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Article: CRISPR Libraries and Whole-Genome Screening to Identify Essential Factors for Viral Infections.

Paiva, Isadora Marques / Damasceno, Samara / Cunha, Thiago Mattar

Advances in experimental medicine and biology

2023 Volume 1429, Page(s) 157–172

Abstract: The CRISPR-Cas9 system has revolutionized genetics and offers a simple and inexpensive way of generating perturbation that results in gene repression, activation, or editing. The advances in this technique make possible the development of CRISPR ... ...

Abstract	The CRISPR-Cas9 system has revolutionized genetics and offers a simple and inexpensive way of generating perturbation that results in gene repression, activation, or editing. The advances in this technique make possible the development of CRISPR libraries which consist of a set of sgRNAs to cause perturbations in several genes in the same cell population. The use of libraries raised the CRISPR-Cas9 technique to a genomic scale and provides a powerful approach for identifying previously unknown molecular mechanisms and pathways involved in a specific phenotype or biological process. More specifically, the CRISPRko libraries (set of sgRNAs for gene knockout) and their high-throughput screenings are widely used in research with viral agents, and it was enlarged even more with the COVID-19 pandemic. With this chapter, we aim to point out how this tool helps in understanding virus-host relationships, such as the mechanisms of virus entry into the cell, the essential factors for its replication, and the cellular pathways involved in the response against the pathogen. The chapter also provided some practical considerations for each step of an experimentation using these tools that include choosing the library and screening type, the target cell, the viral strain, the library amplification and guaranteeing its coverage, the strategies for the gene screening pipeline by bioinformatics, and finally, target validation. To conclude, it was presented a table reviewing the last updates in the research for antiviral therapies using CRISPR libraries.
MeSH term(s)	Humans ; CRISPR-Cas Systems/genetics ; RNA, Guide, CRISPR-Cas Systems ; Pandemics ; COVID-19/genetics ; Virus Diseases/diagnosis ; Virus Diseases/genetics ; Gene Editing
Chemical Substances	RNA, Guide, CRISPR-Cas Systems
Language	English
Publishing date	2023-07-24
Publishing country	United States
Document type	Journal Article
ZDB-ID	410187-X
ISSN	0065-2598
ISSN	0065-2598
DOI	10.1007/978-3-031-33325-5_9
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Article ; Online: Acute ethanol exposure leads to long-term effects on memory, behavior, and transcriptional regulation in the zebrafish brain.

Sartori, Barbara Miranda / Moreira Júnior, Renato Elias / Paiva, Isadora Marques / Moraes, Izabela Barbosa / Murgas, Luis David Solis / Brunialti-Godard, Ana Lúcia

Behavioural brain research

2023 Volume 444, Page(s) 114352

Abstract: Alcohol consumption is associated with alterations in memory and learning processes in humans and animals. In this context, research models such as the zebrafish (Danio rerio) arise as key organisms in behavioral and molecular studies that attempt to ... ...

Abstract	Alcohol consumption is associated with alterations in memory and learning processes in humans and animals. In this context, research models such as the zebrafish (Danio rerio) arise as key organisms in behavioral and molecular studies that attempt to clarify alterations in the Central Nervous System (CNS), like those related to alcohol use. Accordingly, we used the zebrafish as a model to evaluate the effects of ethanol on the learning and memory process, as well as its relationship with behavior and transcriptional regulation of lrfn2, lrrk2, grin1a, and bdnf genes in the brain. To this end, for the memory and learning evaluation, we conducted the Novel Object Recognition test (NOR); for behavior, the Novel Tank test; and for gene transcription, qPCR, after 2 h, 24 h, and 8 days of ethanol exposure. As a result, we noticed in the NOR that after 8 days of ethanol exposure, the control group spent more time exploring the novel object than when compared to 2 h post-exposure, indicating that naturally zebrafish remember familiar objects. In animals in the Treatment group, however, no object recognition behavior was observed, suggesting that alcohol affected the learning and memory processes of the animals and stimulated an anxiolytic effect in them. Regarding transcriptional regulation, 24 h after alcohol exposure, we found hyper-regulation of bdnf and, after 8 days, a hypo-regulation of lrfn2 and lrrk2. To conclude, we demonstrated that ethanol exposure may have influenced learning ability and memory formation in zebrafish, as well as behavior and regulation of gene transcription. These data are relevant for further understanding the application of zebrafish in research associated with ethanol consumption and behavior.
MeSH term(s)	Animals ; Humans ; Ethanol/pharmacology ; Zebrafish/physiology ; Brain-Derived Neurotrophic Factor ; Learning ; Brain ; Behavior, Animal ; Membrane Glycoproteins ; Nerve Tissue Proteins/pharmacology ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2/pharmacology ; Zebrafish Proteins
Chemical Substances	Ethanol (3K9958V90M) ; Brain-Derived Neurotrophic Factor ; LRFN2 protein, human ; Membrane Glycoproteins ; Nerve Tissue Proteins ; LRRK2 protein, zebrafish (EC 2.7.11.1) ; Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 (EC 2.7.11.1) ; Zebrafish Proteins
Language	English
Publishing date	2023-02-24
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	449927-x
ISSN	1872-7549 ; 0166-4328
ISSN (online)	1872-7549
ISSN	0166-4328
DOI	10.1016/j.bbr.2023.114352
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Article ; Online: Macrophage-derived human resistin promotes perivascular adipose tissue dysfunction in experimental inflammatory arthritis.

