In vitro evaluation of antileishmanial activity of Boswellia serrata essential oil nanoliposome.

BACKGROUND: Leishmaniasis poses a significant health risk. OBJECTIVES: This study aimed to evaluate the effects of Boswellia serrata (B. serrata) essential oil nanoliposomes on Leishmania tropica (L. tropica) in vitro. METHODS: A mixture of B. serrata essential oil, phosphatidylcholine and Tween 80 were used to prepare B. serrata essential oil nanoliposomes, followed by drying, hydration and size characterisation. The promastigotes of L. tropica were cultured in Roswell Park Memorial Institute medium (RPMI-1640) containing streptomycin, penicillin and fetal bovine serum. Different concentrations of B. serrata essential and nanoliposomes were tested for their antileishmanial properties by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium Bromide tests (MTT). RESULTS: Results of Dynamic Light Scattering (DLS) for B. serrata nanoliposomes indicate that they are successful at producing nanoliposomes with dimensions of 74.8 nm. At 1 µg/mL dose, B. serrata essence caused 17 ± 1.73% mortality, while B. serrata nanoliposomes induced 26 ± 1.15% mortality. B. serrata essence achieved a mortality of 55 ± 2.88% at 10 µg/mL, whereas B. serrata nanoliposomes demonstrated a mortality of 63.66±0.88% at 10 µg/mL. Furthermore, there was a significant difference between similar concentrations of B. serrata and B. serrata nanoliposomes. The LC50 of B. serrata essential oil is 7.26 µg/mL in the 95% confidence interval (12.13-5.25). The LC90 value of B. serrata essential oil is 129.37 µg/mL in the 95% confidence interval (50.07-852.58). The LC50 value of B. serrata nanoliposome is 4.20 µg/mL in the 95% confidence interval (6.13-3.10). LC90 value for B. serrata nanoliposome is calculated as 91.89 µg/mL in the 95% confidence interval (37.09-583.29). CONCLUSIONS: In vitro experiments have shown that B. serrata oil and the nanoliposome suppress the proliferation of L. tropica promastigotes, which suggests it may be a promising option for treating leishmaniasis.

Pivotal role of permeability-glycoprotein in absorption of praziquantel into Dicrocoelium dendriticum parasite.

Dicrocoelium dendriticum affects the livers of ruminants and causes several deleterious effects on animal health status. The aim of this study was to investigate the role of permeability-glycoprotein (P-gp) in absorption of praziquantel (PZQ) into D. dendriticum flukes by co-incubation with verapamil (VPL), an inhibitor of P-gp, under in vitro conditions. Mature flukes of D. dendriticum were collected from naturally infected sheep livers. The flukes were incubated with different concentrations of PZQ and VPL (50 and 100 µg/ml) in culture media and after several times of exposure (2, 6, 12, and 24 h), the concentration of PZQ absorbed in the parasites was measured by high-performance liquid chromatography. At 2-h post-incubation, the highest concentration of PZQ was noted as 0.92 µg/ml in the flukes treated with 100 µg/ml of each PZQ and VPL. After 24-h of exposure, VPL at all tested concentrations resulted in significant increase in absorption of PZQ into the parasite. Co-incubation of lancet flukes with VPL and PZQ roughly doubled the absorption of PZQ into them. Results of tegumental structures analysis by light microscopy confirmed higher efficacy of combination of VPL and PZQ. In conclusion, co-administration of VPL, especially at the concentration of 100 µg/ml, was able to increase PZQ uptake in Dicrocoelium flukes at all time points of the study.

In vitro toxicity of combination of amitraz and carvacrol on Demodex canis.

Background: Canine generalized demodicosis is an inflammatory skin disease, which usually has time-consuming and frustrating treatments. Aim: The present study aimed to evaluate the acaricidal activity of carvacrol as a green drug and a combination of carvacrol and amitraz on Demodex canis mites and investigate mites' survival lifetime. Methods: Mite samples were collected from a dog affected by demodicosis and had been diagnosed with deep skin scrapings. The motility of mites was examined by using optical microscopy. Samples were tested with 5% of carvacrol, the combination of Carvacrol 5% + Amitraz 0.05%, Amitraz 0.05% as a positive control, and mineral oil as a negative control. The interval time between the adding the test solutions and the moment the last mite ceased was defined as the survival time in the samples and the killing times of mites in each group were compared with each other. Results: It was determined that after administration of a 5% concentration of carvacrol, lethal effects were faster than the combination solution of Carvacrol 5% + Amitraz 0.05%, and the survival times in the control groups were longer than in both treatment groups. Conclusion: Carvacrol, as one of the most important bioactive terpenes, had the most effective in vitro miticidal activity, and it seems that carvacrol alone or in combination with amitraz can be suggested as a possible therapy in the treatment of canine demodicosis.

