scholarly journals Coinfection of Chlamydia spp. and herpesvirus in juvenile farmed Siamese crocodiles (Crocodylus siamensis) in Thailand

2021 ◽  
pp. 1908-1914
Author(s):  
Weena Paungpin ◽  
Metawee Thongdee ◽  
Somjit Chaiwattanarungruengpaisan ◽  
Ladawan Sariya ◽  
Wanna Sirimanapong ◽  
...  
Keyword(s):  
Histopathological Examination ◽  
Tissue Sample ◽  
Phylogenetic Analyses ◽  
Aquatic Animal ◽  
Tissue Samples ◽  
Health Care Unit ◽  
Causative Agents ◽  
Herpesvirus 1 ◽  
Polymerase Chain ◽  
Crocodylus Siamensis

Background and Aim: For a decade, chlamydial and herpesvirus infections have caused significant morbidity and mortality in farmed crocodiles. In September 2017, a total of 160 juvenile freshwater Siamese crocodiles (Crocodylus siamensis) with conjunctivitis/pharyngitis lesions were admitted at the Veterinary Aquatic Animal Research Health Care Unit, Faculty of Veterinary Science, Mahidol University. All crocodiles did not respond well to antibiotics or supportive treatments and died. This study aimed to detect and identify the causative agents associated with conjunctivitis/pharyngitis and fatal outcomes in juvenile farmed Siamese crocodiles. Materials and Methods: A total of 138 pharyngeal and conjunctival swabs and conjunctival scrapes were collected from live crocodiles. All swab and scrape samples were DNA-extracted and amplified by polymerase chain reaction (PCR) using Chlamydiaceae- and herpesvirus-specific primers. Tissue samples (brain, lung, liver, heart, spleen, and intestine) were collected from two representative postmortem animals. All tissue samples were processed for molecular and pathological analyses. Results: PCR examinations identified chlamydial and herpesvirus DNA in 92% (126/138) and 100% (138/138), respectively, of the tested swab and scrape samples. Of those positive samples, 79% (26/33), 67% (4/6), and 98% (97/99) of the pharyngeal swabs, conjunctival swabs, and conjunctival scrapes, respectively, were positive for both chlamydial and herpesvirus DNA. Histopathological examination indicated necrosis and mononuclear cell infiltration in the liver, kidney, and intestine of the affected animals. The intracytoplasmic accumulation of Chlamydia was randomly observed in the examined tissue sample. Moreover, the presence of chlamydial and herpesvirus DNA was also detected in the tissue samples, including the heart, intestine, brain, lung, liver, and spleen, of the affected animals by PCR. Phylogenetic analyses revealed that Chlamydia spp. detected in the juvenile Siamese crocodiles was notably different from other known species in the Chlamydia genus, while the herpesvirus detected in the crocodiles was closely related to crocodyline herpesvirus 1. Conclusion: Based on histopathological and molecular examinations, this report provided the first evidence of coinfection of Chlamydia spp. and crocodyline herpesvirus 1 in juvenile Siamese crocodiles in Thailand.

A Microdissection and Molecular Genotyping Assay to Confirm the Identity of Tissue Floaters in Paraffin-Embedded Tissue Blocks

2003 ◽  
Vol 127 (2) ◽  
pp. 213-217 ◽  
Author(s):  
Jennifer L. Hunt ◽  
Patricia Swalsky ◽  
E. Sasatomi ◽  
Laura Niehouse ◽  
Anke Bakker ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tissue Sample ◽  
Small Samples ◽  
Allelic Loss ◽  
Chain Reaction ◽  
Neoplastic Tissue ◽  
Tissue Samples ◽  
Genotyping Assay ◽  
Tissue Identification ◽  
Polymerase Chain

