Qi-Zhong Zhang

Identification and Effect of Two Flavonoids from Root Bark of Morus alba against Ichthyophthirius multifiliis in Grass Carp

Ichthyophthirius multifiliis (Ich) is an important ciliate that parasitizes gills and skin of freshwater fish and causes massive fish mortality. In this study, two flavonoids (kuwanons G and O) with anti-Ich activity were isolated by bioassay-guided fractionation from the root bark of Morus alba, an important plant for sericulture. The chemical structures of kuwanons G and O were elucidated by spectroscopic analyses. Kuwanons G and O caused 100% mortality of I. multifiliis theronts at the concentration of 2 mg/L and possessed a median effective concentration (EC50) of 0.8 ± 0.04 mg/L against the theronts. In addition, kuwanons G and O significantly reduced the infectivity of I. multifiliis theronts at concentrations of 0.125, 0.25, 0.5, and 1 mg/L. The median lethal concentrations (LC50) of kuwanons G and O to grass carp were 38.0 ± 0.82 and 26.9 ± 0.51 mg/L, which were approximately 50 and 35 times the EC50 for killing theronts. The results indicate that kuwanons G and O have the potential to become safe and effective drugs to control ichthyophthiriasis.

Antiparasitic Effect of Cynatratoside-C from Cynanchum atratum against Ichthyophthirius multifiliis on Grass Carp

Ichthyophthirius multifiliis (Ich), a fish ectoparasite, comprises an important challenge in the aquaculture industry. In this study, a steroidal glycoside, cynatratoside-C, isolated from Cynanchum atratum roots by bioassay-guided fractionation was used to treat I. multifiliis. The cynatratoside-C at 0.25 mg/L demonstrated a 100% mortality of I. multifiliis in vitro after 5 h exposure. The 5 h median effective concentration (EC50) of cynatratoside-C to nonencysted tomonts was 0.083 mg/L. In addition, cynatratoside-C at concentrations of 0.125 and 0.06 mg/L could completely terminate the reproduction of encysted tomonts. The cynatratoside-C at 2 mg/L could cure the infected grass carp within 48 h. The exact mechanism of cynatratoside-C for killing I. multifiliis is unknown, but it manifests itself microscopically through loss of membrane integrity of nonencysted tomonts or through releasing immature theronts from encysted tomonts. The immature theronts finally died before infecting fish. On the basis of these results, cynatratoside-C could be used as a natural anti-I. multifiliis agent.

Parasiticidal effects of Morus alba root bark extracts against Ichthyophthirius multifiliis infecting grass carp

Ichthyophthirius multifiliis (Ich), an important fish parasite, can cause significant losses in aquaculture. To find efficacious drugs to control Ich, the root bark of white mulberry Morus alba was evaluated for its antiprotozoal activity. Bark was powdered and extracted with 1 of 5 organic solvents: petroleum ether, chloroform, ethyl acetate, acetone, or methanol. The extracts were concentrated, dissolved in 0.1% (v/v) DMSO, and used for anti-Ich trials. Acetone and ethyl acetate extracts significantly reduced the survival of Ich tomonts and theronts. In vitro, acetone extract at 25 mg l-1 killed all non-encysted tomonts, at 50 mg l-1 eradicated all encysted tomonts, and at 8 mg l-1 caused mortality of all theronts. Ethyl acetate extract at 50 mg l-1 eliminated all non-encysted tomonts, at 100 mg l-1 killed all encysted tomonts and terminated tomont reproduction, and at 8 mg l-1 killed all theronts. Low concentrations (2 and 4 mg l-1) of acetone and ethyl acetate extracts could not kill all theronts after 4 h exposure, but a significant decrease in theront infectivity was observed following 30 min of pretreatment with the extracts. The 96 h LC(50) values of acetone and ethyl acetate extracts to grass carp were 79.46 and 361.05 mg l-1, i.e. much higher than effective doses for killing Ich theronts (8 mg l-1 for both extracts) and non-encysted tomonts (12.5 and 25 mg l-1, respectively). Thus M. alba extract may be a potential new, safe, and efficacious drug to control Ich.

In vitro and in vivo assessment of the effect of antiprotozoal compounds isolated from Psoralea corylifolia against Ichthyophthirius multifiliis in fish.

