Prognostic Factors in Recurrent Congenital Muscular Torticollis

@#Introduction: Congenital muscular torticollis (CMT), primarily resulting from unilateral shortening and fibrosis of the sternocleidomastoid muscle. One of the common surgical complications is recurrent deformity. However, the associations between unipolar or bipolar release, age of the patient, and the recurrence of the disease are unclear. Therefore, the purpose of this study was to evaluate the factors associated with recurrence after surgery. Materials and Methods: A retrospective review was performed in 47 patients who were diagnosed with CMT and had been treated surgically with unipolar or bipolar release between January 2007 and December 2015. Demographic data (sex, sides, surgical technique, age at time of surgery, period of follow-up, complications and recurrence) were recorded. Results: Forty-seven patients with an average age of 8.7 years old at time of surgery. Twenty-six patients had rightsided muscular torticollis, while 21 had left-sided. The average follow-up time was 2 years (range, 2–4 years). The average age of unipolar release was 8.8 years old (range, 2- 18 years old), while the average age of bipolar release was 8.7 years old (range, 2–13 years old). Recurrence occurred in 11 patients (9 in unipolar and 2 in bipolar release). Sex, side of deformity, type of surgery and age at time of surgery showed no statistically significant as a factor for recurrence rate, however recurrence of unipolar more than bipolar surgery was nearly two times revealing clinical significance. Conclusions: Sex, side of deformity, type of surgery and age at time of surgery were not associated with the recurrence deformity.

Genetic variation of NADH dehydrogenase subunit 1 (nad1) mitochondrial gene sequence in adult Necator americanus hookworms recovered from a female patient in Thailand

@#Two female and one male adult hookworms were recovered from a female patient in Thailand. Based on gross and microscopic morphology, the three hookworms are members of Necator americanus. Phylogenetic reconstruction based on partial NADH dehydrogenase subunit 1 (nad1) mitochondrial gene sequences shows that these hookworms belong to the same genetic lineage as N. americanus adult worm from Zhejiang, China. The male and female hookworms were genetically distinct, belonging to two different nad1-haplotypes. This is the first report targeting the nad1 gene on the identification and genetic characterization of the human hookworms originated from infected patient. The nad1 gene marker is useful for species and higher taxa differentiation of hookworms.

Genetic differentiation of Anisakis species (Nematoda: Anisakidae) in marine fish Priacanthus tayenus from Gulf of Thailand

@#Members of the genus Anisakis are parasitic nematodes of the family Anisakidae. They are potential zoonotic parasites, causing anisakiasis in humans who consume raw or undercooked seafood (fish or squid) infected with the larvae of this nematode. In the present study, anisakid nematodes collected from the marine fish Priacanthus tayenus (Purplespotted big-eye) caught from the Gulf of Thailand were examined morphologically and characterized genetically by DNA sequence analysis. Sequence data from the mitochondrial cytochrome c oxidase subunit II (mtDNA cox2) gene were used to identify these nematodes to species level and to evaluate the phylogenetic relationship among various taxa. All the 15 third-stage larvae of Anisakis nematodes investigated in this study belonged to the same genetic lineage as the A. typica species complex (named here as A. typica sp. T – T for Thailand). Eight mtDNA cox2 haplotypes were revealed in the 15 isolates of this Anisakis from Thailand. The mtDNA cox2 haplotypes of A. typica sp. T from Thailand were genetically distinct from those of the A. typica sensu stricto. Taxonomic description of this A. typica sp. T as a distinct species however awaits the availability of adult specimens.

Geographical distribution and incidence of Angiostrongylus lungworms (Nematoda: Angiostrongylidae) and Geographical distribution and incidence of Angiostrongylus lungworms (Nematoda: Angiostrongylidae) and their rodent hosts in Thailand

