of over-the-counter treatments in eliminating head louse infestations drives a need for health care provider recognition of the limitations of current treatments and for judicious use of treatments that remain effective.

Speed of flea knockdown of spinosad compared to afoxolaner, and of spinosad through 28 days post-treatment in controlled laboratory studies.

BACKGROUND: The speed of flea knockdown by different products and their duration of effectiveness are factors which affect veterinarian prescribing decisions. To further validate the month-long pulicidal effectiveness of spinosad and determine its rate of flea knockdown to that of afoxolaner, three studies were conducted in two laboratories in the United States, utilizing flea infestations from colonies which are regularly refreshed through introduction of locally caught fleas. METHODS: All study assessors were blinded, dogs were ranked by pre-study flea counts and randomized accordingly, and treatments administered on Day 0. All studies included a negative control group; two also included an afoxolaner group. In one study, flea challenges for treated and control dogs (10 per group) were completed 21 and 28 days after treatment and counts were performed 24 h later. In each of two speed-of-knockdown (SOK) studies, 36 dogs were randomized, six dogs per group, to: untreated controls; administered oral afoxolaner (2.6-6.2 mg/kg); or oral spinosad (32.1-59.2 mg/kg). In the SOK studies, live fleas from Day -1 infestations were counted after being combed off at 1 and 3 h after treatment, and after reinfestations on Day 7. RESULTS: There were no treatment-related adverse events. Spinosad was 98.6% effective at 28 days post treatment. For SOK, geometric mean live flea counts for afoxolaner were not different from controls at any assessment. For spinosad, all mean counts were significantly lower than in controls (p ≤ 0.0128) except at 1 h post treatment in both studies. Spinosad was significantly more effective than afoxolaner in both studies at 3 h post treatment (p ≤ 0.0065) and post-Day 7 infestation (p ≤ 0.0054), and at 1 h post treatment (p = 0.0276) and post-Day 7 infestation in one study. CONCLUSIONS: These data validate spinosad's faster onset of flea knockdown than afoxolaner against infestations present at the time of treatment, and faster residual speed of flea knockdown for at least 7 days post treatment, and confirm spinosad's extended residual speed of kill for at least 28 days post treatment.

Reduced Blood Loss and Transfusion Rates: Additional Benefits of Local Infiltration Anaesthesia in Knee Arthroplasty Patients.

We hypothesised that local infiltration of anaesthesia (LIA) containing adrenaline may reduce peri-operative blood loss and transfusion requirements in primary total knee arthroplasty patients (TKA) when compared to simple patient control analgesia (PCA). In this retrospective cohort study there were 22 TKAs in the PCA group and 27 TKAs in the LIA group. There were no drains used or significant differences in demographics between groups. There was a statistically significant difference in blood loss (P = 0.003), between the LIA group (M = 942 ml, CI 829-1055), and the PCA group (M = 1314 ml, CI 1099-1527). Patients receiving PCA were 4.3 times more likely to require blood transfusion. Using the LIA technique reduces blood loss and risk of blood transfusion.

Assessment of owner-administered monthly treatments with oral spinosad or topical spot-on fipronil/(S)-methoprene in controlling fleas and associated pruritus in dogs.

Monitoring of the performance of flea control products under conditions of natural challenge is valuable in assessing continued effectiveness and determining the ongoing relevance of laboratory studies. A multi-clinic, investigator-blinded study was undertaken in client-owned dogs to investigate and compare the flea control provided by 3 consecutive monthly treatments of oral spinosad (SPN) or fipronil/(S)-methoprene topical (FSM) spot-on. The first household dog meeting enrollment criteria and with at least 10 fleas (whole-body flea count) served as the index dog in a household against which primary objectives were set. Stratification was based on pruritus scores at the enrollment visit and on single or multiple pet household. Index pets were randomized to treatment with either SPN or FSM, dispensed on day 0 for at-home administration by owners. All other household dogs and cats, maximum 4 pets per household, were dispensed the same treatment as the index dog (spinetoram was dispensed for cats in SPN households). Subsequent treatments were dispensed when index dogs were returned for whole-body flea counts and pruritus-scoring at visits on days 30 and 60, with final assessments on day 90 (±5 days on each occasion). Primary endpoints were the number of flea-free index dogs in each group one month after the final treatment, the reduction in owner-reported pruritus, and the reduction from baseline mean flea counts. One hundred twenty eight index dogs were enrolled (65 in the SPN arm; 63 in the FSM arm) at 10 clinics in FL (6), NC (2), LA (1), and TX (1). On day 0, geometric mean flea counts were 57.7 (range: 10-1469) and 44.8 (10-717) for the SPN and FSM groups, respectively. On Day 90, 55 of 58 (95%) and 21 of 55 (38%) index dogs completing the study were flea-free in SPN and FSM groups, respectively; mean SPN pruritus scores declined to 0.92 (6.67 on day 0), and to 3.83 (6.33 on day 0) for FSM; geometric mean flea counts (% control) were 0.08 (99.9%) and 5.19 (88.4%), for SPN and FSM groups, respectively. Between-treatment differences were highly statistically significant (p<0.0001). In conclusion, SPN provided reliable flea control in client-owned dogs, regardless of challenge level.