scholarly journals Developmental and Reproductive Outcomes in Male Rats Exposed to Triclosan: Two-Generation Study

2021 ◽  
Vol 12 ◽  
Author(s):  
Bruno Garcia Montagnini ◽  
Simone Forcato ◽  
Karine Vandressa Pernoncine ◽  
Mariana Cunha Monteiro ◽  
Marina Rangel Ferro Pereira ◽  
...  
Keyword(s):  
Endocrine Disruptor ◽  
Wistar Rats ◽  
Corn Oil ◽  
Sperm Quality ◽  
Male Rats ◽  
Control Group ◽  
Sperm Viability ◽  
Male Wistar Rats ◽  
Reproduction Toxicity ◽  
Environmental Compartments

Triclosan (TCS) is a phenolic compound with broad-spectrum antimicrobial action that has been incorporated into a variety of personal care products and other industry segments such as toys, textiles, and plastics. Due to its widespread use, TCS and its derivatives have been detected in several environmental compartments, with potential bioaccumulation and persistence. Indeed, some studies have demonstrated that TCS may act as a potential endocrine disruptor for the reproductive system. In the current study, we are reporting on the results obtained for male rats after a two-generation reproduction toxicity study conducted with TCS. Female and male Wistar rats were treated daily by gavage with TCS at doses of 0.8, 2.4, and 8.0 mg/kg/day or corn oil (control group) over 10 weeks (F0) and over 14 weeks (F1) before mating and then throughout mating, until weaning F2 generations, respectively. TCS exposure decreased sperm viability and motility of F1 rats at the dose of 2.4 mg/kg. The effects of TCS on sperm quality may be related to the exposure window, which includes the programming of reproductive cells that occurs during fetal/neonatal development.

scholarly journals Upregulation of Kiss-1 and Gpr54 Genes Expression in Pituitary of Male Rats Following the Central Administration of Neuropeptide Y

2019 ◽  
Vol 4 (4) ◽  
pp. 137-142
Author(s):  
Vahid Azizi ◽  
Shahrbanoo Oryan ◽  
Homayuon Khazali ◽  
Abdolkarim Hosseini
Keyword(s):  
Gene Expression ◽  
Pituitary Gland ◽  
Neuropeptide Y ◽  
Wistar Rats ◽  
Third Ventricle ◽  
Male Rats ◽  
Control Group ◽  
Central Administration ◽  
Reproductive Axis ◽  
Male Wistar Rats

Introduction: The neuropeptide Y (NPY) in the neural circuits of the hypothalamus has a stimulating effect on reproductive activities in mammals. Kisspeptin (KiSS1) is a quintessential neurotransmitter in the reproductive axis which directly stimulates gonadotropin-releasing hormone neurons in the hypothalamus. The distribution of KiSS1 expressing cells in the pituitary was described previously. Despite earlier reports showing the KiSS1 receptor, G-protein coupled receptor 54 (GPR54) expression in the pituitary, the potential physiological roles of kisspeptin at this gland have remained obscure. Accordingly, this study investigated the role of NPY on the relative expression of Kiss1 and Gpr54 genes in the pituitary gland in male Wistar rats. Methods: In general, 20 male Wistar rats weighing 200-250 g in 4 groups (5 in each group) received saline, NPY (2.3 nM), BIBP3226 (NPY receptor antagonist, 7.8 nM), and NPY+ BIBP3226. Then, they received the simultaneous injection of these molecules through the third ventricle of the brain. Finally, the relative mean expressions of Kiss1 and Gpr54 genes in the anterior pituitary were quantitatively analyzed by the real-time polymerase chain reaction. Results: The central injection of NPY increased the relative mean expressions of Kiss1 and Gpr54 genes in the pituitary gland compared to the control group although the injection of BIBP3226 eradicated these effects. However, the gene expression of Gpr54 in the rats receiving NPY coupled with BIBP3226 in hypophysis in comparison to the group receiving only NPY demonstrated a significant reduction (P<0.05). Conclusion: Overall, the central injection of NPY stimulated the gene expression of Kiss1 and Gpr54 in the pituitary gland.

scholarly journals The Effect of Soy Milk on Mounting Latency, Mounting Frequency, and Reproductive Development in Male Wistar Rats (Rattus Norvegicus)

