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To evaluate the efficacy of and resistance to mebendazole (500 mg) and levamisole (40 or 80 mg), alone or in combination, for the treatment of Ascaris lumbricoides, Trichuris trichiura and hookworm infections on Pemba Island - an area exposed to periodic school-based mebendazole treatment since 1994. A randomized, placebo-controlled trial was carried out in 914 children enrolled from the first and fifth grades of primary schools. Stool samples collected at baseline and 21 days after treatment were examined by the Kato-Katz technique to assess the prevalence and intensity of helminth infection. Efficacies of mebendazole and levamisole as single treatments against intestinal nematode infections were comparable with those in previous trials, but mebendazole treatment of hookworm infections gave significantly lower cure (7.6%) and egg reduction (52.1%) rates than reported in a study undertaken before the beginning of periodic chemotherapy (cure rate, 22.4%; egg reduction rate, 82.4%). Combined treatment with mebendazole and levamisole had a significantly higher efficacy against hookworm infections (cure rate, 26.1%; egg reduction rate, 88.7%) than either drug given alone. No difference in mebendazole efficacy was found in children who had been treated repeatedly compared with those who had not been treated previously. The overall efficacy of mebendazole against hookworm infections after periodic chemotherapy is reduced. The efficacy of benzimidazoles in chemotherapy-based control programmes should be monitored closely. Combined treatment with mebendazole and levamisole may be useful as a tool to delay the development of benzimidazole resistance.

Combination therapy of many anthelmintic drugs has been used to achieve fast animal curing. Q-DRENCH is an oral suspension, containing four different active drugs against GIT worms in sheep, commonly used in Australia and New Zeeland. The anti-parasitic drugs are Albendazole (ALB), Levamisole HCl (LEV), Abamectin (ABA), and Closantel (CLO). The main purpose of this study is to present a new simultaneous stability-indicting HPLC-DAD method for the analysis of the four drugs. The recommended liquid system was 1 mL of Triethylamine/L water, adjusting the pH to 3.5 by glacial acetic acid: acetonitrile solvent (20:80, v/v). Isocratic elusion achieved the desired results of separation at a 2 mL/min flow rate using Zorbax C-18 as a stationary phase. Detection was performed at 210 nm. The linearity ranges were 15.15 to 93.75 μg/mL for ALB, 25 to 150 μg/mL for LEV, 30 to 150 μg/mL for ABA, and 11.7 to 140.63 μg/mL for CLO. Moreover, the final greenness score was 0.62 using the AGREE tool, which reflects the eco-friendly nature. Moreover, the four drugs were determined successfully in the presence of their stressful degradation products. This work presents the first chromatographic method for simultaneous analysis for Q-DRENCH oral suspension drugs in the presence of their stressful degradation products.

Individuals with high Loa loa microfilarial densities (MFD) risk serious adverse events (SAEs) following ivermectin treatment. A single dose of levamisole (LEV) induces a temporary, progressive MFD decrease. This double-blind, randomized, placebo-controlled trial evaluated the safety and efficacy of 3-day and 5-day LEV regimens for the treatment of loiasis in the Republic of the Congo. Participants were randomly assigned to receive placebo (PLA), LEV-3, or LEV-5. Safety was assessed by the occurrence of SAEs and the frequency and severity of AEs. Efficacy was measured by changes in Loa loa MFD. No SAEs were reported, and AE severity was comparable across treatment arms. On day 5, MFD reductions were greater in the LEV-3 and LEV-5 groups compared to PLA ( −1.6%, 29.3%, and 51.4%, respectively; P  < 0.001). By day 7, a higher proportion of participants achieved ≥40% MFD reduction in the LEV-5 group (44.4%) compared to LEV-3 (34.6%) and PLA (8.3%) ( P  = 0.051). Post hoc contrasts confirmed significantly greater MFD reduction with LEV-5 versus LEV-3. These findings suggest that a 5-day LEV regimen is safe and moderately effective for reducing L. loa MFD and may represent a promising alternative for individualised management in loiasis-endemic areas, particularly where onchocerciasis is hypoendemic and coendemic with loiasis. Trial registration: NCT06252961. Loiasis poses significant treatment challenges, particularly in regions where onchocerciasis is hypoendemic and coendemic. Here, the authors demonstrate that a 5-day levamisole regimen is safe and more effective than shorter treatments in reducing Loa loa microfilarial densities, offering an alternative approach for managing loiasis in affected areas.

