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Molecular occurrence of canine babesiosis in rural dog population in Pakistan

Abdullah Sagir Ahmad ; Imran Rashid ; Kamran Ashraf ; Wasim Shehzad ; Matiullah Khan ; Kashif Hussain ; Shahid Hussain Farooqi ; Amjad Khan ; Muhammad Luqman Sohail

Tropical Biomedicine.2018;35(3):593-603.

Canine babesiosis is an important tick-borne protozoal disease of dogs that poses major health problem worldwide. Farm dogs in rural areas are the companion animals, that not only watch the livestock herds but also guard the house of the owners. Each farmer keeps his companion dog to get all the services. In our study, a total of 450 blood samples of farm dogs from three different ecological zones (Southern, Central and Northern regions of the province; Punjab) of Pakistan, were collected to examine through microscopy and PCR. Examination of thin blood smears revealed an overall prevalence of 12.8% (58/450) of canine babesisal parasites. However, PCR analysis revealed 46.8% (211/450) and 7.3% (33/450) samples positive for B. gibsoni and B. vogeli, respectively. The amplicons of 671 bp and 590 bp were amplified for the detection of B. gibsoni and B. vogeli, respectively through PCR. The results of multivariate analysis showed that the occurrence of canine babesiosis is higher in the Central Punjab and younger age of the dogs, while breed and sex of the host were not significantly associated with the occurrence of the disease. Mixed infection of B. gibsoni and B. vogeli was observed only in 3 dogs each in district Kasur and Rawalpindi. Our study is the first report to observe the occurrence of canine babesiosis in rural dogs in Pakistan through PCR.

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Group B streptococcus infection in a sudden unexpected death of infancy – the importance of microbiological investigation at post-mortem

Khalid, N ; Zainun, K.A ; Hisham, S ; Mazan, N.I ; Amin Nordin, S

Tropical Biomedicine.2018;35(3):604-609.

Group B streptococcus (GBS) is a common cause of infection in newborns and in early infants. However, GBS infection in an infant older than three months is infrequently reported in the literature. We reported a case of an apparently well six-month-old infant who died of sudden death due to GBS pneumonia, diagnosed at autopsy. The six-month-old, apparently well male infant was brought in dead to the Emergency Department. He underwent medicolegal autopsy four hours after death, as part of an overall sudden unexpected death in infancy investigation (SUDI). Apart from whitish froth oozing out of both nostrils, he appeared to be well-nourished infant without any deformity, syndromic features or obvious suspicious marks of injury externally. Internal examination showed generalized hyperinflated with patchy consolidation of upper and middle lobes of bilateral lung. Multiple matted mesenteric lymphadenopathy were also detected. Blood and lung tissue specimens collected under aseptic technique yielded growth of GBS. Post-mortem histology from consolidated lungs confirmed pneumonic features while mesenteric lymph nodes showed reactive changes inkeeping with underlying infective process. Death was attributed to GBS pneumonia. This case highlights the importance of a detailed autopsy in sudden unexpected death in infancy (SUDI) and the crucial role of post-mortem microbiological study in such cases. Relevant autopsy protocols that need to be employed during microbiological sampling are briefly discussed.

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Larvicidal and adulticidal activities of castor oil against the dengue vector, Aedes aegypti

Wamaket, N ; Dieng,H ; Komalamisra, N ; Apiwathnasorn, C ; Morales, R.E ; Thanomsub, B.W ; Srisawat,R ; Attrapadung, S

Tropical Biomedicine.2018;35(3):610-618.

Plants contain numerous bioactive compounds that can be used to develop environmentally-safe insect control agents. Castor oil is a vegetable oil derived from the seeds of castor bean and is widely used as an industrial lubricant and medicinal purgative. In search of an alternative natural insecticide, the objective of this study was to evaluate the larvicidal and adulticidal activities of castor oil against the important dengue vector, Ae. aegypti. Larvicidal and adulticidal bioassays were conducted following the World Health Organization methods. Larvicidal activity was observed at castor oil concentrations of 10, 25, 50, 75, and 100 ppm; larval mortality was checked after 48 h of exposure and the lethal concentration (LC) at LC50 and LC90 were 51.38 and 116.26 ppm, respectively. Adulticidal activity was determined by topical application at the concentrations of 1, 5, 10, 15, and 20 µg/ mg on female mosquitoes and the mortality was checked after 24 h of exposure. The effective adulticidal activity was apparent with the LD50 and LD90 values of 6.03 and 25.07 µg/mg against female mosquitoes. The results indicated that castor oil has potential in the practical control of both immature and adult stages of the mosquito vector.