2021 ◽  
Vol 9 (B) ◽  
pp. 670-678
Author(s):  
Nurdiana Nurdiana ◽  
Pradnyawati Chania ◽  
Rifzi Nurvitasari ◽  
Azmiatun Nisa ◽  
Styan Wahyu Diana ◽  
...  
Keyword(s):  
Rattus Norvegicus ◽  
Wistar Rats ◽  
Reproductive Development ◽  
Normal Diet ◽  
Male Rats ◽  
Male Rat ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Soy Milk ◽  
Male Wistar Rats

AIM: This research aims to examine the effects of soy milk on mounting latency (ML), mounting frequency (MF), estrogen levels, androgen-binding protein (ABP) expression, and spermatogenesis in male rats (Rattus norvegicus). METHODS: Twenty-four male wistar rats (Rattus norvegicus) aged 4 weeks were divided into four groups. Control group (given a normal diet), P1; P2; P3 (given the normal diet and soy milk powder at doses of 7.1; 14.2; 21.3 g/KgBW/day, respectively) for 6 weeks. Observation of ML and MF were performed at 9 weeks 5 days of age, and rat surgery was performed at 10 weeks of age. Analysis of estrogen hormone levels was conducted by enzyme-linked immunosorbent assay (ELISA), ABP staining was using immunohistochemistry method, testicular spermatogenesis was observed using histopathological methods, and observation of spermatozoa was performed under the microscope.  RESULTS: The results showed no significant reduction of ML and MF, estrogen levels, and ABP expression (p ≤ 0.256; 0.865; 0.959, respectively) in male rat, but there was a significant decrease in the number, morphology, motility of spermatozoa, and testicular histophatology, (p ≤ 0.000, 0.003, 0.008, 0.000, respectively). CONCLUSION: The administrassion of soy milk in various doses (7.1;14.2;21.3 g/KgBW/day) in male Wistar rats (Rattus norvegicus) had showed significantly difference on histopathological evaluation using Johnson’s scoring system, sperm quantity and quality, while on mounting latency and frequency, estrogen levels, and ABP expressions did not show significantly difference between groups. That describe of isoflavone in soy milk can affect several aspects related to male endocrine and reproductive development.

scholarly journals PENGARUH JAMU DENGAN Tribulus terrestris TERHADAP KUALITAS SPERMA TIKUS WISTAR JANTAN (Rattus norvegicus)

2015 ◽  
Vol 3 (2) ◽  
Author(s):  
Delano Pelealu ◽  
Lydia Tendean ◽  
Benny Wantouw
Keyword(s):  
Experimental Study ◽  
Rattus Norvegicus ◽  
Wistar Rats ◽  
Sperm Morphology ◽  
Sperm Quality ◽  
Control Group ◽  
Tribulus Terrestris ◽  
Normal Morphology ◽  
Normal Sperm ◽  
Male Wistar Rats

