scholarly journals Skin swabbing protocol to collect DNA samples from small-bodied fish species

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 1064
Author(s):  
Ceinwen Tilley ◽  
Iain Barber ◽  
William Norton
Keyword(s):  
Dna Extraction ◽  
Fish Species ◽  
Standard Procedure ◽  
Experimental Models ◽  
Stress Axis ◽  
Small Laboratory ◽  
Current Standard ◽  
Caudal Fin ◽  
Animal Use ◽  
Detailed Protocol

Fish species are commonly used as experimental models in the laboratory. DNA is routinely collected from these animals to permit identification of their genotype. The current standard procedure to sample DNA is fin clipping, which involves anaesthetising individuals and removing a portion of the caudal fin. While fin clipping reliably generates good quality DNA samples for downstream applications, there is evidence that it can alter health and welfare, leading to infection and impacting on the fish’s behaviour. This in turn can result in greater variation in the data collected. In a recent study we adapted a skin swabbing protocol to collect DNA from small-bodied fish, including sticklebacks and zebrafish, without the use of anaesthetics or sharp instruments. A rayon-tipped swab was used to collect mucus from the flank of the fish, which was then used for DNA extraction. We subsequently demonstrated that compared to fin clipping, skin swabbing triggered fewer changes in stress axis activation and behaviour. We also found that data collected from fish that had been swabbed were less variable than data from fish that had been fin clipped, potentially allowing smaller sample sizes in experimental groups after using this technique, and thereby reducing animal use. Here we provide a detailed protocol explaining how to collect DNA samples from small laboratory fish using skin swabs.

scholarly journals Estimates of population parameters and consumption/biomass ratio for fishes in reservoirs, Paraná State, Brazil

2012 ◽  
Vol 10 (1) ◽  
pp. 177-188 ◽  
Author(s):  
Éder André Gubiani ◽  
Luiz Carlos Gomes ◽  
Angelo Antonio Agostinho
Keyword(s):  
Fish Species ◽  
Ecosystem Modeling ◽  
The State ◽  
Natural Mortality ◽  
Population Parameters ◽  
Southern Brazil ◽  
Growth Constant ◽  
Caudal Fin ◽  
Tropical Regions ◽  
Biomass Ratio

Consumption, natural mortality, and growth are important parameters for the construction for both fisheries and ecosystems models. We estimated k (growth constant of the von Bertalanffy's function), Loo 1(asymptotic length), Woo (asymptotic weight), Ar (caudal fin index), M (natural mortality), and Q/B (consumption/biomass ratio) for several fish species inhabiting reservoirs. We explored possible relationships among these parameters for 135 fish species sampled in thirty reservoirs in the State of Paraná, southern Brazil. Maximum length (Lmax) varied from 2.60 to 79.00 cm, Loo from 2.73 to 85.05 cm, Woo from 0.23 to 9,490.26 g, k from 0.01 to 2.38 yr-1, Ar from 0.03 to 3.60, M from 0.16 to 2.82 yr-1, and Q/B from 3.31 to 67.18. Significant correlations were observed between most pairs of parameters, except for Ar and k, Ar and Loo , and for M and Q/B. The estimates presented here may be useful for constructing Ecopath models, both in Brazil and other tropical regions, where the use of ecosystem modeling is growing.

scholarly journals Preferable Transplant Site for Hepatocyte Transplantation in a Rat Model

2021 ◽  
Vol 30 ◽  
pp. 096368972110400
Author(s):  
Hiroyuki Ogasawara ◽  
Akiko Inagaki ◽  
Ibrahim Fathi ◽  
Takehiro Imura ◽  
Hiroki Yamana ◽  
...  
Keyword(s):  
Serum Albumin ◽  
In Vivo Imaging ◽  
Standard Procedure ◽  
Graft Function ◽  
Rat Hepatocytes ◽  
Hepatocyte Transplantation ◽  
Current Standard ◽  
Imaging Evaluation ◽  
Transplant Site

