Histopathology and Molecular Characterization of Microsporidian Parasites in Cultured Shrimp Penaeus Monodon Fabricius 1798 of Bangladesh

Black tiger shrimp (Penaeus monodon Fabricius 1798) cultured in Bangladesh was investigated for the presence of microsporidian parasite. Histological section of hepatopancreas showed a large number of microsporidian spores under light microscopy. Spores under scanning electron microscope appeared oval shapes. Histology of infected shrimps showed severe degeneration of hepatopancreatic tubules. Early and late stage of microsporidian parasites in hepatopancreatic tubules were also observed. DNA extracted from the hepatopancreas of shrimps were subjected to PCR amplification using primers targeting microsporidian SSU rRNA gene. The PCR amplified an expected product of ~328 bp and the sequences showed 81 - 82% identity with the Paranucleospora theridion reported from western Norway in 2008. Further screening of field samples was carried out using EHP-specific primers. DNA extracted from ten hepatopancreas samples of P. monodon were tested and none found to be positive for EHP (Enterocytozoon hepatopenaei). This is the first report for the identification of microsporidian parasites in cultured shrimp along the south-west region of Bangladesh. Asiat. Soc. Bangladesh, Sci. 45(2): 187-196, December 2019

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IDENTIFICATION AND CHARACTERIZATION OF PIGMENTED BACTERIA ISOLATED FROM MALAYSIAN SEAWATER

International Journal of Students Research in Technology & Management ◽

10.18510/ijsrtm.2019.741 ◽

2019 ◽

Vol 7 (4) ◽

pp. 01-08

Author(s):

Nur Afifah Mursyida Zaujan ◽

Mohamad Zohdi Othman ◽

Fatin Najihah Mohd Lutfi ◽

Kamarul Rahim Kamarudin ◽

Hanina Mohd Noor ◽

...

Keyword(s):

Human Health ◽

Pcr Amplification ◽

Fluorescent Light ◽

Rrna Gene ◽

Yellow Orange ◽

Exiguobacterium Profundum ◽

The Stability ◽

Antimicrobial Metabolites ◽

Pigmented Bacteria

Purpose of study: Bacteria can naturally produce pigments that can be useful for various applications as they possess antimicrobial metabolites among other numerous benefits towards the human health. This study was carried out to identify the species of marine bacterial isolates PMA, PM3C1 and PM5C1 exhibiting yellow, orange and green colors respectively. Methodology: The current study is using Polymerase Chain Reaction (PCR) amplification and sequence analysis of their 16S rRNA gene. The stability of pigments extracted from the bacterial samples was also analyzed against different temperature and light conditions. Main Findings: Sequence alignment using BLAST revealed that the yellow, orange, and green-pigmented bacteria have 84% similarity with Staphylococcus aureus, 85% similarity with Exiguobacterium profundum and 95% similarity with Pseudomonas aeruginosa respectively. The green pigment showed major changes in color following exposure to sunlight and fluorescent light, and when incubated at 24°C and 50°C. Exposure to direct sunlight also results in the reduction of color for the yellow and orange extracts, while no effect was observed for both pigments under fluorescent light. Incubation at 50°C results in the reduction of the orange color, while the yellow pigment was observed to be unaffected suggesting its stability at high temperature. Implications: Natural pigments production can provide many advantages including reduction of pollution generation, ease of disposal and other benefits to the human health.

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Specific PCR Detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in Chicken Meat

Journal of Food Protection ◽

10.4315/0362-028x-72.7.1491 ◽

2009 ◽

Vol 72 (7) ◽

pp. 1491-1495 ◽

Cited By ~ 16

Author(s):

DANIELA PENTIMALLI ◽

NICOLETTE PEGELS ◽

TERESA GARCÍA ◽

ROSARIO MARTÍN ◽

ISABEL GONZÁLEZ

Keyword(s):

