CC BY
02023, Scienceline Publication
Worlds Veterinary Journal
World Vet J, 13(1): 144-151, March 25, 2023
DOI: https://dx.doi.org/10.54203/scil.2023.wvj15
The Effect of Shrimp Shell (Litopenaeus vannamei) Extract on Testicular Parameters of Streptozotocin-induced Diabetic Rats
Aniek Prasetyaningsih1 , Yosua Kristian Adi , Abner Amadeuz Wicaksono1 , and Vinsa Cantya Prakasita'
'Department of Biology, Faculty of Biotechnology, Universitas Kristen Duta Wacana, Yogyakarta-55224, Indonesia
2Department of Reproduction and Obstetrics, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta-5528', Indonesia
Corresponding author's email: vinsa.cantya.p@staff.ukdw.ac.id ABSTRACT
Diabetes mellitus (DM) is a chronic metabolic disorder that has become a major health problem worldwide. Reproductive dysfunction is one of the main complications of DM, particularly in men. However, as is known, shrimp shell extract contains nutrients, such as astaxanthin, that affect reproductive traits. The present study aimed to evaluate the effect of shrimp shell extract on the volume, weight, and histological features of the testes of a DM rat model. Fifteen adult male rats were randomly divided into three groups. Group A (n = 5) was a healthy control group, group B (n = 5) was a DM control group, and group C (n = 5) was a DM group treated with shrimp shell extract. Rats in groups B and C were treated with streptozotocin to induce DM. Rats in group C were given shrimp shell extract at 25 mg/kg body weight for 30 consecutive days after DM induction. Testicles were collected and submitted to dimension, weight, and histological examinations. The testicle volume and weight of rats in group C were significantly higher and heavier, respectively, than rats in group B and did not differ from rats in group A. The seminiferous tubule diameter of rats in group C was significantly larger than rats in group B and did not differ from rats in group A. Rats in group B had a lower testicle volume and lighter testicle weight as well as a shorter seminiferous tubule diameter than rats in groups A and C. In conclusion, shrimp shell extract could improve male fertility parameters in a DM rat model. However, the mechanism of action needs to be studied further.
Keywords: Astaxanthin, Diabetes mellitus, Fertility, Seminiferous tubule, Testis INTRODUCTION
Diabetes mellitus (DM) is a chronic metabolic disorder that has become a major health problem worldwide. A previous study reported that in 2015, there were 415 million people with DM worldwide; this number is predicted to rise to over 642 million by 2040 (Ogurtsova et al., 2017). In 2012, there were 1.5 million deaths due to DM (Ogurtsova et al., 2017). This disease could affect the quality of a patient's life due to its many complications. One of the complications is reproductive dysfunction (Shi et al., 2017). It has been recognized that abnormal blood glucose levels can impair reproductive function in men with DM (Maresch et al., 2018). DM can affect reproductive function, including ejaculation, penile erection, fertility, sperm maturation, and spermatogenesis (Ding et al., 2015; Nna et al., 2017). Insulin-based therapy was introduced in the 1920s and is still the primary treatment for patients with type 1 DM and some patients with advanced stages of type 2 DM (Tavares et al., 2018). In addition, there are some antidiabetic drugs for type 2 DM, such as sulfonylureas, meglitinide, biguanides, and thiazolidinediones (Skliros et al., 2016). Studies regarding the effectiveness of existing antidiabetic drugs on the male reproductive system have been carried out in animal models of DM (Adaramoye and Lawal, 2014; Alves et al., 2014; Ayuob et al., 2015; Zaidi et al., 2017; He et al., 2021). Although the use of antidiabetic drugs is relatively safe and they have been widely prescribed in patients with DM, some side effects, such as hypoglycemia, hyperlactatemia, or metabolic acidosis, may still result from long-term use of currently available antidiabetic drugs (Anagnostis et al., 2018; Wang and Hoyte, 2019).
