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Azomite is a hydrated calcium sodium aluminosilicat rich in rare earth elements. To investigate the dietary effects of Azomite on growth, intestine microbiota and morphology, immunohematological changes and disease resistance, seven diets with Azomite supplementation of 0 (the control), 1.0, 2.0, 3.0, 4.0, 5.0 and 6.0 g/kg (A0, A1, A2, A3, A4, A5, A6), were prepared and fed to largemouth bass, Micropterus salmoides (7.96 ± 0.19) for 60 days. The results revealed that the weight gain (WG) increased first and then decreased with the increasing dietary Azomite, and the A2 group presented the highest WG and lowest feed conversion ratio among all the groups. The supplementation of 2.0 g/kg Azomite significantly increased the intestine protease activity, the crude protein of whole body and protein retention (P < 0.05), and high inclusion of Azomite (6.0 g/kg) significantly reduced the lipid retention (P < 0.05). The amounts of red blood cells in A5, A6 groups, white blood cells in A3, A5, A6 groups and lymphocyte in A2-A6 groups were all significantly higher than those in the control group (P < 0.05). In addition, serum superoxide dismutase and catalase activities in A5, A6 groups, and serum alkaline phosphatase and lysozyme activities in A2-A4 groups showed significantly higher values than the control group (P < 0.05). Intestinal microbiota analysis indicated that the Tenericutes abundance was increased, whereas Proteobacteria abundance was decreased in all Azomite supplemented groups. The villus height in A2-A4 groups, and the villus width in A2 group were significantly higher than those of the control group (P < 0.05). The cumulative mortality was reduced by the addition of 2.0-5.0 g/kg Azomite after challenging with A. hydrophila (P < 0.05). In conclusion, proper addition of Azomite in diets improved the growth, intestine morphology, immune response and disease resistance in largemouth bass, and the optimal inclusion was estimated to be 2.0-3.0 g/kg diet., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

There is a current interest from the food packaging, biomedical and agricultural sectors in hybrid materials formed from clays and natural polymeric compounds. However, research investigating the toxicity of vermiculite-cellulose nanocrystal (VERN) hybrid on the testes of Wistar rats is rare. Twenty rats, divided into control and treatment groups, were orally administered distilled water, 5, 10, and 20 mg/kg bw VERN daily for two consecutive weeks. At the termination of experiments, the testicular organo-somatic index, superoxide dismutase, catalase, and gamma-glutamyl transferase activities were not significantly changed by VERN relative to the controls. Contrarily, myeloperoxidase, glutathione peroxidase, and malondialdehyde levels were depleted in the testes of treated rats. Moreso, VERN increased follicle-stimulating and luteinizing hormones, and decreased testosterone levels at the 20 mg/kg dose. Histology of the testes revealed healthy looking Leydig cells at the doses of 5 and 10 mg/kg VERN. Overall, these results indicate that oral exposure of VERN was not overly deleterious to the redox and structural histoarchitecture in the testes of rats., (© 2019 Blackwell Verlag GmbH.)

The present experiment was conducted to evaluate the effect of dietary supplementation with some feed additives (potassium sorbate; Sor, hydrated sodium calcium almuniosilicate; Hsc and L-methionine; L-M) against aflatoxin B1 (AF) toxicity in rabbits. A total of 72 growing rabbits (5-week-old) were distributed into six equal groups (4 replicates with 3 rabbits each). The experimental groups are as follows: control group, AF group (supplemented with AF 0.3 mg/kg diet), AF + Sor group (AF 0.3 mg/kg diet + Sor 2 g/kg diet), AF + Hsc group (AF 0.3 mg/kg diet + Hsc 5 g/kg diet), AF + L-M group (AF 0.3 mg/kg diet + L-M 8 g/kg diet) and AF + Mix group (AF 0.3 mg/kg diet + 2 Sor + 5 Hsc + 8 L-M g/kg diet). Live body weight and weight gain at 13 weeks of age were significantly reduced by AF. Feed intake at 13 weeks of age was decreased in AF, AF + Hsc and AF + Mix compared to the control. AF, AF + Hsc and AF + Mix showed the lowest total antioxidant capacity compared to the control. The highest level of reactive oxygen species and 8-Hydroxy-2-desoxyguanosine was observed in AF group. Using of other supplements with AF increased immunoglobulinM than AF alone. In conclusion, dietary supplementation of Sor, L-M, Hsc or their mixture was effective in reducing the adverse effects of AF on performance, antioxidant and immune status of rabbits with more better improvement obtained by Sor or L-M separately., (© 2019 Blackwell Verlag GmbH.)