The rat lungworm Angiostrongylus cantonensis, a zoonotic parasite, is known to be responsible for eosinophilic meningitis and meningoencephalitis in humans in many countries worldwide. Another congener A. malaysiensis is a potential pathogen. Rodents as natural definitive host of the parasites are abundant and globally widespread. In this study, the prevalence of Angiostrongylus infection in wild rats was investigated in twenty-four provinces of Thailand during the period December 2011 to June 2014. Of the 669 wild rats sampled, 46 (6.88%) were infected with Angiostrongylus lungworms. The rodents harbouring A. cantonensis worms included Bandicota indica, Bandicota savilei, Rattus exulans, Rattus norvegicus, Rattus rattus complex and Rattus tiomanicus, and those harbouring A. malaysiensis were B. savilei, Rattus losea, R. norvegicus and R. rattus complex. No parasite was recovered from Maxomys surifer (n=11), Mus musculus (n=1), Niviventer fulvescens (n=2), Rattus argentiventer (n=4), Rattus nitidus (n=3) and Sundamys muelleri (n=3). In positive rats, the incidence of infection with Angiostrongylus lungworms was variable among host species and provinces. There were also considerable variation in the proportion of male and female worms among rodent hosts and localities. Two hundred and thirty-five of the collected worms were male and 282 were female. The mean worm burden in the positive rats was 11.24 and ranged from 1 to 61. 81.82% (423/517) of the adult worms were morphologically identified as A. cantonensis, and 18.18% (94/517) were A malaysiensis. One R. rattus from Prachuap Khiri Khan had mixed infection of A. cantonensis and A. malaysiensis (10 worms of each species). The overall number of male (202) and female (221) A. cantonensis worms was not significantly different (χ2 = 0.86, 0.50 > P > 0.30). However, the overall number of male (33) and female (61) A. malaysiensis worms was significantly different (χ2 = 8.34, P < 0.01). The present study added one new definitive host (R. tiomanicus) for A. cantonensis and two new definitive hosts (B. savilei and R. losea) for A. malaysiensis in Thailand. Our data update and contribute significantly to existing knowledge of the geographical distribution of A. cantonensis in wild rats in Thailand and confirm the occurrence of A. malaysiensis throughout the country.

Purification of a specific immunodiagnostic Parastrongylus cantonensis antigen by electroelution from SDS-polyacrylamide gels.

A 31-kDa glycoprotein antigen was purified by electrophoresing the crude extract of Parastrongylus cantonensis adult worms in a 12% SDS-polyacrylamide gel, identifying the 31-kDa component with prestained molecular weight standards, cutting the desired gel strip, and then isolating it by electroelution. Antigen fraction of 31 kDa was re-electrophoresed, transferred to a nitrocellulose membrane and found to be reactive with only the sera from patients with parastrongyliasis. No reactive band was observed with the sera from other related parasitic infections, eg, gnathostomiasis, toxocariasis, filariasis, paragonimiasis, cysticercosis and malaria, and the normal healthy control sera. This antigen fraction isolated showed 100% sensitivity and 100% specificity in the enzyme-linked immunosorbent assay (ELISA) for the detection of 31-kDa specific antibody in the sera from patients with parastrongyliasis. The P. cantonensis antigen of 31 kDa has been obtained by this means with a high degree of purity and applied successfully in conventional ELISA for the specific immunodiagnosis of human parastrongyliasis.

Detection of circulating antigens of Parastrongylus cantonensis in human sera by dot-blot ELISA and sandwich ELISA using monoclonal antibody.

A dot-blot ELISA was compared with a previously performed sandwich ELISA for the detection of Parastrongylus cantonensis antigens in sera from patients. Using the same monoclonal antibody and the same sera, 6 of 10 sera (60%) from parastronglyiasis patients were positive in dot-blot ELISA, whereas with sandwich ELISA, 5 of the same patient sera (50%) were positive. The specificity in both assays was 100% using 50 sera from patients with other parasitic diseases; of these, 10 each were from patients with cysticercosis, filariasis, gnathostomiasis, malaria and toxocariasis. The control group consisted of 53 sera from normal health Thais and Malaysians. The sensitivity of the assays was, however, slightly better with dot-blot ELISA and because it is simple, quick and cost-effective, it may be a test of choice for specific diagnosis of human parastrongyliasis.

Detection of circulating antigens of Parastrongylus cantonensis in human sera by sandwich ELISA with specific monoclonal antibody.