Fedoce, Aline G / Veras, Flávio P / Rosa, Marcos H / Schneider, Ayda H / Paiva, Isadora M / Machado, Mirele R / Freitas-Filho, Edismauro G / Silva, Josiane F / Machado, Caio C / Alves-Filho, José C / Cunha, Fernando Q / N Z Ramalho, Leandra / Louzada-Junior, Paulo / Bonavia, Anthony S / Tostes, Rita C

Biochemical pharmacology

2024 Volume 224, Page(s) 116245

Abstract: Cardiovascular disease (CVD) is the leading cause of death in rheumatoid arthritis (RA). Resistin is an adipokine that induces adipose tissue inflammation and activation of monocytes/macrophages via adenylate cyclase-associated protein-1 (CAP1). Resistin ...

Abstract	Cardiovascular disease (CVD) is the leading cause of death in rheumatoid arthritis (RA). Resistin is an adipokine that induces adipose tissue inflammation and activation of monocytes/macrophages via adenylate cyclase-associated protein-1 (CAP1). Resistin levels are increased in RA and might cause perivascular adipose tissue (PVAT) dysfunction, leading to vascular damage and CVD. This study aimed to investigate the role of resistin in promoting PVAT dysfunction by increasing local macrophage and inflammatory cytokines content in antigen-induced arthritis (AIA). Resistin pharmacological effects were assessed by using C57Bl/6J wild-type (WT) mice, humanized resistin mice expressing human resistin in monocytes-macrophages (hRTN
Language	English
Publishing date	2024-04-28
Publishing country	England
Document type	Journal Article
ZDB-ID	208787-x
ISSN	1873-2968 ; 0006-2952
ISSN (online)	1873-2968
ISSN	0006-2952
DOI	10.1016/j.bcp.2024.116245
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Article ; Online: Ethanol Preference Leads to Alterations in Telomere Length, Mitochondria Copy Number, and Antioxidant Enzyme Activity in Zebrafish Brains.

Moraes, Izabela Barbosa / Paiva, Isadora Marques / Moreira-Júnior, Renato Elias / Sartori, Bárbara Miranda / Franco, Rodrigo Rodrigues / Espindola, Foued Salmen / Murgas, Luis David Solis / Brunialti-Godard, Ana Lúcia

Frontiers in bioscience (Landmark edition)

2023 Volume 28, Issue 4, Page(s) 73

Abstract: Background: The motivations for and effects of ethanol consumption vary considerably among individuals, and as such, a significant proportion of the population is prone to substance abuse and its negative consequences in the physical, social, and ... ...