The aim of the QUASAR trial was to determine the size and duration of any survival benefit from adjuvant chemotherapy for patients with colorectal cancer at low risk of recurrence, for whom the indication for such treatment is unclear. After apparently curative resections of colon or rectal cancer, 3239 patients (2963 [91%] with stage II [node negative] disease, 2291 [71%] with colon cancer, median age 63 [IQR 56–68] years) enrolled between May, 1994, and December, 2003, from 150 centres in 19 countries were randomly assigned to receive chemotherapy with fluorouracil and folinic acid (n=1622) or to observation (with chemotherapy considered on recurrence; n=1617). Chemotherapy was delivered as six 5-day courses every 4 weeks or as 30 once-weekly courses of intravenous fluorouracil (370 mg/m 2) with high-dose (175 mg) L-folinic acid or low-dose (25 mg) L-folinic acid. Until 1997, levamisole (12 courses of 450 mg over 3 days repeated every 2 weeks) or placebo was added. After 1997, patients who were assigned to receive chemotherapy were given fluorouracil and low-dose folinic acid only. The primary outcome was all-cause mortality. Analyses were done by intention to treat. This trial is registered with the International Clinical Trial Registry, number ISRCTN82375386. At the time of analysis, 61 (3·8%) patients in the chemotherapy group and 50 (3·1%) in the observation group had missing follow-up. After a median follow-up of 5·5 (range 0–10·6) years, there were 311 deaths in the chemotherapy group and 370 in the observation group; the relative risk of death from any cause with chemotherapy versus observation alone was 0·82 (95% CI 0·70–0·95; p=0·008). There were 293 recurrences in the chemotherapy group and 359 in the observation group; the relative risk of recurrence with chemotherapy versus observation alone was 0·78 (0·67–0·91; p=0·001). Treatment efficacy did not differ significantly by tumour site, stage, sex, age, or chemotherapy schedule. Eight (0·5%) patients in the chemotherapy group and four (0·25%) in the observation group died from non-colorectal cancer causes within 30 weeks of randomisation; only one of these deaths was deemed to be possibly chemotherapy related. Chemotherapy with fluorouracil and folinic acid could improve survival of patients with stage II colorectal cancer, although the absolute improvements are small: assuming 5-year mortality without chemotherapy is 20%, the relative risk of death seen here translates into an absolute improvement in survival of 3·6% (95% CI 1·0–6·0).

Levamisole is an immunomodulatory agent that was used to treat various cancers before being withdrawn from the United States market in 2000 because of adverse effects. Levamisole is currently approved as an antihelminthic agent in veterinary medicine, but is also being used illicitly as a cocaine adulterant. Potential complications associated with use of levamisole-laced cocaine include neutropenia, agranulocytosis, arthralgias, retiform purpura, and skin necrosis. Treatment is primarily supportive, and skin lesions typically resolve with cessation of cocaine use. The incidence of hospitalizations related to use of levamisole-contaminated cocaine continues to increase and clinicians should be aware of the more common clinical manifestations.