Mitigation strategies are vital in minimizing the health and economic risks associated with dairy cattle exposure to aflatoxin (AF). The objective of this study was to determine the effects of a commercially available aluminosilicate clay in a lactation diet on production responses, blood chemistry, and liver inflammatory markers of multiparous lactating Holstein cows during an AF challenge. Sixteen multiparous lactating Holstein cows [body weight (mean ± SD) = 758 ± 76 kg; days in milk = 157 ± 43 d] were assigned to 1 of 4 treatments in a replicated 4 × 4 Latin square design with 21-d periods: no adsorbent and no AF challenge (CON), no adsorbent and an AF challenge (POS), 113 g of aluminosilicate clay top-dressed on the total mixed ration (adsorbent; FloMatrix, PMI Nutritional Additives, Arden Hills, MN) with an AF challenge (F4), or 227 g of adsorbent with an AF challenge (F8). The challenge consisted of 100 μg of AFB 1 /kg of dietary dry matter intake administered orally. For each period, milk was sampled 3× daily from d 14 to 21; blood, feces, and urine were sampled on d 14, 18, and 21; and liver samples were taken on d 18. Liver tissue was assessed for gene expression and histological hepatocyte inflammation. Statistical analysis was preformed using the MIXED and GLIMMIX procedures of SAS (SAS Institute Inc., Cary, NC). Fat-corrected milk (POS = 37.2, F4 = 39.2, and F8 = 38.9 kg/d) increased as concentration of adsorbent in the diet increased. There was a decrease in milk AFM 1 concentration at d 18 as concentration of adsorbent in the diet increased (POS = 0.33, F4 = 0.32, and F8 = 0.27 µg/kg). There was a decrease in AFM 1 concentration in urine (POS = 2.10, F4 = 1.89, and F8 = 1.78 µg/kg) and AFB 1 concentration in feces (POS = 4.68, F4 = 3.44, and F8 = 3.17 µg/kg) as concentration of adsorbent in the diet increased. Cows in CON had greater concentrations of serum cholesterol (202 mg/dL) and plasma superoxide dismutase (2.8 U/mL) compared with cows in POS (196 mg/dL and 2.6 U/mL, respectively). Plasma glutamate dehydrogenase increased as concentration of adsorbent in the diet increased (POS = 37.8, F4 = 39.3, and F8 = 39.1 U/L). The expression of NFKB1 was greater in the liver of cows in POS (0.78) compared with cows in CON (0.70). The expression of MTOR was greater in the liver of cows in CON (1.19) compared with cows in POS (0.96). When compared with cows in CON, cows in POS had greater odds ratio for hepatocyte inflammation (odds ratio = 5.14). In conclusion, the adsorbent used in this study had a positive effect on milk production and hepatocyte inflammation and reduced AF transfer., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Halloysite (Al 2 Si 2 O 5 (OH) 4 ·nH 2 O) nanotubes (HNTs) are natural clay materials and widely applied in many fields due to their natural hollow tubular structures. Many in vitro studies indicate that HNTs exhibit a high level of biocompatibility, however the in vivo toxicity of HNTs remains unclear. The objective of this study was to assess the hepatic toxicity of the purified HNTs in mice via oral route. The purified HNTs were orally administered to mice at 5, 50, and 300 mg/kg body weight (BW) every day for 30 days. Oral administration of HNTs stimulated the growth of the mice at the low dose (5 mg/kg BW) with no liver toxicity, but inhibited the growth of the mice at the middle (50 mg/kg BW) and high (300 mg/kg BW) doses. In addition, oral administration of HNTs at the high dose caused Al accumulation in the liver but had no marked effect on the Si content in the organ. The Al accumulation caused significant oxidative stress in the liver, which induced hepatic dysfunction and histopathologic changes. These findings demonstrated that Al accumulation-induced oxidative stress played an important role in the oral HNTs-caused liver injury., (© 2018 Wiley Periodicals, Inc.)