Abstract: It is known that Tribulus terrestris can improve sperm quality. One of the Indonesian traditional medicine, jamu, contains this Tribulus terrestris. This study aimed to investigate the effect of jamu containing Tribulus terrestris on sperm quality which consisted of concentration, motility, and morphology of spermatozoa of male wistar rats (Rattus norvegicus). This was an experimental study. Nine male wistar rats were randomly divided into 3 groups of 3 rats each: Group P0 (control), group P1 (given 5 mg of Tribulus terrestris), and group P2 (given 10 mg of Tribulus terrestris). This study was conducted for 52 days. The results showed that the spermatozoa concentration of P1 was 49x106 sperm/ml suspension and of P2 53x106 sperm/ml suspension; both were lower than of P0 (59x106 sperm/ml suspension). The sperm motility category (A+B) of P1 was 33% and of P2 (37%); both were lower than of P0 (45%). The motility category A was not found in P1 and P2. The percentage of spermatozoa with normal morphology of P1 was 45% and the abnormal ones was 55%, meanwhile in P2 there was 49% of normal sperm morphology and 51% of abnormal morphology. Conclusion: Jamu containing Tribulus terrestris had no effect in improvement of the sperm quality.Keywords: jamu, Tribulus terrestris, sperm qualityAbstrak: Tribulus terrestris dikenal sebagai bahan yang dapat memperbaiki kualitas sperma. Salah satu jenis jamu yang diproduksi di Indonesia mengandung Tribulus terrestris Penelitian ini bertujuan untuk mengetahui pengaruh jamu dengan Tribulus terrestris terhadap konsentrasi, motilitas, dan morfologi spermatozoa tikus wistar jantan (Rattus norvegicus). Penelitian ini menggunakan metode eksperimental. Sampel 9 ekor tikus wistar jantan (Rattus norvegicus) dibagi menjadi 3 kelompok yakni, kelompok P0 (kontrol); kelompok P1 (pemberian Tribulus terrestris 5 mg); dan kelompok P2 (pemberian Tribulus terrestris 10 mg). Penelitian dilakukan selama 52 hari. Hasil penelitian menunjukkan bahwa konsentrasi spermatozoa P1 49x106 sperma/ml suspensi dan P2 53x106 sperma/ml suspensi lebih rendah dibandingkan P0 59x106 sperma/ml suspensi. Motilitas spermatozoa kategori A+B pada P1 (33%) dan P2 (37%) lebih rendah dibandingkan pada P0 (45%). Tidak ditemukan motilitas kategori A pada P1 dan P2. Morfologi spermatozoa normal pada P1 sebesar 45% dan abnormal 55%. P2 memiliki morfologi spermatozoa normal sebesar 49% dan abnormal 51%. Simpulan: Pemberian jamu dengan Tribulus terrestris tidak meningkatkan kualitas sperma yang mencakup konsentrasi, motilitas, dan morfologi spermatozoa.Kata kunci: jamu, Tribulus terrestris, kualitas sperma

scholarly journals THE EFFECTS OF KEFIR ON THE INFLAMATORY STATUS AND THYROID FUNCTION (EXPERIMENTAL STUDY ON WISTAR RATS AFTER EXPOSED TO CHLORPYRIFOS)

2016 ◽  
Vol 9 (5) ◽  
pp. 165 ◽  
Author(s):  
Rasipin Rasipin ◽  
Edi Dharmana ◽  
Suharyo Hadisaputro ◽  
Suhartono .
Keyword(s):  
Thyroid Function ◽  
Wistar Rats ◽  
Thyroid Dysfunction ◽  
Corn Oil ◽  
Thyroid Stimulating Hormone ◽  
Male Rats ◽  
Tnf Α ◽  
Male Wistar Rats ◽  
Α Level ◽  
Factor Α

ABSTRACTObjective: Goiter is an enlarged thyroid gland remained a health problem in the agricultural areas. Chlorpyrifos (CPF) is a pesticide widely used byfarmers. Previous studies proved that CPF exposure caused thyroid dysfunction. The objective of this study was to evaluate the effects of kefir on theinflammatory status and thyroid function in male Wistar rats after exposed to CPF using biochemical and histopathological assays.Methods: Male rats were divided into 4 groups, i.e., CPF 5+kefir (5 mg/kg+3.6 ml/200 g, respectively), CPF 5 (5 mg/kg), corn oil (CO 1 ml/200 g), andnegative control (NC: Without CPF, CO, and kefir).Results: Kefir supplementation dose 3.6 ml/200 g once a day for 28 days in the rats after exposed to CPF dose 5 mg/kg once a day for 14 days, inCPF 5+kefir as compared to CPF 5: Significantly (p<0.05) decreased serum tumor necrosis factor-α (TNF-α) level; significantly (p<0.01) maintainedserum levels of tumor growth factor-β (TGF-β) and thyroid stimulating hormone (TSH) not to decrease; not significant (p>0.05) decreased the level ofinterleukin-1β, cluster of differentiation-26 expression and level of T serum; not significant (p>0.05) maintained the level of anti-thyroid peroxidasenot to decrease; and not significant (p>0.05) increased the apoptosis index. This study suggests that CPF exposure causes the inflammatory processwhich leads to thyroid dysfunction.4Conclusion: Kefir supplementation significantly decreased the level of TNF-α and maintained the levels of TGF-β and TSH not to decrease, possible toreduce the inflammatory and thyroid dysfunction processes caused by exposure to CPF in experimental animals.Keywords: Kefir, Chlorpyrifos, Inflammation, Thyroid function. 

scholarly journals Attenuation of Carbon Tetrachloride – Induced Hepatoxicity by Dacryodes edulis Seeds Ethanolic Extract in Male Wistar Rats

