scholarly journals Fatal Hymenolepis nana cestodiasis in a ring-tailed lemur (Lemur catta)

2021 ◽  
pp. 030098582110425
Author(s):  
Esther E. V. Crouch ◽  
Charlotte Hollinger ◽  
Stephanie Zec ◽  
Denise McAloose

The cestode Hymenolepis nana is a common parasite of humans and mice. Fecal shedding in the absence of clinical disease has previously been reported in ring-tailed lemurs ( Lemur catta). This report describes fatal, disseminated H. nana cestodiasis infection in an aged ring-tailed lemur in a zoological collection. The parasites were associated with severe multifocal to coalescing and regionally extensive pyogranulomatous hepatitis and moderate multifocal pneumonia. The morphology of the parasites was highly unusual. Profiles were variably sized, ellipsoid to irregularly serpiginous, lined by a thin tegument, and filled with lightly eosinophilic fibrillar stroma and numerous, round basophilic cells. Polymerase chain reaction targeting a portion of the 18S rRNA gene and DNA sequencing of the amplicon showed 100% homology with H. nana.

Author(s):  
Satriya Putra Prakoso ◽  
I Nengah Wirajana ◽  
I Wayan Suarsa

The aim of this research was to amplificate 18S rRNA gene fragment from honey’s metagenomic DNA using Polymerase Chain Reaction (PCR). The honey sample was collected from Seraya Tengah village, Karangasem regency. The best result of primer design from in silico test was continued to in vitro test using PCR method. The optimum conditions for amplification was obtained as follows: pre-denaturation at 95oC for 3 minutes and continued with 30 of amplification cycle (denaturation at 95°C for 1 minutes, annealing at 55°C for 1 minutes and elongation at 72°C for 1 minutes) and the last step continued with extension process at 72°C for 2 minutes. The size of DNA fragment band of amplified product was about 100 bp which obtained from the honey’s metagenomic DNA.


2019 ◽  
Vol 7 (4) ◽  
Author(s):  
Cinzia Daga ◽  
Simona Cau ◽  
Maria Giovanna Tilocca ◽  
Barbara Soro ◽  
Aldo Marongiu ◽  
...  

Fish is one of fourteen allergens that must be highlighted on the label within the ingredients list. It should be noted that the European regulation, is very restrictive to allergens with zero tolerance. Therefore it is important to establish sensitive and specific methods for detecting fish allergen. Applicability to detect and quantify fish allergen by droplet digital polymerase chain reaction (ddPCR) has been evaluated in this work. Genomic DNA of three fish species belonging to the most common fish families were analyzed. PCR primers were designed to amplify a 166 bp region of the 18S rRNA gene. Comparative studies were performed to establish the optimal primer and probe concentrations.  Annealing temperature was determined by using thermal gradient. The results have shown good applicability of the optimized 18S rRNA gene-method to detect and quantify small amounts of the target in all samples analyzed. However, validation studies are needed in order to apply ddPCR technology for routine allergens analysis.  


2021 ◽  
Vol 42 (6) ◽  
pp. 3527-3534
Author(s):  
Mayara Marques Pereira Fernandes ◽  
◽  
Marcelo Renan Serrate Rodrigues ◽  
Jessica Damiana Marinho Valente ◽  
Marcelli Pascoal Nogueira ◽  
...  

This is the first report of Babesia vogeli molecular detection in dogs from the state of Acre, northern Brazil. This study aimed to perform the molecular detection of Babesia vogeli in dogs in the municipality of Rio Branco, Acre. Blood samples were collected from 47 dogs presenting with clinical signs comparable to hemoparasitosis. These were dogs which were attended in veterinary clinics from Rio Branco municipality, Acre. Physical examinations, packed cell volume (PCV) determination, platelet number estimation, hemoparasite investigation in the blood (collected from the pinna and peripheral blood), and polymerase chain reaction (PCR) for piroplasm based on the 18S rRNA gene, were performed. One dog (1/47, 2.1%; CI 95%: 0.1-11.3%) tested positive to Babesia vogeli in the polymerase chain reaction (PCR) assay for piroplasms and the resulting sequence showed 100% identity with Babesia vogeli isolates deposited in GenBank®. Co-infection with Ehrlichia spp. was also observed by direct examination (via blood smear). The clinical and hematological alterations observed in the positive animal were anorexia, dehydration, white mucous membranes, anemia and thrombocytopenia.


