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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. CONCLUSION: Sex, side of deformity, type of surgery and age at time of surgery were not associated with the recurrence deformity.

Bactrocera (Bulladacus) cinnabaria and B. (Bactrocera) propinqua are tephritid fruit flies of the subfamily Dacinae, tribe Dacini. The whole mitogenomes of these two species (first report for the subgenus Bulladacus) possess 37 genes (13 protein-coding genes – PCGs, 2 rRNA and 22 tRNA genes). The mitogenome of B. cinnabaria (15,225 bp) is shorter than that of B. propinqua (15,927 bp), mainly due to the smaller size of the control region and intergenic spacers in B. cinnabaria. Molecular phylogeny based on mitochondrial genes (mt-genes) reveals two clades of the genus Bactrocera: one comprising the subgenus Bactrocera and the other comprising the subgenera Bulladacus, Daculus, Tetradacus and unassigned Bactrocera sp. ‘yunnanensis’. The subgenera represented by two or more taxa are monophyletic. B. (Bulladacus) cinnabaria forms a sister group with the subgenus Tetradacus (B. minax and B. tsuneonis) and B. sp. ‘yunnanensis’, in a clade containing also the basal sister lineage of the subgenus Daculus (B. oleae and B. biguttula). B. propinqua forms a sister group with B. ritsemai and B. limbifera in a subclade containing also B. umbrosa, B. curvifera and B. moluccensis of the monophyletic subgenus Bactrocera. The present study supports the synonymy of B. ruiliensis with B. thailandica. It also shows a high genetic similarity between (a) B. melastomatos and B. rubigina, (b) B. papayae and B. philippinensis, (c) B. dorsalis and B. invadens, (d) B. tryoni and B. neohumeralis, and (e) B. cheni and B. tuberculata; and B. cheni is distinct from and not a synonym of B. tsuneonis or B. lombokensis.

Objective: To measure and compare the incidence of cast-related pressure injuries at the heel between sponge-incorporated long leg cast and multilayer padding cast following orthopedic lower extremity surgery. Materials and Methods: Patients aged 1 to 18 years who were scheduled for elective lower extremity soft tissue surgery were recruited. They were equally randomized into two groups: sponge padding cast (Group 1) and multilayer padding cast (Group 2). A multilayer padding cast, consisting of five layers of Webril®, was applied to the heel and bony prominence areas. At the 2nd and 4th weeks postoperatively, cast-related heel pressure injuries were evaluated using the 2016 National Pressure Ulcer Advisory Panel (NPUAP) staging system. Changes in alignment and other complications were also recorded. Results: 100 patients were recruited. At two weeks postoperatively, the incidence of cast-related heel pressure injuries in Group 1 was 4/46 cases (8.7%), with all cases being stage 1. In Group 2, there were 6/54 cases (11.1%), with 3 cases in stage 1 and 3 cases in deep tissue pressure injury (p=0.479). At four weeks postoperatively, Group 1 had 6/46 cases (13%), with 4/46 cases (8.7%) being stage 1 and 2/46 cases (4.3%) being deep tissue pressure injuries. Group 2 had 6/54 cases (11.1%) (p>0.99), with all cases being stage 1. There was no statistical difference in loss of alignment. No serious complications, such as compartment syndrome or neurovascular injury, were found. Conclusion: The incidence of cast-related heel pressure injury was comparable between sponge padding cast and a multilayer cotton padding cast.

Objective: Distraction osteogenesis (DO) is a well-known technique. The traditional method utilized the lengthening frequency of four times a day (QID). Many mechanical factors may affect the DO outcome. However, the effect of distraction frequency has not been proven clinically. Therefore, we aim to investigate whether the BID and QID lengthening frequency affect the healing index and complications of the DO. Materials and Methods: We retrospectively reviewed patients who had undergone DO from 2010 – 2021. The patient was divided into BID and QID groups. Demographics, lengthening outcomes, and complications between the two groups were compared. We used the inverse probability of treatment weighting (IPTW) to determine the effect of treatment. Results: The median healing index (HI) of the patients whose lengthening was done QID is 41.36 (IQR 32.72 - 67.68) days/cm, and BID is 49.12 (IQR 35.28 - 62.54) days/cm, which did not differ significantly. The Odds ratio of achieving HI < 45 days/cm for patients receiving QID lengthening compare to BID is 1.12 (95% CI 0.31-3.99, p = 0.862 ). The IPTW did not show a difference in average treatment effects between QID and BID lengthening. The rates of minor and major complications were not significantly different between the two groups (26.1% in QID group and 32.1% in BID group). Conclusion: The frequency of QID and BID lengthening results in comparable HI and complications for patients who undergo DO. However, prospective research is needed to evaluate the effect of frequency differences in a clinical setting.