Background Levamisole is less expensive and has a better toxicity profile compared to other steroid sparing agents used in nephrotic syndrome. It has a plasma half-life of 2.0 to 5.6 hours, but is conventionally administered on alternate days. We aimed to assess whether daily levamisole is safe and more effective than standard alternate-day therapy in maintaining remission in children with frequently relapsing or steroid-dependent nephrotic syndrome (FR/SDNS). Methods An open-label randomized controlled trial was conducted in children with FR/SDNS. Group A received daily while Group B received alternate-day levamisole (2–3 mg/kg/dose) for 12 months. Prednisolone was tapered off by 3 months. Patients were monitored for relapses, further steroid requirement, and adverse effects. Results A total of 190 children with FR/SDNS (94 in Group A and 96 in Group B) were analyzed. Sustained remission for 12 months was observed in 36% of Group A and 27% of Group B patients ( p  = 0.18). Numbers completing 12 months in the study were 67% in Group A and 56% in Group B ( p  = 0.13). Time to first relapse, persistent FR/SDNS, and withdrawal due to poor compliance were statistically similar in both groups, while relapse rate and cumulative steroid dosage were significantly lower in Group A compared to Group B ( p  = 0.03 and p  = 0.02, respectively). The incidence of adverse effects was comparable in both groups, with reversible leucopenia and hepatic transaminitis being the commonest. Conclusions Daily levamisole therapy was not superior to alternate-day therapy in maintaining sustained remission over 12 months. Nevertheless, relapse rate and cumulative steroid dosage were significantly lower without increased adverse effects. Graphical abstract A higher resolution version of the Graphical abstract is available as Supplementary information

Pseudomonas aeruginosa ( P. aeruginosa ) is one of the most common ones that harm fish. P. aeruginosa has been regarded as one of the most significant threats to the fishing industry, which also affects public health. Thus, the present investigation was done in two steps; the first step was to examine the prevalence and the antibiogram of P. aeruginosa among Nile tilapia ( Oreochromas niloticus ( O. niloticus )) from aquaculture farms in Kafr El-shiekh Governorate with an emphasis on their antibiotic resistance genes ( BlaTEM, tetA, and sul1 ). The second step was to investigate the effect of levamisole as a feed supplement for tilapia fish on growth performance, immunity, serum biochemistry, and the protective effect against artificial infection with the previously isolated in the first step P. aeruginosa strain. One hundred samples were collected from morbid Nile tilapia fish in the first step. The incidence of P. aeruginosa was 14%. Susceptibility of P. aeruginosa isolates to 9 antimicrobial agents showed that about half of P. aeruginosa isolates were multidrug-resistant (MDR) to (5–6) antibiotics. All of the isolates were sensitive to amikacin, ciprofloxacin, and norfloxacin (100%), and half of them were resistant to azithromycin, amoxicillin with clavulanic, tetracycline, and sulfa with trimethoprim. P. aeruginosa isolates were confirmed diagnosed using the 16S rRNA gene, which was detected in 100% of the tested isolates, and was also evaluated for the presence of antibiotic resistance genes ( blaTEM, tetA, and sul1 ), which were 85.7%, 85.7%, and 100%, respectively. In the second step, a 2-month feeding trial was performed on 160 O. niloticus fish with a weight of 56.75 ± 3 g. Fish were randomly distributed into four groups, each at a rate of 10 fish per aquarium in four replicates, and fed on a diet containing 0.0, 500, 750, and 1000 mg levamisole/kg diet. At the end of the feeding trial, fish were challenged by pathogenic P. aeruginosa, which was isolated in the first step. The results of the in vivo trial showed that levamisole safely improved the growth and immunity of Nile tilapia without side effects on liver function.

Vaccination-induced allergic reactions in bulls raise both body and testicular temperatures, leading to germ cell damage, epididymal dysfunction, accelerated testicular ageing, and increased sperm abnormalities that ultimately degrade semen quality. This study aimed to ameliorate vaccination stress and improve semen quality using meloxicam and levamisole. The present study was conducted at the Artificial Breeding Research Centre, ICAR-NDRI, Karnal, Haryana, with twelve Sahiwal breeding bulls that were divided into four groups: Group I (Control), Group II (Meloxicam), Group III (Levamisole), and Group IV (Meloxicam + Levamisole). Semen parameters were evaluated during pre- and post-vaccination. Treated groups showed significant improvement ( p  < 0.05) in initial progressive motility, live sperm, HOST, and acrosome integrity compared to the control, with the highest improvement observed in the combination group (meloxicam + levamisole). Sperm abnormalities, moribund, dead, apoptotic sperm, and lipid peroxidation were significantly higher in the control group. The meloxicam + levamisole group exhibited significantly lower ( p  < 0.05) sperm abnormalities and lipid peroxidation. Improvement was observed in semen quality with treatment (Meloxicam + levamisole-treated bulls) in percentage of live (+ 53.11%), moribund (− 33.83%), dead (− 36.70%), apoptotic spermatozoa (− 44.01%), and DNA fragmentation index (− 47.07%) in relation to the control groups on the 7th post vaccination day.