1999 ◽  
Vol 62 (6) ◽  
pp. 682-685 ◽  
Author(s):  
KAREN C. JINNEMAN ◽  
JUNE H. WETHERINGTON ◽  
WALTER E. HILL ◽  
CURTIS J. OMIESCINSKI ◽  
ANN M. ADAMS ◽  
...  

An oligonucleotide-ligation assay (OLA) was developed and compared to a restriction fragment length polymorphism (RFLP) test for distinguishing between 294-bp polymerase chain reaction (PCR) amplification products of the 18S rRNA gene from Cyclospora and Eimeria spp. The PCR/OLA correctly distinguished between three Cyclospora, three E. tenella, and one E. mitis strains and the ratio of positive to negative spectrophotometric absorbance (A490) values for each strain ranged from 4.086 to 15.280 (median 9.5). PCR/OLA provides a rapid, reliable, spectrophotometric alternative to PCR/RFLP.


2021 ◽  
pp. 2878-2882
Author(s):  
Sirikanda Thanasuwan ◽  
Anupong Tankrathok

Background and Aim: Fasciola spp. are important foodborne trematodes and waterborne zoonotic parasites that cause health problems and economic losses worldwide, including in Thailand. Fasciola spp. are usually detected by sedimentation or the formalin-ethyl acetate concentration technique (FECT) under microscopy, which is less specific and sensitive. Accurate detection is important to detect real incidence for protection against and elimination of fasciolosis in the area. This study aimed to determine the distribution of Fasciola spp. and compare the specificity and sensitivity of FECT under microscopy to that of polymerase chain reaction (PCR) in cattle feces. Materials and Methods: The study was conducted in Kalasin Province, Thailand. Feces of 46 cattle were investigated for infection with Fasciola spp. To detect infection, FECT under microscopy and PCR amplification of the 28S rRNA gene of Fasciola spp. were used to identify egg parasites. Results: Feces of 16 of 46 (34.78%) cattle were positive for Fasciola spp. using FECT under microscopy, whereas PCR showed that 67.39% (31 of 46) were positive for Fasciola spp. False-negative results were as high as 32.61% when diagnosed under microscopy. Conclusion: This study confirmed the infection of cattle with Fasciola spp. in Kalasin Province, indicating that PCR demonstrated higher sensitivity and specificity when diagnosing infection. FECT under microscopy can still be used as a primary and traditional method for diagnosis. However, relapse cases of Fasciola spp. and Paramphistomum spp. should be diagnosed by microscopy combined with PCR. This is the first report on the molecular distribution of fecal samples in cattle in Kalasin Province.


2019 ◽  
Vol 10 (04) ◽  
pp. 655-661
Author(s):  
Zainab H Abood AL-Asadi

Aspergillosis refers to fungi infections of the respiratory tract caused by Aspergillus species, especially Aspergillus fumigatus. Infection of A. fumigatus was increased in the last few years due to either resistances to antibiotics or the influence of other factors such as other fungal infections. The present study aimed to review the impact of Aspergillus fumigatus in Aspergillosis cases, and study the role of Singleplex PCR for amplification of ITS1, ITS4 of rRNA gene in the detection of fungal isolate. In this study, One hundred sputum samples were collected from patients admitted to the specialize chest and respiratory diseases center / Baghdad who were suffering from respiratory problems. During these studied, molds were isolation and identification based on Conventional method (Direct microscopy by using 10% KOH, and fungal culture was done on Sabouraud Dextrose agar supplemented with chloramphenicol and on Czapek-Dox agar incubated at 37°C and examined for 3-7 days then macroscopic, microscopic examination of the colony by(lactophenol cotton blue stain )and molecular methods by using Polymerase chain reaction (PCR)technique for identification. The 10% KOH examination was positive for 35 cases, while laboratory culturing was positive for 53 cases. Aspergillus sp were isolated from 44(83%) patients; A. fumigatus was isolated in 23 (42. 4%) patients while A. flavus, A. niger, and A. terreus were isolated from 11 (20. 08%), (13. 2%) and 3 (5. 7%) patients respectively, also isolated Penicillium spp. at percentage 1(1. 9%). In this study. The ages of participants ranged from 10-70years with a mean age of 34years, the males were more susceptible to fungal infection, were recorded 35/53 (66. 3), compared to females were 18/53 (33. 96). The infection of fungi was more prevalent in ages 30-40recorded 26(53. 06%) followed by ages 40-50, 13(26. 5), while the lowest infection recorded in the age group 10- 20 years was 2(2. 04%). DNA isolated from twenty-three A. fumigatus isolates was used as a template, and the specific of oligonucleotide primer sequences were used in conventional PCR to detect the presence of internal transcribed spacer region ( ITS) region of the rRNA gene for Aspergillus fumigates. The results of the PCR amplification of the rRNA gene showed that this gene was present in 19 samples out 23 positive samples which isolation with a PCR product size of approximated 385 bp, while 4 samples out 23 positive samples showed negative results for the presence of this gene as indicated by the absence of the PCR products in their relevant lanes. Statistical analysis revealed that the PCR to have a sensitivity of 95. 1% in the detection of Aspergillus fumigatus in Aspergillosis cases. Polymerase chain reaction (PCR) is a rapid, specific, and sensitive method to detect Aspergillus fumigatus in aspergillosis cases of humans.