The complete mitogenomes of fruit flies Zeugodacus (Javadacus) calumniatus, Z. (Javadacus) heinrichi and Z. (Sinodacus) hochii have similar gene order and contain 37 genes and a non-coding region. They share an identical start codon for the respective protein-coding genes (PCGs), an identical TAA stop codon for 11 PCGs, TAG for cob, and an incomplete T stop codon for nad5. The cloverleaf structure of most of the tRNAs is similar in the three Zeugodacus species. Phylogenetic analyses reveal Z. (Parasinodacus) cilifer to be external to two main clades: (A) monophyletic subgenus Zeugodacus; and (B) subgenera Javadacus and Sinodacus. The present results indicate that the taxonomic status of some taxa needs clarification. Z. calumniatus is genetically very similar to Z. tau and is not congruent with its current placement in the munda complex. Z. mukiae NC_067083 is genetically very similar to Z. scutellaris, but differs significantly from Z. mukiae MG683384 of the arisanicus (arisanica) complex. On the other hand, Z. proprediaphorus is genetically distinct from and not a synonym of Z. diaphorus. Z. caudatus sensu stricto from Indonesia forms a sister lineage with Z. diversus, instead of with the Malaysian and Chinese taxa of Z. caudatus sensu lato. A notable incongruence is the sister lineage of Z. (Sinodacus) hochii and Z. (Javadacus) heinrichi among other taxa of subgenus Javadacus. A more extensive taxon sampling, particularly the subgenus Sinodacus (and other subgenera), is needed to clarify/resolve their subgenus status.

Abstract Objectives The sterile exsanguination tourniquet (SET) could be an alternative for providing bloodless surgeries in orthopedic femoral-related surgeries in pediatric patients where the standard pneumatic tourniquet would not be feasible. This randomized-controlled study aimed to evaluate the efficacy of SET in decreasing total perioperative blood loss and blood transfusion. Methods We conducted an unplanned interim analysis of data from a randomized-controlled trial. At the time of the analysis, 31 pediatric patients had been randomly assigned to undergo surgery with the SET application (the SET group, 15 patients) and without the SET application (the control group, 16 patients). An intention-to-treat analysis was performed to evaluate the total perioperative blood loss, postoperative blood transfusion, estimated intraoperative blood loss, total drainage volume, postoperative hemoglobin level, and operative time according to the significance level adjusted for multiplicity (p

Abstract Background Surgical treatment for severe lower limb deformities in patients with hypophosphatemic rickets has shown satisfactory outcomes. However, the rates of recurrence of deformities after surgical correction were high, and studies on predictive factors of recurrence were limited. This study aimed to determine the predictive factors for the recurrence of lower limb deformities after surgical correction in patients with hypophosphatemic rickets, and the effects of each predictor on the recurrence of deformities. Methods We retrospectively reviewed the medical records of 16 patients with hypophosphatemic rickets aged 5–20 years and who had undergone corrective osteotomies between January 2005 and March 2019. Demographic data from the patients, biochemical profiles, and radiographic parameters were collected. Univariable Cox proportional hazard analyses of recurrence were performed. Kaplan–Meier failure estimation curves for deformity recurrences of potential predictors were created. Results A total of 38 bone segments were divided into 2 groups: 8 segments with recurrent deformities and 30 segments without recurrent. The average follow-up time was 5.5 ± 4.6 years. Univariable Cox proportional hazard analyses of recurrence found that an age 10 years (p = 0.02). Conclusions Identifying predictive factors for the recurrence of lower limb deformities after surgical correction in hypophosphatemic rickets can assist in early recognition, proper intervention, and prevention. We found that an age

