Evaluation of the Sprague-Dawley rat as a model for vertical transmission of Brucella abortus

Vertical transmission of Brucella abortus in Sprague-Dawley (SD) rats was verified with microbiologic, serologic, and polymerase chain reaction (PCR) methods. The 38 initially Brucella-free SD rats, weighing 200 to 250 g, were injected subcutaneously with 50 microL of a suspension containing 1 x 10(9) colony-forming units (cfu) of B. abortus biotype 1 Korean isolate. The rats were allowed to mate with uninfected SD rats. The isolate was detected by culture and by AMOS (abortus, melitensis, ovis, suis) PCR in testis tissue of infected male rats and splenic tissue of infected female rats. By 7 d after inoculation, the results of both the rose bengal test (RBT) and the plate agglutination test (PAT) were positive for antibody against B. abortus; the reciprocal antibody titre ranged from 200 to 400 in the 1-mo-old offspring and 800 in their dams. The infected rats directly transmitted Brucella to their breeding partners and offspring. Fetuses of infected dams were found to be infected at 20 d of gestation. These data are discussed in relation to a model for epizootic and zoonotic cases possibly involving wild animals. Additional rigorous experiments are warranted to explore the value of this model in developing measures to prevent congenital brucellosis.     

Immunization of proteins from Toxascaris leonina adult worm inhibits allergic specific Th2 response

Recently, the influence of parasitic infections on the incidence of allergic diseases has become the focus of increased attention. In order to ascertain whether parasite-derived proteins could inhibit the allergic specific Th2 response, we applied excretory-secretory protein (Tl-ES) or total protein (Tl-TP) of the adult worm Toxascaris leonina to asthma model mice prior to or simultaneously with OVA challenge, after which we assessed the OVA-specific Th2 responses. The group subjected to immunization with Tl-ES and Tl-TP (immunized group) evidenced a thinning of the bronchial epithelial and muscle layer, a disruption and shedding of epithelial cells, a reduction in the number of goblet cells, and a reduction in mucus production as compared to the group treated with Tl-ES coupled with OVA challenge (challenge with OVA groups) and the OVA-induced asthma group. The administration of Tl-ES and Tl-TP, regardless of injection time, was shown to inhibit the recruitment of inflammatory cells into the airway, and in particular, macrophages, neutrophils, and lymphocytes were significantly reduced as the result of the parasite proteins. However, the total number of eosinophils was slightly reduced as the result of the administration of parasite proteins. Sensitization and OVA challenge was shown to accelerate the secretion of Th2 cytokines (IL-4 and IL-5) within the lung, but in the immunized groups, those levels were lower. The administration of Tl-TP and OVA challenge group also evidenced a significant reduction in IL-4 levels as compared to the OVA-challenged group. The concentrations of Th2 cytokines in the Tl-ES and OVA challenge group were more similar to those observed in the OVA-challenged group. The concentration of IL-10 and TGF-beta in the lung was decreased substantially in the OVA-only challenge group, but the Tl-TP immunized group exhibited significantly induced IL-10 cytokine. OVA-specific IgG2a, IgG1, and IgE levels in the immunized groups were significantly lower than those detected in the OVA-challenged group. In conclusion, parasite-derived protein is able to inhibit OVA-specific Th2 responses, and in particular, immunization with parasite proteins exerts a more profound protective effect than is seen with the treatment of allergic reactions. The results of our study are encouraging in terms of our further understanding of the molecular basis of immune evasion by nematodes.     

Molecular serotyping and antimicrobial resistance profiles of Actinobacillus pleuropneumoniae isolated from pigs in South Korea

Background: Actinobacillus pleuropneumoniae (APP) causes porcine pleuropneumonia (PP).

Objective: Serotypes and antimicrobial resistance patterns in APP isolates from pigs in Korea were examined.

Methods: Sixty-five APP isolates were genetically serotyped using standard and multiplex PCR (polymerase chain reaction). Antimicrobial susceptibilities were tested using the standardized disk-agar method. PCR was used to detect β-lactam, gentamicin and tetracycline-resistance genes. The random amplified polymorphic DNA (RAPD) patterns were determined by PCR.

Results: Korean pigs predominantly carried APP serotypes 1 and 5. Among 65 isolates, one isolate was sensitive to all 12 antimicrobials tested in this study. Sixty-two isolates was resistant to tetracycline and 53 isolates carried one or five genes including tet(B), tet(A), tet(H), tet(M)/tet(O), tet(C), tet(G) and/or tet(L)-1 markers. Among 64 strains, 9% and 26.6% were resistance to 10 and three or more antimicrobials, respectively. Thirteen different antimicrobial resistance patterns were observed and RAPD analysis revealed a separation of the isolates into two clusters: cluster II (6 strains resistant to 10 antimicrobials) and cluster I (the other 59 strains).

