Creolin® administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom

Carvalho, Lucas Rannier Ribeiro Antonino; Pereira, Helder Camilo da Silva; Silva, Hugo Thyares Fonseca Nascimento Pereira da; Lucena, Ricardo Barbosa de; Guerra, Ricardo Romão

doi:10.1590/0103-8478cr20180699

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PATHOLOGY • Cienc. Rural 49 (5) • 2019 • https://doi.org/10.1590/0103-8478cr20180699 copy

Creolin^® administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom

Creolina ^® administrada por diferentes vias em ratos envenenados experimentalmente com veneno de Bothrops jararaca

Authorship SCIMAGO INSTITUTIONS RANKINGS

ABSTRACT:

This study aimed to evaluate the effects of Creolin^® when administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom. In female Wistar rats, the Bothropic venom was inoculated intramuscularly, and then the rats were either treated with Creolin^® (administered orally, topically, or intramuscularly), or with amixture of venom + Creolin^® intramuscularly. Animals that received Creolin^®, apart from the venom, by oral, topical, or intramuscular routes developed local symptoms and showed laboratory findings similar to those animals that received only the venom. Conversely, animals inoculated with the venom incubated with Creolin^® showed no signs of local venom toxicity (necrosis or hemorrhage) and displayed hematological parameters within the normal range for the species. These results suggest that Creolin^® exhibited an antiophidian effect only when it is mixed with the venom and administered intramuscularly.

Key words:
antivenom; folk medicine; bothropic venom

Authorship

Lucas Rannier Ribeiro Antonino Carvalho

Programa de Pós-graduação em Ciência Animal, Universidade Federal da Paraíba (UFPB), Areia, PB, Brasil.Universidade Federal da Paraíba (UFPB)BrazilAreia, PB, BrazilPrograma de Pós-graduação em Ciência Animal, Universidade Federal da Paraíba (UFPB), Areia, PB, Brasil.

http://orcid.org/0000-0002-5875-8573

Helder Camilo da Silva Pereira

Universidade Federal da Paraíba (UFPB), Curso de Medicina Veterinária, Areia, PB, Brasil.Universidade Federal da Paraíba (UFPB),BrazilAreia, PB, BrazilUniversidade Federal da Paraíba (UFPB), Curso de Medicina Veterinária, Areia, PB, Brasil.

http://orcid.org/0000-0001-9265-8966

Hugo Thyares Fonseca Nascimento Pereira da Silva

Ricardo Barbosa de Lucena

Programa de Pós-graduação em Ciência Animal, Universidade Federal da Paraíba (UFPB), 58397-000, Areia, PB, Brasil. Universidade Federal da Paraíba (UFPB)BrazilAreia, PB, BrazilPrograma de Pós-graduação em Ciência Animal, Universidade Federal da Paraíba (UFPB), 58397-000, Areia, PB, Brasil.

http://orcid.org/0000-0003-1162-1948

Ricardo Romão Guerra ^*E-mail: ricardo@cca.ufpb.br. *Corresponding author.

http://orcid.org/0000-0001-8226-8606

E-mail: ricardo@cca.ufpb.br. ^*Corresponding author.

DECLARATION OF CONFLICT OF INTERESTS

The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

AUTHORS’ CONTRIBUTIONS

All authors contributed equally for the conception and writing of the manuscript. All authors critically revised the manuscript and approved of the final version.

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (1)
Tables (3)