Abstract Background Several types of pelvic osteotomy techniques have been reported and employed by orthopedic surgeons to enhance the approximation of symphyseal diastasis in bladder exstrophy patients. However, there is limited evidence on a long-term follow-up to confirm which osteotomy techniques provide the most suitable and effective outcomes for correcting pelvic deformities. This study aimed to describe the surgical technique of bilateral iliac bayonet osteotomies for correcting pelvic bone without using fixation in bladder exstrophy and to report on the long-term clinical and radiographic outcomes following the bayonet osteotomies. Methods We retrospectively reviewed patients with bladder exstrophy who underwent bilateral iliac bayonet osteotomies with the closure of bladder exstrophy between 1993 and 2022. Clinical outcomes and radiographic pubic symphyseal diastasis measurements were evaluated. From a total of 28 operated cases, eleven were able to attend a special follow-up clinic or were interviewed by telephone by one of the authors with completed charts and recorded data. Results A total of 11 patients (9 female and 2 male) with an average age at operation of 9.14 ± 11.57 months. The average followed-up time was 14.67 ± 9.24 years (0.75–29), with the average modified Harris Hip score being 90.45 ± 1.21. All patients demonstrated decreased pubic symphyseal diastasis distance (2.05 ± 1.13 cm) compared to preoperative (4.58 ± 1.37 cm) without any evidence of nonunion. At the latest follow-up, the average foot progression angle was externally rotated 6.25° ± 4.79° with full hips ROM, and no patients reported abnormal gait, hip pain, limping, or leg length discrepancy. Conclusions Bilateral iliac wings bayonet osteotomies technique demonstrated a safe and successful pubic symphyseal diastasis closure with an improvement both clinically and radiographically. Moreover, it showed good long-term results and excellent patient’s reported outcome scores. Therefore, it would be another effective option for pelvic osteotomy in treating bladder exstrophy patients.

Angiostrongylus cantonensis is the major etiological nematode parasite causing eosinophilic meningitis and/or eosinophilic meningoencephalitis in humans. The rapid global spread of Angiostrongylus cantonensis and the emerging occurrence of the infection have exposed the shortcomings of traditional/conventional diagnostics. This has spurred efforts to develop faster, simpler and more scalable platforms that can be decentralized for point-of-need laboratory testing. By far, the point-of-care immunoassays such as the lateral flow assay (LFA) are the best-placed. In this work, a LFA in the form of an immunochromatographic test device (designated AcAgQuickDx), based on the detection of a circulating Angiostrongylus cantonensis-derived antigen, was established using anti-31 kDa Angiostrongylus cantonensis antibody as the capture reagent and anti-Angiostrongylus cantonensis polyclonal antibody as the indicator reagent. The AcAgQuickDx was evaluated for its diagnostic potential with a total of 20 cerebrospinal fluids (CSF) and 105 serum samples from patients with angiostrongyliasis and other clinically related parasitic diseases, as well as serum samples from normal healthy subjects. Three of the ten CSF samples from serologically confirmed angiostrongyliasis cases and two of the five suspected cases with negative anti-Angiostrongylus cantonensis antibodies showed a positive AcAgQuickDx reaction. Likewise, the AcAgQuickDx was able to detect Angiostrongylus cantonensis specific antigens in four serum samples of the 27 serologically confirmed angiostrongyliasis cases. No positive reaction by AcAgQuickDx was observed in any of the CSF (n = 5) and serum (n = 43) samples with other parasitic infections, or the normal healthy controls (n = 35). The AcAgQuickDx enabled the rapid detection of active/acute Angiostrongylus cantonensis infection. It is easy to use, can be transported at room temperature and does not require refrigeration for long-term stability over a wide range of climate. It can supplement existing diagnostic tests for neuroangiostrongyliasis under clinical or field environments, particularly in remote and resource-poor areas.