2020 ◽  
Vol 11 (2) ◽  
pp. 9490-9500
Keyword(s):  
Body Weight ◽  
Carbon Tetrachloride ◽  
Wistar Rats ◽  
Corn Oil ◽  
Ethanolic Extract ◽  
Seed Extract ◽  
Control Group ◽  
Once Daily ◽  
Male Wistar Rats ◽  
Dacryodes Edulis

Dacryodes edulis (African pear) seed has a long history of use in ethnomedicine. However, there is a paucity of information on its ameliorative potential on hepatic damage. In the present study, the potentials of D. edulis ethanolic seed extract (DET) in attenuating carbon tetrachloride (CCl4)-induced hepatotoxicity in male Wistar rats was assessed. Ninety male Wistar rats were divided into 3 groups (n = 10) designated: control, CCl4, and DET, respectively. Animals in the control group were administered 2 mL/kg body weight corn oil, while those assigned to CCl4 and DET groups were administered i.p. 30% CCl4 in corn oil (1:1 v/v) at 2 mL/kg. Treatment was administered once every 24 h consecutively for 72 h. Thereafter, control and CCl4 groups were treated once daily with 1 mL distilled water by gavage, while DET rats were treated once daily with 1500 mg/kg DET by gavage. At every 7 days interval and for 28 consecutive days, animals were starved overnight, weighed, and six rats from each group were sacrificed under anesthesia to collect blood and liver samples. After 28 days, remarkable improvements in feed intake, body weight, serum levels of hepatic function indicators, and hepatic histology were observed in DET rats compared to CCl4 rats. These results suggest a possible ameliorative potential of D. edulis ethanolic seed extract against CCl4-induced hepatic injury.

scholarly journals THE EFFECTS OF NICOTINE EXPOSURE PER INHALATION TO THE CHANGE OF MOTILITY AND MORPHOLOGY OF THE RAT’S SPERM

2019 ◽  
Vol 26 (2) ◽  
Author(s):  
Puruhito Eko Nugroho ◽  
Fikri Rizaldi ◽  
Wahjoe Djati Soesanto
Keyword(s):  
Wistar Rats ◽  
Male Rats ◽  
Control Group ◽  
Nicotine Exposure ◽  
Motile Sperm ◽  
Normal Morphology ◽  
Sperm Analysis ◽  
Treatment Groups ◽  
Male Wistar Rats ◽  
The Difference

Objective: Smoking are known to have detrimental effect on sperm. The effects are suspiciously come from nicotine as its main pharmacologic component. This study is carried out to analyze the effect of nicotine exposure per inhalation to the difference of motile sperm percentage and normal morphology sperm percentage of male wistar rats. Material & Methods: Thirty male rats were used for this research. They were randomly divided into 5 groups and treated with nicotine inhalation 0.5 mg/kgBW,  1 mg/kgBW, 2 mg/kgBW and 4 mg/kgBW for 30 days while the control rats were not given any treatment. At the end of experiment, sperm analysis was carried out to measure the percentage of motile sperm and percentage of sperm with normal morphology. Results: Percentage of motile sperm significantly decreased (p<0.05) in all treatment groups compared to control group. Comparison between treatment groups revealed that the percentage of motile sperm significantly decreased in 0.5 mg/kgBW vs 2 mg/kgBW, 0.5 mg/kgBW vs 4 mg/kgBW, and 1 mg/kgBW vs 4 mg/kgBW. Percentage of normal morphology sperm decreased significantly (p<0.05) in 1 mg/kgBW, 2 mg/kgBW, and 4 mg/kgBW compared to control groups. Comparison between treatment groups revealed that there were no significant decreased between each treatment groups. Conclusion: This research conclude that nicotine exposure per inhalation can deteriorate the percentage of motile sperm and the percentage of normal morphology sperm of male wistar rats.

scholarly journals Testicular and epididymal histomorphometric assessment in F1 male Wistar rats after gestational Ricinus communis oil exposure

2015 ◽  
Vol 32 (03) ◽  
pp. 187-191 ◽  
Author(s):  
S. Salami ◽  
J. Omirinde ◽  
M. Balogun ◽  
Y. Raji
Keyword(s):  
Wistar Rats ◽  
Ricinus Communis ◽  
Male Rats ◽  
Seminiferous Tubules ◽  
Control Group ◽  
Critical Periods ◽  
Luminal Diameter ◽  
Male Wistar Rats ◽  
And Function ◽  
Tubular Diameter