A specific monoclonal antibody (AW-3C2) as revealed by ELISA was produced against the adult worm antigens of Parastrongylus cantonensis and used in a sandwich ELISA for the detection of circulating antigens in the sera of parastrongyliasis patients and those with other parasitic diseases. A total of 60 sera was used in this study. Of these, 10 each were from patients with parastrongyliasis filariasis, gnathostomiasis, malaria and toxocariasis. The control group consisted of 53 serum samples from normal healthy Thais and Malaysians. The mean +/- optical density (OD) values for the normal Thai and Malaysian groups were 0.126 +/- 0.028 and 0.124 +/- 0.029, respectively. Mean OD values of parastrongyliasis patient group differed significantly from that of the normal groups as well as those of other parasitic infections. Using a cut-off point of mean OD +/- 3SD of the normal control groups as indicating a positive reading, the specificity of the assay with this monoclonal antibody was 100% while the sensitivity was 50%.

Detection of circulating antigens of Parastrongylus cantonensis in human sera by sandwich ELISA with specific monoclonal antibody.

A specific monoclonal antibody (AW-3C2) as revealed by ELISA was produced against the adult worm antigens of Parastrongylus cantonensis and used in a sandwich ELISA for the detection of circulating antigens in the sera of parastrongyliasis patients and those with other parasitic diseases. A total of 60 sera was used in this study. Of these, 10 each were from patients with parastrongyliasis, cysticercosis, filariasis, gnathostomiasis, malaria and toxocariasis. The control group consisted of 53 serum samples from normal healthy Thais and Malaysians. The mean +/- optical density (OD) values for the normal Thai and Malaysian groups were 0.126 +/- 0.028 and 0.124 +/- 0.029, respectively. The mean OD values of the parastrongyliasis patient group differed significantly from that of the normal groups as well as those of other parasitic infections. Using a cut-off point of OD +/- 3SD of the control groups as indicating a positive reading, the specificity of the assay with this monoclonal antibody was 100% while the sensitivity was 50%.

Parasitology proficiency testing in the quality assessment programme in Thailand.

Review of participant performance in the quality assessment programme from 1988 to 1991 shows that laboratories have been able to identify most common helminths and protozoa in multiple fecal samples. Ability has improved during the period reviewed. Proficiency in identifying the diagnostic stages of helminths is considerably higher than in protozoa. Analyses of results have shown that participant laboratories correctly identified approximately 64 per cent of intestinal helminths and 40 per cent of intestinal protozoa. Correct identification of blood parasites was achieved by an average of 50 per cent of the laboratories. Their rate of achievement was 10 per cent higher than in the last period reviewed. There is still room for improvement and well-trained personnel should lead to better performance.

Identification of fecal parasites in the quality assessment programme for the year 1984-1987, in Thailand.

Review of data from the quality assessment programme of the DHLQC for the years 1984-1987 shows that some laboratories still have difficulty identifying fecal parasites; particularly the protozoa and the rare parasites although percentages of correct identification have improved in recent years. Helminth ova or larvae in fecal specimens were correctly identified by approximately 55 per cent of the laboratories. Correct identification of intestinal protozoan cysts, however, was achieved by an average of only 29 per cent of the participants.

In vivo and in vitro studies of tinidazole in Trichomonas vaginalis infection.

A total 787 patients with complaints of vaginal discharges were examined, of which 106 patients were found to be positive for Trichomonas vaginalis by the normal saline method, and 114 patients were positive by the culture method. In the in vivo series, 55 patients were treated with a single dose of 1.8 gm of tinidazole, and with a follow-up examination 7 days there after. For the in vitro study 55 swab specimens were cultured to study the tinidazole sensitivity of Trichomonas vaginalis. The survival of Trichomonas vaginalis at each concentration of tinidazole was presented as a cumulative frequency. At the concentration of 1 microgram/ml, none of the organisms were killed; but at the concentration of 6 micrograms/ml, the mortality rate was 100%. In the in vivo series a 100% cure rate was obtained with tinidazole with no side effects, and in vitro series the mean minimum trichomonicidal concentration (MCC) at 24 hours was 3.76 +/- 1.25 micrograms/ml, with the range of 2--6 micrograms/ml. These studies revealed the sensitivity of Trichomonas vaginalis to tinidazole in Thailand and that tinidazole at a single oral dose of 1.8 gm perferably after a meal is effective, safe and cheap for the treatment of Trichomonas vaginalis.