• Title/Summary/Keyword: molecular prevalence

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Prevalence and Molecular Characterization of ESBL Producing Enterobacteriaceae from Highly Polluted Stretch of River Yamuna, India

  • Siddiqui, Kehkashan;Mondal, Aftab Hossain;Siddiqui, Mohammad Tahir;Azam, Mudsser;Haq., Qazi Mohd. Rizwanul
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.135-144
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    • 2018
  • The rapid increase in number and diversity of Extended Spectrum ${\beta}$-Lactamases (ESBLs) producing Enterobacteriaceae in natural aquatic environment is a major health concern worldwide. This study investigates abundance and distribution of ESBL producing multidrug resistant Enterobacteriaceae and molecular characterization of ESBL genes among isolates from highly polluted stretch of river Yamuna, India. Water samples were collected from ten different sites distributed across Delhi stretch of river Yamuna, during 2014-15. A total of 506 non duplicate Enterobacteriaceae isolates were obtained. Phenotypic detection of ESBL production and antibiotic sensitivity for 15 different antibiotics were performed according to CLSI guidelines (Clinical and Laboratory Standard Institute, 2015). A subset of ESBL positive Enterobacteriaceae isolates were identified by 16S rRNA gene and screened for ESBL genes, such as $bla_{CTX-M}$, $bla_{TEM}$ and $bla_{OXA}$. Out of 506 non-duplicate bacterial isolates obtained, 175 (34.58%) were positive for ESBL production. Susceptibility pattern for fifteen antibiotics used in this study revealed higher resistance to cefazolin, rifampicin and ampicillin. A high proportion (76.57%) of ESBL positive isolates showed multidrug resistance phenotype, with MAR index of 0.39 at Buddha Vihar and Old Delhi Railway bridge sampling site. Identification and PCR based characterization of ESBL genes revealed the prevalence of $bla_{CTX-M}$ and $bla_{TEM}$ genes to be 88.33% and 61.66%, respectively. Co-occurrence of $bla_{CTX-M}$ and $bla_{TEM}$ genes was detected in 58.33% of the resistant bacteria. The $bla_{OXA}$ gene was not detected in any isolates. This study highlights deteriorating condition of urban aquatic environment due to rising level of ESBL producing Enterobacteriaceae with multidrug resistance phenotype.

Molecular Prevalence of Acarapis Mite Infestations in Honey Bees in Korea

  • Ahn, Ah-Jin;Ahn, Kyu-Sung;Noh, Jin-Hyeong;Kim, Young-Ha;Yoo, Mi-Sun;Kang, Seung-Won;Yu, Do-Hyeon;Shin, Sung Shik
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.315-320
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    • 2015
  • Acarapis mites, including Acarapis woodi, Acarapis externus, and Acarapis dorsalis, are parasites of bees which can cause severe damage to the bee industry by destroying colonies and decreasing honey production. All 3 species are prevalent throughout many countries including UK, USA, Iran, Turkey, China, and Japan. Based on previous reports of Acarapis mites occurring in northeast Asia, including China and Japan, we investigated a survey of Acarapis mite infestations in honey bees in Korean apiaries. A total of 99 colonies of Apis mellifera were sampled from 5 provinces. The head and thorax of 20 bees from each colony were removed for DNA extraction. PCR assays were performed with 3 primer sets, including T, A, and K primers. Results indicated that 42.4% (42/99) of samples were Acarapis-positive by PCR assay which were sequenced to identify species. Each sequence showed 92.6-99.3% homology with reference sequences. Based on the homology, the number of colonies infected with A. dorsalis was 32 which showed the highest infection rate among the 3 species, while the number of colonies infected with A. externus and A. woodi was 9 and 1, respectively. However, none of the Acarapis mites were morphologically detected. This result could be explained that all apiaries in the survey used acaricides against bee mites such as Varroa destructor and Tropilaelaps clareae which also affect against Acarapis mites. Based on this study, it is highly probable that Acarapis mites as well as Varroa and Tropilaelaps could be prevalent in Korean apiaries.

Occurrence and Molecular Identification of Anisakis Dujardin, 1845 from Marine Fish in Southern Makassar Strait, Indonesia

  • Anshary, Hilal;Sriwulan, Sriwulan;Freeman, Mark A.;Ogawa, Kazuo
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.9-19
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    • 2014
  • Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica AB517571/DQ116427. The difference of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait.