Pcr Amplification ◽

Chicken Meat ◽

Pcr Analysis ◽

Rrna Gene ◽

Pcr Assay ◽

Gyra Gene ◽

Specific Primers ◽

Specific Pcr ◽

Arcobacter Butzleri ◽

Species Specific

An enrichment PCR assay using species-specific primers was developed for the detection of Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter skirrowii, and Arcobacter cibarius in chicken meat. Primers for A. cryaerophilus, A. skirrowii, and A. cibarius were designed based on the gyrA gene to amplify nucleic acid fragments of 212, 257, and 145 bp, respectively. The A. butzleri–specific primers were designed flanking a 203-bp DNA fragment in the 16S rRNA gene. The specificity of the four primer pairs was assessed by PCR analysis of DNA from a panel of Arcobacter species, related Campylobacter, Helicobacter species, and other food bacteria. The applicability of the method was then validated by testing 42 fresh retail-purchased chicken samples in the PCR assay. An 18-h selective preenrichment step followed by PCR amplification with the four Arcobacter primer sets revealed the presence of Arcobacter spp. in 85.7% of the retail chicken samples analyzed. A. butzleri was the only species present in 50% of the samples, and 35.7% of the samples were positive for both A. butzleri and A. cryaerophilus. A. skirrowii and A. cibarius were not detected in any of the chicken samples analyzed. The enrichment PCR assay developed is a specific and rapid alternative for the survey of Arcobacter contamination in meat.

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Characterization of Diazotrophs Containing Mo-Independent Nitrogenases, Isolated from Diverse Natural Environments

Applied and Environmental Microbiology ◽

10.1128/aem.02694-07 ◽

2008 ◽

Vol 74 (11) ◽

pp. 3471-3480 ◽

Cited By ~ 47

Author(s):

Doris A. Betancourt ◽

Telisa M. Loveless ◽

James W. Brown ◽

Paul E. Bishop

Keyword(s):

Azotobacter Vinelandii ◽

Wood Chip ◽

Pcr Amplification ◽

Rrna Gene ◽

Natural Environments ◽

Gene Sequence Analysis ◽

Nitrogenase Genes ◽

Free Media ◽

Aerobic And Anaerobic

ABSTRACT Molybdenum-independent nitrogenases were first described in the nitrogen-fixing bacterium Azotobacter vinelandii and have since been described in other diazotrophic bacteria. Previously, we reported the isolation of seven diazotrophs with Mo-independent nitrogenases from aquatic environments. In the present study, we extend these results to include diazotrophs isolated from wood chip mulch, soil, “paraffin dirt,” and sediments from mangrove swamps. Mo-deficient, N-free media under both aerobic and anaerobic conditions were used for the isolations. A total of 26 isolates were genetically and physiologically characterized. Their phylogenetic placement was determined using 16S rRNA gene sequence analysis. Most of the isolates are members of the gamma subdivision of the class Proteobacteria and appear to be specifically related to fluorescent pseudomonads and azotobacteria. Two other isolates, AN1 and LPF4, are closely related to Enterobacter spp. and Paenibacillus spp., respectively. PCR and/or Southern hybridization were used to detect the presence of nitrogenase genes in the isolates. PCR amplification of vnfG and anfG was used to detect the genetic potential for the expression of the vanadium-containing nitrogenase and the iron-only nitrogenase in the isolates. This study demonstrates that diazotrophs with Mo-independent nitrogenases can be readily isolated from diverse natural environments.

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CHARACTERIZATION OF BACTERIAL ISOLATES FOR SUSTAINABLE RICE BLAST CONTROL

Revista Caatinga ◽

10.1590/1983-21252020v33n313rc ◽

2020 ◽

Vol 33 (3) ◽

pp. 702-712

Author(s):

BÁRBARA ESTEVAM DE MELO MARTINS ◽

AMANDA ABDALLAH CHAIBUB ◽

MARCIO VINICIUS DE CARVALHO BARROS CORTÊS ◽

VALÁCIA LEMES DA SILVA LOBO ◽

MARTA CRISTINA CORSI DE FILIPPI

Keyword(s):