Intraportal injection is regarded as the current standard procedure of hepatocyte transplantation (HTx). In islet transplantation, which shares many aspects with HTx, recent studies have clarified that instant blood-mediated inflammatory reaction (IBMIR), characterized by strong innate immune responses, can cause poor engraftment, so other transplant sites to avoid such a reaction have been established. Although IBMIR was reported to occur in HTx, few reports have evaluated alternative transplant sites for HTx. In this study, we sought to determine the optimum transplant site for HTx. Rat hepatocytes (1.0 × 107) were transplanted at the 9 transplant sites (intraportal (IPO), intrasplenic (IS), liver parenchyma, subcutaneous, intraperitoneal, renal subcapsular, muscle, inguinal subcutaneous white adipose tissue, and omentum) of analbuminemic rats. The serum albumin levels, immunohistochemical staining (albumin, TUNEL, and BrdU), and in vivo imaging of the grafts were evaluated. The serum albumin levels of the IPO group were significantly higher than those of the other groups ( p < .0001). The BrdU-positive hepatocyte ratio of liver in the IS group (0.9% ± 0.2%) was comparable to that of the IPO group (0.9% ± 0.3%) and tended to be higher than that of the spleen in the IS group (0.5% ± 0.1%, p = .16). Considering the in vivo imaging evaluation and the influence of splenectomy, the graft function in the IS group may be almost entirely achieved by hepatocytes that have migrated to the liver. The present study clearly showed that the intraportal injection procedure is more efficient than other procedures for performing HTx

scholarly journals Randomized controlled Phase III trial to evaluate omentum preserving gastrectomy for patients with advanced gastric cancer (JCOG1711, ROAD-GC)

2020 ◽  
Vol 50 (11) ◽  
pp. 1321-1324
Author(s):  
Yuya Sato ◽  
Takanobu Yamada ◽  
Takaki Yoshikawa ◽  
Ryunosuke Machida ◽  
Junki Mizusawa ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Advanced Gastric Cancer ◽  
Standard Procedure ◽  
Locally Advanced ◽  
Operation Time ◽  
Phase Iii ◽  
Current Standard ◽  
Phase Iii Trial ◽  
D2 Lymph Node Dissection ◽  
Randomized Controlled

Abstract Gastrectomy with omentectomy and D2 lymph node dissection is the current standard procedure for locally advanced gastric cancer. However, some retrospective studies have reported that omentectomy increased post-operative abdominal complications but provided no survival advantage over omentum preservation. Therefore, we plan a randomized controlled phase III trial to confirm the non-inferiority of omentum preservation compared with omentectomy in patients with cT3 (SS) or cT4a (SE) gastric cancer. A total of 1050 patients will be enrolled from 62 institutions over a period of 6.5 years. The primary end point is relapse-free survival, and the secondary end points are overall survival, blood loss, operation time and adverse events. This trial has been registered at the UMIN Clinical Trials Registry as UMIN000036253.

Comparison of DNA Extraction and PCR Setup Methods for Use in High-Throughput DNA Barcoding of Fish Species

2014 ◽  
Vol 7 (10) ◽  
pp. 1950-1959 ◽  
Author(s):  
Rosalee S. Hellberg ◽  
Michael D. Kawalek ◽  
Khanh T. Van ◽  
Yuelian Shen ◽  
Donna M. Williams-Hill
Keyword(s):  
Dna Barcoding ◽  
Dna Extraction ◽  
High Throughput ◽  
Fish Species

scholarly journals Cellular analysis in bronchoalveolar lavage: inherent limitations of current standard procedure

2017 ◽  
Vol 49 (6) ◽  
pp. 1601844 ◽  
Author(s):  
Benjamin-Alexander Bollmann ◽  
Benjamin Seeliger ◽  
Nora Drick ◽  
Tobias Welte ◽  
Jens T. Gottlieb ◽  
...  
Keyword(s):  
Bronchoalveolar Lavage ◽  
Standard Procedure ◽  
Current Standard ◽  
Cellular Analysis

scholarly journals Major Characteristics of Experimental Models of Abdominal Adhesions