Dietary Supplementation with Eugenol Nanoemulsion Alleviates the Negative Effects of Experimental Coccidiosis on Broiler Chicken's Health and Growth Performance.

The present study investigated the protective efficacy of dietary supplementation with clove essential oil (CEO), its main constituent eugenol (EUG), and their nanoformulated emulsions (Nano-CEO and Nano-EUG) against experimental coccidiosis in broiler chickens. To this aim, various parameters (oocyst number per gram of excreta (OPG), daily weight gain (DWG), daily feed intake (DFI), feed conversion ratio (FCR), serum concentrations of total proteins (TP), albumin (ALB), globulins (GLB), triglycerides (TG), cholesterol (CHO) and glucose (GLU), serum activity of superoxide dismutase (SOD), glutathione s-transferase (GST), and glutathione peroxidase (GPx)] were compared among groups receiving CEO supplemented feed (CEO), Nano-CEO supplemented feed (Nano-CEO), EUG supplemented feed (EUG), Nano-EUG supplemented feed (Nano-EUG), diclazuril supplemented feed (standard treatment, ST), or basal diet [diseased control (d-CON) and healthy control (h-CON)), from days 1-42. Chickens of all groups, except h-CON, were challenged with mixed Eimeria spp. at 14 days of age. Coccidiosis development in d-CON was associated with impaired productivity (lower DWG and higher DFI and FCR relative to h-CON; p < 0.05) and altered serum biochemistry (decreased TP, ALB, and GLB concentrations and SOD, GST, and GPx activities relative to h-CON; p < 0.05). ST effectively controlled coccidiosis infection by significantly decreasing OPG values compared with d-CON (p < 0.05) and maintaining zootechnical and serum biochemical parameters at levels close to (DWG, FCR; p < 0.05) or not different from (DFI, TP, ALB, GLB, SOD, GST, and GPx) those of h-CON. Among the phytogenic supplemented (PS) groups, all showed decreased OPG values compared with d-CON (p < 0.05), with the lowest value being measured in Nano-EUG. All PS groups showed better values of DFI and FCR than d-CON (p < 0.05), but only in Nano-EUG were these parameters, along with DWG, not different from those of ST. Furthermore, Nano-EUG was the only PS group having all serum biochemical values not different (or even slightly improved) relative to ST and h-CON. In conclusion, the tested PS diets, especially Nano-EUG, can limit the deleterious effects of coccidiosis in broiler chickens, due to anticoccidial activity and possibly their reported antioxidant and anti-inflammatory properties, thereby representing a potential green alternative to synthetic anticoccidials.

Laboratory and field efficacy of terpene combinations (carvacrol, thymol and menthol) against the poultry red mite (Dermanyssus gallinae).

In this study, we evaluated the efficacy of combinations of the terpenes carvacrol, thymol, and menthol on Dermanyssus gallinae under in vitro (direct contact and vapour efficacy assays) and in-use conditions. Combinations investigated in vitro included carvacrol-menthol, thymol-menthol, carvacrol-thymol, and carvacrol-thymol-menthol. Results from contact efficacy studies showed that carvacrol-thymol-menthol was the most efficacious compared to the other combinations (P < 0.05), killing 100 % of the mites when at a concentration of 0.5 µg/mL. Thymol-menthol showed the weakest acaricidal activity with an approximate 80 % mortality rate at a concentration of 2 µg/mL. Based on the results of the vapour efficacy assay, all tested combinations appear to exert their acaricidal efficacy not only through direct contact, but also by fumigant action. A subsequent in-field efficacy investigation in a cage system laying farm was carried out to confirm the miticidal activity of the carvacrol-thymol-menthol combination. A formulation based on the triple combination of carvacrol-thymol-menthol diluted 1:40 in water was sprayed, with 1-week interval, in an 8000-laying hen farm, in presence of hens. The number of mites were counted by monitoring traps for 21 days through the study. The number of mites in the treated group decreased until day 5 after the first spray, but increased on days 5 and 7. However, after the second spray on day 7, the number of mites showed a significant decrease on day 10. This terpene-based combination which showed promising in vitro and in field miticidal activity can be a step forward in greening the future of pest management, also producing residue-free eggs.