Rice Blast ◽

Pcr Amplification ◽

Morphological Characterization ◽

Bacterial Isolates ◽

Specific Primers ◽

Leaf Blast ◽

Fluorescent Pigment ◽

Gram Staining ◽

Bacillus Spp

ABSTRACT Rice blast (Magnaporthe oryzae) limits rice (Oryza sativa) grain yields worldwide. The objective of this investigation was to morphologically, biochemically, and molecularly characterize six bacterial isolates, BRM 32109, BRM 32110, BRM 32111, BRM 32112, BRM 32113, and BRM 32114, and to determine their potential as antagonists to M. oryzae. Morphological characterization was based on colony formation and color, Gram staining, and fluorescent pigment production. Biochemical studies were based on cellulase, chitinase, phosphatase, indoleacetic acid, and siderophore production, as well as biofilm formation. The molecular identification used specific primers for PCR amplification of the 16S rRNA region, followed by sequencing. The antagonism studies involved three experiments, which had randomized designs. Two of them were conducted in laboratory conditions, pairing bacterial colonies and M. oryzae, using bacterial filtrates, and the third was conducted in greenhouse conditions. BRM 32111 and BRM 32112 were identified as Pseudomonas sp., BRM 32113 as Burkholderia sp., BRM 32114 as Serratia sp., and BRM 32110 and BRM 32109 as Bacillus spp. BRM 32112, BRM 32111, and BRM 32113 inhibited the colony of M. oryzae by 68%, 65%, and 48%, respectively. The bacterial suspensions of the BRM 32111, BRM 32112, and BRM 3212 filtrates suppressed leaf blast by 81.0, 79.2, and 66.3%, respectively. BRM 32111 and BRM 32112 were determined to be antagonists of M. oryzae and were found to solubilize phosphate, produce siderophores and cellulose, form biofilms, and suppress leaf blast. These isolates should be further investigated as potential biological control agents for leaf blast control.

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Genetic characterization of Blastocystis from wild animals in Sichuan Wolong National Natural Reserve, southwestern of China-Zoonotic potential

10.21203/rs.3.rs-471433/v2 ◽

2021 ◽

Author(s):

Shanyu Chen ◽

Wangyu Meng ◽

Ziyao Zhou ◽

Lei Deng ◽

Xiaogang Shi ◽

...

Keyword(s):

Pcr Amplification ◽

Human Infection ◽

Rrna Gene ◽

Zoonotic Potential ◽

Wild Animals ◽

Genetic Characteristics ◽

Natural Reserve ◽

Faecal Samples ◽

Wide Range

Abstract Background Blastocystis, a highly prevalent eukaryotic parasite, has been identified in a wide range of hosts, including humans, domestic and wild animals. Many animals are potential sources of Blastocystis infection for humans, while few information about the prevalence of Blastocystis in wild animals have being documented. Therefore, the present study was designed to investigate the prevalence and subtypes of Blastocystis in wild animals of Sichuan Wolong National Natural Reserve, southwestern of China, so as to assess the zoonotic potential of these animals. Methods A total of 300 faecal samples were collected from 27 wildlife species in three areas of Sichuan Wolong National Natural Reserve in southwestern China. The subtype (ST) genetic characteristics and prevalence of Blastocystis were determined by PCR amplification of the barcode region (a fragment of ∼600 bp) of the SSU rRNA gene, and phylogenetic analysis were further performed to determine the genetic characteristics of Blastocystis subtypes. Results 30 of 300 faecal samples (10.0%) were Blastocystis-positive. The highest prevalence of Blastocystis was found in Yinchanggou (18.3%), which was significantly higher than that in Niutoushan (7.5%), and Genda (5.5%) (P < 0.05). Specifically, the highest prevalence of Blastocystis was found in primates (20.0%, 1/5), followed by rodentia 14.3% (1/7), artiodactyla 13.1% (26/198), carnivora 2.3% (2/87), galliformes 0% (0/3). Sequence analysis showed 5 subtypes (ST1, ST3, ST5, ST13, and ST14), with ST13 and ST14 as the predominant subtype (33.3%, 10/30), followed by ST1 (20.0%, 6/30). Conclusions To the best of our knowledge, this is the first molecular investigation on Blastocystis infection in wild animals in southwestern of China. ST1, ST3, and ST5 were identified in both humans and wild animals, suggesting that these wild animals may be potential reservoirs of Blastocystis for human infection.

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Identification and molecular characterization of Wolbachia strains in natural populations of Aedes albopictus in China

Parasites & Vectors ◽

10.1186/s13071-020-3899-4 ◽

2020 ◽

Vol 13 (1) ◽

Cited By ~ 3

Author(s):

Yaping Hu ◽

Zhiyong Xi ◽

Xiaobo Liu ◽

Jun Wang ◽

Yuhong Guo ◽

...