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Genotyping of Mycobacterium tuberculosis isolates from northwest Iran for determination on the mechanism of transmission

Mahdavipoor, B ; Asgharzadeh, M ; Hajibonabi, F ; Rashedi, J ; Pourostadi, M ; Bahador, T.N ; Asadi, N ; Kafil, H.S ; Barhaghi, M.H.S

Tropical Biomedicine.2018;35(3):619-626.

Planning to control tuberculosis requires identification of dominant strains in the region, transmission patterns and risk factors that are possible by using molecular genotyping techniques. The aim of this study is to determine the transmission of tuberculosis in the northwest of Iran in order to better understand the spread of disease in northwest of Iran. In this study, 194 positive mycobacterium cultivars in northwest of Iran were investigated using exact tandem repeat-variable number tandem repeats (ETR-VNTR) method. The ETR-VNTR method was identified 55 different patterns in 194 isolates, which contained 25 clusters and 30 unique patterns, and the largest cluster had 33 isolates, and discriminatory power of ETRVNTR method was determined 0.9322 in the examined samples. There are strains of Mycobacterium tuberculosis located in the northwest of Iran that infect people, and ETRVNTR method can be used as a first-line method to examine the dynamics of tuberculosis transmission.

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Therapeutic use of Lucilia sericata maggot in controlling bacterial bio-burden in Rat wound model

Borkataki, S ; Katoch, R ; Goswami, P ; Bhat, A. ; Bhardwaj, H.R ; Chakraborty, D ; Chandrawathani, P

Tropical Biomedicine.2018;35(3):627-638.

Delayed wound healing due to extraneous bacterial contamination, antibacterial resistance and other associated factors are of great concern in dealing patients having chronically infected wound. Medicinal properties of certain maggots of Calliphoridae family are known for its effective wound debridement therapy. The objective of the study was to evaluate the wound healing potential of maggots of Lucilia sericata in an experimentally infected cutaneous wound model in Wistar rat. The study was carried out by using male Wistar rats (n=48) by creating excisional wounds and later contaminated with mixed population of gram positive and gram-negative bacteria. Animals were divided randomly in to four groups with 12 individuals each, being denominated as control, antibiotic treated, maggot treated, and antibiotic plus maggot combination treated group. Ten pre-sterilized maggots were applied per centimetre square wound bed for 24 hours. Different wound kinetics in L. sericata maggot treated wounds revealed significant reduction in wound area with maximum contraction, early elimination of bacterial bioburden as compared to group of infected control and group of rats receiving only antibiotic treatment. The histopathological examination of wounded tissue of maggot treated groups showed early and better epithelialization, collagenation and neovascularization with complete healing of wound in two weeks. The maggot effects on healing when used singly or in combination with antibiotic were recorded to be similar. The results of the present study clearly demonstrate that the maggots of L. sericata possesses a definite antibacterial action along with removal of dead tissues and effectively reduced the bacterial bio-burden in infected wound and induced wound healing quickly.

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Aedes albopictus in urban and forested areas of Malaysia: A study of mitochondrial sequence variation using the CO1 marker

Adilah-Amrannudin, N ; Hamsidi, M ; Ismail, N.-A ; Dom, N.C ; Ismail, R ; Ahmad, A.H ; Mastuki, M.F ; Yusoff, F.H.M, F.H.M ; Adam, N.F.M ; Camalxaman, S.N

Tropical Biomedicine.2018;35(3):639-652.

This study explores the use of a long fragment of the mitochondrial cytochrome oxidase subunit 1 (CO1) marker to elucidate the genetic diversity of Aedes albopictus sampled from urban and forested regions in Peninsular and East Malaysia. A total of 36 samples were collected from 5 localities from which its genetic variability was analysed. 33 distinct mtDNA haplotypes were identified following the amplification and sequencing of the concatenated CO1 gene. The analysed region of the CO1 gene identified substantial levels of genetic diversity among mosquitoes in urban populations and revealed unique genealogical relationships between local isolates as revealed in the haplotype network. This study highlights the reliability of the long CO1 fragment to identify genetic divergence of Aedes albopictus which can be utilized in forthcoming studies.

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Efficacies of five edible mushroom extracts as odor baits for resting boxes to attract mosquito vectors: A field study in Samut Songkhram Province, Thailand

Chaiphongpachara, T. ; Padidpoo, O. ; Chansukh, K.K. ; Sumruayphol, S.

Tropical Biomedicine.2018;35(3):653-633.