Objective To determine the radiographic outcomes following dome or wedge‐shaped proximal tibial osteotomy in the management of infantile Blount disease with a particular interest in sagittal alignment of the knee joint. Method Medical records of patients with Langenskiöld stage 2 Blount disease (aged ≤5 years) who underwent surgical correction between January 2005 and November 2019 were retrospectively identified. Patients with metabolic bone disease, bone tumors, prior traumatic fractures, congenital anomalies, inadequate plain films, and incomplete medical documents were excluded. Patient characteristics (e.g. age, gender, and body mass index [BMI]) and surgical characteristics (e.g. side, type of surgery, and follow‐up times) were recorded. Antero–posterior (AP) and lateral knee radiographs were analyzed. Data were categorized by surgical technique as dome‐shaped proximal tibial osteotomy or wedge‐shaped proximal tibial osteotomy. The femorotibial angle (FTA) was used to evaluate the correction angle in varus deformities. Sagittal alignment of the lower limbs using the posterior tibial slope (PTS) angle was measured postoperatively at 3, 6, 12, and 24 months, and at the final follow‐up visit. Results The present study included 72 surgeries of 46 patients who had undergone proximal tibial osteotomy. Twenty‐nine (63%) were male. The mean age of patients at the time of surgery was 34.50 months (range, 26–47). The mean weight was 23.11 ± 4.98 kg (mean ± SD); the mean height was 95.33 ± 6.36 cm, and the mean BMI was 25.32 ± 4.36 kg/m2. The mean duration of follow up was 4.77 ± 2.78 years. Sixty‐four patients (88.90%) received dome‐shaped proximal osteotomy of the tibia, while 8 (11.10%) received wedge‐shaped proximal osteotomy of the tibia. The average FTA of the total correction measured was 29.32° ± 7.98°. The demographic data of the two groups were not significantly different for gender, age, BMI, side follow‐up times, and the total correction of varus deformities. In the dome‐shaped osteotomy group, the mean correction of the FTA was 29.59° ± 7.45°. The mean degree of the PTS angle was 6.50° at 3 months, 6.38° at 6 months, 5.32° at 12 months, 5.17° at 24 months, and 5.53° at the final follow‐up visit. In the wedge‐shaped proximal tibial osteotomy group, the mean correction of the FTA was 27.25° ± 11.77°. The PTS was 6.00° at 3 months, 7.50° at 6 months, 7.00° at 12 months, 5.40° at 24 months, and 5.57° at the final follow‐up visit. No significant difference was observed in the radiological outcome between surgical techniques. Conclusion Dome and wedge‐shaped proximal tibial osteotomies did not demonstrate significant differences in the PTS angle in children with Blount disease.

Objectives The aims of this paper were: (i) to examine the intra‐observer and inter‐observer reliability of the shaft‐condylar angle (SCA) and the lateral capitellohumeral angle (LCHA); (ii) to study the influence of experience level on the inter‐observer and intra‐observer reliability; and (iii) to determine the influence of the the age of the patients on reliability. Method A retrospective cohort study was conducted. The study reviewed 81 elbow radiographs. The patients were aged between 2 and 13 years. All the images taken between 2000 and 2017 were independently measured by a senior pediatric orthopaedic surgeon, a pediatric orthopaedic surgeon, a pediatric orthopaedic fellow, an orthopaedic chief resident, a general practitioner, and a pediatric orthopaedic research assistant. Measurement was performed two times within a 2‐week interval. Inexperienced observers (general practitioner and research assistant) were supervised by senior pediatric orthopaedic surgeons for at least 30 radiographs before performing the measurement. Inclusion criteria were as follows: (i) age 2–13 years; and (ii) no previous elbow fracture. Exclusion criteria: elbow radiographs do not show true lateral view. The intraclass correlation coefficient (ICC) was used to calculate the reliability. Results The mean values of SCA and LCHA were 43° and 48°, respectively. For SCA, intra‐observer reliability was excellent (ICC = 0.85) for one observer, good (range = 0.73–0.76) for three observers, and moderate (0.59) for one observer. Inter‐observer reliability was moderate (0.48, 0.58), whereas the reliability categorized by age group showed excellent agreement (0.88–0.94). For LCHA, intra‐observer reliability was excellent (0.84–0.89) for three observers and good (0.66–0.80) for two observers. Inter‐observer reliability was moderate (0.44–0.45). Conversely, the reliability classified by age group showed excellent agreement (0.83–0.91). Conclusion Intra‐observer reliability for LCHA and SCA were excellent to good for most observers. Inter‐observer reliability was moderate for LCHA and SCA. Reliability classified by age group showed excellent to good agreement. Reliability was influenced by the level of experience, especially for non‐medical staff.