Many studies have concerned the role of natural products on DM and male reproductive functions (Tran et al., 2020; Swelum et al., 2021; Thikekar et al., 2021; Fu et al., 2022). Some researchers have used whole plants or part of plant extracts, such as Chlorophytum borivilianum (root), Amaranthus spinosus (stem), Danae racemosa (leaves), and Nigella sativa (seeds), while others have used just specific compounds such as phenols, flavonoids, and flavanones isolated from plants (Nna et al., 2017). However, most of those natural products are not included in the food ingredients consumed daily by people. In this study, Litopenaeus vannamei shell extract was used in an animal model of DM to evaluate its effect on male reproductive organs. Litopenaeus vannamei is one of the most widely cultivated shrimp species besides Penaeus monodon and Penaeus chinensis (FAO, 2014). Shrimp is commercialized as seafood and is usually sold whole or sometimes only the meat of shrimp. Thus, shrimp shells are abundant in solid waste and underutilized in the food industry. Shrimp shells still contain some nutrients, such as minerals, proteins, chitin, and
ISSN 2322-4568
A R
c n
t < e 5 d :
O rvj
O 2
2 O
3 2
0
1 —
G
HH
2 A L
A R T
HH
C L E
144
chitosan (Cavalcanti et al., 2016). Chitin has a variety of biological and biomedical uses, including tissue healing. Chitosan is also known for its potential therapeutic effects, including anti-inflammatory, antioxidant, antidiarrheal, and anti-Alzheimer's disease effects (Satitsri and Muanprasat, 2020). Chitosan can enhance the size of the antral follicle, the number of endometrial arterioles, and the endometrial thickness of female rats exposed to lead acetate (Purwitasari et al., 2019). Although mammals do not have endogenous chitin (Ohno et al., 2013), a previous study demonstrated that chitosan, a derivative of a natural carbohydrate biopolymer derived from chitin deacetylation, could improve sperm count and the motility of progressive sperm of lead-acetate-induced rats (Marianti et al., 2020). This suggests that chitin and/or chitosan may be involved in the male reproductive system. In addition, shrimp shell extract contains astaxanthin, which is known to have good effects on the male reproductive system in some species, such as rainbow trout and discus (Ahmadi et al., 2006; Haque et al., 2023). Astaxanthin could have a protective effect on sperm mitochondrial function and also ameliorate testicular heat stress and reproductive poison damage (Liu et al., 2016). However, research concerning the effect of astaxanthin supplementation via shrimp shell extract on the male reproductive organs in the DM condition is still limited. The present study aimed to evaluate the effect of shrimp shell extract on the volume, weight, and histological features of the testicular organ of a DM rat model.
MATERIALS AND METHODS
Ethical approval
The experimental protocols carried out in this study had been approved by Universitas Kristen Duta Wacana (UKDW) Medical Research Ethics Committee with Ethical Clearance Certificate Number: 1265/C.16/FK/2021.
Shrimp shell extraction
Fresh shrimp (L. vannamei) was obtained from a fish market located on the south beach of Java, Indonesia, during the rainy season in July 2021. Shrimp shell extraction was carried out at the Biotechnology for Health Laboratory (Indonesia). Shrimp shells were separated from the flesh manually and washed with running water. Shrimp shells were dried using an oven at 40°C for approximately a day. Then, shrimp shells were ground into powder. Dried shrimp shell powder was subjected to a 3-day extraction using a maceration method as described by Najoan et al. (2021). Ethanol (70%) was used as the solvent at a 1:10 (v/v) ratio with water. The maceration was repeated two times for 3 days, respectively. Subsequently, evaporation was carried out using a rotary evaporator at 5 rpm and 40°C, and then continued in an oven at 40°C until the consistency was like a paste (Najoan et al., 2021).
Experimental animals
A total of 15 male Wistar rats (Rattus norvegicus) aged 8-12 weeks with an average body weight of 190 g from the Faculty of Biology, Universitas Gadjah Mada, Indonesia, were used in this study. Before the experiment began, acclimatization was carried out for 7 days. The rats were given access to water and food ad libitum during this period. The rats were maintained in five plastic boxes, each with three rats at room temperature in a tropical environment with a 12-h photoperiod. After acclimatization, the rats were divided randomly into three groups. Group A (n = 5) was a healthy control group, group B (n = 5) was a DM control group, and group C (n = 5) was a DM group treated with shrimp shell extract. Group A did not receive any treatment during the experiment. In groups B and C, DM was induced by intraperitoneally injecting 50 mg/kg body weight (BW) of streptozotocin (STZ) (Cayman Chemical, USA) diluted in citrate buffer. The STZ dosage for DM induction was previously described by Suman et al. (2016). Three days after DM induction, a blood sample was collected via the caudal vena cava to measure blood glucose levels using glucose meters (OneTouch, USA). The diabetic condition was proven by high blood glucose levels (> 150 mg/dL) (Furman, 2021). After inducing DM, rats in group C were given shrimp shell extract at the dosage of 25 mg/kg BW, and rats in group B were given 1 ml of sterile water as a placebo. The shrimp shell extract dosage was chosen according to a previous study by Wisaksono et al. (2021). Sterile water for the rats in group B and shrimp shell extract for the rats in group C was administered orally via gavage for 30 consecutive days.
Sample collection and histology slide preparation
After 30 days of oral treatment using shrimp shell extract, all rats were euthanized for sample collection. Before euthanasia, rats were anesthetized using tiletamine and zolazepam (Zoletil, Virbac, India) at 20 mg/kg BW (Limprasutr et al., 2021). After the rat was fully anesthetized, indicated by the absence of a pedal reflex (Sivula and Suckow, 2018), it was euthanized by cutting the respiratory tract and carotid vessel in the cervix. The testicles were removed from their scrotum and fixed using a 10% formalin solution. The testicles were measured for dimension and weight before being processed for histological staining. Testicle tissue was trimmed, processed with paraffin, and cut at 5 ^m thickness. The tissue slides were placed on the slide warmer for 30 minutes. Subsequently, tissue slides were deparaffinized using xylene and rehydrated using a graded series of alcohol. Haematoxylin and eosin (HE) staining was performed before the
145
dehydration and clearing processes. Then, the slide was mounted with a cover slip. Histological examination was carried out using a light microscope (Olympus, Tokyo, Japan) with 40* magnification.
