Aims: Cholera epidemics have been occurred in Malaysia since 1991 till 2003 which can be proved from the records by the Infectious Diseases Division of the Ministry of Health. Moreover, there were also course of cholera epidemics from the year 1994 to 2003 which had been happened in Sarawak. Cholera outbreaks in Malaysia mostly caused by the El Tor O1 Vibrio cholerae serogroup. The aims of this study were to detect the presence of V. cholerae in clinical and environmental samples (n=28) from Limbang, Sarawak by collaboration with Sarawak Government Hospital and to detect the toxin genes from the isolates. Methodology and results: All the isolates were sub-cultured in alkaline peptone water (APW). The boiled-cell method was used for DNA extraction. The total DNA extracted was amplified by polymerase chain reaction (PCR). Two types of PCR were used in this study which are 16S rRNA PCR and multiplex PCR. The results obtained from the study found out that 16 out of 28 (57.14%) samples were confirmed to be V. cholerae species. Four primers specific for V. cholerae were used in multiplex PCR (O1 type, O139 type, ctxA and ctxAB) to confirm the species type and the toxin genes. All samples shown positive for V. cholerae O1 serotype and 100% positive to all genes for the identification of ctxA and ctxAB genes. Conclusion, significance and impact of study From this study, it showed that multiplex PCR can be used for research purposes in molecular genetics field involving cholera outbreak.
Vibrio cholerae--genetics ; Cholera Toxin

No abstract available.
Calcium* ; Cholera Toxin* ; Cholera* ; Cytoplasm* ; Humans* ; T-Lymphocytes*

We used cholera toxin B subunit (CTB) as a neural tracer to localize motor neuronal cell bodies innervating the digastric muscle. After CTB injection into the left anterior belly, CTB-labelled motor neuronal cell bodies were found in caudal half of the left and right trigeminal nucleus, the left and right facial nucleus, the accessory facial nucleus and the accessory trigeminal nucleus in pons. The total number of CTB-labelled motor neuronal cell bodies were 1,179+/-119.5 in the left pons and 246+/-61.8 in the right pons after CTB injections into the left anterior belly of digastric muscle. After CTB injection into left posterior belly, CTB-labelled motor neuronal cell bodies were found only in the left ventral part of accessory facial nucleus in caudal pons and the total number of CTB-labelled motor neuronal cell bodies were 270+/-29.3.
Animals ; Cholera Toxin* ; Cholera* ; Motor Neurons* ; Pons ; Rats* ; Trigeminal Nuclei

The author evaluated the optimal concentration of 3 compositions of TIC medium which has used as the melanacytes culture medium. The concentrations of placental extract and bovine pituitary extract, which have the ability to promote proliferation of melanocytes, were evaluated also. Modified TIC medium with above 5 components of evaluated concentration was very effective in melanocytes culture. The results were as follows : l. 12-0-tetradecanoyl-phorbol-13-acetate (TPA) showed effective melanocytes proliferating activity at the concentration of 30ngml (p(0.05) 2. Isobutylmet:hyl xanthine (IBMX) showed effective melanocytes proliferating activity at the concentration of 0.3mM (p(0.05) 3. Cholera toxin (CT) showed effective melanocytes proliferating activity at the concentration of )OnM (p(0.05) 4. Two percentages of placental extract in culture medium showed effective melanocytes proliferating activity. S. Two percentages of bovine pituitary extract in culture medium showed effective melanocytes proliferating activity. 6. Placental extract and isobutylmethyl xanthine proved to have high melanocytes proliferating activity. 7. Melanocytes proliferated rapidly on modified TIC medium (Proliferation doubling time . about 43 hours) 8. The peak time of melanocytes proliferation (7.2 X 10/cm) was observed on the seventh day of culture, From this data, this culture system can be recommended as a new melanocytes culture.
Cholera Toxin ; Humans* ; Melanocytes* ; Tics ; Xanthine

Background: Only strains of V cholerae 01 that produce cholera toxin have been associated with epidemics and pandemics in the past; therefore, production of cholera toxin has become an important marker for identifying isolates with the potential to cause epidemics. The Reversed Passive Latex Agglutination test (RPLA) is a semi-quantitative determination of CT or LT in culture fluid. \r\n', u'Objective: To determine the cholera toxin by RPLA in order to evaluate the severity of the epidemic and find an interventional solution.\r\n', u'Subjects and methods: The technique of RPLA enables soluble antigen such as bacterial toxins to be detected in an agglutination assay. A total of 44 strains of V. cholerae 01 were tested for the production of the cholera toxin by RPLA.\r\n', u' Results and conclusion: The concentration of cholera toxin was determined from strains isolated in 2007 much higher than that from the strains isolated in 2004. These results can explain the severe cholera endemic in 2007 in comparison with the endemic in 2004. The RPLA test is a simple and reliable method for determining cholera toxin and suitable for epidemiologic studies on cholera. \r\n', u'
Reversed passive latex agglutination ; cholera toxin