The study was conducted to examine the effect of supplementing bentonite clay with or without a Saccharomyces cerevisiae fermentation product (SCFP; 19 g of NutriTek + 16 g of MetaShield, both from Diamond V, Cedar Rapids, IA) on the performance and health of dairy cows challenged with aflatoxin B 1 (AFB 1 ). Twenty-four lactating Holstein cows (64 ± 11 d in milk) were stratified by parity and milk production and randomly assigned to 1 of 4 treatment sequences. The experiment had a balanced 4 × 4 Latin square design with 6 replicate squares, four 33-d periods, and a 5-d washout interval between periods. Cows were fed a total mixed ration containing 36.1% corn silage, 8.3% alfalfa hay, and 55.6% concentrate (dry matter basis). Treatments were (1) control (no additives), (2) toxin (T; 1,725 µg of AFB 1 /head per day), (3) T + clay (CL; 200 g/head per day; top-dressed), and (4) CL+SCFP (CL+SCFP; 35 g/head per day; top-dressed). Cows were adapted to diets from d 1 to 25 (predosing period) and then orally dosed with AFB 1 from d 26 to 30 (dosing period), and AFB 1 was withdrawn from d 31 to 33 (withdrawal period). Milk samples were collected twice daily from d 21 to 33, and plasma was sampled on d 25 and 30 before the morning feeding. Transfer of ingested AFB 1 into milk aflatoxin M 1 (AFM 1 ) was greater in T than in CL or CL+SCFP (1.65 vs. 1.01 and 0.94%, respectively) from d 26 to 30. The CL and CL+SCFP treatments reduced milk AFM 1 concentration compared with T (0.45 and 0.40 vs. 0.75 µg/kg, respectively), and, unlike T, both CL and CL+SCFP lowered AFM 1 concentrations below the US Food and Drug Administration action level (0.5 µg/kg). Milk yield tended to be greater during the dosing period in cows fed CL+SCFP compared with T (39.7 vs. 37.7 kg/d). Compared with that for T, plasma glutamic oxaloacetic transaminase concentration, indicative of aflatoxicosis and liver damage, was reduced by CL (85.9 vs. 95.2 U/L) and numerically reduced by CL+SCFP (87.9 vs. 95.2 U/L). Dietary CL and CL+SCFP reduced transfer of dietary AFB 1 to milk and milk AFM 1 concentration. Only CL prevented the increase in glutamic oxaloacetic transaminase concentration, and only CL+SCFP prevented the decrease in milk yield caused by AFB 1 ingestion., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Populations consuming aflatoxin (AF) and fumonisin (FN)-contaminated foods may be at increased risk for hepatocellular carcinoma (HCC) and developmental disorders; consequently, development of intervention strategies to reduce AF/FN-induced liver disease and adverse health effects in humans could be very useful. Calcium montmorillonite clay (NovaSil) has been shown to absorb AF in vitro, in multiple animal models, as well as in human studies. In the present study, we aimed to evaluate whether uniform particle size NovaSil (UPSN) possessed an ability to modulate the co-carcinogenic potentials of aflatoxin B 1 (AFB 1 ) and fumonisin B 1 (FB 1 ) in F344 rats. Sequential treatment of FB 1 following AFB 1 synergistically induces preneoplastic alterations as well as liver damage, indicating that AFB 1 acts as an initiator while FB 1 as a promoter in the carcinogenesis model, confirming findings from previous studies. The enterosorbent agent UPSN clay at dose of up to 0.5% in diet was shown to be effective in modulating the toxicity and carcinogenicity of co-exposure to AFB 1 and FB 1 , as demonstrated by significant reduction in number and size of hepatic GST-P + foci, in alterations indicative of liver toxicity, and in levels of AFB 1 and FB 1 biomarkers., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The effect of Sodium Bentonite (SB) enriched diet on growth performance, innate immune response, and disease resistance in stinging catfish, Heteropneustes fossilis against Aeromonas hydrophila is reported. The infected fish fed with 5% SB had the maximum weight gain diet (PWG %) and specific growth rate (SGR %) were 26% and 29% when compared to 14% and 17% with 10% diet. Similarly the phagocytic activity increased significantly when infected fish were fed with 5% or 10% SB diets during the experimental period; the complement, respiratory burst and lysozyme activities were also significantly enhanced on weeks 2 and 4. The lower cumulative mortality (10% and 15%) was observed when the infected fish were fed with 5% and 10% SB diets for 30 days. The results suggest that the infected H. fossilis after administration of 5% and 10% SB enriched diets for 30 days had significantly improved growth performance, innate immunity, and disease resistance against A. hydrophilla. Hence, sodium bentonite can be used as a feed additive to stimulate immunity and for disease resistance in the effective production of economically valuable freshwater catfish, H. fossilis., (Crown Copyright © 2018. Published by Elsevier Ltd. All rights reserved.)

A presence of mycotoxins in feed is one of the most alarming issues in the poultry feed industry. Ochratoxins, produced by several Aspergillus and Penicillium species, are important mycotoxin regarding the health status of poultry birds. Ochratoxins are further classified into to several subtypes (A, B, C, etc) depending on their chemical structures, but ochratoxin A (OTA) is considered the most important and toxic. Bentonite clay, belonging to phyllosilicates and formed from weathering of volcanic ashes, has adsorbent ability for several mycotoxins. The present study was designed to study the effects of bentonite clay upon OTA-induced immunosuppression in broiler chicks. For this, 480 day-old broiler chicks were procured from a local hatchery and then different combinations of OTA (0.15, 0.3, or 1.0 mg/kg) and bentonite clay (5, 10, and 20 g/kg) were incorporated into their feed. At 13, 30, and 42 days of age, parameters such as antibody responses to sheep red blood cells, in situ lymphoproliferative responses to mitogen (PHA-P), and in situ phagocytic activity (i.e., via carbon clearance) were determined respectively. The results indicated there was a significant reduction of total antibody and immunoglobulin titres, lymphoproliferative responses, and phagocytic potential in OTA-treated birds, suggesting clear immunosuppression by OTA in birds in a dose-dependent manner. These results were also significantly lower in all combination groups (OTA with bentonite clay), suggesting few to no effects of feeding bentonite clay upon OTA- induced alterations in different immune parameters.

Purpose: To study the safety and biocompatibility of Laponite clay (LAP) within an intravitreal and suprachoroidal administration in rabbit eyes., Methods: Thirty-two New Zealand albino rabbits were divided into two experimental groups to test intravitreal (IVT group) and suprachoroidal (SCS group) administration of a 100-μl and 50-μl Laponite suspension respectively. Following injection, the eyes were monitored by ocular tonometry, slit-lamp eye examination and indirect ophthalmoscopy, at 24 h, 1, 4, 12, and 14 weeks post administration. Histological examination was also performed to determine whether any ocular pathological change had occurred. Throughout the study, LAP presence in vitreous was estimated by complexometric titration with ethylenediaminetetraacetic acid (EDTA), taking advantage of the Laponite high content of magnesium ions., Results: Neither significant differences in the intraocular pressure, nor relevant ocular complications were found in the two experimental groups after LAP administration. The histology of the retina remained unchanged. LAP presence in vitreous could be indirectly confirmed by complexometric titration until 14 weeks post administration in eyes of IVT group., Conclusion: Laponite could be considered as a vehicle for potential clinical use in ocular drug administration, due to its proven ocular biocompatibility and its transparency in gel state.