Data collection
The dimensions of the testicle, including length (l), width (w), and thickness (t) were measured using a vernier calliper. These dimensions were used to calculate the volume of the testicle by using the formula of volume (v) for ellipsoid (v = [tc/6] * l * w * t) (Van der Plas et al., 2013). The testicle weight was measured with a digital scale (Camry Scale, USA). The seminiferous tubule diameter was determined from histology slides. Three photomicrographs were taken for each histology slide using a digital camera connected to a light microscope. The objective lens was 4* magnification, and the total magnification was 40*. The diameter of 10 seminiferous tubules in each photomicrograph was measured using the Image Raster 3.0 software (Optilab, AZ 85012, USA). Finally, the mean seminiferous tubule for each sample was calculated.
Data analysis
Statistical analyses were conducted using SAS version 9.4 (SAS Inst. Cary, NC, USA.). The data are presented as mean ± standard error. Descriptive statistics were analyzed by using the MEANS Procedure. The testicle volume, testicle weight, and seminiferous tubule diameter of each group were analyzed by multiple analyses of variance using the generalized linear model procedure. Least-squares means were obtained from each group of the variables and were compared by using Tukey-Kramer adjustment for multiple comparisons. Correlation analysis between seminiferous tubule diameter and testicle volume, and testicle weight was carried out using Pearson correlation analysis of SAS. For all the statistical analyses, p < 0.05 was considered statistically significant.
RESULTS
There were 14 rats included at the end of this study. One rat from group C was excluded due to inferiority within the group. The average testicle volume and testicle weight of the rats differed significantly between the three groups (p < 0.05, Table 1). The testicle volume of rats in group B was significantly lower compared with the testicle volume of rats in groups A and C (p < 0.05). The rats in group C had the highest testicle volume, but it was not significantly different compared with the testicle volume of the rats in group A (p > 0.05). The testicle weight of rats in group B was significantly lower compared with the testicle weight of the rats in group A and group C (p < 0.05; Table 1). The rats in group C had the highest testicle weight, but it was not significantly different compared with the testicle weight of the rats in group A (p > 0.05).
The average seminiferous tubule diameter of the rats differed significantly between the three groups (p < 0.05) (Table 1). The seminiferous tubule diameter of the rats in group B was significantly shorter than that of the rats in groups A and C (p < 0.05). The rats in group C had the largest seminiferous tubule diameter, but it was not significantly different compared with the seminiferous tubule diameter of the rats in group A (p > 0.05). Pearson correlation analysis showed a strong and significant relationship between the seminiferous tubule diameter and testicle volume, and testicle weight (p < 0.05, Table 2).Histological observation showed that the rats in groups A and C had rounder seminiferous tubules compared with the rats in group B. Some grooved surfaces of seminiferous tubules could be observed in the rats in group B (Figure 1, black arrows).
Table 1. Testicular parameters in a diabetic rat model treated with shrimp shell (Litopenaeus vannamei) extract in Indonesia
Testicular parameters Group A Group B Group C
Number of samples 5 5 4
Testicle volume (cm3) 1.07±0.07a 0.54±0.07b 1.21±0.08a
Testicle weight (g) 1.17±0.08a 0.62±0.08b 1.26±0.09a
Seminiferous tubule diameter (^m) 329±9a 240±9b 339±10a
The values are presented as the mean ± standard error. Group A: healthy control group; group B: Diabetes mellitus control group; group C: Diabetes
mellitus group treated with shrimp shell extract. Different superscripts indicate significant differences in rows (p < 0.05).
Table 2. Pearson correlation analysis of seminiferous tubule diameter, testicle volume, and testicle weight in a diabetic rat model treated with shrimp shell (Litopenaeus vannamei) extract in Indonesia
Seminiferous tubule diameter
Variables Correlation coefficient (r) p value
Testicle volume 0.940 <0.001
Testicle weight 0.937 <0.001
146
Figure 1. Histological changes in seminiferous tubules of diabetic rats. A-E: The healthy control rats (Group A); F-J: The diabetes mellitus (DM) rats (Group B); K-N: The diabetic rats treated with shrimp shell extract (Group C).
147
DISCUSSION
The protocols for STZ-induced insulin deficiency and hyperglycemia in mice and rats have been well established (Furman, 2021). Suman et al. (2016) used the same STZ dosage to induce type 2 DM in combination with a high-fat diet. STZ injection increases glucose, insulin, free fatty acid, and triglyceride concentrations. A single intraperitoneal injection of a low STZ dose (30 mg/kg BW) in adult male Wistar rats affects pancreatic p-cells as well as the reproductive system via its diabetogenic effect (Omolaoye et al., 2018). Researchers have also reported adverse effects of STZ-induced DM on the male reproductive system in experimental animals (Omolaoye et al., 2018; Kotian et al., 2019; Sampannang et al., 2020). Maresch et al. (2019) demonstrated two major pathways of hyperglycemia-induced organ damage in the testis and epididymis, namely the diacylglycerol-protein kinase C pathway and the polyol pathway. The present study demonstrated that compared with the rats in group A, the rats in group B showed a significant decrease (p < 0.05) in the testicle volume, testicle weight, and seminiferous tubule diameter after 30 days of STZ injection from 1.07 cm3, 1.17 g, and 329 ^m, respectively, to 0.54 cm3, 0.62 g, and 240 ^m, respectively.