Background Reproductive efficiency in sheep is influenced by complex interactions between the endocrine and immune systems, particularly during early pregnancy. Immunomodulators such as levamisole are known to enhance immune responses and have been investigated for their potential to improve reproductive outcomes in various animal species. However, the effects of levamisole administered at the time of breeding on both pregnancy rates and key immunoendocrine markers in ewes remain unclear. This study aimed to evaluate the effects of levamisole, administered at varying doses during breeding, on pregnancy rates and selected immune and endocrine parameters in ewes. A total of 30 ewes were randomly assigned to three groups ( n  = 10 each): a control group received subcutaneous saline; the second group received levamisole at 2.5 mg/kg; and the third group received 7.5 mg/kg, all administered at the time of breeding. Blood samples were collected on days 0, 10, and 20 post-breeding. Levels of IFN-γ, IL-2, TNF-α, progesterone, activin A, activin B, follistatin, and total immunoglobulin (Ig) were measured using commercial ELISA kits. Results Pregnancy rates did not differ statistically between the groups ( p  > 0.05). However, on day 10, pregnant ewes in the 2.5 mg/kg group showed significantly higher activin A levels ( p  < 0.05). Activin B levels were elevated in pregnant ewes in both the control group (day 10) and the 7.5 mg/kg group (day 20) ( p  < 0.05). Follistatin levels were higher on day 10 in pregnant control animals ( p  < 0.05). IFN-γ and IL-2 levels increased in pregnant animals, particularly in control and 7.5 mg/kg groups, respectively. Progesterone levels peaked on day 10 in all groups. Conclusions While the measured immune and endocrine parameters varied, these changes did not impact the pregnancy rates.

Background A novel way to study the species composition and diversity of nematode parasites in livestock is to perform deep sequencing on composite samples containing a mixture of different species. Herein we describe for the first time the nematode community structures (nemabiomes) inhabiting Swedish sheep and how these are/were affected by host age and recent anthelmintic treatments. Methods A total of 158 fecal samples were collected ( n  = 35 in 2007 and n  = 123 in 2013–2016) and cultured from groups of sheep on 61 commercial farms in the south-central part of the country where most animals are grazed. Among the samples, 2 × 44 (56%) were paired collections from the same groups pre- and post-treatment with anthelmintics such as macrocyclic lactones, benzimidazoles or levamisole. Samples were analyzed for their nemabiome using the PacBio platform followed by bioinformatic sequence analysis with SCATA. Species richness and diversity were calculated and analyzed in R. Results Nematode ITS2 sequences were found in all larval culture samples except two, even though the fecal egg counts were below the McMaster threshold in 20 samples. Sequencing yielded, on average, 1008 sequences per sample. In total, 16 operational taxonomical units (OTU), all with ≥ 98 % identity to sequences in the NCBI database, were recognized. The OTUs found represented nematode species of which ten are commonly associated with sheep. Multiple species were identified in all pre-anthelmintic treatment larval culture samples. No effects on nematode diversity were found in relation to host age. On the other hand, recent anthelmintic treatment lowered species richness, especially after use of ivermectin and albendazole. Interestingly, despite zero egg counts after use of levamisole, these samples still contained nematode DNA and especially H. contortus . Conclusions Our findings provide evidence that nemabiome analysis combined with diversity index analysis provides an objective methodology in the study of the efficacy of anthelmintic treatment as both high and low abundant species were detected. Graphical Abstract