Abstract	Background: The motivations for and effects of ethanol consumption vary considerably among individuals, and as such, a significant proportion of the population is prone to substance abuse and its negative consequences in the physical, social, and psychological spheres. In a biological context, the characterization of these phenotypes provides clues for understanding the neurological complexity associated with ethanol abuse behavior. Therefore, the objective of this research was to characterize four ethanol preference phenotypes described in zebrafish: Light, Heavy, Inflexible, and Negative Reinforcement. Methods: To do this, we evaluated the telomere length, mtDNA copy number using real-time quantitative PCR (qPCR), and the activity of these antioxidant enzymes: catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the brain, and the interactions between these biomarkers. Changes observed in these parameters were associated with ethanol consumption and alcohol abuse. Results: The Heavy, Inflexible, and Negative Reinforcement phenotypes showed ethanol preference. This was particularly the case with the Inflexible phenotype, which was the group with the greatest ethanol preference. These three phenotypes showed telomere shortening as well as high SOD/CAT and/or GPx activities, while the Heavy phenotype also showed an increase in the mtDNA copy number. However, the Light phenotype, containing individuals without ethanol preference, did not demonstrate any changes in the analyzed parameters even after being exposed to the drug. Additionally, the PCA analysis showed a tendency to cluster the Light and Control groups differently from the other ethanol preference phenotypes. There was also a negative correlation between the results of the relative telomere length and SOD and CAT activity, providing further evidence of the biological relationship between these parameters. Conclusions: Our results showed differential molecular and biochemistry patterns in individuals with ethanol preference, suggesting that the molecular and biochemical basis of alcohol abuse behavior extends beyond its harmful physiological effects, but rather is correlated with preference phenotypes.
MeSH term(s)	Animals ; Antioxidants/pharmacology ; Zebrafish/genetics ; Zebrafish/metabolism ; Alcoholism ; DNA Copy Number Variations ; Catalase/genetics ; Catalase/metabolism ; Catalase/pharmacology ; Superoxide Dismutase/genetics ; Superoxide Dismutase/metabolism ; Ethanol ; Brain/metabolism ; Mitochondria/metabolism ; DNA, Mitochondrial/genetics ; Telomere/genetics ; Telomere/metabolism ; Oxidative Stress
Chemical Substances	Antioxidants ; Catalase (EC 1.11.1.6) ; Superoxide Dismutase (EC 1.15.1.1) ; Ethanol (3K9958V90M) ; DNA, Mitochondrial
Language	English
Publishing date	2023-05-01
Publishing country	Singapore
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2704569-9
ISSN	2768-6698 ; 2768-6698
ISSN (online)	2768-6698
ISSN	2768-6698
DOI	10.31083/j.fbl2804073
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Article: Melatonin administration attenuates acute stress by inducing sleep state in zebrafish (Danio rerio)

Lunkes, Luciana Crepaldi / Paiva, Isadora Marques / Egger, Renata Catão / Braga, Weslley Fernandes / Alvarez-Leite, Jacqueline Isaura / da Cunha Barreto-Vianna, André Rodrigues / Murgas, Luis David Solis

Comparative biochemistry and physiology. 2021 Aug., v. 246

2021

Abstract: Melatonin plays a fundamental homeostatic role in basic biological functions, and an anti-stress role has been also proposed for this hormone. This study aimed to evaluate hormonal, enzymatic and behavioral parameters of zebrafish that received ... ...

Abstract	Melatonin plays a fundamental homeostatic role in basic biological functions, and an anti-stress role has been also proposed for this hormone. This study aimed to evaluate hormonal, enzymatic and behavioral parameters of zebrafish that received administration of melatonin and were submitted to acute stress. A total of 120 wild-type zebrafish were divided into five groups: naïve control (N), negative control group (Stress/C), positive control treated with diazepam (Stress/Diaz), treatment with melatonin at dose 1 (Stress/Melt. 1) and treatment with melatonin at dose 2 (Stress/Melt. 2). The exposure to treatments (diazepam or melatonin) was performed prior to the acute stress protocol, based on a chase by a fishing net during 5 min followed by exposure to the air for 1 min. The body cortisol levels were assessed, as well as oxidative stress (thiobarbituric acid reactive substances, reactive species of oxygen and antioxidant activity), and fish behavior (open field test). Melatonin was able to modulate acute stress effects on zebrafish by inhibiting cortisol increasing levels, reducing locomotor parameters, inducing a sleep state, reducing lipid peroxidation and stimulating antioxidant enzymatic activity.
Keywords	Danio rerio ; air ; antioxidant activity ; cortisol ; diazepam ; enzyme activity ; fish behavior ; lipid peroxidation ; melatonin ; oxidative stress ; reactive oxygen species ; sleep
Language	English
Dates of publication	2021-08
Publishing place	Elsevier Inc.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	189285-x
ISSN	0306-4492 ; 0742-8413 ; 1532-0456
ISSN	0306-4492 ; 0742-8413 ; 1532-0456
DOI	10.1016/j.cbpc.2021.109044
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Article ; Online: Bee pollen in zebrafish diet affects intestinal microbiota composition and skin cutaneous melanoma development.