Shrimp shells are a waste product in the food industry. However, some nutrients contained in shrimp shells are still useful, such as chitosan (de Queiroz et al., 2017). N,0-Carboxymethyl chitosan, a derivative of chitosan, can be used to increase fiber contents; the resilience of food storage; and the stability of nutrients, including lowering the levels of dry substances, lowering the ash content, increasing the protein content, maintaining the fat content, and increasing the level of nitrogen-free extract (Kusuma et al., 2015). Nadapdap et al. (2014) demonstrated that supplementation with chitosan derived from shrimp shells could improve sperm count, normal sperm morphology, sperm motility, and sperm viability of Wistar rats treated with lead. The possible mechanism for this is that chitosan binds to the lead and forms bonds that make it hydrophilic and thus excretable via urine, thus reducing the male reproductive side effects of lead. However, the direct mechanism of action of chitosan on the male reproductive system is still unclear. Abd El-Hakim et al. (2020) demonstrated that a combination of chitosan-stabilized selenium nanoparticles and metformin could increase sperm motility, sperm viability, and sperm concentration and reduce sperm abnormality in an STZ-induced DM rat model. This suggests that chitosan may act as a delivery agent for the other substance. In addition, astaxanthin can be found in the shrimp shell when extracted using a maceration method with 70% ethanol as the solvent (Wisaksono et al., 2021). Astaxanthin is a xanthophyll carotenoid found in various microorganisms, marine animals, and crustaceans, including shrimp shells (Higuera-Ciapara et al., 2006; Ambati et al., 2014; Wisaksono et al., 2021). Astaxanthin has many biological activities and health benefits, such as antioxidant, anti-lipid peroxidation, anti-inflammatory, and anticancer activities; cardiovascular disease prevention; and immunomodulation (Visioli and Artaria, 2017; Faraone et al., 2020; Fouad et al., 2021). Martinez-Alvarez et al. (2020) stated that the use of astaxanthin and astaxanthin-containing lipid extracts as a food ingredient might have a double function: a technological function because they can provide foods with attractive reddish color and a bioactive function (for example, antioxidant activity) when consumed. Moreover, astaxanthin is safe to consume daily at a dosage ranging from 2 to 24 mg (Brendler and Williamson, 2019).
The use of astaxanthin in DM has been studied by many researchers (Feng et al., 2020; Landon et al., 2020; Ahriyasna et al., 2021; Wisaksono et al., 2021). However, the reproductive aspect in such studies has not been evaluated. The present study revealed that shrimp shell extract could protect STZ-induced rats from testicular damage. This was denoted by the improvement in testicle volume, testicle weight, and seminiferous tubule diameter in STZ-induced rats that were supplemented with shrimp shell extract for 30 days. However, the effect of shrimp shell extract on sperm parameters and reproductive hormones still needs to be clarified. Baskovic et al. (2021) reported that intraperitoneal injection of astaxanthin has a favorable effect on histological morphometric testicular parameters (mean seminiferous tubule diameter, mean seminiferous lumen diameter, epithelial height, tubular area, luminal area, and Johnsen score) in testicular torsion/detorsion-induced rats. This effect is mediated by the antioxidant activity of astaxanthin (Demir et al., 2022). Astaxanthin supplementation of 50-100 mg/kg feed for 6 weeks to improve reproductive performance has been reported in many studies in various species such as Nodipecten nodosus (Linnaeus, 1758), Procambarus clarkia, and layer breeder roosters (Suhnel et al., 2014; Zhenhua et al., 2020; Gao et al., 2021). Wisaksono et al. (2021) reported that supplementation with shrimp shell extract could reduce blood glucose levels in STZ-induced rats. This strengthens the notion that the mechanism by which shrimp shell extract protects STZ-induced rats from reproductive organ damage not only comes from its astaxanthin content, which has bioactive activity but might also be caused by lowering hyperglycemia.
It has been reported that long-term hyperglycemia can increase levels of reactive oxygen species and advanced glycation end products, inhibits endothelial nitric oxide synthase metabolism, and decrease endothelial synthesis and the release of nitric oxide, which leads to erectile dysfunction in patients with DM (He et al., 2021). In addition, hyperglycemia interferes with gonadotropin-releasing hormone secretion, thus reducing gonadotropin and prolactin secretion, which in turn leads to a significant decrease in testosterone secretion from Leydig cells and ultimately to spermatogenesis disorders (He et al., 2021).