Pharmacokinetics and therapeutic efficacies of fenbendazole in comparison with levamisole in helminth-infected Caspian turtles (Mauremys caspica).

The pharmacokinetics and bioavailability of fenbendazole and levamisole were determined in Caspian turtles after a single intravenous (i.v.) and subcutaneous (s.c.) administration. Thirty turtles diagnosed as naturally infected with Serpinema microcephalus and Falcaustra armenica nematodes received fenbendazole (50 mg/kg) or levamisole (10 mg/kg) by i.v. and s.c. administrations. Blood samples were collected at time 0, 0.125, 0.25, 0.5, 1, 2, 4, 8, 12, 24, and 48 h after drug administration. Plasma drug concentrations were determined by a validated high-performance liquid chromatography method. Data were analyzed by noncompartmental methods. The mean elimination half-life of levamisole was 5.16 h and 12.03 h for i.v. and s.c. routes, respectively, and for fenbendazole, the mean elimination half-life was 25.38 h (i.v.) and 29.77 h (s.c.). The total clearance and volume of distribution at steady-state for levamisole and fenbendazole following i.v. administration were 0.22, 0.44 ml/g/h, and 1.06 and 7.35 ml/g, respectively. For the s.c. route, the peak plasma concentration of levamisole and fenbendazole was 10.53 and 5.24 µg/mL, respectively. The s.c. bioavailability of levamisole and fenbendazole was complete. Considering high anthelmintic efficacy and bioavailability after s.c. administration of levamisole and fenbendazole, and the absence of adverse effects, this route of administration is an easy and efficacious way of treating nematodes in Caspian turtles.

Pharmacokinetics and therapeutic efficacy of levamisole in Ascaridia galli experimentally infected ducks.

This study investigated the pharmacokinetics and therapeutic efficacy of levamisole (LVS) after intravenous (i.v.) and oral administrations to healthy and Ascaridia galli-infected ducks by developing an infection model. Twenty-four two-week old ducklings were experimentally infected with A. galli. The ducks were monitored for the development of infection and after 8 weeks they were administered with LVS at a single dose of 30 mg/kg by oral or i.v. administration. Sixteen healthy ducks were subjected to the same treatment and served as control. Serial blood samples were taken for LVS determination with HPLC-UV and pharmacokinetic analysis was carried out based on the non-compartmental approach. The LVS therapeutic efficacy was determined 1 week post drug administration by intestinal worm count at necropsy. In vivo data on development of ascariasis in ducks showed that 8 weeks post inoculation the number of eggs per gram of feces reached at least 100 in each bird. After a single dose of LVS, no parasites were recovered upon necropsy. Results of the pharmacokinetic study showed no statistical differences between infected and non-infected birds for both routes of administration. The mean oral bioavailability was slightly below 50% in both experimental groups. In conclusion, the pharmacokinetics of LVS in ducks was not affected by experimentally-induced ascariasis. A single dose of LVS was found to be efficient against experimental ascariasis in ducks induced by in field isolates of A. galli.

Antitrichomonal activity of nanoemulsion of carvacrol on Trichomonas galline: formulation development and in vitro characterization.

In recent years, increasing attention has been paid in veterinary medicine to find novel natural resources to reduce the use of synthetic drugs, avoid side effects, and for better compliance of the animals' owners. Metronidazole has been used for many years in the treatment of birds' trichomonosis. Carvacrol is a terpenoid and several biologic activities was attributed to it. The present study developed and characterized a carvacrol nanoemulsion (NanoCAV) and investigated its antitrichomonal activity on Trichomonas gallinae, the causative agent of pigeon trichomonosis, under in vitro condition and compared it with carvacrol (CAV) and the standard antitrichomonal dug, metronidazole (MTZ). Additionally, cytotoxicity of the developed formulation to the fibroblast cell line was evaluated. The NanoCAV mean size and surface charge were 80.5 nm and -31.2 mv, respectively. No significant cytotoxicity was observed for the NanoCAV. Incorporation efficiency of NanoCAV was measured as 75%. Results of antitrichomonal activity assay showed 12 h fifty percent lethal concentrations of 0.39 and 0.27 µg/ml for CAV and NanoCAV, respectively. The NanoCAV based on in vitro activity and low cytotoxicity, can be further studied for its efficacy and safety profile in the pigeons.

