Selamectin increases cisplatin sensitivity by inhibiting cisplatin-resistant genes expression and autophagy in uveal melanoma.

Cisplatin resistance is the main reason for uveal melanoma (UM) treatment failure. Thus, developing strategy that increasing cisplatin sensitivity is needed. In this study, we performed drug repositioning analysis with the Connectivity Map database using a panel of previously identified cisplatin sensitivity-associated genes and cisplatin resistance-associated genes as the signature and obtained the antiparasitic drug selamectin. We demonstrated that the selamectin and cisplatin combination showed a synergistic effect on inhibiting UM cell growth. Experiments in tumor-bearing nude mice further showed that selamectin and cisplatin have synergistic effects in reducing tumor growth. Previous studies have linked increased autophagy with tumor resistance to chemotherapy. We found that selamectin inhibited the expression of the autophagy-related gene ATG9B, thus reducing autophagy. The cisplatin resistance-associated genes PDGFRB, DUSP1, MAST1 and IL11 were significantly downregulated in UM cells treated with selamectin. In summary, our study shows that selamectin enhanced the sensitivity of UM to cisplatin, through the mechanism of inhibiting cisplatin resistance-associated gene expression and autophagy. These findings may provide a new strategy for the treatment of UM.

A rapid evidence assessment of the potential risk to the environment presented by active ingredients in the UK's most commonly sold companion animal parasiticides.

A number of parasiticides are commercially available as companion animal treatments to protect against parasite infestation and are sold in large volumes. These treatments are not intended to enter the wider environment but may be washed off or excreted by treated animals and have ecotoxic impacts. A systematic literature review was conducted to identify the existing evidence for the toxicity of the six most used parasiticides in the UK: imidacloprid, fipronil, fluralaner, afoxolaner, selamectin, and flumethrin. A total of 17,207 published articles were screened, with 690 included in the final evidence synthesis. All parasiticides displayed higher toxicity towards invertebrates than vertebrates, enabling their use as companion animal treatments. Extensive evidence exists of ecotoxicity for imidacloprid and fipronil, but this focuses on exposure via agricultural use and is not representative of environmental exposure that results from use in companion animal treatments, especially in urban greenspace. Little to no evidence exists for the ecotoxicity of the remaining parasiticides. Despite heavy usage, there is currently insufficient evidence to understand the environmental risk posed by these veterinary treatments and further studies are urgently needed to quantify the levels and characterise the routes of environmental exposure, as well as identifying any resulting environmental harm.

The Veterinary Anti-Parasitic Selamectin Is a Novel Inhibitor of the Mycobacterium tuberculosis DprE1 Enzyme.

Avermectins are macrocyclic lactones with anthelmintic activity. Recently, they were found to be effective against Mycobacterium tuberculosis, which accounts for one third of the worldwide deaths from antimicrobial resistance. However, their anti-mycobacterial mode of action remains to be elucidated. The activity of selamectin was determined against a panel of M. tuberculosis mutants. Two strains carrying mutations in DprE1, the decaprenylphosphoryl-ß-D-ribose oxidase involved in the synthesis of mycobacterial arabinogalactan, were more susceptible to selamectin. Biochemical assays against the Mycobacterium smegmatis DprE1 protein confirmed this finding, and docking studies predicted a binding site in a loop that included Leu275. Sequence alignment revealed variants in this position among mycobacterial species, with the size and hydrophobicity of the residue correlating with their MIC values; M. smegmatis DprE1 variants carrying these point mutations validated the docking predictions. However, the correlation was not confirmed when M. smegmatis mutant strains were constructed and MIC phenotypic assays performed. Likewise, metabolic labeling of selamectin-treated M. smegmatis and M. tuberculosis cells with 14C-labeled acetate did not reveal the expected lipid profile associated with DprE1 inhibition. Together, our results confirm the in vitro interactions of selamectin and DprE1 but suggest that selamectin could be a multi-target anti-mycobacterial compound.

Structural mechanism underlying the differential effects of ivermectin and moxidectin on the C. elegans glutamate-gated chloride channel GLC-2.