148
CONCLUSION
In conclusion, supplementation of shrimp shell extract in STZ-induced rats could improve testicle volume, testicle weight, and seminiferous tubule diameter, which are fertility parameters in males. Shrimp shells are a waste product of the food industry that might be useful in preventing reproductive problems in patients with DM in the future. However, the mechanism of action in reproductive health, especially in pathological conditions, needs to be studied further.
DECLARATIONS
Acknowledgments
This study was supported by the Faculty of Biotechnology, Universitas Kristen Duta Wacana, Yogyakarta-55224, Indonesia.
Authors' contribution
Aniek Prasetyaningsih and Vinsa Cantya Prakasita designed the research project and obtained the funding. Aniek Prasetyaningsih, Abner Amadeuz Wicaksono, and Yosua Kristian Adi conducted the experiments and collected the samples. Vinsa Cantya Prakasita analyzed the data and prepared the manuscript. All authors read and contributed to evaluating the manuscript.
Competing interests
The authors have not declared any conflict of interest.
Ethical consideration
Plagiarism, consent to publish, misconduct, data fabrication and/or falsification, double publication and/or submission, and redundancy have been checked by the authors.
Funding
This research was funded by an LPPM UKDW Featured Research Grant and a Faculty of Biotechnology UKDW Research Grant 2021.
REFERENCES
Abd El-Hakim YM, Abdel-Rahman MA, Khater SI, Hamed Arisha A, Metwally MMM, Nassan MA, and Hassan ME (2020). Chitosan-stabilized selenium nanoparticles and metformin synergistically rescue testicular oxidative damage and steroidogenesis-related genes dysregulation in high-fat diet/streptozotocin-induced diabetic rats. Antioxidants, 10(1): 17. DOI: https://www.doi.org/10.3390/antiox10010017
Adaramoye OA and Lawal SO (2014). Effect of kolaviron, a biflavonoid complex from Garcinia kola seeds, on the antioxidant, hormonal and spermatogenic indices of diabetic male rats. International Journal of Andrologia, 46(8): 878-886. DOI: https://www.doi.org/10.1111/and.12160
Ahmadi MR, Bazyar AA, Safi S, Ytrestoyl T, and Bjerkeng B (2006). Effects of dietary astaxanthin supplementation on reproductive characteristics of rainbow trout (Oncorhynchus mykiss). Journal of Applied Ichthyology, 22(5): 388-394. DOI: https://www.doi.org/10.1111/j.1439-0426.2006.00770.x
Ahriyasna R, Agustini TW, Djamiatun K, and Primal D (2021). The improvement of insulin resistance and the antioxidant capacity in type 2 diabetes mellitus rats with whiteleg shrimp shell powder (Litopenaeus vannamei). Slovak Journal of Food Sciences, 15: 703-711. DOI: https://www.doi.org/10.5219/1684
Alves MG, Martins AD, Vaz CV, Correia S, Moreira PI, Oliveira PF, and Socorro S (2014). Metformin and male reproduction: Effects on Sertoli cell metabolism. British Journal of Pharmacology, 171(4): 1033-1042. DOI: https://www.doi.org/10.1111/bph.12522
Ambati RR, Phang SM, Ravi S, and Aswathanarayana RG (2014). Astaxanthin: Sources, extraction, stability, biological activities and its commercial applications-A review. Marine Drugs, 12(1): 128-152. DOI: https://www.doi.org/10.3390/md12010128
Anagnostis P, Siolos P, Christou K, Gkekas NK, Kosmidou N, Athyros VG, and Karagiannis A (2018). The effect of antidiabetic medications on the cardiovascular system: A critical appraisal of current data. Hormones, 17: 83-95. DOI: https://www.doi .org/10.1007/s42000-018-0017-5
Ayuob NN, Murad HAS, and Ali SS (2015). Impaired expression of sex hormone receptors in male reproductive organs of diabetic rat in response to oral antidiabetic drugs. Folia Histochemica Et Cytobiologica, 53(1): 35-48. DOI: https://www.doi .org/10.5603/FHC.a2015.0005
Baskovic M, Bojanac AK, Sincic N, Peric MH, Krsnik D, and Jezek D (2021). The effect of astaxanthin on testicular torsion-detorsion injury in rats -detailed morphometric evaluation of histological sections. Journal of Pediatric Urology, 17(4): 439.E1-439.E12. DOI: https://www.doi.org/10.1016/j.jpurol.2021.03.020
Brendler T and Williamson EM (2019). Astaxanthin: How much is too much? A safety review. Phytotherapy Research, 33(12): 3090-3111. DOI: https://www.doi .org/10.1002/ptr.6514