Objective This study was aimed to find the radiographic parameter predicting recurrence of stage 2 Blount's disease. Method We retrospectively reviewed radiographs of 82 legs from 49 patients diagnosed with stage 2 Blount's disease by Langenskiöld classification who had failed brace treatment and underwent valgus osteotomy between 1998 to 2016. Age ranged from 26 to 47 months. The metaphyseal–diaphyseal angle was measured preoperatively. The medial metaphyseal slope of the proximal tibia and femorotibial angle were measured preoperatively and 3, 6, 12, and 24 months postoperatively in both non‐recurrence (group 1) and recurrence (group 2) group. The receiver operating characteristic curve calculated using MedCalc software was used to determine the medial metaphyseal slope predicting risk for recurrence. Statistical analysis was performed using SPSS software. Results The mean follow‐up time was 4.83 ± 0.38 years. The mean age was 34.57 ± 5.76 in group 1 and 33.2 ± 1.48 in group 2 (P = 0.258). The mean preoperative metaphyseal slope was 62.39° ± 9.75° in group 1 and 73.22° ± 6.59° in group 2 (P = 0.02). The mean preoperative femorotibial angle (FTA) was −14.31° ± 8.25° in group 1 and −18.89° ± 7.74° in group 2 (P = 0.1). The mean preoperative metaphyseal diaphyseal angle (MDA) was 14.75° ± 4.21° in group 1 and 20.11° ±5.16° in group 2 (P = 0.001). Demographic data including age, gender, weight, height, and body mass index showed no statistically significant difference between both groups. Out of 82 legs, 9 (10.97%) had recurrence. Preoperatively, the metaphyseal–diaphyseal angle showed statistical significance between both groups. The medial metaphyseal slope showed statistically significant difference between group 1 and group 2 at 3, 6, 12, and 24 months postoperatively. The receiver operating characteristic curve showed that a medial metaphyseal slope more than 70° at 12 months (sensitivity 88.89% and specificity 69.86%) and more than 62° at 24 months postoperatively (sensitivity 100%, specificity 52.3%) was a predictor for recurrence of stage 2 Blount's disease. Conclusion Medial metaphyseal slope more than 62° over the 24‐month follow‐up was associated with recurrence of varus deformity.

Objective: Congenital muscular torticollis, characterized by unilateral shortening of the ternocleidomastoid muscle that results in the head tilting towards the affected side, is the third most common musculoskeletal anomaly. Subjective assessment using the Lee scoring system has been the most common method for evaluating the surgical outcomes. The aim of this study was to investigate the efficacy of a novel assessment tool for scoring of functional outcomes after surgical release in congenital muscular torticollis. Methods: This study enrolled the patients aged 5-15 years who were diagnosed with congenital muscular torticollis and treated by surgical release at the Department of Orthopaedic Surgery, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand during 1998-2011. An image processing program was used to measure all evaluated parameters. Craniofacial asymmetry, neck rotation deficits, neck flexion/extension deficits, and degree of head-tilting were graded as poor, fair, good, or excellent by 2 observers. The results of the Lee scoring system and the new scoring system were then compared. Results: Nineteen patients (7 males, 12 females) with a mean age of 7.2±2.57 years were included. Photographs of the patients were evaluated and categorized by outcome score using both outcome scoring systems. All intraclass correlation coefficients were higher than 0.6 for both scoring systems, which suggests good inter-operator reliability (p