Di Chiacchio, Isabela M / Gómez-Abenza, Elena / Paiva, Isadora M / de Abreu, Danilo J M / Rodríguez-Vidal, Juan Francisco / Carvalho, Elisângela E N / Carvalho, Stephan M / Solis-Murgas, Luis David / Mulero, Victoriano

Scientific reports

2022 Volume 12, Issue 1, Page(s) 9998

Abstract: Bee pollen is recommended as dietary supplement due to immunostimulating functions including antioxidant, anti-inflammatory and anti-carcinogenic properties. Nevertheless, the effectiveness of such properties is still not well understood. As diet can be ... ...

Abstract	Bee pollen is recommended as dietary supplement due to immunostimulating functions including antioxidant, anti-inflammatory and anti-carcinogenic properties. Nevertheless, the effectiveness of such properties is still not well understood. As diet can be associated with animal performance, microbiota modulation and potentially factor for cancer, this study aimed to analyze if bee pollen could influence growth, gut microbial and skin cutaneous melanoma development in zebrafish. Control diets based on commercial flakes and Artemia were compared with the same diet supplemented with bee pollen. Fish weight gain, increased length, intestinal bacteria metagenomics analysis, serum amyloid A gene expression and cutaneous melanoma transplantation assays were performed. Bee pollen affected microbiota composition and melanoma development. Differential abundance revealed higher abundance in the control group for Aeromonadaceae family, Aeromonas and Pseudomonas genus, A. sobria, A. schubertii, A. jandaei and P. alcaligenes species compared with pollen diet group. Pollen group presented higher abundance for Chromobacterium genus and for Gemmobacter aquaticus, Flavobacterium succinicans and Bifidobacterium breve compared with control group. Unexpectedly, fish fed with bee pollen showed higher tumor growth rate and larger tumor size than control group. This is the first study to report intestinal microbial changes and no protective cancer properties after bee pollen administration.
MeSH term(s)	Animals ; Bees ; Diet ; Gastrointestinal Microbiome ; Melanoma/etiology ; Pollen ; Skin Neoplasms/etiology ; Zebrafish ; Melanoma, Cutaneous Malignant
Language	English
Publishing date	2022-06-15
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-022-14245-3
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Article ; Online: Nephrotic Syndrome and Renin-angiotensin System: Pathophysiological Role and Therapeutic Potential.

Dos Anjos, Alessandra Aguiar / de Paiva, Isadora Tucci / Simões Lima, Giovanna Letícia / da Silva Filha, Roberta / Fróes, Brunna Pinto E / Brant Pinheiro, Sérgio Veloso / Silva, Ana Cristina Simões E

Current molecular pharmacology

2022 Volume 16, Issue 4, Page(s) 465–474

Abstract: Idiopathic Nephrotic Syndrome (INS) is the most frequent etiology of glomerulopathy in pediatric patients and one of the most common causes of chronic kidney disease (CKD) and end-stage renal disease (ESRD) in this population. In this review, we aimed to ...

Abstract	Idiopathic Nephrotic Syndrome (INS) is the most frequent etiology of glomerulopathy in pediatric patients and one of the most common causes of chronic kidney disease (CKD) and end-stage renal disease (ESRD) in this population. In this review, we aimed to summarize evidence on the pathophysiological role and therapeutic potential of the Renin-Angiotensin System (RAS) molecules for the control of proteinuria and for delaying the onset of CKD in patients with INS. This is a narrative review in which the databases PubMed, Web of Science, and Sci- ELO were searched for articles about INS and RAS. We selected articles that evaluated the pathophysiological role of RAS and the effects of the alternative RAS axis as a potential therapy for INS. Several studies using rodent models of nephropathies showed that the treatment with activators of the Angiotensin-Converting Enzyme 2 (ACE2) and with Mas receptor agonists reduces proteinuria and improves kidney tissue damage. Another recent paper showed that the reduction of urinary ACE2 levels in children with INS correlates with proteinuria and higher concentrations of inflammatory cytokines, although data with pediatric patients are still limited. The molecules of the alternative RAS axis comprise a wide spectrum, not yet fully explored, of potential pharmacological targets for kidney diseases. The effects of ACE2 activators and receptor Mas agonists show promising results that can be useful for nephropathies including INS.
MeSH term(s)	Humans ; Angiotensin-Converting Enzyme 2 ; Nephrotic Syndrome/drug therapy ; Proteinuria ; Renal Insufficiency, Chronic/drug therapy ; Renin-Angiotensin System/physiology
Chemical Substances	Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
Language	English
Publishing date	2022-06-20
Publishing country	United Arab Emirates
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ISSN	1874-4702
ISSN (online)	1874-4702
DOI	10.2174/1874467215666220616152312
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Article ; Online: Synonymous mutation rs1129293 is associated with PIK3CG expression and PI3Kγ activation in patients with chronic Chagas cardiomyopathy.