BACKGROUND AND PURPOSE: Nematode glutamate-gated chloride channels (GluCls) are targets of ivermectin (IVM) and moxidectin (MOX), structurally dissimilar macrocyclic lactone (ML) anthelmintics. IVM and MOX possess different pharmacokinetics and efficacy profiles but are thought to have the same binding site, through which they allosterically activate GluCls, apart from the GLC-2 receptor, which is antagonized by IVM. Our goal was to determine GLC-2 sensitivity to MOX, investigate residues involved in antagonism of GLC-2, and to identify differences in receptor-level pharmacology between IVM and MOX. EXPERIMENTAL APPROACH: Two-electrode voltage clamp electrophysiology was used to study the pharmacology of Caenorhabditis elegans GLC-2 receptors heterologously expressed in Xenopus laevis oocytes. In silico homology modeling identified Cel-GLC-2 residues Met291 and Gln292 at the IVM binding site that differ from other GluCls; we mutated these residues to those found in ML-sensitive GluCls, and those of filarial nematode GLC-2. KEY RESULTS: We discovered that MOX inhibits wild-type C. elegans GLC-2 receptors roughly 10-fold more potently than IVM, and with greater maximal inhibition of glutamate activation (MOX = 86.9 ± 2.5%; IVM = 57.8 ± 5.9%). IVM was converted into an agonist in the Met291Gln mutant, but MOX remained an antagonist. Glutamate responses were abrogated in a Met291Leu Gln292Thr double mutant (mimicking filarial nematode GLC-2), but MOX and IVM were converted into positive allosteric modulators of glutamate at this construct. CONCLUSIONS AND IMPLICATIONS: Our data provides new insights into differences in receptor-level pharmacology between IVM and MOX and identify residues responsible for ML antagonism of GLC-2.

Repurposing Avermectins and Milbemycins against Mycobacteroides abscessus and Other Nontuberculous Mycobacteria.

Infections caused by nontuberculous mycobacteria (NTM) are increasing worldwide, resulting in a new global health concern. NTM treatment is complex and requires combinations of several drugs for lengthy periods. In spite of this, NTM disease is often associated with poor treatment outcomes. The anti-parasitic family of macrocyclic lactones (ML) (divided in two subfamilies: avermectins and milbemycins) was previously described as having activity against mycobacteria, including Mycobacterium tuberculosis, Mycobacterium ulcerans, and Mycobacterium marinum, among others. Here, we aimed to characterize the in vitro anti-mycobacterial activity of ML against a wide range of NTM species, including Mycobacteroides abscessus. For this, Minimum Inhibitory Concentration (MIC) values of eight ML were determined against 80 strains belonging to nine different NTM species. Macrocyclic lactones showed variable ranges of anti-mycobacterial activity that were compound and species-dependent. Milbemycin oxime was the most active compound, displaying broad-spectrum activity with MIC lower than 8 mg/L. Time kill assays confirmed MIC data and showed bactericidal and sterilizing activity of some compounds. Macrocyclic lactones are available in many formulations and have been extensively used in veterinary and human medicine with suitable pharmacokinetics and safety properties. This information could be exploited to explore repurposing of anti-helminthics for NTM therapy.

Case series: Efficacy of a formulation containing selamectin and sarolaner against naturally acquired mite infestations (Demodex sp., Ornithonyssus bacoti) in degus (Octodon degus).

Ectoparasite infestations are not common in degus. Two cases are presented here where use of Stronghold® Plus/Revolution® Plus (selamectin and sarolaner topical solution) was successfully administered to a degu (Octodon degus) for treatment of naturally-occurring mite infesations. Selamectin (Stronghold®/Revolution®) has been demonstrated to be effective against naturally-occurring mite infections in dogs and selamectin is approved for use in dogs for the treatment of sarcoptic mange (caused by Sarcoptes scabiei) at a dose of 6 mg/kg. In the first case, a 2.6-years-old female degu housed in a group with four other degus was presented with pruritic skin reactions, restlessness and hairloss. Mites morphologically similar to Demodex sp. were detected in the deep skin scrapings. All four degus were treated with Stronghold® Plus/Revolution® Plus (30 mg/kg selamectin and 5 mg/kg sarolaner) once a week for a total of six treatments. The spot-on was administered topically on the dorsal cervical region. Following treatment the degu presenting with clinical signs showed a rapid improvement with the pruritus and overall dermatitis resolving within 2 weeks of treatment. Skin scrapes and microscopic examination of epidermal debris collected from the affected degu were negative for mites from day 14 onwards. In the second case, a group of four 4-6.5-years-old female and male degus that were housed together were infested with Ornithonyssus bacoti. All animals were treated with 30 mg/kg selamectin and 5 mg/kg sarolaner in four total weekly doses. One week later no living mites were found on the patients or in their environment. The four degus improved visibly, and within three weeks of treatment the skin lesions associated with the infestation subsided. The antiparasiticides showed a satisfactory efficacy and were well tolerated (n = 9 animals treated in a total).

Successful treatment of otodemodicosis due to Demodex cati with sarolaner/selamectin topical solution in a cat.