Background: There were very few options available for distal foot and toe defects that required a vascularized flap for coverage. As such, the use of a free flap was often justified in this region of the foot. The use of perforator flaps has created a new subset of local tissue transfer alternatives that increases the potential that the difficulties associated with microvascular tissue transfers could be avoided. The first dorsal metatarsal artery (FDMA) perforator flap was one variant of this new type of tissue transfer. The aim of this report was to describe our experience using FDMA perforator flap to cover great toe defect. Methods: A standard FDMA flap from the dorsum of the foot was raised in reversed fashion based on the distal communicating branch or “perforator” from plantar foot circulation in two patients with great toe defect. Results: Salvage of the great toe was achieved in both patients. FDMA perforator flap achieved both reconstructive goals, and the donor site closure can be successfully performed without tension. Donor site healing was achieved in both patients with no associated complications by the 1-year follow-up. Conclusions: FDMA flap can be successfully used as a local flap to cover distal foot and toe wounds. However, direct donor site closure can be problematic and may need skin graft.

Background: Metatarsus adductus (MA) is a common pediatric foot deformity. Current recommendations suggest observation until 4–6 months, then casting if the deformity persists. Based on our review of the literatures, no randomized controlled trial has been conducted to study the effectiveness of parental stretching in the correction of MA in newborn. Material and methods: Ninety-four newborn feet that were diagnosed as MA by clinical examination were enrolled. Feet were randomized into two groups: observation group and stretching group. Outcome measurements were performed to compare success rate between groups. Results: According to Pearson’s χ 2 test, there were no statistically significant differences between groups with regard to the overall success of the parental stretching program ( p = 0.191). There was also no significant difference between groups for mild degree or moderate-to-severe degree ( p = 0.134, p = 0.274, respectively). A more rapid success rate was observed in the stretching group at the first month follow-up, but rate of improvement then decreased. The stretching group tended to have a lower success rate compared to the observation group in moderate-to-severe feet, but the difference was not statistically significant. Conclusions: Parental stretching program found no benefit over observation group in this study. Parental stretching program should not be applied for newborn babies with moderate-to-severe MA as the result from the study appeared to have lower success rate compared to observation group. Observe until 4–6 months, then corrective casting for the persisting deformity is recommended.

The tephritid fruit fly Zeugodacus tau (Walker) is a polyphagous fruit pest of economic importance in Asia. Studies based on genetic markers indicate that it forms a species complex. We report here (1) the complete mitogenome of Z. tau from Malaysia and comparison with that of China as well as the mitogenome of other congeners, and (2) the relationship of Z. tau taxa from different geographical regions based on sequences of cytochrome c oxidase subunit I gene. The complete mitogenome of Z. tau had a total length of 15631 bp for the Malaysian specimen (ZT3) and 15835 bp for the China specimen (ZT1), with similar gene order comprising 37 genes (13 protein-coding genes-PCGs, 2 rRNA genes, and 22 tRNA genes) and a non-coding A + T-rich control region (D-loop). Based on 13 PCGs and 15 mt-genes, Z. tau NC_027290 (China) and Z. tau ZT1 (China) formed a sister group in the lineage containing also Z. tau ZT3 (Malaysia). Phylogenetic analysis based on partial sequences of cox1 gene indicates that the taxa from China, Japan, Laos, Malaysia, Bangladesh, India, Sri Lanka, and Z. tau sp. A from Thailand belong to Z. tau sensu stricto. A complete cox1 gene (or 13 PCGs or 15 mt-genes) instead of partial sequence is more appropriate for determining phylogenetic relationship.