Silva, Maria Cláudia / Fuzo, Carlos Alessandro / Marques Paiva, Isadora / Lopes Bibó, Naira / Tavares de Oliveira, Maykon / da Silva Soares, Hellen Anastácia / Chevillard, Christophe / Kalil, Jorge / Cunha-Neto, Edecio / Cunha, Thiago Mattar / Silva, João Santana

Immunobiology

2022 Volume 227, Issue 5, Page(s) 152242

Abstract: Single nucleotide polymorphisms (SNPs) that do not change the composition of amino acids and cause synonymous mutations (sSNPs) were previously considered to lack any functional roles. However, sSNPs have recently been shown to interfere with protein ... ...

Abstract	Single nucleotide polymorphisms (SNPs) that do not change the composition of amino acids and cause synonymous mutations (sSNPs) were previously considered to lack any functional roles. However, sSNPs have recently been shown to interfere with protein expression owing to a myriad of factors related to the regulation of transcription, mRNA stability, and protein translation processes. In patients with Chagas disease, the presence of the synonymous mutation rs1129293 in phosphatidylinositol-4,5-bisphosphate 3-kinase gamma (PIK3CG) gene contributes to the development of the chronic Chagas cardiomyopathy (CCC), instead of the digestive or asymptomatic forms. In this study, we aimed to investigate whether rs1129293 is associated with the transcription of PIK3CG mRNA and its activity by quantifying AKT phosphorylation in the heart samples of 26 chagasic patients with CCC. Our results showed an association between rs1129293 and decreased PIK3CG mRNA expression levels in the cardiac tissues of patients with CCC. The phosphorylation levels of AKT, the protein target of PI3K, were also reduced in patients with this mutation, but were not correlated with PI3KCG mRNA expression levels. Moreover, bioinformatics analysis showed that rs1129293 and other SNPs in linkage disequilibrium (LD) were associated with the transcriptional regulatory elements, post-transcriptional modifications, and cell-specific splicing expression of PIK3CG mRNA. Therefore, our data demonstrates that the synonymous SNP rs1129293 is capable of affecting the PIK3CG mRNA expression and PI3Kγ activation.
MeSH term(s)	Chagas Cardiomyopathy/genetics ; Class Ib Phosphatidylinositol 3-Kinase/genetics ; Humans ; Phosphatidylinositol 3-Kinases ; Polymorphism, Single Nucleotide ; Proto-Oncogene Proteins c-akt ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Silent Mutation
Chemical Substances	RNA, Messenger ; Class Ib Phosphatidylinositol 3-Kinase (EC 2.7.1.137) ; PIK3CG protein, human (EC 2.7.1.137) ; PIK3R3 protein, human (EC 2.7.1.137) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1)
Language	English
Publishing date	2022-07-09
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	563292-4
ISSN	1878-3279 ; 0171-2985
ISSN (online)	1878-3279
ISSN	0171-2985
DOI	10.1016/j.imbio.2022.152242
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Article: Bee pollen as a dietary supplement for fish: Effect on the reproductive performance of zebrafish and the immunological response of their offspring

Di Chiacchio, Isabela M. / Paiva, Isadora M. / de Abreu, Danilo J.M. / Carvalho, Elisângela E.N. / Martínez, Pedro J. / Carvalho, Stephan M. / Mulero, Victoriano / Murgas, Luis David S.

Fish & shellfish immunology. 2021 Dec., v. 119

2021

Abstract: Bee pollen, a natural resource collected by bees, is rich in many nutrients, therefore it may represent a useful dietary supplement. Different uses of bee pollen are proposed due to its beneficial health properties, which includes the capacity to improve ...