CASE SUMMARY: A 10-year-old spayed female Russian Blue cat was presented with a 3-month history of excessive otic discharge and scratching, only involving the right ear. Other than a moderate amount of ceruminous exudate present within the right ear on video-otoscopic examination, there were no other cutaneous abnormalities. The cat was deemed to be otherwise in good health based on physical examination and several laboratory profiles. A diagnosis of otodemodicosis was determined due to the presence of a large number of Demodex cati mites retrieved from cerumen. Treatment consisted only of monthly topical application of sarolaner/selamectin to the nape of the neck with a marked reduction in mite counts and otic pruritus after a single dose. Complete resolution was achieved after a total of four doses. RELEVANCE AND NOVEL INFORMATION: This is the first report to describe the resolution of mite infestation owing to D cati after treatment with a sarolaner-containing spot-on product. In addition, to the best of the author's knowledge, this is the first report of any isoxazoline product used in the successful treatment of demodicosis affecting the ear canal. In general, there is a lack of reports describing safe and effective treatments for feline otodemodicosis. Topically applied sarolaner/selamectin resulted in resolution of mites while avoiding any potential ototoxic events from medications applied directly into the ear, and provided a treatment that was easier to apply than oral or injectable macrocyclic lactones.

Ivermectin, a potential anticancer drug derived from an antiparasitic drug.

Ivermectin is a macrolide antiparasitic drug with a 16-membered ring that is widely used for the treatment of many parasitic diseases such as river blindness, elephantiasis and scabies. Satoshi omura and William C. Campbell won the 2015 Nobel Prize in Physiology or Medicine for the discovery of the excellent efficacy of ivermectin against parasitic diseases. Recently, ivermectin has been reported to inhibit the proliferation of several tumor cells by regulating multiple signaling pathways. This suggests that ivermectin may be an anticancer drug with great potential. Here, we reviewed the related mechanisms by which ivermectin inhibited the development of different cancers and promoted programmed cell death and discussed the prospects for the clinical application of ivermectin as an anticancer drug for neoplasm therapy.

Unexpected effects of ivermectin and selamectin on inducible CreER activity in mice.

BACKGROUND: Anti-parasitics are frequently used in research animal facilities to treat a multitude of common infections, with pinworms and fur mites being amongst the most common. Ivermectin and selamectin are common oral and topical treatments for these infections, respectively. Although commonly thought to be innocuous to both the research animals and any transgenic elements that the animals may carry, evidence exists that ivermectin is capable of activating the recombinase activity of at least one CreER. The goal of the current study was to determine if there was an effect of either anti-parasitic agent on the activity of CreER proteins in transgenic mice. CASE PRESENTATION: We analyzed the offspring of transgenic mice exposed to either ivermectin or selamectin during pregnancy and nursing. Through analysis of reporter genes co-expressed with multiple, independently generated transgenic CreER drivers, we report here that ivermectin and selamectin both alter recombinase activity and thus may have unintended consequences on gene inactivation studies in mice. CONCLUSIONS: Although the mechanisms by which ivermectin and selamectin affect CreER activity in the offspring of treated dams remain unclear, the implications are important nonetheless. Treatment of pregnant transgenic mice with these anti-parasitics has the potential to alter transgene activity in the offspring. Special considerations should be made when planning treatment of transgenic mice with either of these pharmacologics.

Demodicose felina em Santa Catarina, Brasil / Feline demodicosis in Santa Catarina state, Brazil

A demodicose felina é considerada uma dermatopatia rara e pode ser causada pelos ácaros Demodex cati, Demodex gatoi e uma terceira espécie ainda não nomeada. Foi atendido um felino adulto apresentando prurido intenso há 9 meses e histórico de tratamento com cefalexina e prednisolona, com piora progressiva. Ao exame físico, havia alopecia, hiperqueratose, escoriações e eritema em cabeça, pescoço, região lombossacra, cauda e membros pélvicos, além da presença de pulgas. Para puliciose, foram prescritos selamectina spot on a cada 30 dias e uso de amitraz no ambiente a cada sete dias e, para controle da infecção secundária pelas escoriações, foram recomendados banhos semanais com clorexidine. Realizaram-se raspado de pele profundo e arrancamento de pelos para tricograma e exame parasitológico de pele, respectivamente, com diagnósticos de demodicose por Demodex cati, e dermatite micótica associada a infecção bacteriana secundária. O tratamento foi modificado para uso de selamectina a cada 2 semanas, mas tutor não retornou e informou, após vários meses, ter feito terapia com selamectina apenas a cada 30 dias e descontinuidade dos banhos. Não foi possível associar a demodicose, para este felino, a outras comorbidades e acredita-se que a apresentação generalizada da doença tenha se dado pelo prurido causado pela puliciose.