2020 ◽  
Vol 8 (10) ◽  
pp. 1578 ◽  
Author(s):  
Massimo Iorizzo ◽  
Gianfranco Pannella ◽  
Silvia Jane Lombardi ◽  
Sonia Ganassi ◽  
Bruno Testa ◽  
...  

Lactic acid bacteria could positively affect the health of honey bees, including nutritional supplementation, immune system development and pathogen colonization resistance. Based on these considerations the present study evaluated predominant Lactic Acid Bacteria (LAB) species from beebread as well as from the social stomach and midgut of Apis mellifera ligustica honey bee foragers. In detail, for each compartment, the diversity in species and biotypes was ascertained through multiple culture-dependent approaches, consisting of Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE), 16S rRNA gene sequencing and Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). The study of a lactic acid bacteria community, performed with PCR-DGGE and sequence analysis targeting the V1–V3 region of the 16S rRNA gene (rDNA), highlighted the presence of a few species, including Apilactobacillus kunkeei, Lactiplantibacillus plantarum, Fructobacillus fructosus, Levilactobacillus brevis and Lactobacillus delbrueckii subsp. lactis. Depending on the different compartments, diverse levels of biodiversity in species were found. Particularly, a very low inter-species biodiversity was detected in the midgut that was prevalently dominated by the presence of Apilactobacillus kunkeei. On the other hand, the beebread was characterized by a reasonable biodiversity showing the presence of five species and the predominance of Apilactobacillus kunkeei, Lactiplantibacillus plantarum and Fructobacillus fructosus. The RAPD-PCR analysis performed on the three predominant species allowed the differentiation into several biotypes for each species. Moreover, a relationship between biotypes and compartments has been detected and each biotype was able to express a specific biochemical profile. The biotypes that populated the social stomach and midgut were able to metabolize sugars considered toxic for bees while those isolated from beebread could contribute to release useful compounds with functional properties. Based on this knowledge, new biotechnological approaches could be developed to improve the health of honey bees and the quality of bee products.


2002 ◽  
Vol 65 (5) ◽  
pp. 780-785 ◽  
Author(s):  
IRENE V. WESLEY ◽  
KAREN M. HARMON ◽  
JAMES S. DICKSON ◽  
ANN RAMOS SCHWARTZ

A multiplex polymerase chain reaction was developed to simultaneously identify Listeria monocytogenes and species of the genus Listeria. Two sets of primers were used, with the first amplifying a 938-bp region of the 16S rRNA gene that is highly conserved in all Listeria species and the second amplifying a 174-bp region of the listeriolysin (hlyA) gene of L. monocytogenes. Thus, isolates of Listeria spp. yield a single 938-bp product, whereas L. monocytogenes isolates yield both the 938-bp product and a 174-bp product. The specificity of the assay was verified with all six Listeria species and 11 serotypes of L. monocytogenes, as well as nonrelated bacteria. The multiplex PCR assay was used to determine the incidence of Listeria spp., especially L. monocytogenes, in mechanically separated turkey samples (n = 150 samples). L. monocytogenes strains were selected by using the University of Vermont two-step enrichment protocol and plating to selective Palcam agar. The multiplex PCR assay was used for verification of presumptive Listeria colonies. Approximately 38% of mechanically separated turkey samples (57 of 150) yielded L. monocytogenes; an additional 18% of these samples (27 of 150) harbored other Listeria spp. Fifty-one percent (29 of 57) of the L. monocytogenes isolates were of serogroup 1, 44% (25 of 57) were of serogroup 4, and 2% (1 of 57) were assigned to serogroups other than 1 and 4.


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