Ameliorative effect of Psidium guajava essential oil supplemented feed on chicken experimental coccidiosis.

Present study investigated the therapeutic efficacy of Psidium guajava essential oil (EO) on chicken experimental coccidiosis in comparison to the diclazuril. Seventy-five 1-day-old Ross 308 broiler chickens were allocated into 5 groups: CEO1: received EO at 1 mg kg-1 of feed; CEO5: received EO at 5 mg kg-1 of feed; CT: received diclazuril as standard treatment; CNT: received only basal diet; NC: control chickens; all of the groups except NC were challenged with mixed Eimeria spp. on d14 and received supplemented diet from d1 to d42. Zootechnical records and oocyst per gram (OPG) of feces samples were analyzed on weekly basis. On the last day of the study, blood samples were taken to measure serum concentrations of biochemical parameters and also activities of antioxidant enzymes. ß-caryophyllene and α-pinene were determined as major constituents of the EO. On the 3rd, 4th, and 5th weeks, a significant difference was noted in feed conversion ratio (FCR) between CEO1, CEO5, and CT in comparison to NC and CNT chickens (p < 0.05). No significant difference was observed in OPG between CEO1 and CEO5 (p > 0.05); however, CT showed a lower number of OPG relative to EO supplemented groups (p < 0.05). The highest serum activity of glutathione peroxidase was observed in CEO5 which was higher than other groups (p < 0.05). Results of the present study showed that supplementation of P. guajava EO especially at 5 mg kg-1 of feed may have beneficial effects on prevention of coccidiosis and improvement of health in broilers.

Therapeutic efficacy of triclabendazole in comparison to combination of triclabendazole and levamisole in sheep naturally infected with Fasciola sp.

Fascioliasis is an important parasitic disease which affects productivity of ruminants, and imposes significant economic losses. Benzimidazoles are effective in the treatment of fascioliasis; however, there are several reports on benzimidazoles resistant flukes. Combinational therapy is an approach to delay the emergence of resistant flukes. The present study aimed to compare the efficacy of triclabendazole (TBZ) and combination of triclabendazole and levamisole (TBZ + LVM) in the treatment of sheep naturally infected with Fasciola sp. For this purpose, 40 ewes infected with Fasciola sp. in three groups received TBZ, and TBZ + LVM, or remained untreated as CON. Fecal egg count (FEC), fecal egg count reduction (FECR), liver enzymes activity, albumin, globulin, and total protein levels were measured on day 0, 7, 14, and 28 post treatments. Obtained results showed that treatment with TBZ and TBZ + LVM resulted in significant reduction in FEC (P < 0.05), and FECR reached to values of higher than 90% on 28 day post treatment. The FEC for TBZ + LMV on day 7 and 14 were 12.25 ± 3.82 and 3.08 ± 1.03, respectively which was significantly lower in comparison to TBZ and CON (P < 0.05). Efficacy of TBZ + LMV was higher than TBZ on day 7 and 14 post treatment; however, no significant difference was observed on 28 day. The liver enzyme activities on days 7 and 14 were lower in the TBZ + LVM sheep in comparison to the TBZ and CON. Treatment with TBZ or TBZ + LVM resulted in an increase in albumin and a decrease in globulin. Over all, the present study clarified the importance of combinational therapy, and demonstrated that combination of TBZ + LVM resulted in higher efficacy and earlier improvement of liver conditions in sheep naturally infected with Fasciola sp.

In vitro toxicity of cinnamaldehyde and nanoemulsion of cinnamaldehyde on protoscoleces of hydatid cyst.