Keyword(s):

Aedes Albopictus ◽

Dengue Infection ◽

Natural Populations ◽

Pcr Amplification ◽

Surface Protein ◽

Multiple Infection ◽

Rrna Gene ◽

Dengue Outbreaks ◽

Surface Protein Gene

Abstract Background Aedes albopictus is naturally infected with Wolbachia spp., maternally transmitted bacteria that influence the reproduction of hosts. However, little is known regarding the prevalence of infection, multiple infection status, and the relationship between Wolbachia density and dengue outbreaks in different regions. Here, we assessed Wolbachia infection in natural populations of Ae. albopictus in China and compared Wolbachia density between regions with similar climates, without dengue and with either imported or local dengue. Results To explore the prevalence of Wolbachia infection, Wolbachia DNA was detected in mosquito samples via PCR amplification of the 16S rRNA gene and the surface protein gene wsp. We found that 93.36% of Ae. albopictus in China were positive for Wolbachia. After sequencing gatB, coxA, hcpA, ftsZ, fbpA and wsp genes of Wolbachia strains, we identified a new sequence type (ST) of wAlbB (464/465). Phylogenetic analysis indicated that wAlbA and wAlbB strains formed a cluster with strains from other mosquitoes in a wsp-based maximum likelihood (ML) tree. However, in a ML tree based on multilocus sequence typing (MLST), wAlbB STs (464/465) did not form a cluster with Wolbachia strains from other mosquitoes. To better understand the association between Wolbachia spp. and dengue infection, the prevalence of Wolbachia in Ae. albopictus from different regions (containing local dengue cases, imported dengue cases and no dengue cases) was determined. We found that the prevalence of Wolbachia was lower in regions with only imported dengue cases. Conclusions The natural prevalence of Wolbachia infections in China was much lower than in other countries or regions. The phylogenetic relationships among Wolbachia spp. isolated from field-collected Ae. albopictus reflected the presence of dominant and stable strains. However, wAlbB (464/465) and Wolbachia strains did not form a clade with Wolbachia strains from other mosquitoes. Moreover, lower densities of Wolbachia in regions with only imported dengue cases suggest a relationship between fluctuations in Wolbachia density in field-collected Ae. albopictus and the potential for dengue invasion into these regions.

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Characterization of a Novel Spirochete Associated with the Hydrothermal Vent Polychaete Annelid, Alvinella pompejana

Applied and Environmental Microbiology ◽

10.1128/aem.67.1.110-117.2001 ◽

2001 ◽

Vol 67 (1) ◽

pp. 110-117 ◽

Cited By ~ 38

Author(s):

Barbara J. Campbell ◽

S. Craig Cary

Keyword(s):

High Temperature ◽

Hydrothermal Vent ◽

Denaturing Gradient Gel Electrophoresis ◽

Pcr Amplification ◽

Dorsal Surface ◽

Specific Primers ◽

Gradient Gel Electrophoresis ◽

Polychaetous Annelid ◽

Alvinella Pompejana

ABSTRACT A highly integrated, morphologically diverse bacterial community is associated with the dorsal surface of Alvinella pompejana, a polychaetous annelid that inhabits active high-temperature deep-sea hydrothermal vent sites along the East Pacific Rise (EPR). Analysis of a previously prepared bacterial 16S ribosomal DNA (rDNA) library identified a spirochete most closely related to an endosymbiont of the oligochete Olavius loisae. This spirochete phylotype (spirochete A) comprised only 2.2% of the 16S rDNA clone library but appeared to be much more dominant when the same sample was analyzed by denaturing gradient gel electrophoresis (DGGE) and the terminal restriction fragment length polymorphism procedure (12 to 18%). PCR amplification of the community with spirochete-specific primers used in conjunction with DGGE analysis identified two spirochete phylotypes. The first spirochete was identical to spirochete A but was present in only one A. pompejana specimen. The second spirochete (spirochete B) was 84.5% similar to spirochete A and, more interestingly, was present in the epibiont communities of all of theA. pompejana specimens sampled throughout the geographic range of the worm (13°N to 32°S along the EPR). The sequence variation of the spirochete B phylotype was less than 3% for the range of A. pompejana specimens tested, suggesting that a single spirochete species was present in the A. pompejanaepibiotic community. Additional analysis of the environments surrounding the worm revealed that spirochetes are a ubiquitous component of high-temperature vents and may play an important role in this unique ecosystem.

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Isolation and Molecular Characterization of Riemerella anatipestifer from Domesticated Ducks of Assam, India

Indian Journal of Animal Research ◽

10.18805/ijar.b-4295 ◽

2021 ◽

Author(s):

M.K. Doley ◽

S. Das ◽

R.K. Sharma ◽

P. Borah ◽

D.K. Sarma ◽

...