Bactrocera latifrons is a serious pest of solanaceous fruits and Bactrocera umbrosa is a pest of Artocarpus fruits, while Bactrocera melastomatos infests the fruit of Melastomataceae. They are members of the subgenus Bactrocera. We report here the complete mitochondrial genome of these fruit flies determined by next-generation sequencing and their phylogeny with other taxa of the subgenus Bactrocera. The whole mitogenomes of these three species possessed 37 genes namely, 13 protein-coding genes (PCGs), 2 rRNA and 22 tRNA genes. The mitogenome of B. latifrons (15,977 bp) was longer than those of B. melastomatos (15,954 bp) and B. umbrosa (15,898 bp). This difference can be attributed to the size of the intergenic spacers (283 bp in B. latifrons, 261 bp in B. melastomatos, and 211 bp in B. umbrosa). Most of the PCGs in the three species have an identical start codon, except for atp8 (adenosine triphosphate synthase protein 8), which had an ATG instead of GTG in B. umbrosa, whilst the nad3 (NADH dehydrogenase subunit 3) and nad6 (NADH dehydrogenase subunit 6) genes were characterized by an ATC instead of ATT in B. melastomatos. The three species had identical stop codon for the respective PCGs. In B. latifrons and B. melastomatos, the TΨC (thymidine-pseudouridine-cytidine)-loop was absent in trnF (phenylalanine) and DHU (dihydrouracil)-loop was absent in trnS1 (serine S1). In B. umbrosa, trnN (asparagine), trnC (cysteine) and trnF lacked the TψC-loop, while trnS1 lacked the DHU-stem. Molecular phylogeny based on 13 PCGs was in general concordant with 15 mitochondrial genes (13 PCGs and 2 rRNA genes), with B. latifrons and B. umbrosa forming a sister group basal to the other species of the subgenus Bactrocera which was monophyletic. The whole mitogenomes will serve as a useful dataset for studying the genetics, systematics and phylogenetic relationships of the many species of Bactrocera genus in particular, and tephritid fruit flies in general.

Bactrocera caudata is a pest of pumpkin flower. Specimens of B. caudata from the northern hemisphere (mainland Asia) and southern hemisphere (Indonesia) were analysed using the partial DNA sequences of the nuclear 28S rRNA and internal transcribed spacer region 2 (ITS-2) genes, and the mitochondrial cytochrome c oxidase subunit I (COI), cytochrome c oxidase subunit II (COII) and 16S rRNA genes. The COI, COII, 16S rDNA and concatenated COI+COII+16S and COI+COII+16S+28S+ITS-2 nucleotide sequences revealed that B. caudata from the northern hemisphere (Peninsular Malaysia, East Malaysia, Thailand) was distinctly different from the southern hemisphere (Indonesia: Java, Bali and Lombok), without common haplotype between them. Phylogenetic analysis revealed two distinct clades (northern and southern hemispheres), indicating distinct genetic lineage. The uncorrected 'p' distance for the concatenated COI+COII+16S nucleotide sequences between the taxa from the northern and southern hemispheres ('p' = 4.46-4.94%) was several folds higher than the 'p' distance for the taxa in the northern hemisphere ('p' = 0.00-0.77%) and the southern hemisphere ('p' = 0.00%). This distinct difference was also reflected by concatenated COI+COII+16S+28S+ITS-2 nucleotide sequences with an uncorrected 'p' distance of 2.34-2.69% between the taxa of northern and southern hemispheres. In accordance with the type locality the Indonesian taxa belong to the nominal species. Thus the taxa from the northern hemisphere, if they were to constitute a cryptic species of the B. caudata species complex based on molecular data, need to be formally described as a new species. The Thailand and Malaysian B. caudata populations in the northern hemisphere showed distinct genetic structure and phylogeographic pattern.

Angiostrongylus costaricensis is a zoonotic parasitic nematode that causes abdominal or intestinal angiostrongyliasis in humans. It is endemic to the Americas. Although the mitochondrial genome of the Brazil taxon has been published, there is no available mitochondrial genome data on the Costa Rica taxon. We report here the complete mitochondrial genome of the Costa Rica taxon and its genetic differentiation from the Brazil taxon. The whole mitochondrial genome was obtained from next-generation sequencing of genomic DNA. It had a total length of 13,652 bp, comprising 36 genes (12 protein-coding genes-PCGs, 2 rRNA and 22 tRNA genes) and a control region (A + T rich non-coding region). It is longer than that of the Brazil taxon (13,585 bp). The larger mitogenome size of the Costa Rica taxon is due to the size of the control region as the Brazil taxon has a shorter length (265 bp) than the Costa Rica taxon (318 bp). The size of 6 PCGs and the start codon for ATP6, CYTB and NAD5 genes are different between the Costa Rica and Brazil taxa. Additionally, the two taxa differ in the stop codon of 6 PCGs. Molecular phylogeny based on 12 PCGs was concordant with two rRNA, 22 tRNA and 36 mitochondrial genes. The two taxa have a genetic distance of p = 16.2% based on 12 PCGs, p = 15.3% based on 36 mitochondrial genes, p = 13.1% based on 2 rRNA genes and p = 10.7% based on 22 tRNA genes, indicating status of sibling species. The Costa Rica and Brazil taxa of A. costaricensis are proposed to be accorded specific status as members of a species complex.