2020 ◽  
Vol 13 (2) ◽  
pp. 157-162
Author(s):  
Boris Yushkov ◽  
Alexey Sarapultsev ◽  
German Sarapultsev
Keyword(s):  
Abdominal Cavity ◽  
Surgical Techniques ◽  
Experimental Models ◽  
The Body ◽  
Laboratory Animals ◽  
Small Laboratory ◽  
Medical Sciences ◽  
Abdominal Adhesions ◽  
Therapeutic Approaches ◽  
Modeling Techniques

The search for preventive and treatment methods for adhesions of the abdominal cavity and adhesive disease is one of the most important tasks of pharmaceutical and medical sciences; and the solution is based on experimental research studies involving animals. However, the varietyof adhesion modeling techniques, as well as specific features of experimental animals imply considerable difficulties to such research. The aim of the review was to describe and systematize experimental models of the adhesive process in the peritoneum applicable to small laboratory animals. The authors identify major models of adhesion induction, emphasizing the species differences of small laboratory animals that could affect the interpretation and extrapolation of the data obtained. The authors have proven that since adhesion is a complete product of the body inflammatory response to tissue damage, the treatment of adhesions should be solely based on surgical techniques, while therapeutic approaches might only prevent, slow down or reduce the intensity of adhesion processes.

scholarly journals Organ and tissue damage related to SARS-CoV-2: the biological model for experimental (preclinical) trials on golden hamsters Mesocricetus auratus

2021 ◽  
Vol 10 (4) ◽  
pp. 25-34
Author(s):  
S.V. Chepur ◽  
◽  
M.A. Tyunin ◽  
V.A. Myasnikov ◽  
I.I. Alekseeva ◽  
...  
Keyword(s):  
Infectious Disease ◽  
Structural Damage ◽  
Standard Procedure ◽  
Mesocricetus Auratus ◽  
Experimental Models ◽  
Biological Model ◽  
Internal Organs ◽  
Golden Hamsters ◽  
Changes Over Time ◽  
Over Time

Introduction. Amid the pandemic of the new coronavirus infection COVID-19, experimental models for screening both newly developed vaccines and drugs, and the already registered active pharmaceutical in-gredients tested for new indications are in high demand. The registration of changes in biometric and mor-phological parameters that are significant for the investigative pathology can be an optimal screening tool for antiviral drugs and vaccines to treat COVID-19. In this regard, the purpose of this study was to describe the changes over time of organ and tissue changes using a biological model of a SARS-CoV-2-associated infection in the golden hamsters Mesocricetus auratus. Materials and methods. We performed the study in golden hamsters Mesocricetus auratus weighing 80–100 g. The animals were infected through intranasal administration of a culture of SARS-CoV-2 virus containing 4 × 104 TCID50/mL. We recorded the animals’ weight before the infection and during the next 14 days and measured the specific gravity of the internal organs and the degree of their moisture saturation on days 3, 7, and 14 after the infection. Additionally, tissue samples were fixed in 10% neutral formalin; the preparation of histological specimens was performed according to the standard procedure. Statistical data processing was carried out using non-parametric tests. Results. In the model of SARS-CoV-2 infection in Mesocricetus auratus, we observed the manifestation of the infectious disease after 3 days. The most pronounced pathological changes in the overall health status of the animals and in the histology of internal organs were seen 7 days after the infection. We determined the weight loss and significant deviations in gravimetric coefficients of lungs, heart, spleen, and kidneys to be the indicators revealing the infectious disease course changes over time. Histologic evaluation showed typical changes in the SARS-CoV-2-associated visceral damage: the formation of polymorphonuclear cell infiltrates in the lung interalveolar septa and patchy dystrophic changes in the neurocytes of the brain screen-type centers with the demyelination of the commissural nerve guides. The detected pathological manifestations corresponded in time to an increase in the virus replicative activity in the lungs. Conclusion. The experimental model of the SARS-CoV-2-associated infection in golden hamsters Mesocricetus auratus can be recommended for screening experimental (preclinical) studies of the promising drugs’ efficacy for pathogenetic and etiotropic therapy of COVID-19. Keywords: SARS-CoV-2, golden hamsters, morphometry, organ and tissue structural damage