Highlights • Two antiprotozoal compounds from P. corylifolia were isolated and identified.• In vitro antiprotozoal efficacies of isopsoralen and psoralidin were evaluated.• Psoralidin had a detrimental effect on I. multifiliis trophont in situ.• The action mechanism against I. multifiliis trophont in situ was investigated.

Serum Fractalkine and Interferon-Gamma Inducible Protein-10 Concentrations Are Early Detection Markers for Acute Renal Allograft Rejection

OBJECTIVE The aims of this study were to determine if characterization of serum concentrations of interferon-gamma inducible protein-10 (IP-10), fractalkine, and their receptors (CXCR3 and CX3CR1) were predictive of acute allograft rejection in kidney transplant recipients. METHODS Kidney transplant recipients (n = 52) were enrolled in this study and divide into either the acute rejection (AR, n = 15) or non-acute rejection (NAR, n = 35) groups. Serum samples from recipients were collected 1 day prior to transplantation and on days 1, 3, 5, 7, and 9 post-transplantation. The accuracy of chemokine concentrations for predicting acute rejection episodes was evaluated using receiver operator characteristic (ROC) curves. RESULTS AR was diagnosed in 15 patients based on histologic changes to renal biopsies. AR patients had significantly higher serum fractalkine, CXCR1, IP-10, and CXCR3 levels compared to levels observed in the NAR group and healthy controls. Fractalkine and IP-10 had the largest area under the ROC curve at 0.86 (95% confidence interval: 0.77-0.96). Following steroid therapy, chemokine levels decreased, which may serve to predict the therapeutic response to steroid therapy. CONCLUSION Measuring serum levels of fractalkine, IP-10, and their receptors (especially the fractalkine/IP-10 combination) may serve as a noninvasive approach for the early diagnosis of renal allograft rejection.

Anthelmintic activity of saikosaponins a and d from radix bupleuri against Dactylogyrus spp. infecting goldfish.

Disease caused by the parasitic helminths Dactylogyrus spp. results in significant economic damage to the aquaculture industry. Treatment using common chemicals (e.g. formalin) is usually dissatisfactory due to environmental problems, risk of residues, toxicity to fish, and the possibility of anthelmintic resistance. The search for an alternative drug is thus becoming more urgent. This study was designed to evaluate in vivo the anthelmintic efficacy of total saponin (TS), saikosaponin a (SSa), and saikosaponin d (SSd) from radix bupleuri (i.e. the dried root of Bupleurum sp.) based on our previous screening works, with the aim of determining which has commercial potential. Results showed that median effective concentration (EC₅₀) values for TS, SSa, and SSd were 2.01, 1.46, and 0.74 mg l⁻¹, respectively. The acute toxicities against goldfish Carassius auratus for TS, SSa, and SSd were also determined, with median lethal concentration (LC₅₀) of 8.99, 11.20, and 1.54 mg l-1, respectively. The resulting therapeutic indices (TIs) indicated that SSa (TI = 7.67) is a potential therapeutic agent for treating Dactylogyrus infection.

Cynatratoside-C efficacy against theronts of Ichthyophthirius multifiliis, and toxicity tests on grass carp and mammal blood cells.

Infection by Ichthyophthirius multifiliis, a ciliated protozoan parasite, results in high fish mortality and causes severe economic losses in aquaculture. To find new, efficient anti-I. multifiliis agents, cynatratoside-C was isolated from Cynanchum atratum by bioassay-guided fractionation in a previous study. The present study investigated the anti-theront activity, determined the toxicity of cynatratoside-C to grass carp Ctenopharyngodon idellus and mammalian blood cells, and evaluated the protection of cynatratoside-C against I. multifiliis theront infection in grass carp. Results showed that all theronts were killed by 0.25 mg l-1 of cynatratoside-C in 186.7 ± 5.8 min. Cynatratoside-C at 0.25 mg l-1 was effective in treating infected grass carp and protecting naive fish from I. multifiliis infestation. The 96 h median lethal concentration (LC50) of cynatratoside-C to grass carp and 4 h median effective concentration (EC50) of cynatratoside-C to theront were 46.8 and 0.088 mg l-1, respectively. In addition, the hemolysis assay demonstrated that cynatratoside-C had no cytotoxicity to rabbit red blood cells. Therefore, cynatratoside-C could be a safe and effective potential parasiticide for controlling I. multifiliis.