Cerebrospinal fluid (CSF) samples from clinically diagnosed patients with detectable Angiostrongylus canto-nensis-specific antibodies (n = 10), patients with clinically suspected cases that tested negative for A. cantonensis-an-tibodies (n = 5) and patients with cerebral gnathostomiasis (n = 2) and neurocysticercosis (n = 2) were examined by a single-step polymerase chain reaction (PCR) method using the AC primers for the 66-kDa native protein gene. The PCR method detected A. cantonensis DNA in CSF samples from four of 10 serologically confirmed angiostrongyliasis cases. The PCR results were negative for the remaining CSF samples. The nucleotide sequences of three positive CSF-PCR samples shared 98.8-99.2% similarity with the reference sequence of A. cantonensis. These results indicate the potential application of this PCR assay with clinical CSF samples for additional support in the confirmation of eosinophilic meningitis due to A. cantonensis.

The fruit fly Bactrocera caudata is a pest species of economic importance in Asia. Its larvae feed on the flowers of Cucurbitaceae such as Cucurbita moschata. To-date it is distinguished from related species based on morphological characters. Specimens of B. caudata from Peninsular Malaysia and Indonesia (Bali and Lombok) were analysed using the partial DNA sequences of cytochrome c oxidase subunit I (COI) and 16S rRNA genes. Both gene sequences revealed that B. caudata from Peninsular Malaysia was distinctly different from B. caudata of Bali and Lombok, without common haplotype between them. Phylogenetic analysis revealed two distinct clades, indicating distinct genetic lineage. The uncorrected 'p' distance for COI sequences between B. caudata of Malaysia-Thailand-China and B. caudata of Bali-Lombok was 5.65%, for 16S sequences from 2.76 to 2.99%, and for combined COI and 16S sequences 4.45 to 4.46%. The 'p' values are distinctly different from intraspecific 'p' distance (0-0.23%). Both the B. caudata lineages are distinctly separated from related species in the subgenus Zeugodacus - B. ascita, B. scutellata, B. ishigakiensis, B. diaphora, B. tau, B. cucurbitae, and B. depressa. Molecular phylogenetic analysis indicates that the B. caudata lineages are closely related to B. ascita sp. B, and form a clade with B. scutellata, B. ishigakiensis, B. diaphora and B. ascita sp. A. This study provides additional baseline for the phylogenetic relationships of Bactrocera fruit flies of the subgenus Zeugodacus. Both the COI and 16S genes could be useful markers for the molecular differentiation and phylogenetic analysis of tephritid fruit flies.

Human gnathostomiasis is a parasitic disease caused by Gnathostoma nematode infection. A rapid, reliable, and practical immunoassay, named dot immuno-gold filtration assay (DIGFA), was developed to supporting clinical diagnosis of gnathostomiasis. The practical tool detected anti-Gnathostoma-specific IgG4 in human serum using crude extract of third-stage larvae as antigen. The result of the test was shown by anti-human IgG4 monoclonal antibody conjugated colloidal gold. The sensitivity and specificity of the test were both 100% for detection in human sera from patients with gnathostomiasis (13/13) and from healthy negative controls (50/50), respectively. Cross-reactivity with heterogonous serum samples from patients with other helminthiases ranged from 0 (trichinosis, paragonimiasis, clonorchiasis, schistosomiasis, and cysticercosis) to 25.0% (sparganosis), with an average of 6.3% (7/112). Moreover, specific IgG4 antibodies diminished at 6 months after treatment. This study showed that DIGFA for the detection of specific IgG4 in human sera could be a promising tool for the diagnosis of gnathostomiasis and useful for evaluating therapeutic effects. [ABSTRACT FROM AUTHOR]