Cystic echinococcosis is a major parasitic and zoonotic disease and surgery is the most common treatment of this disease which carries the risk of intraoperative leakage and recurrence. Using scolicidal agent to inactivate cyst contents reduces the risk of recurrence. Considering side effects of available scolicidals and growing interests on natural pharmaceuticals, the present study aimed to evaluate toxicity of cinnamaldehyde (CA), the main component of cinnamon essential oil, and a developed nanoemulsion of cinnamaldehyde (nano-CA) on protoscoleces of hydatid cyst. Nanoemulsion was prepared by the low energy system and characterized by dynamic light scattering to confirm dimensions. For evaluation of scolicidal effects, serial dilutions of CA and nano-CA were mixed with protoscolices suspension and mortality were recorded at 10, 30, and 60 minutes by eosin exclusion test. Albendazole was used as the positive control. The mean diameter of nano-CA was characterized as equal to 88.5 nm, and poly dispersity index was 0.09. After 30 min of treatment, nano-CA, at 50 µg/ml, killed 99.33% of protoscoleces. At the same time point and concentration, CA only caused mortality rate of 26.18%. 30 min-LC50 value of 369.39 µg/ml was obtained for CA, while after 30 min of exposure, nano-CA showed promising rapid activity with LC50 value of 3.22 µg/ml. Nano formulation significantly increased scolicidal activity of CA probably by increasing penetration and tegumental disorganization of protoscoleces. Further in vivo safety studies are needed to introduce nano-CA as a clinically applicable scolicidal agent.

Development and characterization of monoterpene loaded microemulsions as novel scolicidal agents.

Cystic echinococcosis (CE) is one of the most important zoonotic diseases. The primary treatment is surgery and chemical sterilization of the parasitic layers by injection of a scolicidal agent. Available scolicidals possess side effects, and may cause postoperative complications. Several studies reported the scolicidal properties of monoterpene phenols and alcohols such as carvacrol, thymol, and geraniol. The present study aimed to develop, characterize, and assess monoterpene loaded microemulsions as novel green scolicidals products. For this purpose, microemulsions composing 0.37%, 0.75%, and 1.5% of monoterpenoid(s), thymol, carvacrol, and geraniol, alone or in binary or ternary mixtures were formulated. Samples were analyzed by visual inspection, polarizing optical microscope, and dynamic light scattering (DLS). The stability of the samples was evaluated up to a 3-month storage. For the scolicidal bioassay, samples at different concentrations of 200, 100, 50, 25, and 10 µg/ml were added to wells containing 104 viable protoscoleces and mortality rates were recorded at 2, 5, 10, and 20 min after exposure. Results of the present study showed that microemulsions formulated with 0.75% of pure carvacrol or the binary mixture of thymol and carvacrol at 0.375% are promising scolicidal agents.

Detection of Malaria Parasites and Other Haemosporidia in Migratory and Native Birds in Mazandaran and Golestan Provinces, Iran.

Background: A variety of haemoprotozoa including Plasmodium, Haemoproteus and Leucocytozoon cause infections in birds and are transmitted by some known vectors. These parasites cause anemia, low appetite, weakness and ultimately death in birds. The present study was aimed to determine these parasites, in birds of Mazandaran and Golestan provinces in Iran. Methods: The project was performed on 340 live birds in 2016. The samples were collected from February to September 2016, from each bird, two thin and thick blood smears were prepared and the remaining blood about 1ml was kept in EDTA-containing tubes for molecular studies. The slides were stained with 10% Giemsa, then examined microscopically. About ten percent of the negative samples were considered for Polymerase Chain Reaction (PCR) technique, using specific primers to diagnose Plasmodium and Haemoproteus spp. Electrophoresis was done for PCR products and relevant bands to the parasites were identified based on the size. The considered birds belonged to ducks, chickens, roosters, and pigeons. Results: From 340 microscopically examined blood samples 32 (9.5%) samples were positive. Twenty-five (7.35%) of them were infected with the genus Haemoproteus. Seven samples (14%) out of 50 microscopically negative samples were found as Haemoproteus or Plasmodium spp when PCR technique was employed. Conclusion: This study revealed the existence of malaria parasites and other haemosporidia in birds in Iran. Employing molecular methods (PCR examination) could detect more infections.

Peganum harmala and Nigella sativa: anti-leishmanial activity against Leishmania major promastigotes and amastigotes: in vitro and ex vivo experiment.