Abstract Introduction: Fetal programming hypothesis presupposes that stimulus or insult acting during critical periods of uterine growth and development do alter tissue structure and function. In the present study changes in histomorphometric integrity of the testes and epididymis in adult F1 male rats maternally exposed to Ricinus communis oil (RCO) at different gestation periods were assessed. Materials and Methods: Therapeutic dose of RCO 950 mg/kg BW was administered to pregnant Wistar rats at gestation days GD 1-7, 7-14, 14-21 and 1-21 respectively. Testes and epididymis of adult male F1 offspring were then harvested for Histomorphometry assessment under Light microscope. Seminiferous tubular diameter (STD), seminiferous luminal diameter (SLD) and Seminiferous epithelial height (EH) of both peripheral and central seminiferous tubules were measured in the testes. Epididymal tubular diameter (ETD), epididymal luminal diameter (ELD) and epididymal epithelial height (EEH) were measured in epididymis. Results: Results were expressed as the mean ± SEM and significance taken at p<0.05. STD significantly (p<0.001) increased in F1 males from GD1-7, 7-14, 14-21 relative to control group. SEH significantly (p<0.001) decreased in F1 males from GD1-7,7-14 and 1-21. SLD increased significantly (p<0.001) in F1males from GD1-7, 7-14, 14-21 and 1-21. ETD significantly (p<0.01) increased in F1 males from GD1-7, 7-14, 14-21. EEH significantly (p<0.001) increased in F1 males from GD1-7 and GD7-14. ELD however increased significantly (p<0.001) only in F1 males from GD1-7. Conclusion: Maternal RCO exposure at different gestation periods impaired negatively histomorphometry of the testis and epididymis in male offspring.

scholarly journals Anti-fertility effect of hydro-alcoholic extract of fennel (Foeniculum vulgare Mill) seed in male Wistar rats

2016 ◽  
Vol 60 (3) ◽  
pp. 357-363 ◽  
Author(s):  
Esrafil Mansouri ◽  
Majid Asadi-Samani ◽  
Wesam Kooti ◽  
Maryam Ghasemiboroon ◽  
Damoon Ashtary-Larky ◽  
...  
Keyword(s):  
Wistar Rats ◽  
Sperm Count ◽  
Seed Extract ◽  
Male Rats ◽  
Control Group ◽  
Alcoholic Extract ◽  
Foeniculum Vulgare ◽  
Fertility Effect ◽  
Male Wistar Rats ◽  
The Right

AbstractIntroduction: The study aimed to investigate the anti-fertility effect of fennel (Foeniculim vulgare Mill) seed extract in male rats.Material and Methods: Forty Wistar rats were divided into five equal groups. The control group received distilled water and the experimental groups were orally administered 1 ml of hydro-alcoholic extract of fennel seed in four doses of 35, 70, 140, and 280 mg/kg/b.w. daily for 60 days. After the last gavage, the rats were anaesthetised and the caudal part of the right epididymis was used for sperm counting. After fixation of the testes, microscopic sections were prepared and histological changes were evaluated.Results: The number of spermatogonia after doses of 140 and 280 mg/kg and Sertoli cells after a dose of 140 mg/kg decreased significantly as compared with the control group (P < 0.05). The number of primary spermatocytes and sperm count decreased significantly in the experimental groups (70, 140, and 280 mg/kg) when compared to the control group (P < 0.05). Furthermore, thickening of the basement membrane, cell apoptosis, and irregular arrangement of the germinal epithelium were observed in the experimental groups.Conclusion: Hydro-alcoholic fennel seed extract at these doses could reduce reproductivity and has anti-fertility activity in male rats.

scholarly journals Histomorphological Effects of Cottonseed Oil on Testes in Adult Male Wistar Rats

2018 ◽  
Vol 25 (4) ◽  
Author(s):  
Lilian Ebele Chris-Ozoko ◽  
Onome Okpe ◽  
Wilson Josiah Iju ◽  
John Chukwuma Oyem
Keyword(s):  
Adult Male ◽  
Wistar Rats ◽  
Cottonseed Oil ◽  
Male Rats ◽  
Seminiferous Tubules ◽  
Control Group ◽  
Luminal Diameter ◽  
Orogastric Tube ◽  
Group I ◽  
Male Wistar Rats