Three experiments were conducted to determine the effect of increasing concentrations of a smectite clay on toxin binding capacity, ruminal fermentation, diet digestibility, and growth of feedlot cattle. In Exp. 1, 72 Angus × Simmental steers were blocked by BW (395 ± 9.9 kg) and randomly allotted to 3 treatments (4 pens/treatment and 6 steers/pen) to determine the effects of increasing amounts of clay (0, 1, or 2%) on performance. The clay was top-dressed on an 80% concentrate diet at a rate of 0, 113, or 226 g/steer daily to achieve the 0, 1, and 2% treatments, respectively. Steers were slaughtered at a target BW of 606 kg. In Exp. 2, 6 steers (596 ± 22.2 kg initial BW) were randomly allotted to the same 3 treatments in a replicated 3 × 3 Latin square design (21-d periods) to determine the effects of increasing amounts of clay on ruminal pH, VFA, and nutrient digestibility. In Exp. 3, 150 mg of clay was incubated in 10 mL of rumen fluid with 3 incremental concentrations (6 replicates per concentration) of aflatoxin B (AFB) or ergotamine tartate (ET) to determine binding capacity. During the first 33-d period, there was a quadratic effect of clay on ADG ( < 0.01) and G:F ( < 0.01), increasing from 0 to 1% clay and then decreasing from 1 to 2% clay. However, during the second 30-d period, clay linearly decreased ADG and G:F ( ≤ 0.03) and overall ADG, DMI, and G:F were not impacted ( ≥ 0.46). Clay linearly decreased marbling score ( = 0.05). Hepatic enzyme activity did not differ among treatments on d 0 or at slaughter ( ≥ 0.15). Clay linearly decreased ruminal lactate and propionate, linearly increased formate and the acetate:propionate ratio ( ≤ 0.04), and tended ( = 0.07) to linearly increase butyrate. Clay tended to linearly increase ( = 0.06) OM and CP apparent digestibility. Ruminal pH, urine pH, and other digestibility measures did not differ among treatments ( ≥ 0.15). Clay was able to effectively bind AFB and ET at concentrations above the normal physiological range (52 and 520 μg/mL), but proportional adsorption was decreased to 35.5 and 91.1% at 5,200 μg/mL ( < 0.01) for AFB and ET, respectively. In conclusion, clay effectively binds ruminal toxins, decreases ruminal lactate, and improves performance only during adaptation to a high-concentrate feedlot diet.

Quaternized carboxymethyl chitosan/organic rectorite (QCMC/OREC) nanocomposites were rapidly prepared by intercalating QCMC into the layer of OREC under microwave irradiation. And then copper-chelate QCMC/OREC (QCMC/OREC-Cu) nanocomposites were obtained by mixing QCMC/OREC with CuSO4 solution. XRD and TEM results indicated that QCMC/OREC nanocomposites were obtained and QCMC was dispersed in the interlayer of OREC. Besides, FT-IR results revealed that the hydrogen bonds and electrostatic interaction in QCMC/OREC-Cu were both stronger than those in QCMC/OREC because of introducing the Cu(2+). The thermogravimetric analysis showed that the thermal stability of QCMC/OREC-Cu nanocomposites was higher than QCMC and QCMC/OREC. Algae inhibition assay revealed that QCMC/OREC-Cu nanocomposites had stronger antifouling activity than original QCMC and QCMC/OREC. This work provides important basis for developing novel antifouling materials., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Oral supplementation of clay has been reported to function as buffer in dairy cows. However, its effects on rumen, blood, and fecal pH have varied among studies. Our objective was to determine the effects of 3 concentrations of dietary clay supplementation after a grain challenge. Ten multiparous rumen-cannulated Holstein cows [body weight (mean ± standard deviation)=648±12kg] with 142±130 (60 to 502) days in milk were assigned to 1 of 5 treatments in a replicated 5×5 Latin square design balanced to measure carryover effects. Periods (21d) were divided into an adaptation phase (d 1 to 18, with regular total mixed ration fed ad libitum) and a measurement phase (d 19 to 21). Feed was restricted on d 18 to 75% of the average of the total mixed ration fed from d 15 to 17 (dry matter basis), and on d 19 cows received a grain