Four Cases of Taenia saginata Infection with an Analysis of COX1 Gene

  • Cho, Jaeeun;Jung, Bong-Kwang;Lim, Hyemi;Kim, Min-Jae;Yooyen, Thanapon;Lee, Dongmin;Eom, Keeseon S.;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.79-83
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    • 2014
  • Human taeniases had been not uncommon in the Republic of Korea (=Korea) until the 1980s. The prevalence decreased and a national survey in 2004 revealed no Taenia egg positive cases. However, a subsequent national survey in 2012 showed 0.04% (10 cases) prevalence of Taenia spp. eggs suggesting its resurgence in Korea. We recently encountered 4 cases of Taenia saginata infection who had symptoms of taeniasis that included discharge of proglottids. We obtained several proglottids from each case. Because the morphological features of T. saginata are almost indistinguishable from those of Taenia asiatica, molecular analyses using the PCR-RFLP and DNA sequencing of the cytochrome c oxidase subunit 1 (cox1) were performed to identify the species. The PCR-RFLP patterns of all of the 4 specimens were consistent with T. saginata, and the cox1 gene sequence showed 99.8-100% identity with that of T. saginata reported previously from Korea, Japan, China, and Cambodia. All of the 4 patients had the history of travel abroad but its relation with contracting taeniasis was unclear. Our findings may suggest resurgence of T. saginata infection among people in Korea.

Seroprevalence and molecular detection of Mycoplasma gallisepticum and M. synoviae infection in broiler breeder in Jeonbuk providence, Korea (전북지역 육용종계에서 Mycoplasma gallisepticum과 M. synoviae의 항체 및 유전자 양성률 조사)

  • Kwak, Kilhan;Lee, Heungiae;Yuk, Hyunsu;Lee, Jaeuk;Lee, Kwanho;Yi, Young-Joo;Lee, Sang-Myeong
    • Korean Journal of Veterinary Service
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    • v.39 no.2
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    • pp.101-105
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    • 2016
  • The present study investigated serological and molecular prevalence of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infection in unvaccinated broiler breeder farms in Jeonbuk providence. An enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) had been used to determine antibody titers against MG and MS, and genome of these pathogens, respectively. Seventy five percent of farms were seropositive for MG and 94% of farms were seropositive for MS. In addition, the rate of antibody positive flocks against MG were 65.3% (32/49), while the rate of positive flocks against MS were 84.2% (80/95). The geometric mean antibody titers were $802.2{\pm}626$ and $27,726.7{\pm}2426$ against MG and MS, respectively. Interestingly, none of samples was positive for MG genome by PCR, while 94% (farms), 82% (flocks) and 62.6% (broiler breeder) were positive for MS genome by PCR. These findings suggest that the prevalence of MG or MS infection could be higher than expected. Thus, strict prevention program including vaccination and environmental sanitation should be implemented to avoid disease transmission from breeder to broilers as well as transmission among broilers.

Molecular Characterization of Fluoroquinolone Resistant Escherichia coli Isolates from Chickens in Korea (닭에서 동정된 플르오르퀴놀론 내성 대장균 균주의 분자생물학적 성상에 관한 연구)

  • Sung, Ji-Youn;Oh, Ji-Eun
    • Journal of Digital Convergence
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    • v.14 no.4
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    • pp.371-378
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    • 2016
  • An aim of current study was to investigate the prevalence and the mechanism of quinolone-resistance in E. coli isolates obtained from chicken cecum in Korea. In addition, multilocus sequence typing (MLST) was also performed for the molecular characterization of E. coli isolates. In an antimicrobial susceptibility test by the disk diffusion method, the 63.5% (54/85) of E. coli isolates showed the resistance to quinolone group of antimicrobial agents. All of the 54 E. coli isolates showing resistant to quinolone group had sense mutations in gyrA gene and point mutations at the $57^{th}$, $80^{th}$, or $84^{th}$ residues in parC gene were detected in 90.7% of the isolates. Interestingly, E. coli ST was closely related to amino acid substitutions in parE gene. Our results indicated that the long-term use of antimicrobial agents in food-producing animals was strongly associated with a prevalence of antimicrobial resistance in commensal Enterobacteriaceae, suggesting the need for continuous surveillance and monitoring of antimicrobial resistant determinants in bacterial isolates from food animals.