Leishmaniosis is one of the most important vectors borne disease that is endemic in tropical and subtropical areas. There are many approved treatment for different types of leishmaniosis but all are with some adverse side effects that limited its uses. Here, we attempt to evaluate in vitro and ex vivo anti-leishmanial activities of Peganum harmala (P. harmala) and N. sativa (Nigella sativa) on promastigotes and amastigotes of L. major. The plants were extracted by maceration method and prepared in concentrations of 7.8, 3.9, 1.9, and 0.9 µg. L. major were cultured in RPMI-1640 medium alone and in J774 cell line separately. The extracts at different concentrations were assessed against promastigote (in vitro assay) and amastigotes (ex vivo assay) of L. major for 72 h at 22 and 37°C, respectively. In current work, N. sativa at highest concentration (7.8 µg/ml) showed 54.4 and 60% anti-leishmanial activity with IC50 of 5.3 and 3.278 µg/ml, respectively. Also, P. harmala at highest concentration (7.8 µg/ml) showed 68.9 and 58.6% antileishmanial activity with IC50 of 2.4 µg/ml for both of them, respectively. The SI value was 38.22 for N. sativa, 25.9 for P. harmala, 19.4 for Amphotericin B, and 16.33 for Glucantime. The results of our study indicated that N. sativa and P. harmala are effective against L. major promastigotes and amastigotes and could be consider as an alternative treatments for leishmaniosis. Therefore, it is recommended that further studies be performed to confirm the efficacy and evaluate the toxicity of the herbal extracts.

Antitrichomonal activity of metronidazole-loaded lactoferrin nanoparticles in pigeon trichomoniasis.

In recent years, increasing attention has been paid to the novel drug delivery systems to reduce the dose of the drug and avoid side effects. Metronidazole has been used for many years in the treatment of anaerobic bacterial and protozoal infections. Nanolactoferrin, a newly developed antibacterial agent originated from lactoferrin, is applied both as an active therapeutic and a drug nanocarrier. The present study describes the development and characterization of metronidazole-loaded lactoferrin nanoparticles (nano-MTZ) as well as reports their antitrichomonal activity on Trichomonas gallinae, the protozoal causative agent of pigeon trichomoniasis. The activity of the nano-MTZ is compared with the regular metronidazole formulation (MTZ) under in vitro and in vivo conditions. Additionally, cytotoxicity of the nano-MTZ to fibroblast cell line and possible hepatotoxicity in treated pigeons were evaluated. Nano-MTZ was prepared based on the thermal treatment method and the average size and surface charge of the dispersion were 30.6 nm and - 44.6 mv, respectively. No significant cytotoxicity was noted for the nano-MTZ in comparison to the MTZ. Loading efficiency in nano-MTZ was calculated as 55%. In vitro susceptibility results demonstrated 24 h 90% lethal concentration values of 4.23 and 6.64 µg/mL for MTZ and nano-MTZ, respectively. Oral treatment of the pigeons experimentally infected with T. gallinae resulted in the earlier eradication of the infection in the nano-MTZ-treated pigeons. No adverse effects on the liver function have been observed for the nano-MTZ. These findings suggest that nanolactoferrin is a promising platform for the development of novel MTZ formulations with improved antitrichomonal activity.

Pharmacokinetics and pharmacodynamics of single and multiple-dose levamisole in belugas (Huso huso): Main focus on immunity responses.

The pharmacokinetics of levamisole were determined in the belugas after single intravascular (IV), and single and multiple-dose oral by feed administrations. Also, the effect of levamisole (LVM) on the stress and immune responses of belugas were assessed. One hundred-fourteen healthy belugas in 4 different groups received single LVM administration at the doses of 50 and 100 mg/kg via IV and oral routes. A separate group of 24 belugas were administered oral LVM at the dose of 100 mg/kg for 5 days. Blood samples were collected at different time points after administrations to measure plasma concentrations of LVM by a validated high-performance liquid chromatography (HPLC) assay. For immunological evaluations, a total of 126 belugas received 50 and 100 mg/kg LVM via medicated feed for 5 days or served as the control without any medication; blood samples were recovered on day 0, 1, 3, 5, 7, 10, and 14 to measure hemolytic activity of the complement system (HAC50), serum lysozyme activity, serum antibacterial activity, glucose, cortisol, total protein, albumin and C3 contents. In the single-dose administration, quantified LVM concentrations were dose-dependent and the oral bioavailability was in the range of 43.2-49.6%. In the multiple-dose administration, the peak plasma concentration at the steady state was 45.2 mg/ml, and accumulation ratio was calculated as 3.6. In the immunological study, LVM especially at the dose of 100 mg/kg increased HAC50, lysozyme and antibacterial activity in the sera of treated fish. No significant effect of LVM on glucose and albumin content was observed, but cortisol levels decreased and C3 content was increased, more significantly by LVM at the dose of 100 mg/kg. Our results indicate that LVM is well absorbed after oral administration and reached to concentrations that can affect stress indicators and improve immune responses in belugas.