Cavalcanti ASRRM, Rosa MEC, Cavalcanti C, and Lisboa HM (2016). Seasonality study of Penaeus vannamei shrimp shells from aquaculture. Revista Brasileira de Produtos Agroindustriais, 18: 487-493. DOI: http://www.doi.org/10.15871/1517-8595/rbpa.v18nespp487-493
Demir S, Kazaz IO, Kerimoglu G, Demir EA, Colak F, Yilmaz S, and Mentese A (2022). Astaxanthin protects testicular tissue against torsion/detorsion-induced injury via suppressing endoplasmic reticulum stress in rats. Journal of Investigative Surgery, 35(5): 1044-1049. DOI: https://www.doi.org/10.1080/08941939.2021.1995540
149
de Queiroz AR, Lia FB, de Oliveira LVA, de Farias RRI, Lima E, da Silva LRJ, Penich CCA, and Lia FMV (2017). Preparation and characterization of chitosan obtained from shells of shrimp (Litopenaeus vannamei Boone). Marine Drugs, 15(5): 141. DOI: https://www.doi.org/10.3390/md15050141
Ding GL, Liu Y, Liu ME, Pan JX, Guo MX, Sheng JZ, and Huang HF (2015). The effects of diabetes on male fertility and epigenetic regulation during spermatogenesis. Asian Journal of Andrology, 17(6): 948-953. DOI: https://www.doi.org/10.4103/1008-682X.150844
Food and agriculture organization (FAO) (2014). The state of world fisheries and aquaculture: Opportunities and challenges. Food and agriculture organization of the United Nations, Fisheries e Aquaculture Department., Rome. Available at: https://www.fao.org/3/i3720e/i3720e.pdf
Faraone I, Sinisgalli C, Ostuni A, Armentano MF, Carmosino M, Milella L, Russo D, Labanca F, and Khan H (2020). Astaxanthin anticancer effects are mediated through multiple molecular mechanisms: A systematic review. Pharmacological Research, 155: 104689. DOI: https://www.doi.org/10.1016/j.phrs.2020.104689
Feng W, Wang Y, Guo N, Huang P, and Mi Y (2020). Effects of astaxanthin on inflammation and insulin resistance in a mouse model of gestational diabetes mellitus. Dose-Response, 18(2): 1559325820926765. DOI: https://www.doi.org/10.1177/1559325820926765
Fouad MA, Sayed-Ahmed MM, Huwait EA, Hafez FH, and Osman AMM (2021). Epigenetic immunomodulatory effect of eugenol and astaxanthin on doxorubicin cytotoxicity in hormonal positive breast cancer cells. BMC Pharmacology and Toxicology, 22: 8. DOI: https://www.doi .org/10.1186/s40360-021 -00473-2
Fu Y, Yuan P, Zheng Y, Gao L, Wei Y, Chen Y, Li P, Ruan Y, Zheng X, and Feng W (2022). Ephedra herb reduces adriamycin-induced testicular toxicity by upregulating the gonadotropin-releasing hormone signalling pathway. Biomedicine & Pharmacotherapy, 150: 113061. DOI: https://www.doi.org/10.1016/j.biopha.2022.113061
Furman BL (2021). Streptozotocin-induced diabetic models in mice and rats. Current Protocols, 1: e78. DOI: https://www.doi.org/10.1002/cpz1.78
Gao S, Heng N, Liu F, Guo Y, Chen Y, Wang L, Ni H, Sheng X, Wang X, Xing K et al. (2021). Natural astaxanthin enhanced antioxidant capacity and improved semen quality through the MAPK/Nrf2 pathway in aging layer breeder roosters. Journal of Animal Science and Biotechnology, 12: 112. DOI: https://www.doi.org/10. 1186/s40104-021-00633-8
Haque R, Sawant PB, Sardar P, Varghese T, Xavier KAM, Chadha NK, Sundaray JK, Haldar C, Jana P, and Pattanaik SS (2023). Shrimp shell waste-derived astaxanthin in synergistic combination with its commercial variant augments gonadal maturation and upregulates vitellogenin gene expression of discus (Symphysodon aequifasciatus). Aquaculture, 562: 738828. DOI: https://www.doi.org/10.1016/j.aquaculture.2022.738828
He Z, Yin G, Li QQ, Zeng Q, and Duan J (2021). Diabetes mellitus causes male reproductive dysfunction: A review of the evidence and mechanisms. In Vivo, 35(5): 2503-2511. DOI: https://www.doi.org/10.21873/invivo.12531
Higuera-Ciapara I, Félix-Valenzuela L, and Goycoolea FM (2006). Astaxanthin: A review of its chemistry and applications. Critical Reviews in Food Science and Nutrition, 46(2): 185-196. DOI: https://www.doi.org/10.1080/10408690590957188
Hu Y, Ding B, Shen Y, Yan RN, Li FF, Sun R, Jing T, Lee KO, and Ma JH (2021). Rapid changes in serum testosterone in men with newly diagnosed type 2 diabetes with intensive insulin and metformin. Diabetes Care, 44(4): 1059-1061. DOI: https://www.doi.org/10.2337/dc20-1558
Kotian SR, Kumar A, Mallik SB, Bhat NP, Souza AD, and Pandey AK (2019). Effect of diabetes on the male reproductive system-a histomorphological study. Journal of Morphology Sciences, 36(1): 17-23. DOI: http://www.doi.org/10.1055/s-0039-1683405