Feline demodicosis is considered a rare dermatopathy and can be caused by Demodex cati, Demodex gatoi and a third species not yet named. An adult male feline was attended with severe pruritus for 9 months and a history of treatment with cephalexin and prednisolone, with progressive worsening. On physical examination, there was alopecia, hyperkeratosis, abrasions and erythema on the head, neck, lumbosacral region, tail and pelvic limbs, in addition to the presence of fleas. For pulicosis, selamectin spot on was prescribed every 30 days and use of amitraz in the environment every seven days. In order to control secondary infection, weekly baths with chlorhexidine were recommended. Deep skin scraping and hair plucking were performed for trichogram and parasitological skin examination, respectively, with diagnoses of demodicosis by Demodex cati, and mycotic dermatitis associated with secondary bacterial infection. The treatment was modified to use selamectin every 2 weeks, but the tutor did not return and reported, after several months, that he had done therapy with selamectin only every 30 days and discontinued baths. For this feline, it was not possible to associate demodicosis with other comorbidities. It is believed that the generalized presentation of the disease occurred due to the pruritus caused by pulicosis.

Feline demodicosis in Santa Catarina state, Brazil / Demodicose felina em Santa Catarina, Brasil

Feline demodicosis is considered a rare dermatopathy and can be caused by Demodex cati, Demodex gatoi and a third species not yet named. An adult male feline was attended with severe pruritus for 9 months and a history of treatment with cephalexin and prednisolone, with progressive worsening. On physical examination, there was alopecia, hyperkeratosis, abrasions and erythema on the head, neck, lumbosacral region, tail and pelvic limbs, in addition to the presence of fleas. For pulicosis, selamectin spot on was prescribed every 30 days and use of amitraz in the environment every seven days. In order to control secondary infection, weekly baths with chlorhexidine were recommended. Deep skin scraping and hair plucking were performed for trichogram and parasitological skin examination, respectively, with diagnoses of demodicosis by Demodex cati, and mycotic dermatitis associated with secondary bacterial infection. The treatment was modified to use selamectin every 2 weeks, but the tutor did not return and reported, after several months, that he had done therapy with selamectin only every 30 days and discontinued baths. For this feline, it was not possible to associate demodicosis with other comorbidities. It is believed that the generalized presentation of the disease occurred due to the pruritus caused by pulicosis.

A demodicose felina é considerada uma dermatopatia rara e pode ser causada pelos ácaros Demodex cati,Demodex gatoi e uma terceira espécie ainda não nomeada. Foi atendido um felino adulto apresentando prurido intenso há 9 meses e histórico de tratamento com cefalexina e prednisolona, com piora progressiva. Ao exame físico, havia alopecia, hiperqueratose, escoriações e eritema em cabeça, pescoço, região lombossacra, cauda e membros pélvicos, além da presença de pulgas. Para puliciose, foram prescritos selamectina spot on a cada 30 dias e uso de amitraz no ambiente a cada sete dias e, para controle da infecção secundária pelas escoriações, foram recomendados banhos semanais com clorexidine. Realizaram-se raspado de pele profundo e arrancamento de pelos para tricograma e exame parasitológico de pele, respectivamente, com diagnósticos de demodicose por Demodex cati, e dermatite micótica associada a infecção bacteriana secundária. O tratamento foi modificado para uso de selamectina a cada 2 semanas, mas tutor não retornou e informou, após vários meses, ter feito terapia com selamectina apenas a cada 30 dias e descontinuidade dos banhos. Não foi possível associar a demodicose, para este felino, a outras comorbidades e acredita-se que a apresentação generalizada da doença tenha se dado pelo prurido causado pela puliciose.

Evaluation of a topical sarolaner-selamectin combination to control flea populations on naturally infested cats in private residences in West Central Florida.

Historic data show that home flea infestations can be managed by treating all animals on the premises with a highly effective flea control product. The use of effective products has also been shown to reduce pruritus and minimize dermatologic lesions in both cats and dogs. Therefore, an in-home study was conducted in West Central Florida USA to evaluate the efficacy of a topically applied selamectin-sarolaner formulation to control fleas in naturally infested cats over a 12-week period. Thirty-seven cats in 21 households were treated once monthly with the selamectin-sarolaner topical solution. In the topical fluralaner treatment (positive control) group, forty-three cats in 20 households were treated once on day 0. A combined total of thirty dogs in both groups were treated once monthly with oral sarolaner. Fleas on cats were counted by flea combing, fleas on dogs were counted using visual area counts and fleas in the indoor premises were assessed using intermittent-light flea traps. Blinded-assessments of feline dermatologic lesions (modified-SCORFAD) were conducted monthly by a boarded-dermatologist and pruritus severity was evaluated by pet owners. Three consecutive monthly treatments of selamectin-sarolaner reduced flea populations on cats by 96.3 % within 7 days and by 100% from week 6 to the end of the 12-week study. The topical application of fluralaner reduced flea populations by 98.1 % within 7 days and efficacy reached 100% by week 12. At the end of the study, fleas were completely eradicated (from cats, dogs and homes) in every home regardless of treatment group. Owner reported cat pruritus was reduced by > 87 % in both treatment groups by week 12. Significant improvements in dermatologic lesion scores (> 81 %) were achieved by both products by the end of the study. Monthly applications of topical selamectin-sarolaner or topical fluralaner to cats living in the heavy flea challenge environment of West Central Florida USA were effective in eradicating flea infestations, reducing pruritus and improving dermatologic lesions.