Cottonseed oil is an extract from the seeds of cotton plant, that is rich in protein, carbohydrate and fiber.          The objective of the research was to study the effect of cottonseed oil on the histology of the testis.          Materials and methods. Twelve adult male Wistar rats weighing 200 – 250g, were randomly sorted into four groups (I - IV), each containing three animals. Cottonseed oil was administered orally to the experimental animals by means of an orogastric tube. The rats in Group I (the control group) received standard rat pellet and oral normal saline ad libitum only. The animals in Groups II, III, and IV received 0.2 ml, 0.6 ml and 1.0 ml of cotton seed oil per body weight respectively. On the 21st day, the animals were sacrificed by cervical dislocation. The testes were excised and, then, fixed with appropriate fixatives for routine tissue processing.          Results. The results revealed an increase in the luminal diameter of seminiferous tubules, a decrease in the Leydig cell population, the disorientation of spermatogenic series and their detachment from the germinal epithelium.          Conclusions. The present study demonstrated that oral administration of cottonseed oil to male rats displayed a deleterious effect on their testis by disrupting spermatogenesis and the histological architecture of the testis.

scholarly journals Evaluating the Effects of Noise Pollution on the Levels of Blood Cortisol, Testosterone, and Thyroid in Male Wistar Rats

Health Scope ◽  
2021 ◽  
Vol 10 (3) ◽  
Author(s):  
Seyed-Hosein Abtahi-Eivary ◽  
Ali Tajpoor ◽  
Ali Firoozi ◽  
Shahrzad Mehrzad ◽  
Mohammad Hosein Beheshti
Keyword(s):  
Wistar Rats ◽  
Statistical Tests ◽  
Noise Pollution ◽  
Well Being ◽  
Serum Levels ◽  
Sound Level ◽  
Male Rats ◽  
Control Group ◽  
Male Wistar Rats ◽  
Commercial Kits

Background: Noise pollution is a global problem causing changes in the secretion of various hormones and consequently affecting social well-being and quality of life in cities. Objectives: This study aimed to investigate the effect of noise pollution on the levels of testosterone, thyroid, and cortisol hormones in male rats. Methods: In this experimental study, a total of 70 male Wistar rats (200 - 250 g) were randomly assigned into one control and six experimental groups, with 10 rats in each group. Experimental groups were exposed to noise with different intensity (dB) and time (min) as follows: (I) 60 dB, 30 min; (II) 60 dB, 60 min; (III) 85 dB, 30 min; (IV) 85 dB, 60 min; (V) 110 dB, 30 min; (VI) 110 dB, 60 min; (VII) controls. Animals in the experimental groups were exposed to noise in an acoustic chamber designed for this purpose for 50 days. The Noise.exe software was used to generate noise, and the sound level meter (model TES 1358) was used to determine the accuracy of the intensity and frequency of sound. To determine plasma levels of the hormones, appropriate research and commercial kits were used, which were based on the ELISA method. To determine the concentration of hormones other than TSH, human assay kits were used. All statistical tests were performed in SPSS software version 21. Results: Serum levels of cortisol in the 110-dB (30 and 60 min), 65-dB (60 min), and 85-dB (60 min) groups were significantly higher than the control group (P ≤ 0.05). Also, cortisol levels in the 65-dB and 85-dB (30 minutes) groups were higher than the control group; however, the increase was not significant (P > 0.05). The levels of T4, T3, and TSH in the 60-dB and 85-dB groups were significantly lower than in the control group (P ≤ 0.05). The serum levels of T4, T3, and TSH hormones in the 110-dB group were insignificantly lower than the control group (P > 0.05). The serum level of testosterone in the 110-dB group was significantly lower than the control group (P ≤ 0.05). The mean serum levels of testosterone in the 65-dB and 85-dB groups were insignificantly lower than the control group (P > 0.05). Conclusions: Based on this study, exposure to noise pollution increased cortisol secretion and decreased T4, T3, TSH, and testosterone levels in rats. As this hormonal imbalance may create direct and indirect effects, studies and strategies are recommended to control the imbalance of hormones in the polluted environments.

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