Kusuma HS, Al-sa'bani AF, and Darmokoesoemo H (2015). N,O-carboxymethyl chitosan: An innovation in new natural preservative from shrimp shell waste with a nutritional value and health orientation. Procedia Food Science, 3: 35-51. DOI: https://www.doi.org/10.1016/j.profoo.2015.01.004
Landon R, Gueguen V, Petite H, Letourneur D, Pavon-Djavid G, and Anagnostou F (2020). Impact of astaxanthin on diabetes pathogenesis and chronic complications. Marine Drugs, 18(7): 357. DOI: https://www.doi.org/10.3390/md18070357
Limprasutr V, Sharp P, Jampachaisri K, Pacharinsak C, and Durongphongtorn S (2021). Tiletamine/zolazepam and dexmedetomidine with tramadol provide effective general anesthesia in rats. Animal Models and Experimental Medicine, 4(1): 40-46. https://www.doi.org/10.1002/ame2.12143
Liu W, Kang XF, and Shang XJ (2016). Astaxanthin in male reproduction: Advances in studies. Zhonghua Nan Ke Xue (National Journal of Andrology), 22(10): 938-943. Available at: https://pubmed.ncbi.nlm.nih.gov/29278478/
Marianti A, Isnaeni W, Setiati N, and Sumadi S (2020). Effects of chitosan on sperm quality of lead acetate-induced rats. Journal of Physics: Conference Series, 1567: 032061. Available at: https://iopscience.iop.org/article/10.1088/1742-6596/1567/3/032061/pdf
Maresch CC, Stute DC, Fleming T, Lin J, Hammes HP, and Linn T (2019). Hyperglycemia induces spermatogenic disruption via major pathways of diabetes pathogenesis. Scientific Reports, 9: 13074. DOI: https://www.doi.org/10.1038/s41598-019-49600-4
Maresch CC, Stute DC, Alves MG, Oliveira PF, de Kretser DM, and Linn T (2018). Diabetes-induced hyperglycemia impairs male reproductive function: A systematic review. Human Reproduction Update, 24(1): 86-105. DOI: https://www. doi.org/10.1093/humupd/ dmx03 3
Martínez-Álvarez Ó, Calvo MM, and Gómez-Estaca J (2020). Recent advances in astaxanthin micro/nanoencapsulation to improve its stability and functionality as a food ingredient. Marine Drugs, 18(8): 406. DOI: https://www.doi.org/10.3390/md18080406
Nadapdap TP, Lutan D, Arsyad KHM, and Ilyas S (2014). Influence of chitosan from shrimp skin to quality and quantity of sperm of albino rats after administration of lead. Andrology, 3(1): 1000114. Available at: https://www.longdom.org/open-access/influence-of-chitosan-from-shrimp-skin-to-quality-and-quantity-of-sperm-of-albino-rats-after-administration-of-lead-2167-0250-3-114.pdf
Najoan GC, Prasetyaningsih A, Prakasita VC, Wicaksono AA, and Rahardjo D (2021). Anti-inflammatory activity test of astaxanthin extract from Litopenaeus vannamei shrimp waste against the number of neutrophils and lymphocytes in white rats (Rattus norvegicus) injected with carrageenin. Scholars Academic Journal of Biosciences, 9(5): 123-129. Available at: https://saspublishers.com/media/articles/SAJB_95_123-129 lrOi3ie.pdf
Nna VU, Bakar AB, and Mohamed M (2017). Diabetes mellitus-induced male reproductive impairment: The role of natural products: A review. Journal of Applied Pharmaceutical Science, 7(9): 233-242. DOI: http://www.doi.org/10.7324/JAPS.2017.70932
Ohno M, Togashi Y, Tsuda K, Okawa K, Kamaya M, Sakaguchi M, Sugahara Y, and Oyama F (2013). Quantification of chitinase mRNA levels in human and mouse tissues by real-time PCR: Species-specific expression of acidic mammalian chitinase in stomach tissues. PLoS One, 8(6): e67399. DOI: https://www.doi.org/10.1371/journal.pone.0067399
Omolaoye TS, Skosana BT, and du Plessis SS (2018). Diabetes mellitus-induction: Effect of different streptozotocin doses on male reproductive parameters. Acta Histochemica, 120(2): 103-109. DOI: https://www.doi.org/10.1016/j.acthis.2017.12.005
Ogurtsova K, da Rocha Fernandes JD, Huang Y, Linnenkamp U, Guariguata L, Cho NH, Cavan D, Shaw JE, and Makaroff LE (2017). IDF diabetes atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Research and Clinical Practice, 128: 40-50. DOI: https://www.doi.org/10.1016/j.diabres.2017.03.024
Purwitasari AA, Rozifa AW, Irawan DD, Kalsum U, Ratnawati R, Nurdiana N, and Anita KW (2019). Effect of chitosan on histology of reproductive organs of female Wistar rats (Rattus norvegicus) exposed to acetate lead. Jurnal Kedokteran Brawijaya, 30(4): 259-266. DOI: https://www.doi.org/10.21776/ub.jkb.2019.030.04.5