Safety and efficacy of a new spot-on formulation of selamectin plus sarolaner in the treatment and control of naturally occurring flea infestations in cats presented as veterinary patients in Australia.

BACKGROUND: The safety and efficacy of a new spot-on formulation of selamectin plus sarolaner were evaluated for the treatment and control of natural flea infestations on cats in two non-randomised, multi-centre clinical trials conducted in 8 different locations in Queensland, Australia. METHODS: One hundred and four cats from 65 different households were enrolled across the two studies. Demographic characteristics of cats in the two studies were similar. The new spot-on formulation of selamectin and sarolaner was administered topically once a month for 3 consecutive months at a minimum dosage of 6 mg/kg selamectin (dose range 6-12 mg/kg) plus 1 mg/kg sarolaner (dose range 1-2 mg/kg). Cats were dosed on Days 0 (pre-treatment), 30 and 60 and physical examinations and flea counts were conducted on Days 0, 30, 60 and 90. Efficacy assessments were based on the percentage reduction in live flea counts post-treatment compared to Day 0. RESULTS: In Study A, at enrolment, primary cats had flea counts ranging from 6 to 107 (arithmetic mean 21.0). The selamectin and sarolaner spot-on formulation resulted in arithmetic mean efficacy of 98.0%, 100% and 100% on Days 30, 60 and 90, respectively. In Study B, at enrolment, primary cats had flea counts ranging from 6 to 22 (arithmetic mean 10.0). The selamectin and sarolaner spot-on formulation resulted in arithmetic mean efficacy of 99.7%, 100% and 100% on Days 30, 60 and 90, respectively. CONCLUSIONS: The new spot-on formulation of selamectin plus sarolaner topically administered at monthly intervals at the minimum dosage of 6.0 mg/kg selamectin and 1.0 mg/kg sarolaner was safe and highly effective against natural infestations of fleas under a range of geographical conditions, representative of both tropical and subtropical regions of Australia.

The efficacy of a topical formulation of selamectin plus sarolaner in preventing the development of a macrocyclic lactone-resistant strain of Dirofilaria immitis in cats.

Revolution®/Stronghold® Plus, a topical endectocide incorporating 6 mg/kg selamectin plus 1 mg/kg sarolaner, is approved for use in cats to prevent heartworm disease. The efficacy of selamectin has not previously been evaluated against any macrocyclic lactone (ML)-resistant heartworm strains in cats for prevention of heartworm disease. In this study, an experimental combination formulation of selamectin (6 mg/kg) plus sarolaner (2 mg/kg) was assessed for preventing the development of a ML-resistant strain of Dirofilaria immitis in cats. Forty purpose-bred domestic shorted-haired cats (20 males; 20 females) from 7-9 months of age and negative for heartworm antigen prior to study inclusion were used. On Day -30, cats were inoculated with 100 D. immitis L3 (ZoeMO strain) subcutaneously in the inguinal area. Cats were randomly allocated to one of the following four treatments with associated dosing regimens: T01 (vehicle-treated control on Days 0, 28, and 56), T02 (single dose of selamectin plus sarolaner combination on Day 0 only), T03 (selamectin plus sarolaner combination on Days 0, 28, and 56) or T04 (single dose of selamectin on Day 0 only). All treatments were administered topically in an isopropyl alcohol-based formulation. Selamectin was administered at 6 mg/kg in both standalone and combination formulations. Sarolaner was administered at 2 mg/kg. Cats were necropsied on Day ∼145 (∼175 days post infection) and adult worms were counted. Nine of ten cats in the control group (T01) were infected with adult worms (range, 1-23; geometric mean, 3.5). In contrast, all cats in T03 had zero heartworms. Only two cats in T02 (0-3; 0.2) and a single cat in the T04 (0-1; 0.1) had heartworms. Compared to T01 (control cats), all treated cats had significantly (p < 0.0001) reduced worm burdens, with treatment efficacies of 100% (T03), 93.5% (T02) and 98% (T04). A topical combination of selamectin (6 mg/kg) plus sarolaner (2 mg/kg) was 100% efficacious in preventing the development of an ML-resistant strain of D. immitis (ZoeMO) in cats when administered as three consecutive monthly treatments. A single dose was highly (93.5%) but incompletely effective.