Effective trapping of adult mosquitoes in the wild can reduce the spread of deadly human pathogens, such as Plasmodium species causing malaria. The efficacy of this strategy depends on the capacity of the trap to attract and retain insects, and specific odorants such as octenol in mushrooms are strong attractants for mosquitoes. In this study, we assessed the efficiency of a resting box baited with five different extracts from local edible mushrooms, Pleurotus ostreatus, Thaeogyroporus porentosus, Volvariella volvacea, Pleurotus sajorcaju, and Lentinus edodes, for attracting mosquito vectors in Samut Songkhram Province, Thailand. Extracts were used in identical “resting box” at 50, 100, and 200 mg/mL per box. Compared to the unbaited resting box (control), only traps containing 200 mg/mL of L. edodes extract captured significantly more mosquitoes (16.00 ± 3.61 vs. 7.00 ± 1.00 per resting box per night, p<0.05). Attraction efficacy did not increase progressively with tested amount for most extracts. These findings indicate that L. edodes extract can be used as an inexpensive, non-toxic, and locally sourced attractant to increase the efficacy of mosquito control.

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Seroprevalence of Toxoplasma gondii infection in sheep from northern China

Gao, Y. ; Guo, H.P ; Adjou Moumouni, P.F ; Sun, M. ; Liu, M.M ; Efstratiou, A ; Lee, S.H. ; Wang, G.B. ; Li, J.X. ; Li, Y.C. ; Ringo, A.E. ; Galon, E. ; Masatani, T. ; Du, J.G. ; Xuan, X.N.

Tropical Biomedicine.2018;35(3):664-668.

Toxoplasma gondii is an important zoonotic parasite causing significant health problems to humans and animals. In recent years, a number of investigations about the seroprevalence of T. gondii in China have been reported, but little is known on the prevalence of toxoplasmosis in sheep in northern China. In the present study, a total of 288 sheep serum samples were collected from Inner Mongolia, Heilongjiang, Jilin and Hebei provinces of northern China for T. gondii antibody survey using a latex agglutination test (LAT). Of these, 87 (30.2%) serum samples were positive for antibodies to T. gondii, and the antibody titres ranged from 1:64 to 1:1,024. Seroprevalence of T. gondii infection in sheep was 17.1% in Inner Mongolia, 33.8% in Heilongjiang, 24.6% in Jilin and 46.3% in Hebei. Age and rearing system significantly affected seropositivity. The present survey indicates antibodies to T. gondii are widely prevalent in sheep in northern China, which may cause public health problems in these provinces.

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

Eamsobhana, P. ; Yong, H.S. ; Song, S.L. ; Tungtrongchitr, A. ; Roongruangchai, K.

Tropical Biomedicine.2018;35(3):669-677.

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.

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A preliminary study of platelet hyperactivity in the chronic indeterminate phase of Chagas’ disease

Flavio Rojas Castillejos ; Laura Perez-Campos Mayoral ; Gabriel Mayoral Andrade ; Maria Teresa Hernandez-Huerta ; Socorro Pina-Canseco ; Ruth Martinez Cruz ; Efrain Herrera Colmenares ; Eduardo Perez-Campos Mayoral ; Paz Maria Salazar ; Martha Bucio Torres ; Margarita Cabrera Bravo ; Margarito Martinez Cruz ; Carlos Matí ; as Cervantes ; Roxana Diaz Albarraz ; Joel Lopez Matias ; Gabriela Ines Rios Arias ; Gema Hrnandez Bernardino ; Ernesto Perez Matus ; Rosalinda Mendez Trujillo ; Luis Manuel Sanchez Navarro ; Alma Dolores Perez Santiago ; Eduardo Perez Campos

Tropical Biomedicine.2018;35(3):678-683.

The chronic indeterminate phase of Chagas’ disease is asymptomatic despite positive test results for antibodies specific to Trypanosoma cruzi. CD62P-APC (P-selectin) and PAC-1 FITC (GpIIb/IIIa) may improve diagnosis as biomarkers of platelet activity. Nine asymptomatic seropositive subjects, previously untreated, were selected from a blood bank within a year of Chagas’ disease detection, in addition to a control group of four. All subjects were evaluated by flow cytometry for CD62P, PAC-1 and CD41, and in a complementary study, by Tissue Doppler Echocardiography for isovolumic relaxation times (IVRT) and E/A ratios. The subjects were classified as positive or negative for CD62P and PAC-1 by a cut off obtained from their mean±2SD. For IVRT and E/A ratios, cut offs were obtained from the American Society of Echocardiography and the European Association of Cardiovascular Imaging recommendations. Fisher’s exact test was used for associated findings. Pre-test and post-test probability, sensitivity, specificity, positive and negative predictive values and likelihood ratios were calculated. Abnormalities were expressed as platelet hyperactivity and ventricular dysfunction in CD62P, PAC-1, IVRT and E/A ratios. CD62P appears to have greater sensitivity (0.75) and PAC-1, more accurate specificity (0.75), which may explain thrombotic events in Chagas’ disease. We recommend the use of CD62P and PAC-1 as biomarkers of platelet hyperactivity in patients in the chronic indeterminate phase of Chagas’ disease.

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