150
Sampannang A, Arun S, Burawat J, Sukhorum W, and Iamsaard S (2020). Comparison of male reproductive parameters in mice with type 1 and type 2 diabetes. Clinical and Experimental Reproductive Medicine, 47(1): 20-33. DOI: https://www.doi.org/10.5653/cerm.2020.00388
Satitsri S and Muanprasat C (2O2O). Chitin and chitosan derivatives as biomaterial resources for biological and biomedical applications. Molecules, 25(24): 5961. DOI: https://www.doi.org/10.3390/molecules25245961
Shi GJ, Li ZM, Zheng J, Chen J, Han XX, Wu J, Li GY, Chang Q, Li YX, and Yu JQ (2017). Diabetes associated with male reproductive system damages: Onset of presentation, pathophysiological mechanisms and drug intervention. Biomedicine & Pharmacotherapy, 90: 562-574. DOI: https://www.doi.org/10.1016/j.biopha.2017.03.074
Sivula CP and Suckow MA (2018). Euthanasia. Management of animal care and use programs in research, education, and testing, 2nd Edition. CRC Press/Taylor & Francis., Boca Raton. https://www.ncbi.nlm.nih.gov/books/NBK500441/
Skliros NP, Vlachopoulos C, and Tousoulis D (2016). Treatment of diabetes: Crossing to the other side. Hellenic Journal of Cardiology, 57(5): 304310. DOI: https://www.doi.org/10.1016/j.hjc.2016.07.002
Suman RK, Mohanty IR, Borde MK, Maheshwari U, and Deshmukh YA (2016). Development of an experimental model of diabetes co-existing with metabolic syndrome in rats. Advances in Pharmacological and Pharmaceutical Sciences, 2016: 9463476. DOI: https://www.doi.org/10.1155/2016/9463476
Suhnel S, Lagreze F, Pereira A, Silva F, Gurney-Smith H, Bercht M, Maraschin M, Magalhaes A, and Ferreira J (2014). Effects of astaxanthin on reproductive success in the tropical scallop nodipecten nodosus (Linnaeus, 1758). Journal of Shellfish Research, 33(1): 89-98. DOI: http://www.doi.org/10.2983/035.033.0111
Swelum AA, Hashem NM, Abdelnour SA, Taha AE, Ohran H, Khafaga AF, El-Tarabilu KA, and El-Hack MEA (2021). Effects of phytogenic feed additives on the reproductive performance of animals. Saudi Journal of Biological Sciences, 28(10): 5816-5822. DOI: https://www.doi.org/10.1016/i.sibs.2021.06.045
Tavares RS, Escada-Rebelo S, Silva AF, Sousa MI, Ramalho-Santos J, and Amaral S (2018). Antidiabetic therapies and male reproductive function: Where do we stand?. Reproduction, 155(1): R13-R37. DOI: https://www.doi.org/10.1530/REP-17-0390
Thikekar AK, Thomas AB, and Chitlange SS (2021). Herb-drug interactions in diabetes mellitus: A review based on pre-clinical and clinical data. Phytotherapy Research, 35(9): 4763-4781. DOI: https://www.doi.org/10.1002/ptr.7108
Tran N, Pham B, and Le L (2020). Bioactive compounds in antidiabetic plants: from herbal medicine to modern drug discovery. Biology, 9(9): 252. DOI: https://www.doi.org/10.3390/biology9090252
Van der Plas EM, Zijp GW, Froeling FMJA, Van Der Voort-Doedens LM, Vries AM, Goede J, and Hack WWM (2013). Long-term testicular volume after orchiopexy at diagnosis of acquired undescended testis. The Journal of Urology, 190, 257-262. DOI: http://www.doi.org/10.1016/ijuro.2013.02.004
Visioli F and Artaria C (2017). Astaxanthin in cardiovascular health and disease: Mechanisms of action, therapeutic merits, and knowledge gaps. Food & Function, 8(1): 39-63. DOI: https://www.doi.org/10.1039/C6FO01721E
Wang GS and Hoyte C (2019). Review of biguanide (metformin) toxicity. Journal of Intensive Care Medicine, 34(11-12): 863-876. DOI: https://www.doi.org/10.1177/0885066618793385
Wisaksono AA, Prasetyaningsih A, Prakasita VC, and Najoan GC (2021). Utilization of Littopenaeus vannamei shrimp shell extract as a blood sugar reducing alternative. Scholar Academic Journal of Biosciences, 9(7): 175-181. Available at: https://saspublishers.com/media/articles/SAJB 97 175-181 c.pdf
Zaidi A, Khan M, Sharif A, Shakir L, Irshad A, Ali A, and Shaheryar Z (2017). Comparative study of sperm motility in metformin-using and insulin-dependent diabetics. Biomedical Research and Therapy, 4(6): 1388-1399. DOI: https://www.doi.org/10.15419/bmrat.v4i06.180
Zhenhua An, Yang H, Liu X, and Zhangv Y (2020). Effects of astaxanthin on the immune response and reproduction of Procambarus clarkii stressed with microcystin-leucine-arginine. Fisheries Science, 86: 759-766. DOI: https://www.doi.org/10.1007/s12562-020-01434-0
151