In Vitro Scolicidal Activity of the Sesquiterpenes Isofuranodiene, α-Bisabolol and Farnesol on Echinococcus granulosus Protoscoleces.

Cystic echinococcosis (CE) remains an important challenge both in humans and animals. There is no safe and suitable remedy for CE, so the discovery of new compounds with promising scolicidal effects, particularly from herbal sources, is of great importance for therapeutic uses in the treatment and prevention of CE reappearance. Sesquiterpenes are C15 organic compounds made up of three isoprene units and mostly occurring as fragrant components of essential oils. They are of economic importance for the cosmetic and pharmaceutical industry, and recently attracted the attention of the scientific community for their remarkable parasiticidal properties. In the present study, we have focused on three known sesquiterpenes, isofuranodiene (IFD), α-bisabolol (BSB), and farnesol (FOH), as important phytoconstituents of the essential oils of wild celery (Smyrnium olusatrum), chamomile (Matricaria chamomilla), and acacia farnese (Vachellia farnesiana), respectively. Protoscoleces were recovered from fertile hydatid cysts and were exposed to different concentrations of the three tested compounds for different exposure times. The viability of protoscoleces was confirmed by 0.1% eosin staining. Results of scolicidal activity evaluations showed that IFD possessed the best effect against Echinococcus granulosus protoscoleces (LC50 and LC90 values of 8.87 and 25.48 µg/mL, respectively), followed by BSB (LC50 of 103.2 µg/mL) and FOH (LC50 of 113.68 µg/mL). The overall toxicity of IFD differed significantly from those of FOH and BSB, while there was no significant difference in toxicity between the latter compounds (p > 0.05). The present study showed that IFD seems to be a promising scolicidal agent and can be further tested to become a candidate for CE treatment.

First Report of Joyeuxiella spp. Infection in Eurasian Lynx and Its Histopathology Study from Iran: A Case Report.

According to the last information of IUCN Red List, Eurasian lynx has been endangered since 2010. The population of this animal is heavily affected by infectious parasites. Two adult Eurasian lynx (one male and one female) were illegally hunted and found dead in Parvar National Park, Semnan Province, Iran. After the autopsy, the tissue and parasite samples were collected from gastrointestinal tract and transferred to 70% alcohol. Samples were recovered and identified as Joyeuxiella spp. Sonsino, 1889. Tissue samples were taken from the place of sticking of parasites on the intestinal wall, for histopathological examination, and were transferred to 10% buffered formalin. Following routine processes and H&E staining, the slides were examined microscopically. Main histopathological observations were epithelial hyperplasia and destruction, inflammatory cell infiltration in mucosa and submucosa of jejunum. To the best of the author's knowledge, this is the first report and histopathological study of Joyeuxiella spp. in the world in Eurasian lynx.

Pharmacokinetics of levamisole after intramuscular and oral administrations to Caspian salmon (Salmo trutta caspius).

The pharmacokinetic parameters of levamisole were determined in the Caspian salmon after intramuscular (IM), oral by gavage, and oral by feed administrations. Eighty-one healthy fish in three different groups received levamisole at the dose of 25 mg/fish by each route. Blood samples were collected at time points of 0, 0.5, 1, 2, 4, 6, 12, 14, and 24 hr after administrations. Plasma levamisole concentrations were measured by a validated high-performance liquid chromatography (HPLC) assay and were analyzed using a noncompartmental approach. The mean terminal half-life was 4.56, 3.95, and 2.91 hr for IM, gavage and feed routes, respectively. The peak plasma concentration for IM, gavage, and feed routes of levamisole were 35.53, 4.63, and 8.36 µg/ml, respectively, at the time of 0.25 for IM, and 1 hr for gavage and feed. The relative bioavailability for gavage and feed routes was 54.80 and 69.30. The similar bioavailability for gavage and feed might be indicative of similar efficacy for these routes of administrations. Further studies are warranted to evaluate the absolute oral bioavailability and the effective dose in Caspian salmon.