challenge. The challenge consisted of 20% finely ground wheat administered into the rumen via a rumen cannula, based on the average dry matter intake obtained on d 15 to 17. Treatments were POS (no clay plus a grain challenge), 3different concentrations of clay (0.5, 1, or 2% of dietary dry matter intake), and control (C; no clay and no grain challenge). Statistical analysis was performed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Contrasts 1 (POS vs. C) and 2 (POS vs. the average of 0.5, 1, or 2%) were compared, along with linear and quadratic treatment effects. Rumen, fecal, and blood pH, along with blood metabolites, were measured at 0, 4, 8, 12, 16, 20, 24, 36, and 48h relative to the grain challenge. Cows fed POS had lower rumen pH [(mean ± standard error) 6.03±0.06] than cows fed C (6.20±0.06). Cow fed POS had lower fecal pH (6.14±0.04) than cows fed C (6.38±0.04). We observed a linear treatment effect for rumen pH and fecal pH. Fecal pH (6.22±0.04) was higher for cows fed clay (contrast 2) then for cows fed POS (6.14±0.04). We also observed a treatment difference (contrast 2) for negative incremental area under the curve, pH below 5.6 × h/d, (0.5% clay=7.93±0.83, 1% clay=8.56±0.83, and 2% clay=7.79±0.83) compared with POS (11.0±0.83). Cows fed clay tended to have higher milk yield (0.5% clay=28.8±3.4kg, 1% clay=30.2±3.4kg, and 2% clay=29.1±3.4kg, contrast 2), and had higher 3.5% fat-corrected milk (0.5% clay=29.9±3.5kg, 1% clay=34.1±3.5kg, and 2% clay=33.1±3.4kg), and higher energy-corrected milk (0.5% clay=29.1±3.3kg, 1% clay=32.8±3.4kg, and 2% clay=31.6±3.3kg) than cows fed POS (27.7±3.4kg, 28.0±3.4kg, 27.7±3.3kg, respectively). In conclusion, cows fed clay had higher rumen pH, energy-corrected milk, fat-corrected milk, and a trend for milk yield than cows fed POS., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background and Introduction: Previous studies of soil eating, or geophagia, among pregnant women in sub-Saharan Africa indicate the practice is widespread. Various explanations have been explored to explain the global phenomenon of soil eating, with the most compelling explanation focused on clay's ability to prevent or treat intestinal infection. The urban South African context for clay eating is not well understood. This paper explores clay consumption amongst pregnant migrants who are nationals of countries where clay consumption may be common., Methods: I conducted in-depth interviews with a purposively selected group of Somali, Congolese, and Zimbabwean women (n = 23). Interviews included questions broadly related to maternal and infant nutrition. In addition, I conducted nine focus group discussions (n = 48) with adult Somali, Congolese (DRC), and Zimbabwean men (N = 3) and women (N = 6), segregated by country of origin and gender. This paper focuses specifically on responses related to geophagia., Results: While Somali women did not report consuming clay or charcoal, Congolese and Zimbabwean participants self-reported commonly consuming clay during pregnancy, and at times also when not pregnant. Despite having heard public health messaging that discouraged the practice, participants largely did not describe this consumption in terms of health, but rather in terms of craving and habit. Participants described continued consumption of clay in South Africa, and the only reason for ceasing consumption was in cases of severe constipation., Discussion: The widespread consumption of clay soil by Congolese and Zimbabwean women during pregnancy may be a mechanism through which identity was reasserted and reproduced in a foreign country. Participants' emphasis on clay consumption seemed related to the absence or expense of other craved foods, and perhaps also to feelings of loss in Cape Town., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Three experiments were conducted showing rats' pica behavior (kaolin clay intake) due to running in activity wheels. The amount of kaolin consumed was a positive function of the available time of voluntary running (20, 40, or 60 min), although this relationship was blunted by a descending (i.e., 60 → 40 → 