Schistura syngkai, a new fish species (Teleostei: Nemacheilidae) from Meghalaya, northeast India

Zootaxa ◽  
2019 ◽  
Vol 4701 (2) ◽  
pp. 185-191
Author(s):  
HRISHIKESH CHOUDHURY ◽  
D. KHLUR B. MUKHIM ◽  
ABHINIT DEY ◽  
DEISAKEE P. WARBAH ◽  
DANDADHAR SARMA
Keyword(s):  
Sexual Dimorphism ◽  
Fish Species ◽  
Lateral Line ◽  
Northeast India ◽  
Stone Loach ◽  
Caudal Fin ◽  
Eye Diameter ◽  
New Fish Species

Schistura syngkai, a new stone loach, is described from the Twahdidoh Stream of Wahblei River (Surma-Meghna drainage) in Meghalaya, northeast India. It is unique among its Indian congeners in having a prominent dark-brown to blackish mid-lateral stripe about an eye diameter or more in width, overlain on 12–18 vertically-elongate black blotches on a golden-brown to amber body. Additionally, the species possesses an incomplete lateral line and a slightly emarginate caudal fin, and exhibits no apparent sexual dimorphism. 

scholarly journals Laparoscopic therapy of the coeliac artery compression syndrome: a critical analysis of the current standard procedure

2020 ◽  
Vol 102 (2) ◽  
pp. 104-109 ◽  
Author(s):  
M Sahm ◽  
R Otto ◽  
M Pross ◽  
T Scholbach ◽  
R Mantke
Keyword(s):  
Flow Velocity ◽  
Doppler Ultrasonography ◽  
Clinical Symptoms ◽  
Standard Procedure ◽  
Symptomatic Relief ◽  
Current Standard ◽  
Median Arcuate Ligament ◽  
Coeliac Artery ◽  
Arcuate Ligament

Introduction Median arcuate ligament syndrome has been known anatomically for approximately 100 years and results from a compression of the coeliac axis by fibrous attachment of the diaphragmatic crura. Owing to the rarity of the disease and limited available data, many aspects of treatment are controversial. Currently, laparoscopic decompression is considered by several authors as standard surgical procedure. We present an analysis of the clinical routine of MALS therapy. Methods We conducted a prospective observational trial in patients with MALS between March 2016 and August 2018, in which clinical symptoms, diagnostic evaluation, procedures with complication analysis and follow-up data were recorded. Results A total of 18 patients (12 female, 6 male) with MALS, aged between 15 and 65 years, were included in this study. All patients presented with long-standing abdominal pain. Preoperative Doppler ultrasonography showed a flow velocity of the coeliac artery averaging 289.9cm/second in mid-position of the diaphragm, 285.9cm/second in expiration and 199.0cm/second in inspiration. All operated patients underwent laparoscopic decompression; two patients received an angiographic intervention. Postoperatively, a significant decrease of the flow velocity in mid-position of the diaphragm was detected (P = 0.018). At follow-up after 5.2 months, 50.0% of the patients were pain-free, 37.5% reported symptomatic relief and 12.5% showed evidence for a recurrence. Conclusion MALS is challenging both diagnostically and therapeutically. Laparoscopy with release of the median arcuate ligament is an essential part of the therapy and can be confirmed by Doppler ultrasonography. Disease outcome is also influenced by several predictive factors.

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