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Figure 1
Macroscopic and microscopic evaluation of the right hind limbs of Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^®. A1) Photographic image of the right-side face of the limb inoculated only with Creolin^® intramuscularly. A2) Photomicrography of the inoculation region of Creolin^® with evident neutrophil infiltrate (^*). Hematoxylin-Eosin staining. Bar: 200 micrometers. B1) Photographic image of the right-side face of the limb inoculated only with bothropic poison intramuscularly presenting friable appearance, edema, and hemorrhage. B2) Photomicrography of the venom inoculation region with diffuse hemorrhage (arrows). Hematoxylin-Eosin staining. Bar: 100 micrometers. C1) Photographic image of the right-side face of the limb inoculated with bothropic venom and then Creolin^® intramuscularly, presenting friable appearance, edema, and hemorrhage similar to inoculation only with the venom. C2) Photomicrography of the venom inoculation region and the Creolin^® injected intramuscularly with diffuse hemorrhage (arrows). Hematoxylin-Eosin staining. Bar: 200 micrometers. D1) Photographic image of the right-side face of the limb inoculated with the venom + Creolin^® mixture intramuscularly. D2) Photomicrography of the inoculation region of the venom + Creolin^® mixture injected intramuscularly with the presence of neutrophil infiltrate (^*). Hematoxylin-eosin staining. Bar: 100 micrometers.

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Table 1
Identified changes and their time intervals in a group of Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes.

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Table 2
Renal and muscular histopathological changes in Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes after eight hours of inoculation.

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Table 3
Hematological values of Wistar rats, eight hours after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes (^*). Values expressed as mean ± standard deviation.

Figure 1 Macroscopic and microscopic evaluation of the right hind limbs of Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^®. A1) Photographic image of the right-side face of the limb inoculated only with Creolin^® intramuscularly. A2) Photomicrography of the inoculation region of Creolin^® with evident neutrophil infiltrate (^). Hematoxylin-Eosin staining. Bar: 200 micrometers. B1) Photographic image of the right-side face of the limb inoculated only with bothropic poison intramuscularly presenting friable appearance, edema, and hemorrhage. B2) Photomicrography of the venom inoculation region with diffuse hemorrhage (arrows). Hematoxylin-Eosin staining. Bar: 100 micrometers. C1) Photographic image of the right-side face of the limb inoculated with bothropic venom and then Creolin^® intramuscularly, presenting friable appearance, edema, and hemorrhage similar to inoculation only with the venom. C2) Photomicrography of the venom inoculation region and the Creolin^® injected intramuscularly with diffuse hemorrhage (arrows). Hematoxylin-Eosin staining. Bar: 200 micrometers. D1) Photographic image of the right-side face of the limb inoculated with the venom + Creolin^® mixture intramuscularly. D2) Photomicrography of the inoculation region of the venom + Creolin^® mixture injected intramuscularly with the presence of neutrophil infiltrate (^). Hematoxylin-eosin staining. Bar: 100 micrometers.

Table 1 Identified changes and their time intervals in a group of Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes.

Changes	Control	imC	Bj	Bj+voC	Bj+vtC	Bj+imC	imBj+C
Claudication	Absent	0 h-3 h	0 h-8 h	0 h-8 h	0 h-8 h	0 h-8 h	0 h-8 h
Prostration	Absent	Absent	0 h-5 h	0 h-4 h	0 h-3 h	0 h-4 h	0 h-4 h
Myoclonus	Absent	Absent	Absent	0 h-2 h	0 h-2 h	Absent	0 h-2 h
Bleeding	Absent	Absent	1 h-8 h	2 h-8 h	2 h-8 h	01-8 h	Absent
Pain	0 h-1 h	0 h-2 h	0 h-8 h	0 h-8 h	0 h-8 h	0 h-8 h	0 h-8 h
Dark urine	Absent	Absent	1 h-8 h	2 h-5 h	2 h-5 h	2 h-5 h	2 h-3 h
Anorexia	0 h-1 h	0 h-3 h	0 h-5 h	0 h-5 h	0 h-3 h	0 h-4 h	0 h-5 h
Death	0/5	0/5	0/5	0/5	2/5	0/5	0/5

Control: sterile saline solution; Bj: only intramuscular bothropic venom; imC: only intramuscular Creolin^®; Bj+voC: bothropic venom followed by oral Creolin^® treatment; Bj+vtC: bothropic venom followed by topical Creolin^® treatment; Bj+imC: bothropic venom and followed by intramuscular injection of Creolin^®; imBj+C:intramuscular injection of a mixture of Creolin^® and bothropic venom.