Repurposing of clinically approved drugs for treatment of coronavirus disease 2019 in a 2019-novel coronavirus (2019-nCoV) related coronavirus model / 中华医学杂志(英文版)

Background@#Medicines for the treatment of 2019-novel coronavirus (2019-nCoV) infections are urgently needed. However, drug screening using live 2019-nCoV requires high-level biosafety facilities, which imposes an obstacle for those without such facilities or 2019-novel coronavirus (2019-nCoV). This study aims to repurpose the clinically approved drugs for the treatment of coronavirus disease 2019 (COVID-19) in a 2019-nCoV related coronavirus model.@*Methods@#A 2019-nCoV related pangolin coronavirus GX_P2V/pangolin/2017/ Guangxi was described. Whether GX_P2X uses angiotensin-converting enzyme 2 (ACE2) as the cell receptor was investigated by using small interfering RNA (siRNA) -mediated silencing of ACE2. The pangolin coronavirus model was used to identify drug candidates for treating 2019-nCoV infection. Two libraries of 2406 clinically approved drugs were screened for their ability to inhibit cytopathic effects on Vero E6 cells by GX_P2X infection. The antiviral activities and antiviral mechanisms of potential drugs were further investigated. Viral yields of RNAs and infectious particles were quantified by quantitative real-time polymerase chain reaction (qRT-PCR) and plaque assay, respectively.@*Results@#The spike protein of coronavirus GX_P2V shares 92.2% amino acid identity with that of 2019-nCoV isolate Wuhan-hu-1, and uses ACE2 as the receptor for infection just like 2019-nCoV. Three drugs-cepharanthine (CEP), selamectin and mefloquine hydrochloride exhibited complete inhibition of cytopathic effects in cell culture at 10 μmol/L. CEP demonstrated the most potent inhibition of GX_P2V infection, with a concentration for 50% of maximal effect [EC50] of 0.98 μmol/L. The viral RNA yield in cells treated with 10 μmol/L CEP was 15,393-fold lower than in cells without CEP treatment ([6.48±0.02]×10-4 vs. 1.00±0.12, t=150.38, P<0.001) at 72 h post-infection (p.i.). Plaque assays found no production of live viruses in media containing 10 μmol/L CEP at 48 h p.i. Furthermore, we found CEP has potent antiviral activities against both viral entry (1.00±0.37 vs. 0.46±0.12, t=2.42, P<0.05) and viral replication (1.00±0.43 vs. [6.18±0.95]×10-4, t=3.98, P<0.05).@*Conclusions@#Our pangolin coronavirus GX_P2V is a workable model for 2019-nCoV research. CEP, selamectin and mefloquine hydrochloride are potential drugs for treating 2019-nCoV infection. Our results strongly suggest that CEP is a wide-spectrum inhibitor of pan-betacoronavirus, and clinical trial of CEP for treatment of 2019-nCoV infection is warranted.

Efficacy of single topical treatment of Selamectin (Revolution®) against Ancylostoma ceylanicum in experimentally infected cats.

Ancylostoma ceylanicum, a species of hookworm, is a common parasite of dogs and cats in the Asia-Pacific region. The objective of this study was to determine the efficacy of a single topical treatment of selamectin (Revolution®) against A. ceylanicum in experimentally infected cats. 12 kittens were injected with 300 infective stage larvae (L3) of A. ceylanicum by the subcutaneous route. Faecal samples were examined daily (days 7-15) for the presence of hookworm eggs. On day 18 kittens were stratified by faecal egg count and randomly allocated in equal numbers to control (n = 6) and treatment (n = 6) groups. Faecal egg counts were then performed daily (days 14-27) or every 3 days (days 28-51) until the end of the study and compared between the treated and control groups. Kittens in the treatment group were administered a single topical dose of selamectin (Revolution®), per label recommendations (6 mg/kg), on day 19. Kittens in the control group were not treated. At 4 days post-treatment, no hookworm eggs were detected in the treated group. Faecal samples from this group remained negative throughout the study, the treatment efficacy was 100% egg reduction (P < .0001). Average faecal egg counts remained high (558 ±â€¯231 eggs per gram) in the untreated control group until the end of the study period. In conclusion, a single topical treatment of selamectin (Revolution®) at the recommended dose was highly efficacious against infection with A. ceylanicum in cats.