The application of the cholera vaccine is one of the cholera prevention and control strategies. Cholera vaccines stimulate mucosal immune to play the role of antibacteria and antitoxin. When the cholera toxin B subunit is added in the cholera vaccine, it could also defend against some diarrhea associated pathogens by cross-protection. Oral inactivated cholera vaccines are commercially available now. The oral live vaccine candidates are under development. The development of cholera vaccine is not only on the technical aspect, based on the situations of epidemic areas and population, cost, storage and transportation condition should also be considered. Though the argument on the use of cholera vaccine in epidemic areas and population in high risk existed previously, its vaccination has reached agreement now based on the clinical trials and evaluations during epidemic period.
Administration, Oral ; Cholera ; Cholera Toxin ; Cholera Vaccines ; Cross Protection ; Diarrhea ; Humans ; Vaccination ; Vaccines, Attenuated ; Vaccines, Inactivated

Though the existence of glutamate-immunoreactive(GL-IR) neurons has been suggested in the nucleus ambiguous(NA) by immunocytochemistry, information regarding the distribution of neurons containing glutamate as a neurotransmitter has been to be elucidated. The author focused on distribution and morphology of GL-IR neurons in the NA, which were compared with cholera toxin beta-subunit(CTB) labeled neurons after its injection to the nodose ganglion(NG) in the cat. The results showed that the majority of neurons in the NA were immunoreactive to excitatory neurotransmitter glutamate, and they seemed to be distributed evenly without any special area of predilection or grouping pattern. The cellular shape was predominantly multipolar. GL-IR neurons showed some similarity in morphology and distribution pattern with CTB labeled cells.
Animals ; Cats* ; Cholera Toxin* ; Cholera* ; Glutamic Acid ; Immunohistochemistry ; Larynx ; Motor Neurons ; Neurons* ; Neurotransmitter Agents ; Nodose Ganglion*

In the rat brain stem, neurons innervating the sublingual and submandibular gland were investigated by means of cholera toxin B subunit (CTB) and pseudorabies virus (PRV). Injection of CTB into the sublingual gland and PRV into the submandibular gland, neural tracer labeled neurons showed similar positions in central nervous system with PRV into the sublingual gland and CTB into the submandibular gland. CTB labeled-neurons were observed in superior salivatory nucleus, PRV labeled-neurons in superior salivatory nucleus and reticular nucleus. CTB was more fine tracer than PRV for observation of superior salivatory nucleus. The size of CTB labeled-neurons is larger in submandibular gland than in sublingual gland. The size of PRV labeled-neurons were nearly the same after injection to submandibular or sublingual gland. No neurons were labeled together with CTB and PRV. Neurons innervating sublingual and submandibular gland were localized independently in superior salivatory nucleus. These results provided a neuroanatomical data of the neurons innervating the sublingual and submandibular gland in the superior salivatory nucleus.
Animals ; Brain Stem ; Central Nervous System ; Cholera Toxin* ; Cholera* ; Herpesvirus 1, Suid* ; Neurons ; Pseudorabies* ; Rats ; Sublingual Gland ; Submandibular Gland*

In the present study, the effect of intrathecal (i.t.) or intracerebroventricular (i.c.v.) administration with cholera toxin (CTX) on the blood glucose level was examined in ICR mice. The i.t. treatment with CTX alone for 24 h dose-dependently increased the blood glucose level. However, i.c.v. treatment with CTX for 24 h did not affect the blood glucose level. When mice were orally fed with D-glucose (2 g/kg), the blood glucose level reached to a maximum level at 30 min and almost returned to the control level at 120 min after D-glucose feeding. I.c.v. pretreatment with CTX increased the blood glucose level in a potentiative manner, whereas i.t. pretreatment with CTX increased the blood glucose level in an additive manner in a D-glucose fed group. In addition, the blood glucose level was increased in formalin-induced pain animal model. I.c.v. pretreatment with CTX enhanced the blood glucose level in a potentiative manner in formalin-induced pain animal model. On the other hand, i.t. pretreatment with CTX increased the blood glucose level in an additive manner in formalin-induced pain animal model. Our results suggest that CTX administered supraspinally or spinally differentially modulates the regulation of the blood glucose level in D-glucose fed model as well as in formalin-induced pain model.
Animals ; Blood Glucose ; Brain ; Cholera ; Cholera Toxin ; Glucose ; Hand ; Mice ; Mice, Inbred ICR ; Models, Animal ; Spinal Cord

Animals ; Antibodies, Bacterial ; blood ; Cholera Toxin ; immunology ; Cholera Vaccines ; immunology ; Fimbriae, Bacterial ; immunology ; Mice ; Vibrio cholerae O139 ; immunology