Simplex PCR Assay for Detection of blaTEM and gyrA Genes, Antimicrobial Susceptibility Pattern and Plasmid Profile of Salmonella spp. Isolated from Stool and Raw Meat Samples in Niger State, Nigeria

  • Musa, Dickson A.;Aremu, Kolawole H.;Ajayi, Abraham;Smith, Stella I.
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.230-235
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    • 2020
  • The global evolution of antibiotic resistance has threatened the efficacy of available treatment options with ravaging impacts observed in developing countries. As a result, investigations into the prevalence of antibiotic resistance and the role of plasmids are crucial. In this study, we investigated the presence and distribution of blaTEM and gyrA genes, plasmid profiles, and the antimicrobial susceptibility pattern of Salmonella strains isolated from raw meat and stool sources across Niger State, Nigeria. Ninety-eight samples, comprising 72 raw meat and 26 stool samples, were screened for Salmonella spp. The antimicrobial susceptibility of Salmonella isolates to 10 commonly used antimicrobial agents was determined using the KirbyBauer disc diffusion method. Isolates were further analyzed for plasmids, in addition to PCR amplification of beta-lactamase (blaTEM) and gyrA genes. A total of 31 Salmonella spp. were isolated, with 22 from raw meat (70.97%) and 9 from stool (29.03%). Salmonella spp. with multiple resistance patterns to ceftazidime, cefuroxime, ceftriaxone, erythromycin, ampicillin, cloxacillin, and gentamicin were detected. Ofloxacin and ciprofloxacin were found to be the most effective among the antibiotics tested, with 67.7% and 93.5% susceptible isolates, respectively. Nine (29.03%) isolates harbored plasmids with molecular sizes ranging between 6557 bp and 23137 bp. PCR amplification of gyrA was detected in 1 (3.23%) of the 31 isolates while 28 isolates (90.32%) were positive for blaTEM. This study shows the incidence of antibiotic resistance in Salmonella isolates and the possible role of plasmids; it also highlights the prevalence of ampicillin resistance in this local population.

Molecular Detection and Phylogenetic Analysis of Anaplasma phagocytophilum in Horses in Korea

  • Seo, Min-Goo;Ouh, In-Ohk;Choi, Eunsang;Kwon, Oh-Deog;Kwak, Dongmi
    • Parasites, Hosts and Diseases
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    • v.56 no.6
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    • pp.559-565
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    • 2018
  • The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

Molecular Prevalence of Cryptosporidium spp. in Breeding Kennel Dogs

  • Itoh, Naoyuki;Tanaka, Hazuki;Iijima, Yuko;Kameshima, Satoshi;Kimura, Yuya
    • Parasites, Hosts and Diseases
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    • v.57 no.2
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    • pp.197-200
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    • 2019
  • Cryptosporidium is a common intestinal protozoan that can lead to diarrhea in humans and dogs. The predominant species of infection are C. hominis and C. parvum in humans, and C. canis in dogs. However, C. canis can infect immunocompromised humans. Considering the close contact with humans, dogs have the potential to be reservoirs for human cryptosporidiosis. Breeding kennels are the major supply source of puppies for pet shops. The present study is to determine the molecular prevalence and characteristics of Cryptosporidium spp. found in breeding kennel dogs. A total of 314 fecal samples were collected from young and adult dogs kept in 5 breeding kennels. A polymerase chain reaction targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. To determine the species, the DNA sequences were compared to GenBank data. Overall, 21.0% of the fecal samples were positive for Cryptosporidium spp. infection. Cryptosporidium spp. was detected in all 5 facilities. A sequencing analysis demonstrated that all isolates shared 99-100% similarity with C. canis. The results suggest that Cryptosporidium spp. infection is present at a high-level in breeding kennel dogs. However, because dominant species in this survey was C. canis, the importance of breeding kennel dogs as reservoirs for Cryptosporidium spp. transmission to humans is likely to be low in Japan.

Prevalence and molecular analysis of glucose-6-phosphate dehydrogenase deficiency in Chin State, Myanmar

  • Ja Moon Aung;Zin Moon;Dorene VanBik;Sylvatrie-Danne Dinzouna-Boutamba;Sanghyun Lee;Zau Ring;Dong-Il Chung;Yeonchul Hong;Youn-Kyoung Goo
    • Parasites, Hosts and Diseases
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    • v.61 no.2
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    • pp.154-162
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    • 2023
  • Glucose-6-phosphate dehydrogenase (G6PD) deficiency is caused by X-linked recessive disorderliness. It induces severe anemia when a patient with G6PD deficiency is exposed to oxidative stress that occurs with administration of an antimalarial drug, primaquine. The distribution of G6PD deficiency remains unknown while primaquine has been used for malaria treatment in Myanmar. This study aimed to investigate the prevalence of G6PD deficiency and its variants in Chin State, Myanmar. Among 322 participants, 18 (11 males and 7 females) demonstrated a G6PD deficiency. Orissa variant was dominant in the molecular analysis. This would be related to neighboring Indian and Bangladeshi population, in which Orissa variant was also reported as the main mutation type. The screening test for G6PD deficiency before primaquine treatment appears to be important in Myanmar.