Abstract Context.—A recurring problem in surgical pathology practice is specimen mix-up and floater contamination. While many cases can be resolved histologically, a significant number remain unclear and may have serious clinical and medicolegal implications. Objectives.—To design a microdissection and genotyping assay to identify contaminating floater tissues in paraffin-embedded tissues that is optimized for small samples, and to use the assay to resolve a series of clinical cases with floater tissues. Materials and Methods.—Twenty-one cases of possible tissue floater contamination in paraffin-embedded tissue blocks were included. Using 4 unstained, 4-μm-thick histologic sections, multiple sites were microdissected under direct visualization either by hand or by laser capture microdissection. Nonneoplastic and neoplastic tissues were sampled. Polymerase chain reaction was performed for a panel of 10 polymorphic microsatellite markers at 1p34, 3p26, 5q21, 9p21, 10q23, and 17p13. Allele size and content were analyzed semiquantitatively by fluorescent capillary electrophoresis, and the genotypes for the tissues in the paraffin-embedded tissue blocks were compared for identity. Results.—Tissue identification was successful in all cases, despite small tissue sample size and fixation effects. Comparative analysis of neoplastic tissue floaters and the presumptive source tumor was performed when possible to control for possible allelic loss or microsatellite instability. Conclusions.—Microdissection and genotyping are effective and reliable means to objectively resolve problems of possible floater contamination. Even minute tissue samples provide sufficient DNA template for polymerase chain reaction microsatellite analysis. Because of the potential clinical implications of floaters, we recommend that all suspected floaters that would change a diagnosis from benign to malignant be subjected to genotyping assay to confirm the identity of the floater tissue.

scholarly journals Molecular characterization and phylogenetic analysis of fowl adenovirus serotype-4 from Guangdong Province, China

2020 ◽  
Vol 13 (5) ◽  
pp. 981-986
Author(s):  
Fu Yuming ◽  
Yuan Sheng ◽  
Deng Wenyu ◽  
Chi Shihong ◽  
Li Wenfeng ◽  
...  
Keyword(s):  
Phylogenetic Analyses ◽  
Hypervariable Region ◽  
Guangdong Province ◽  
Hexon Gene ◽  
Adenovirus Serotype ◽  
Tissue Samples ◽  
Hydropericardium Syndrome ◽  
Fowl Adenovirus ◽  
Polymerase Chain ◽  
Fowl Adenovirus Serotype 4

Aim: Our aim in this study was to isolate potentially novel strains of fowl adenovirus serotype-4 (FAdV-4) that is currently circulating in broiler chicken flocks in Guangdong Province, China, and to compare nucleotide and amino acid (AA) sequences of their respective hexon genes. Materials and Methods: The experiment was carried out on poultry farms experiencing outbreaks of FAdV-4-associated hydropericardium syndrome (HPS). Tissue samples from the hearts and livers of deceased chickens were screened for FAdV-4 infection using hexon gene-specific polymerase chain reaction (PCR). Results: New virus isolates were used to infect 7-day-old chicks, which went onto reproduce typical HPS signs. The hypervariable region of the FAdV-4 hexon gene was PCR-amplified and sequenced. The hexon nucleotide and deduced AA sequence identities were 99.8-99.9% and 99.5-99.8%, respectively, among the four novel isolates. In addition, the new isolates were 97-100% and 96.4-99.9% identical to the nucleotide and deduced AA sequences, respectively, of FAdV-4 hexon genes available in the National Center for Biotechnology Information GenBank database. Phylogenetic analyses, based on the hexon gene sequence, revealed that the new isolates, clustered with FAdV-C; the FAdV-A, FAdV-B, FAdV-D, and FAdV-E viruses, were more distantly related. Conclusion: New FAdV-4 isolates from Guangdong Province are similar to those identified in other regions of the world. This information provides critical insight into HPS epidemiology and provides a perspective for monitoring outbreaks and developing strategies for disease prevention.

Biomedical applications: Pitfalls in the practice

1994 ◽  
Vol 52 ◽  
pp. 378-379
Author(s):  
J. D. Shelburne ◽  
Peter Ingram ◽  
Victor L. Roggli ◽  
Ann LeFurgey
Keyword(s):  
Operating Room ◽  
Liquid Nitrogen ◽  
Lung Tissue ◽  
Biomedical Applications ◽  
Microprobe Analysis ◽  
Tissue Sample ◽  
Cellular Level ◽  
Tissue Samples ◽  
Asbestos Fibers

At present most medical microprobe analysis is conducted on insoluble particulates such as asbestos fibers in lung tissue. Cryotechniques are not necessary for this type of specimen. Insoluble particulates can be processed conventionally. Nevertheless, it is important to emphasize that conventional processing is unacceptable for specimens in which electrolyte distributions in tissues are sought. It is necessary to flash-freeze in order to preserve the integrity of electrolyte distributions at the subcellular and cellular level. Ideally, biopsies should be flash-frozen in the operating room rather than being frozen several minutes later in a histology laboratory. Electrolytes will move during such a long delay. While flammable cryogens such as propane obviously cannot be used in an operating room, liquid nitrogen-cooled slam-freezing devices or guns may be permitted, and are the best way to achieve an artifact-free, accurate tissue sample which truly reflects the in vivo state. Unfortunately, the importance of cryofixation is often not understood. Investigators bring tissue samples fixed in glutaraldehyde to a microprobe laboratory with a request for microprobe analysis for electrolytes.