Abstract	Bee pollen, a natural resource collected by bees, is rich in many nutrients, therefore it may represent a useful dietary supplement. Different uses of bee pollen are proposed due to its beneficial health properties, which includes the capacity to improve animal performance and promote immunostimulation. Animal nutrition can directly affect adults and their offspring, and larval stage is a critical moment for fish due to high mortality related to immune challenges. Thus, the present study attempted to evaluate the effects of adding bee pollen to a zebrafish diet, specifically, analyzing the effects on reproduction and immunity transference to descendants. Zebrafish adults received control diets based on commercial flakes and live food Artemia sp. nauplii or bee pollen-supplemented diets, administered three times a day, at the same time. The animals received the diets over 60 d, and throughout this period, they were tested for: egg production per female, total number of eggs, embryo viability rate, larval survival rate after exposure to spring viremia of carp virus and to Salmonella enterica serovar Typhimurium, and larval neutrophil recruitment after tail wounding. Bee pollen supplementation failed to improve egg production and embryo viability, and was unable to substitute flakes in zebrafish breeders. Instead, the offspring of breeders fed with bee pollen supplemented diets showed longer survival upon virus exposure and higher neutrophil migration to wounds. These results indicate that bee pollen can influence vertical immunity through important mechanisms related to offspring immunity in the early stages, when larval immune system is not fully developed.
Keywords	Artemia ; Carp sprivivirus ; Danio rerio ; Salmonella Typhimurium ; animal nutrition ; animal performance ; bee pollen ; bees ; dietary supplements ; egg production ; females ; fish ; immune response ; nauplii ; neutrophils ; progeny ; reproductive performance ; shellfish ; survival rate ; tail ; viability ; viruses
Language	English
Dates of publication	2021-12
Size	p. 300-307.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	1067738-0
ISSN	1095-9947 ; 1050-4648
ISSN (online)	1095-9947
ISSN	1050-4648
DOI	10.1016/j.fsi.2021.10.012
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Article: Sex identification of the ornamental amazon fish Astronotus ocellatus by videoceloscopy and gonadal biopsy

Mansur, Victor Ferreira Ribeiro / Melo, Naiara / Di Chiacchio, Isabela Martins / de Lima Assis, Isadora / Machado, Gilmara Junqueira / Paiva, Isadora Marques / de Carvalho, Aline Ferreira Souza / Pereira, Rodrigo Norberto / Solis Murgas, Luis David

Animal reproduction science. 2021 July, v. 230

2021

Abstract: This study was conducted to evaluate and validate the efficacy and safety of videoceloscopy and gonadal biopsy as sexing methods for the A. ocellatus. A total of 31 adult individuals were used. Florfenicol (50 mg/kg) and morphine (5 mg/kg) were ... ...

Abstract	This study was conducted to evaluate and validate the efficacy and safety of videoceloscopy and gonadal biopsy as sexing methods for the A. ocellatus. A total of 31 adult individuals were used. Florfenicol (50 mg/kg) and morphine (5 mg/kg) were administered intramuscularly during the pre-surgical period. Animals were maintained in a supine position preceding a ventral midline incision and endoscope optics were then utilized for gonad visualization and sex identification. A gonadal fragment was collected using laparoscopic forceps and conditioned in 10 % formalin. To suture the cavity, polyamide yarn was used in a simple and continuous pattern. At 15 days subsequent to surgery, healing was evaluated, and the stitches were removed. Videoceloscopy accuracy and gonadal biopsy effectiveness were 97 % and 83 %, respectively. Total time devoted in the videoceloscopy, gonadal biopsy and surgery was longer for animals identified as males compared to females The survival rate was 100 %. There were differences regarding food consumption at 24 and 36 h post-surgery when compared to control specimens (pre-surgical) Regarding position in the water column, differences were observed at 24 and 72 h after surgery when compared individually to the control specimens. There were differences for interaction behavior at 24, 36 and 60 h, and regarding search for hiding places at 12 and 24 h after surgery in relation to the control specimens. The applied videoceloscopy and gonadal biopsy surgical techniques are, therefore, effective and safe for A. ocellatus sexing procedures.
Keywords	Astronotus ; adults ; animal reproduction ; biopsy ; fish ; florfenicol ; food consumption ; formalin ; gonads ; morphine ; optics ; polyamides ; survival rate
Language	English
Dates of publication	2021-07
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	429674-6
ISSN	1873-2232 ; 0378-4320
ISSN (online)	1873-2232
ISSN	0378-4320
DOI	10.1016/j.anireprosci.2021.106780
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