Keyword(s):

16S Rrna ◽

16S Rrna Gene ◽

Molecular Characterization ◽

Molecular Identification ◽

Rrna Gene ◽

Eastern Region ◽

Riemerella Anatipestifer ◽

Field Samples ◽

Apparently Healthy

Background: Riemerella anatipestifer (R. anatipestifer) is a gram negative, microaerophilic, non-motile, bipolar bacteria. High genetic diversity and molecular differentiation were reported among field isolates. Although the bacterium causes one of the most economically important duck diseases in the north-eastern region of India, little work has been done on isolation, identification and molecular characterization of the bacteria. Hence, the present investigation was undertaken with a view to characterize the R. anatipestifer isolates from ducks of Assam.Methods: Phenotypic and molecular identification of R. anatipestifer isolates from domesticated ducks of Assam, India were carried out during the period from February, 2019 to January 2020. A total of 624 samples (Ocular swab, throat swab, liver, spleen, kidney, brain, heart, lung) from ducks comprising of apparently healthy, ailing and dead ducks were collected from five districts of Assam, India were processed to isolate and identify the bacteria. The tentative identification of the bacteria was done based on phenotypic characteristics viz., colony morphology, growth characteristics and biochemical reactions. All the phenotypically positive isolates were further subjected to molecular identification based on PCR assay targeting 16S rRNA gene and ERIC sequence.Result: The bacteria could be isolated from different field samples. The highest percentage of the samples that yielded the bacteria are from brain (76%) followed by spleen (74%) of dead ducks and less number of ocular swab (33%) from apparently healthy ducks were found positive. Sequencing of the amplified product of the selected R. anatipestifer isolates targeting 16S rRNA gene revealed homology percentage of 96.5-100%. Further, sequences representing five geographical locations were submitted to NCBI gene bank. Phylogenetic studies of the isolates indicated that there is prevalence of at least two genetically different strains of R. anatipestifer in the study area. The study suggested that the R. anatipestifer infection is endemic in Assam causing varying rate of morbidity (39%) and mortality (53%) and molecular based confirmation is necessary besides phenotypic identification.

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Prevalence and Genetic Characterization of Cryptosporidium Infection in Java Sparrows (Lonchura oryzivora) in Northern China

BioMed Research International ◽

10.1155/2017/2318476 ◽

2017 ◽

Vol 2017 ◽

pp. 1-4 ◽

Cited By ~ 2

Author(s):

Qiu-Xia Yao ◽

Xiao-Xuan Zhang ◽

Kai Chen ◽

Jian-Gang Ma ◽

Wen-Bin Zheng ◽

...

Keyword(s):

Pcr Amplification ◽

Northern China ◽

Small Subunit ◽

Rrna Gene ◽

Ssu Rrna ◽

Ssu Rrna Gene ◽

Wide Range ◽

Baseline Information ◽

And Control

Cryptosporidiosis is a cosmopolitan parasitosis that affects a wide range of hosts including birds. As information concerning Cryptosporidium in birds is limited, the present study examined the prevalence and genotypes of Cryptosporidium in Java sparrows in Beijing and Shangqiu, northern China. Three hundred and fifty fecal samples were collected from Java sparrows (Lonchura oryzivora, 225 white Java sparrows and 125 gray Java sparrows) in Beijing and Shangqiu in October 2015, and the samples were examined by PCR amplification of the small subunit ribosomal RNA (SSU rRNA) gene. The overall Cryptosporidium prevalence is 13.42% (47/350), with 16.44% (37/225) in white Java sparrows and 8.00% (10/125) in gray Java sparrows. Cryptosporidium prevalence was 9.82% (16/163) in Java sparrows from Beijing and 16.58% (31/187) in Java sparrows from Shangqiu. The prevalence of Cryptosporidium in females and males was 40.63% (26/64) and 7.34% (21/286), respectively. The Cryptosporidium prevalence in Java sparrows of different ages varied from 10.47% to 16.33%. Sequence analysis of the SSU rRNA gene revealed that all the samples represented C. baileyi. This is the first report of Cryptosporidium in gray Java sparrows in China, which extend the host range for C. baileyi. These results provide baseline information for further studies of molecular epidemiology and control of Cryptosporidium infection in poultry in China.

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