Table 2 Renal and muscular histopathological changes in Wistar rats after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes after eight hours of inoculation.

Renal Changes	Control	imC	Bj	Bj+voC	Bj+vtC	Bj+imC	imBj+C
Granular cylinders	-	-	+++	+++	+++	+++	++
Hyaline cylinders	-	-	++	+++	+++	+++	+
Hydropic degeneration	-	-	+	+	+	+	-
Necrosis of tubules	-	-	++	+	-	+	-
Muscular Changes	Control	imC	Bj	Bj+voC	Bj+vtC	Bj+imC	imBj+C
Bleeding	-	-	+++	+++	+++	++	-
Edema	-	++	+++	+++	++	++	+
Neutrophilic infiltrate	-	+++	-	-	++	+	+++
Hyaline degeneration	-	-	+	+	-	+	-

Control: sterile saline solution; Bj: only intramuscular bothropic venom; imC: only intramuscular Creolin^®; Bj+voC: bothropic venom followed by oral Creolin^® treatment; Bj+vtC: bothropic venom followed by topical Creolin^® treatment; Bj+imC: bothropic venom and followed by intramuscular injection of Creolin^®; imBj+C:intramuscular injection ofa mixture of Creolin^® and bothropic venom.

Table 3 Hematological values of Wistar rats, eight hours after experimental bothropic poisoning (Bothrops jararaca) and treatment with Creolin^® by different access routes (^*). Values expressed as mean ± standard deviation.

Hemogram	Control (n=5)	imC (n=4)	Bj (n=5)	Bj+voC (n=4)	Bj+vtC (n=3)	Bj+imC (n=5)	imBj+C (n=4)
Leukocytes (x10³/μL)	5.2±0.9b	7.8±1.7ab	9.5±1.3a	6.9±0.7ab	5.2±0.9b	7.3±2.3ab	6.2±0.2b
Blood Cells (x10⁶/μL)	7.2±0.4ab	7.5±0.7a	6.6±0.5abc	4.6±0.4d	4.9±0.2d	6.0±0.3c	6.3±0.2bc
Hemoglobin (g/dL)	15.2±1.0a	15.8±0.5a	13.4±1.0b	9.5±0.9c	9.2±0.4c	12.2±0.6b	13.1±0.3b
Hematocrit (%)	42.0±2.6ab	45.2±1.2a	37.3±2.6c	26.7±3.0d	28.1±0.2d	34.1±1.8c	38.0±1.0bc
Plasma Protein (g/dL)	6.4±0.2a	6.7±0.3a	5.2±0.3c	5.6±0.1bc	5.3±0.3bc	5.2±0.3c	6.0±0.1ab
Fibrinogen (mg/dL)	225.0±125.8a	225.0±50.0a	100.0±89.4a	100.0±115.5a	66.6±115.5a	140.0±260.8a	160.0±167.3a
Platelets (10³/µL)	704.3±135.3a	840.5±82.1a	13.6±6.9b	77.2±61.7b	88.3±31.3b	11.0±4.0b	838.0±209.6a

^*Mean in the same line, followed by distinct letters differed by Tukey test and p<0.05. Control: sterile saline solution; Bj: only intramuscular bothropic venom; imC: only intramuscular Creolin^®; Bj+voC: bothropic venom followed by oral Creolin^® treatment; Bj+vtC: bothropic venom followed by topical Creolin^® treatment; Bj+imC: bothropic venom and followed by intramuscular injection of Creolin^®; imBj+C: intramuscular injection of a mixture of Creolin^® and bothropic venom.

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Creolin® administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom

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Mensagem

[1] E-mail: ricardo@cca.ufpb.br. ^*Corresponding author.

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Creolin® administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom

Creolina ® administrada por diferentes vias em ratos envenenados experimentalmente com veneno de Bothrops jararaca

ABSTRACT:

Creolin^® administered by different pathways in rats experimentally poisoned with Bothrops jararaca venom

Creolina ^® administrada por diferentes vias em ratos envenenados experimentalmente com veneno de Bothrops jararaca