Conclusion: Results show that APP serotypes 1 and 5 are the most common in Korea, and multi-drug resistant strains are prevalent. RAPD analysis demonstrated that six isolates resistant to 10 antimicrobials belonged to the same cluster.     

Differentiation of hamster liver oval cell following Clonorchis sinensis infection

Oval cells which appear in the liver after hepatic injuries are suspected to be progenitor cells for both hepatocytes and bile duct cells. Oval cell isolated from the livers of the hamsters treated with diethylnitrosamine and 2-acetylaminofluorene and infected with Clonorchis sinensis (CS). cultured for 2 weeks and evaluated for differentiation and plasticity by electron microscopy and immunohistochemistry. In the CS-uninfected group, glycogen granules and peroxisomes were noted in the cells that were cultured for 2 weeks. Starting at 1 week postculture, immunoreactivity of the cells to cytokeratin 19 markedly decreased but that to albumin and alpha-fetoprotein gradually increased. This means that oval cells isolated from hamsters that were not infected with CS differentiated toward hepatocyte lineage. However, in the CS-infected group, cultured cells contained numerous rough endoplasmic reticulum and showed immunoreactivity that was generally in reverse to that of CS-uninfected group, meaning that cells isolated following CS infection were primed by CS and differentiated toward bile duct cell lineage. The results of this study suggested that oval cells are indeed bipolar progenitor cells for hepatocytes and bile duct cells and can differentiate toward either lineage depending upon the priming factor.     

Accumulation of Cadmium and Antioxidant and Hormonal Responses in the Indian Major Carp <Emphasis Type="Italic">Cirrhinus mrigala</Emphasis> During Acute and Sublethal Exposure

Acute (24 h) and sublethal (35 days) effects of cadmium chloride (CdCl₂) were examined in Cirrhinus mrigala using various endpoints (accumulation pattern, thyroid hormones (THs), and antioxidants). The mean concentrations of CdCl₂ for 24 and 96 h were found to be 35.974 and 22.387 mg L^?l, respectively. LC50 concentration of CdCl₂ for 24 h (35.97 mg L^?l) was used for the acute study. For the sublethal studies, fish were exposed to 3.59 mg L^?1 (Treatment I) and 7.19 mg L^?1 (Treatment II) corresponding to 1/10th and 1/5th of 24 h LC50 of the CdCl₂. During acute exposure, higher accumulation of CdCl₂ was noticed in the gill, liver, and kidney of C. mrigala, which is found in the order gill > liver > kidney tissues. Similarly, in sublethal treatments (Treatment I and II), a concentration and time-dependent increase of CdCl₂ accumulation was noticed in the order of gill > liver > kidney. GSH, GST, and GPx activities were found to be relatively lower from the treated groups in both acute and sublethal treatments. However, LPO activity was significantly increased in CdCl₂-treated fish C. mrigala. Further, plasma T₃ reduction was more pronounced than T₄ in acute study. During sublethal treatments, both T₄ and T₃ levels showed a continuous decrease as the exposure period extended. All the values in this study were statically significant (P < 0.01 and P < 0.05).     

Anthocyanins from black soybean seed coats inhibit UVB-induced inflammatory cylooxygenase-2 gene expression and PGE2 production through regulation of the nuclear factor-kappaB and phosphatidylinositol 3-kinase/Akt pathway

Ultraviolet (UV) radiation can cause inflammatory changes and may further contribute to skin carcinogenesis. Anthocyanins are known to be powerful antioxidants that help protect plants from UV damage. Recently, we isolated anthocyanins from black soybean [Glycine max (L.) Merr] seed coats. Thus, we investigated the protective effect of anthocyanins from black soybean seed coats on UVB radiation-induced inflammatory responses and the molecular mechanism responsible for regulation of apoptosis and inflammatory responses. Anthocyanins inhibited UVB-induced cylooxygenase-2 (COX-2) and PGE 2 production through a nuclear factor-kappaB-dependent pathway and regulation of the PI3 kinase/Akt pathway activated by UVB in a human keratinocyte cell line, HaCaT. Topical application of anthocyanins prior to UVB irradiation of hairless mice also inhibited induction of COX-2 and PGE 2. In conclusion, it is suggested that anthocyanins from the seed coat of black soybeans can be used as a useful drug to modulate oxidative disorders including UVB-induced inflammation.     