Comparative efficacy of topical treatments with Revolution® Plus (selamectin and sarolaner) and Bravecto® for Cats (fluralaner) against Ixodes scapularis ticks on cats.

The efficacy of three consecutive monthly treatments with a novel topical product (Revolution® Plus/Stronghold® Plus, Zoetis) containing selamectin in combination with the isoxazoline, sarolaner, was compared with that of another topical isoxazoline, fluralaner [Bravecto® (fluralaner topical solution) for Cats, Merck] against Ixodes scapularis ticks on cats. Twenty-four cats were ranked by pre-treatment tick counts to form groups of three and were randomly allocated to be treated with placebo, the minimum label dosage of Revolution® Plus (6 mg/kg selamectin plus 1 mg/kg sarolaner) or the minimum label dosage of Bravecto® for Cats (40 mg/kg fluralaner) within the groups. On Days 0, 30, and 60, each cat in the placebo and Revolution® Plus-treated groups was treated topically, whereas cats in the Bravecto® for Cats-treated group were treated topically once on Day 0 with fluralaner and, subsequently, these animals were treated with the placebo on Days 30 and 60 to maintain masking. Doses were calculated based on weight to provide the minimum label dosage for each product; the calculated volume of product to be administered was rounded off to the nearest 0.1 mL. The selamectin plus sarolaner-treated cats received effective dosages of 5.29-7.12 mg/kg selamectin and 0.88-1.19 mg/kg sarolaner, while the fluralaner cats received dosages of 35.21-43.16 mg/kg fluralaner. Cats were infested with approximately 50 unfed viable adult I. scapularis ticks on Days 5, 12, 26, 40, 54, 68, 82, and 88. Efficacy was assessed at 48 h after each infestation. There were no adverse reactions to any treatment during the study. The placebo-treated cats maintained adequate tick infestations throughout the study. Three monthly treatments with selamectin plus sarolaner (Revolution® Plus) resulted in high and consistent efficacy against I. scapularis for up to 30 days after each treatment. Based on geometric means, efficacy was ≥99.1% at all time points assessed. Treatment with fluralaner (Bravecto® for Cats) provided high and consistent efficacy of ≥99.3% up to Day 70. On Day 84, efficacy was 90.1%; however, cats from which ticks were recovered on Day 84 had received approximately 4%-12% less than the minimum dosage of 40 mg/kg fluralaner. Three consecutive monthly treatments with Revolution® Plus or a single treatment with Bravecto® for Cats provided >90% control of I. scapularis ticks over a 12-week time period.

Efficacy and safety of a new topical formulation containing selamectin and sarolaner in the prevention of heartworm disease and the treatment of roundworm infection in cats presented as veterinary patients in Japan.

The efficacy and safety of a new topical formulation containing selamectin plus sarolaner (Revolution® Plus / Stronghold® Plus, Zoetis) was evaluated for the prevention of heartworm (Dirofilaria immitis) disease and the treatment of roundworm infection in cats enrolled as veterinary patients in two field studies conducted in Japan. In the heartworm field study, 91 cats negative for D. immitis antigen and anti-D. immitis antibody were enrolled and received 9 monthly topical treatments with selamectin plus sarolaner during the period of April to December 2015. Efficacy was assessed by testing post-treatment blood samples collected 8, 12, and 15 months after initiation of treatment for the presence of D. immitis antigen and anti-D. immitis antibody. Eighty-seven cats completed the entire study and were included in the determination of efficacy. No D. immitis antigen or anti-D. immitis antibody were detected in any of the post-treatment samples. In the roundworm field study, completed in the period from April to November 2015, 64 cats with ≥100 roundworm eggs per gram (EPG) of feces were enrolled and allocated randomly in a 1:1 ratio, to receive either selamectin plus sarolaner or emodepside plus praziquantel (Profender®, Bayer). Treatments were administered topically on Days 0 and 30, and efficacy was assessed by fecal EPG counts conducted on Days 14, 30, and 60. All cats completed the entire study. At enrollment, all cats were infected with Toxocara cati. Compared to pre-treatment, geometric mean T. cati EPG counts on Days 14, 30, and 60 were reduced by >99.9% in both treatment groups. There were no treatment-related adverse events in either study. Monthly topical administration of Revolution® Plus / Stronghold® Plus providing a minimum of 6 mg/kg selamectin and 1 mg/kg sarolaner was safe and effective in the prevention of heartworm disease and the treatment of roundworm infection in cats enrolled as veterinary patients in Japan.