STUDIES ON SOME BACTERIAL DISEASES IN OREOCHROMIS NILOTICUS WITH SPECIAL REFERENCES TO TREATMENT

2018 ◽  
pp. 39-47
Author(s):  
Viola Zaki ◽  
Ahmed EL-gamal ◽  
Yasmin Reyad
Keyword(s):  
Oreochromis Niloticus ◽  
Bacterial Infections ◽  
Antimicrobial Agents ◽  
Histopathological Examination ◽  
Klebsiella Oxytoca ◽  
Bacterial Isolates ◽  
Bacterial Strains ◽  
Tissue Samples ◽  
Hydropic Degeneration ◽  
Total Prevalence

he present research carried out to study the common bacterial infections in Oreochromis niloticus (Nile tilapia) in Manzala area at Dakahlia governorate and possible antimicrobial agents used for treatment. A total number of 400 fish were randomly collected from Manzala private farms at Dakahlia governorate and subjected to the clinical, bacteriological and histopathological examination. The highest prevalence of bacterial isolates during the whole period of examination of naturally infected O.niloticus was recorded for A.hydrophila (22.66%), followed by V.alginolyticus (19.01%), V.parahemolyticus (13.80%), Streptococcus spp. (12.24%), A.caviae (11.72%), V.cholera (10.16%), A.salmonicida (7.55%), while the lowest prevalence was recorded for Klebsiella oxytoca (2.86%). The seasonal highest total prevalence of bacterial isolates from examined naturally infected O. niloticus was recorded in spring (30.21%), followed by autumn (28.39%), then summer (22.40%) and the lowest prevalence was recorded in winter (19.01%). Histopathological findings of the tissue samples which collected from different organs of naturally infected O.niloticus revealed that spleen show marked hemosiderosis and sever hemorrhage, gills showsever congestion of lamellar capillaries with marked aneurysm, necrosis and hemorrhage of lamellar epithelium and liver show sever hydropic degeneration and necrosis of hepatocytes, Ciprofloxacin was the most effective antibiotic against all isolated bacterial strains

scholarly journals Molecular Identification of Trypanosoma evansi Isolated from Arabian Camels (Camelus dromedarius) in Riyadh and Al-Qassim, Saudi Arabia

Animals ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 1149
Author(s):  
Dina M. Metwally ◽  
Isra M. Al-Turaiki ◽  
Najwa Altwaijry ◽  
Samia Q. Alghamdi ◽  
Abdullah D. Alanazi
Keyword(s):  
Saudi Arabia ◽  
Infection Rate ◽  
Ribosomal Rna ◽  
Phylogenetic Analyses ◽  
Trypanosoma Evansi ◽  
Camelus Dromedarius ◽  
18S Ribosomal Rna ◽  
18S Ribosomal Rna Gene ◽  
Polymerase Chain ◽  
Pcr Targeting

We analyzed the blood from 400 one-humped camels, Camelus dromedarius (C. dromedarius), in Riyadh and Al-Qassim, Saudi Arabia to determine if they were infected with the parasite Trypanosoma spp. Polymerase chain reaction (PCR) targeting the internal transcribed spacer 1 (ITS1) gene was used to detect the prevalence of Trypanosoma spp. in the camels. Trypanosoma evansi (T. evansi) was detected in 79 of 200 camels in Riyadh, an infection rate of 39.5%, and in 92 of 200 camels in Al-Qassim, an infection rate of 46%. Sequence and phylogenetic analyses revealed that the isolated T. evansi was closely related to the T. evansi that was detected in C. dromedarius in Egypt and the T. evansi strain B15.1 18S ribosomal RNA gene identified from buffalo in Thailand. A BLAST search revealed that the sequences are also similar to those of T. evansi from beef cattle in Thailand and to T. brucei B8/18 18S ribosomal RNA from pigs in Nigeria.