Using Synchronous Fluorescence Technique as a Water Quality Monitoring Tool for an Urban River

This paper presents a study performed to evaluate the feasibility of implementing a series of heating rods within a zero-valent iron permeable reactive barrier (ZVI-PRB) to enhance the conventional design of PRBs, and thus to improve the barrier’s removal efficiency, and reduce construction and reactive material costs. A numerical modeling approach is undertaken where a hypothetical case is introduced and typical site values are assigned to it. The amount of electrical energy required to heat the barrier from an initial temperature, T ₁, to a final temperature, T ₂ is assessed. Through different simulation cases, the effect of (1) initial temperature of the groundwater entering the PRB, (2) different amounts of total heat injected, (3) distribution of the heating rods, and (4) different types of porous media, on the temperature increase within the PRB are evaluated. Results show that the amount of energy required to increase the temperature of our hypothetical case by 10°C is approximately 935 W, which could be provided by renewable energies such as solar. It is explained that there are multiple benefits for increasing the temperature within the PRB. Apart from boosting reaction rates, temperature increase can reduce water viscosity and increase flow within the barrier, which broadens a barrier’s catch area. It can enhance solubility of gases to reduce blockage caused by gas generation. An upward flow is formed inside the wall which can also help reduce gas blockage. By implementing heating rods, design widths are reduced, which could potentially reduce the materials (thus cost) used and/or justify the use of more expensive reactive material in PRBs. Heating the PRB was found to generate a heat plume of 20 m length downstream of the PRB after one year, which would increase biodegradation of the residual contaminants leaving the PRB within this zone. Overall, it is concluded that implementing the proposed technology within a ZVI-PRB can be justified, and is found to be beneficial in many aspects.     

Lidocaine hydrochloride—sodium bicarbonate as an anesthetic for soft-shelled turtle Pelodiscus sinensis

ABSTRACT:   Attempts were made to understand how the different sizes (mean body weight of 4.1 ± 0.8 g for small and 182.6 ± 23.7 g for large) of the soft-shelled turtle Pelodiscus sinensis are affected by different temperature (25°C or 30°C), and different concentrations (700, 1000 and 1300 p.p.m.) of anesthetic lidocaine hydrochloride–sodium bicarbonate. The soft-shelled turtles were fed to satiation once daily. Three-way anova design (size × temperature × concentration of lidocaine hydrochloride) with duplicate was used. Twenty each of large and small soft-shelled turtles were distributed into six 30-L tanks. Exposure time of the soft-shelled turtle was affected by all factors (temperature, concentration and size). Exposure time of the soft-shelled turtle for anesthetizing decreased with increase in temperature and in concentration of lidocaine hydrochloride, and decrease in size. Recovery time for the soft-shelled turtle was also affected by all factors. Recovery time of the soft-shelled turtle increased with increase in temperature, concentration of lidocaine hydrochloride, and size. According to these results, lidocaine hydrochloride (1000 p.p.m.)–sodium bicarbonate seemed an effective anesthetic for sedating and handling the soft-shelled turtle. However, when an anesthetic is recommended for wide use, especially on commercially important edible organisms, the time organism takes to metabolize and excrete anesthetic chemicals from its body to a safe level must be given to allow fitness for human consumption.     

Automatic submerging and surfacing performances of model submersible fish cage system operated by air control

An automatic, submersible fish cage system using air control was developed and a set of model experiments were conducted to examine the automatic submerging characteristics of the cage. The components of the fish cage consist of a rigid frame assembly with 6 variable ballast tanks and 6 fixed ballast tanks. The variable ballast tanks were used to change the buoyancy characteristics of the system by air control so that the fish cage can either be placed at the surface or submerged. The cage is free to move vertically within a water column by adjusting the weight and the buoyancy with an air control system. The model of this system, with dimensions of 2.20 m in diameter and length and 1.04 m in net cage depth, was constructed to be 1/10 the size of the full-scale system. In the model experiments, the submerging and surfacing characteristics of the cage were regulated with measurements from a water-pressure gauge and a gyroscope incorporated into the automatic control system. Model tests were performed in a still water tank and a large wave tank to develop the algorithm required to control the cage system and to verify the ability of the automatic submersion mechanism to function. The control system was designed so that when the variable ballast tanks were flooded with water, the model descended. To raise the system, compressed air is injected into the tanks by opening the main evacuation valve on the manifold. After the required amount of compressed air is supplied, the main evacuation valves can be shut and as a result, the fish cage becomes buoyant. Measured performance results in a still water tank are then compared with calculations from a previously developed numerical technique. The submerging and surfacing characteristics of the fish cage were relatively similar to the measurements obtained with the physical model experiments using air control. The cage was submerged to a target depth when incidence wave heights were higher than the critical wave height and raised when little wave actions were detected in a wave tank. On the other hand, the cage was placed at the surface when incidence wave heights were the same as the critical wave height or lower.