Efficacy of three consecutive monthly doses of a topical formulation of selamectin and sarolaner (Revolution® Plus/Stronghold® Plus) compared with a single dose of fluralaner (Bravecto® for Cats) against induced infestations of Ctenocephalides felis on cats.

In a controlled laboratory study, the efficacy against fleas, Ctenocephalides felis, of a single treatment of fluralaner topical solution (Bravecto® for Cats, Merck) was compared with that of three consecutive monthly topical treatments with selamectin and sarolaner (Revolution® Plus, Zoetis). Twenty-four domestic short hair cats were ranked based on host suitability flea counts to form groups of three and were randomly assigned within group to one of three treatments. The first group received a topical treatment with (a) placebo (vehicle control for Revolution® Plus) on Days 0, 30, and 60, (b) 6 mg/kg selamectin and 1 mg/kg sarolaner on Days 0, 30, and 60, or (c) 40 mg/kg fluralaner on Day 0 and placebo (vehicle control for Revolution® Plus) on Days 30 and 60. Because doses were rounded off, the selamectin plus sarolaner-treated cats received effective dosages of 5.25-6.60 mg/kg selamectin and 0.88-1.10 mg/kg sarolaner, while the fluralaner-treated cats received dosages of 34.71-43.08 mg/kg fluralaner. All cats were infested with 100 (±5) fleas on Day -1 and at biweekly intervals after that, from Day 13 to Day 89. Flea comb counts were conducted 24 hours after treatment or after re-infestation. There were no adverse events related to treatment during the study. Except for a single cat from which 20 fleas were recovered on Day 90, all other placebo-treated cats had at least 48 fleas at each count, indicating adequacy of infestation of the controls. Based on geometric mean live flea counts, three consecutive monthly treatments with Revolution® Plus resulted in consistent and high efficacy of ≥98.6% compared with placebo throughout the study. A single treatment with Bravecto® for Cats provided consistent and high efficacy of ≥94.6% on all count days during a period of 12 weeks, the approved duration of efficacy for the product. Based on the efficacy results of the study, both products were equivalent in their ability to control fleas on cats. Use of Bravecto® for Cats every 12 weeks or the consecutive monthly use of Revolution® Plus is expected to provide extended high residual kill over the respective labeled durations of efficacy of the two products.

Efficacy of a new topical formulation of selamectin plus sarolaner for the control of fleas and ticks infesting cats in Japan.

The efficacy of a single application of a new topical formulation containing selamectin plus sarolaner (Revolution® Plus / Stronghold® Plus, Zoetis) was evaluated against fleas and ticks infesting cats enrolled as veterinary patients in two field studies conducted in Japan and against Haemaphysalis longicornis ticks on cats in a laboratory study. In the laboratory study, sixteen cats were ranked based on pre-treatment tick counts and allocated randomly to treatment on Day 0 with either selamectin plus sarolaner or placebo. Cats were infested with adult H. longicornis on Days -2, 5, 12, 19, 26 and 33. Efficacy relative to placebo was based on live attached tick counts conducted 48 h after treatment and subsequent re-infestations. Selamectin plus sarolaner reduced live, attached H. longicornis counts by 96.4% within 48 h of treatment, and by ≥91.7% within 48 h of weekly re-infestation for 35 days, based on arithmetic means. In the field studies, 67 client-owned cats harboring six or more live fleas and 63 cats harboring four or more live attached ticks were enrolled to evaluate selamectin plus sarolaner for efficacy and safety compared with a registered product. Cats were allocated randomly to treatment with selamectin plus sarolaner or fipronil plus (S)-methoprene based on order of presentation. Treatment was administered once on Day 0 and efficacy was assessed by parasite counts conducted on Days 14 and 30 compared to the pre-treatment count. In the flea field study, live flea counts on Days 14 and 30 were reduced by 99.5% and 99.9% in the selamectin plus sarolaner group, and by 97.6% and 98.6% in the fipronil plus (S)-methoprene group, based on least squares mean percentage reductions. Clinical signs typically associated with flea allergy dermatitis improved following treatment. In the tick field study, live tick counts on Days 14 and 30 were reduced by 97.5% and 97.7% in the selamectin plus sarolaner group, and by 91.5% and 93.4% in the fipronil plus (S)-methoprene group, based on least squares mean percentage reductions. Selamectin plus sarolaner was determined to be non-inferior to fipronil plus (S)-methoprene in both field studies. There were no treatment-related adverse events in any study. A single topical dose of Revolution® Plus / Stronghold® Plus providing a minimum dosage of 6.0 mg/kg selamectin and 1.0 mg/kg sarolaner was confirmed to be effective against H. longicornis ticks on cats for one month and safe and effective in the treatment of fleas and ticks on cats enrolled as veterinary patients in Japan.