scholarly journals The Dutch National TissueArchive Portal enables efficient, consistent, and transparent procurement of diagnostic tissue samples for scientific use

2021 ◽  
Author(s):  
Robin Verjans ◽  
Annette H. Bruggink ◽  
Robby Kibbelaar ◽  
Jos Bart ◽  
Aletta Debernardi ◽  
...  
Keyword(s):  
The Netherlands ◽  
Tissue Sample ◽  
Ffpe Tissue ◽  
Tissue Samples ◽  
Internet Portal ◽  
Formalin Fixed Paraffin ◽  
Practical Support ◽  
Formalin Fixed Paraffin Embedded ◽  
The Rich ◽  
Formalin Fixed

AbstractBiobanks play a crucial role in enabling biomedical research by facilitating scientific use of valuable human biomaterials. The PALGA foundation—a nationwide network and registry of histo- and cytopathology in the Netherlands—was established to promote the provision of data within and between pathology departments, and to make the resulting knowledge available for healthcare. Apart from the pathology data, we aimed to utilize PALGA’s nationwide network to find and access the rich wealth of Formalin-Fixed Paraffin-Embedded (FFPE) tissue samples for scientific use.  We implemented the Dutch National TissueArchive Portal (DNTP) to utilize PALGA’s nationwide network for requesting FFPE tissue samples. The DNTP consists of (1) a centrally organized internet portal to improve the assessing, processing, harmonization, and monitoring of the procurement process, while (2) dedicated HUB-employees provide practical support at peripheral pathology departments. Since incorporation of the DNTP, both the number of filed requests for FFPE tissue samples and the amount of HUB-mediated support increased 55 and 29% respectively. In line, the sample procurement duration time decreased significantly (− 47%). These findings indicate that implementation of the DNTP improved the frequency, efficiency, and transparency of FFPE tissue sample procurement for research in the Netherlands. To conclude, the need for biological resources is growing persistently to enable precision medicine. Here, we access PALGA’s national, pathology network by implementation of the DNTP to allow for efficient, consistent, and transparent exchange of FFPE tissue samples for research across the Netherlands.

scholarly journals Occurrence of abortions induced by Neospora caninum in dairy cattle from Santa Catarina, southern Brazil

2017 ◽  
Vol 26 (3) ◽  
pp. 292-298 ◽  
Author(s):  
Cesar Augusto Barbosa de Macedo ◽  
Madlaine Frigo Silveira Barbosa de Macedo ◽  
Ana Carolina Miura ◽  
Alessandra Taroda ◽  
Sergio Tosi Cardim ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Dairy Cows ◽  
High Frequency ◽  
Neospora Caninum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Southern Brazil ◽  
Chain Reaction ◽  
Santa Catarina ◽  
Tissue Samples ◽  
Polymerase Chain

Abstract The aim of the present study was to investigate the occurrence of N. caninum associated with abortions of dairy cattle from Santa Catarina state, southern Brazil by using enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and polymerase chain reaction (PCR). Blood from dairy cows that aborted along with intrathoracic fluid and tissue samples (brain, heart, liver, and lung) from their fetuses were collected and used for serology; PCR, histopathological, and immunohistochemistry (IHC) evaluations were also conducted. Twenty-one cows (51.2%) out of 41, and eight fetuses (26.7%) out of 30 were ELISA (HerdCheck, IDEXX) positive for N. caninum. Dams > 36 months of age had a higher risk of being serum positive than younger animals. PCR and IHC revealed that 38.8% (14/36) and 25.0% (9/36) of the fetuses were positive for N. caninum, respectively for each of the tests. Seropositive cows had a higher frequency of fetuses that were also positive by either intrathoracic fluid, PCR, or IHC. In summary, the present study observed a high frequency of N. caninum in abortions from dairy cows from southern Brazil, with a higher N. caninum prevalence found in cows that were older than 36 months. In addition, serology, PCR, and IHC should be used all together for better diagnosis of neosporosis in cattle.

scholarly journals Development of Quantitative Real-Time Polymerase Chain Reaction for Detection of and Discrimination between Erysipelothrix Rhusiopathiae and Other Erysipelothrix Species

2009 ◽  
Vol 21 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Ho To ◽  
Tomohiro Koyama ◽  
Shinya Nagai ◽  
Kotaro Tuchiya ◽  
Tetsuo Nunoya
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Limit Of Detection ◽  
Enrichment Culture ◽  
Bacterial Species ◽  
Erysipelothrix Rhusiopathiae ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Practical Applications ◽  
Polymerase Chain

