Figure 1. UV–Vis spectroscopy for the synthesized silver nanoparticles.

Figure 1. UV–Vis spectroscopy for the synthesized silver nanoparticles.

Figure 2. FTIR spectroscopy for the synthesized silver nanoparticles.

Figure 2. FTIR spectroscopy for the synthesized silver nanoparticles.

Figure 3. (a) SEM analysis, (b) EDX analysis for the synthesized silver nanoparticles.

Figure 3. (a) SEM analysis, (b) EDX analysis for the synthesized silver nanoparticles.

Figure 4. TEM analysis for the synthesized silver nanoparticles.

Figure 4. TEM analysis for the synthesized silver nanoparticles.

Figure 5. XRD analysis for the synthesized silver nanoparticles.

Figure 5. XRD analysis for the synthesized silver nanoparticles.

Figure 6. Zeta Potential analysis for the synthesized silver nanoparticles.

Figure 6. Zeta Potential analysis for the synthesized silver nanoparticles.

Figure 7. Phenotypic changes observed in earthworms exposed to silver nanoparticles. (a) control; (b) exposed to 10 µg; (c) exposed to 20 µg (Black arrow indicates the brownish pink changes).

Figure 7. Phenotypic changes observed in earthworms exposed to silver nanoparticles. (a) control; (b) exposed to 10 µg; (c) exposed to 20 µg (Black arrow indicates the brownish pink changes).

Figure 8. Phenotypic changes observed in zebrafish exposed to silver nanoparticles. (a) control; (b) exposed to 10 µg; (c) exposed to 20 µg.

Figure 8. Phenotypic changes observed in zebrafish exposed to silver nanoparticles. (a) control; (b) exposed to 10 µg; (c) exposed to 20 µg.

Figure 9. Silver nanoparticle accumulation study using ICP–OES in earthworms.

Figure 9. Silver nanoparticle accumulation study using ICP–OES in earthworms.

Figure 10. Silver nanoparticle accumulation study using ICP–OES in zebrafishes.

Figure 10. Silver nanoparticle accumulation study using ICP–OES in zebrafishes.

Figure 11. Histology images of foregut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. G—Gut, E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM, green denotes arrow disruption of gut.

Figure 11. Histology images of foregut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. G—Gut, E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM, green denotes arrow disruption of gut.

Figure 12. Histology images of midgut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. G—Gut, E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM, green denotes arrow disruption of gut.

Figure 12. Histology images of midgut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. G—Gut, E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM, green denotes arrow disruption of gut.

Figure 13. Histology images of hindgut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM.

Figure 13. Histology images of hindgut region of earthworm—Eudrilus eugeniae (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day. E—Epidermis, LM—Longitudinal Muscle, CM—Circular Muscle. Black arrow denotes lipofuscin, red arrow denotes erosion of epidermis, blue arrow denotes interstitial space, yellow arrow denotes fibrosis of CM.

Figure 14. Histology images of eyes of Zebrafish—Danio rerio (10X Magnification) (a) 10th day Control, (b) 20th day Control, (c) exposed to 10 µg at 20th day, (d) 30th day Control. L–Lens, V–Vitreous.

Figure 14. Histology images of eyes of Zebrafish—Danio rerio (10X Magnification) (a) 10th day Control, (b) 20th day Control, (c) exposed to 10 µg at 20th day, (d) 30th day Control. L–Lens, V–Vitreous.

Figure 15. Histology images of gills of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 15. Histology images of gills of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 16. Histology images of intestines of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 16. Histology images of intestines of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 17. Histology images of liver of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 17. Histology images of liver of Zebrafish—Danio rerio (10X Magnification) (a) 10th day control, (b) exposed to 10 µg at 10th day, (c) exposed to 20 µg at 10th day; (d) 20th day control, (e) exposed to 10 µg at 20th day, (f) exposed to 20 µg at 20th day; (g) 30th day control, (h) exposed to 10 µg at 30th day, (i) exposed to 20 µg at 30th day.

Figure 18. Microscopic images of Zebrafish—Danio rerio embryos (a) control at 0th h, (b) exposed to 10 µg at 0th h, (c) exposed to 20 µg at 0th h, (d) control at 24 hpf, (e) exposed to 10 µg at 24 hpf, (f) exposed to 20 µg at 24 hpf, (g) control at 48 hpf, (h) exposed to 10 µg at 48 hpf, (i) exposed to 20 µg at 48 hpf. White arrow denotes dead embryos (coagulation); Yellow arrows denote undeveloped embryos.

Figure 18. Microscopic images of Zebrafish—Danio rerio embryos (a) control at 0th h, (b) exposed to 10 µg at 0th h, (c) exposed to 20 µg at 0th h, (d) control at 24 hpf, (e) exposed to 10 µg at 24 hpf, (f) exposed to 20 µg at 24 hpf, (g) control at 48 hpf, (h) exposed to 10 µg at 48 hpf, (i) exposed to 20 µg at 48 hpf. White arrow denotes dead embryos (coagulation); Yellow arrows denote undeveloped embryos.

Table 1. Phenotypic changes observed in earthworms exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Table 1. Phenotypic changes observed in earthworms exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Parameter AnalyzedControl10 µg20 µg BeforeAfterBeforeAfterBeforeAfterColor changeBrownish blackBrownish blackBrownish blackBrownish pinkBrownish blackBrownish pinkBehavioral changeNilNilNilNilNilNil

Table 2. Total count of earthworms exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Table 2. Total count of earthworms exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Conc.Before10th day20th day30th dayControl1022275110 µg1017221420 µg1017139

Table 3. Phenotypic changes observed in zebrafish exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Table 3. Phenotypic changes observed in zebrafish exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Parameter AnalyzedControl10 µg20 µg BeforeAfterBeforeAfterBeforeAfterColor changeNilNilNilYellowish tint in the gills and finsNilYellowish tint in the gills and finsBehavioral changeActiveActiveActiveWeak and slowActiveWeak and slowGrowthNormalNormalNormalInhibition in growthNormalInhibition in growth

Table 4. Total count of zebrafishes exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Table 4. Total count of zebrafishes exposed to silver nanoparticles synthesized using Bauhinia purpurea gum.

Conc.Before10th Day20th Day30th DayControl1010101010 µg10109920 µg10988

Table 5. Apical observations observed for zebrafish embryos.

Table 5. Apical observations observed for zebrafish embryos.

Control10 µg20 µg0th h24 hpf48 hpf0th h24 hpf48 hpf0th h24 hpf48 hpfHatchingNANA+++NANA++NANA+Coagulated embryos-+--+++-+++Somite formation+++++++++HeartbeatNANA++NANA++NANA+