20 min) test series of execution (Experiment 1). Pica was also generated by forced running in a motorized wheel for 60 min as a positive function of the speed of wheel rotations at 98, 185, or 365 m/h, independent of the order of execution (Experiment 2). Voluntary running generated more pica than did forced running at 80 m/h, although the distance travelled in the former condition was 27% lesser than that in the latter condition (Experiment 3). Because kaolin intake is regarded as a reliable measure of nausea in rats, these results show that wheel running, either voluntary or forced, induces nausea in rats., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

High contamination by aflatoxin B1 (AFB1) has been detected in Beja province (Tunisia) in many dairy products and animal feed, which has resulted in many tons of cereals and cereals being removed from the market, causing economic loss. While removal represents a means of reducing risk, exposures still occur. Studies have increasingly focused on means of AFB1 biodegradation/elimination using lactic acid bacteria and clay mineral. In the study here, Lactobacillus paracasei BEJ01 (LP) and montmorilonite clay (MT) were used to reduce the physio-/immunotoxicologic disorders that could develop in rats that underwent AFB1 exposures for a total of 7 consecutive days. The results indicated that rats treated with AFB1 (80 μg/kg BW) alone had significant decreases in lymphocytes in their blood (including B-lymphocytes, CD3(+), CD4(+), and CD8(+) T-lymphocyte subtypes, and NK cells), immunoglobulins (IgA and IgG) and pro-inflammatory cytokines; these rats also had altered oxidative stress status. In contrast, in rats treated with LP + MT (2 × 10(9) cfu/ml [∼ 2 mg/kg] + 0.5 mg MT/kg BW) for a total of 7 days before, concurrent with or after AFB1 treatment, there was a significant blockade/mitigation of each AFB1-impacted parameter. Moreover, treatment with the mixture at any point in relation to AFB1 treatment expectedly caused enhanced TNFα and IL-1β expression relative to control values; all other parameters were comparable to values noted in control rats. Alone, the mixture had no impact on host parameters. From the results here it may be concluded the the LP + MT mixture was effective in protecting these hosts against AFB1-induced immunologic/physiologic disorders and that LP + MT could prevent and/or mitigate AFB1 toxicities in vivo.

South Texas currently has the highest incidence of hepatocellular carcinoma (HCC) in the United States, a disease that disproportionately affects Latino populations in the region. Aflatoxin B1 (AFB1) is a potent liver carcinogen that has been shown to be present in a variety of foods in the United States, including corn and corn products. Importantly, it is a dietary risk factor contributing to a higher incidence of HCC in populations frequently consuming AFB1-contaminated diets. In a randomised double-blind placebo controlled trial, we evaluated the effects of a 3-month administration of ACCS100 (refined calcium montmorillonite clay) on serum AFB1-lysine adduct (AFB-Lys) level and serum biochemistry in 234 healthy men and women residing in Bexar and Medina counties, Texas. Participants recruited from 2012 to 2014 received either a placebo, 1.5 g or 3 g ACCS100 each day for 3 months, and no treatment during the fourth month. Adverse event rates were similar across treatment groups and no significant differences were observed for serum biochemistry and haematology parameters. Differences in levels of AFB-Lys at 1, 3 and 4 months were compared between placebo and active treatment groups. Although serum AFB-Lys levels were decreased by month 3 for both treatment groups, the low dose was the only treatment that was significant (p = 0.0005). In conclusion, the observed effect in the low-dose treatment group suggests that the use of ACCS100 may be a viable strategy to reduce dietary AFB1 bioavailability during aflatoxin outbreaks and potentially in populations chronically exposed to this carcinogen., Competing Interests: Statement No potential conflict of interest was reported by the authors.