Quantitative real-time polymerase chain reaction (qPCR) assays were developed and validated in combination with enrichment culture for the detection and discrimination of Erysipelothrix rhusiopathiae and other Erysipelothrix species from tissue samples. The targets for SYBR green qPCR assays were the 16S ribosomal RNA gene for Erysipelothrix species and a gene involved in capsular formation for E. rhusiopathiae. The specificity of the assays was assessed with Erysipelothrix species and other related bacterial species. The limit of detection was found to be 5 colony-forming units per reaction. Amplification of DNA extracted from spleen and joint samples spiked with increasing quantities of Erysipelothrix cells was shown to be equally sensitive to DNA extracted from a pure bacterial culture. The assays were evaluated with 88 tissue samples from 3 experimentally infected pigs and 50 mice and with 36 tissue samples from 3 naturally infected pigs and 11 noninfected pigs. Results were compared with those of direct qPCR and conventional culture. The qPCR after enrichment increased the diagnostic sensitivity over that of culture and qPCR, thereby significantly reducing the total time taken for the detection of E. rhusiopathiae and other Erysipelothrix species. Therefore, this technique could be used for practical applications.

scholarly journals Assessing viability of a minimally invasive autopsy technique in ascertaining the probable cause of death in patients who were SARS CoV19 positive at the time of their demise

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Ajay H. Bhandarwar ◽  
Girish D. Bakhshi ◽  
Eham Arora ◽  
Nikhil Dhimole ◽  
Sanjay R. Bijwe ◽  
...  
Keyword(s):  
Minimally Invasive ◽  
Cause Of Death ◽  
Histopathological Examination ◽  
Health Hazard ◽  
World Health ◽  
Invasive Technique ◽  
Tissue Samples ◽  
Conventional Autopsy ◽  
Minimally Invasive Autopsy ◽  
Health Organization

Abstract Background SARS CoV-19 was declared as a pandemic by the World Health Organization (WHO), raising up challenges on various levels ranging from therapeutics to diagnostics. The conventional autopsy technique may pose a health hazard to health care workers. A minimally invasive autopsy technique can diminish this hazard. Materials and methods Between August and November 2020, 51 patients who were suffering from Covid-19 at the time of their demise were included. A novel minimally invasive ultrasound-guided technique for procuring tissue samples of major organs was employed which were thereafter subject to histopathological examination. A detailed review of the course in hospital was noted. An analysis was performed to correlate the cause of death ascertained from our minimally invasive technique with the cause of death ascertained clinically. Results There was adequate tissue sampling in 45 cases, where the minimally invasive autopsy technique confirmed the cause of death in all 45 cases (100%) and made it more specific in 5 cases (11.11%). Conclusion Minimally Invasive Autopsy is an easily reproducible technique which has the potential to strengthen the probable the cause of death with reasonable certainty while ensuring safety and ethics.

Seasonal pattern and genotype distribution of sapovirus infection in Japan, 2003–2009

2011 ◽  
Vol 140 (1) ◽  
pp. 74-77 ◽  
Author(s):  
S. K. DEY ◽  
O. PHATHAMMAVONG ◽  
T. D. NGUYEN ◽  
A. THONGPRACHUM ◽  
W. CHAN-IT ◽  
...  
Keyword(s):  
Seasonal Pattern ◽  
Age Groups ◽  
Cold Season ◽  
Viral Gastroenteritis ◽  
Genotype Distribution ◽  
Chain Reaction ◽  
The Family ◽  
Causative Agents ◽  
Polymerase Chain ◽  
Predominant Strain

SUMMARYSapovirus, a member of the family Caliciviridae, is one of the major causative agents of viral gastroenteritis affecting all age groups. A total of 3232 faecal specimens collected from infants and children with gastroenteritis in five different regions of Japan during 2003–2009 were examined for sapovirus by reverse transcription–polymerase chain reaction. Sapoviruses were detected in 131 (4·05%) patients with the peak observed mainly in the cold season (November–March) in Japan during 2003–2009. During the last 6 years, sapovirus GI/1 was the predominant strain in Japan followed by GIV, GII/3, GII/6, GII/2, GII/12 and GI, respectively.

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