Aims: To investigate the mechanisms leading to an increase in the prevalence of blaCMY -2 conferring resistance to ceftiofur in pigs receiving a feed medicated with chlortetracycline and penicillin, and to examine the effect of supplementation with a clay mineral on this phenomenon., Methods and Results: In 138 blaCMY -2 -positive Escherichia coli isolates from faeces of pigs receiving feed supplemented or not with 2% clinoptilolite, from day 2 to day 28 after weaning, isolates from the two groups differed significantly with respect to their phylogenetic group: phylotype A predominated in the supplemented group, whereas phylotypes B1 and D predominated in the control group, as determined by PCR. In 36 representative isolates, pulsed-field gel electrophoresis and antimicrobial susceptibility testing revealed that the blaCMY -2 -positive E. coli isolates were polyclonal with diverse antimicrobial resistance patterns and blaCMY -2 -carrying plasmids of incompatibility (Inc) groups, A/C, I1 and ColE were observed in transformants as detected by PCR. Enterobacter cloacae possessing blaCMY -2 -carrying IncA/C plasmids were found in the pens before introduction of this batch of pigs. The blaCMY -2 -positive E. coli isolates were more clonally diverse in the control group than the supplemented group., Conclusions: The blaCMY -2 gene appears to have spread both horizontally and clonally in this batch of pigs and may have spread from previous batches of pigs via plasmids carried by Ent. cloacae and expanded in animals of the present batch in the presence of the selection pressure due to administration of chlortetracycline and penicillin in the feed. Feed supplementation may have an effect on clonal diversity of blaCMY -2 -positive isolates., Significance and Impact of the Study: Implementation of improved hygiene measures, decreased administration of certain antimicrobials on farm and feed supplementation with certain ingredients may limit antimicrobial resistance spread between and within batches of animals., (© 2016 The Society for Applied Microbiology.)

Mica, an aluminosilicate mineral, has been proven to possess anti-tumor and immunostimulatory effects. However, its efficacy and mechanisms in treating various types of tumor are less verified and the mechanistic link between anti-tumor and immunostimulatory effects has not been elucidated. We sought to investigate the therapeutic effect of STB-HO (mica nanoparticles) against one of the most prevalent cancers, the breast cancer. STB-HO was orally administered into MCF-7 xenograft model or directly added to culture media and tumor growth was monitored. STB-HO administration exhibited significant suppressive effects on the growth of MCF-7 cells in vivo, whereas STB-HO did not affect the proliferation and apoptosis of MCF-7 cells in vitro. To address this discrepancy between in vivo and in vitro results, we investigated the effects of STB-HO treatment on the interaction of MCF-7 cells with macrophages, dendritic cells (DCs) and natural killer (NK) cells, which constitute the cellular composition of tumor microenvironment. Importantly, STB-HO not only increased the susceptibility of MCF-7 cells to immune cells, but also stimulated the immunocytes to eliminate cancer cells. In conclusion, our study highlights the possible role of STB-HO in the suppression of MCF-7 cell growth via the regulation of interactions between tumor cells and anti-tumor immune cells.

Gamma-aminobutyric acid (GABA) is a key neurotransmitter where it usually inhibits impulse transmission. GABA release blockage or postsynaptic reaction were determined to provoke epileptic convulsions. The aim of the present study was the development of brain-targeted, nanosized, nontoxic, biocompatible, highly specific formulations. Incorporation of GABA into halloysite nanotubes (HNT) was performed using different methods. Particle size, zeta potential and pH measurements, morphological, thermal, XRD, FTIR analyses and GABA quantification by validated HPLC method were used for the characterization of the systems prepared. Release pattern of GABA from the nanotubes was determined using a dialysis membrane. Following successful incorporation of GABA into HNTs for brain delivery, nanotube formulation coded HNT-GABA H1 was selected for in vivo studies. Smaller particle size with narrow size distribution, possible HNT-GABA interaction indicated by thermal, XRD and FTIR analyses and prolonged release were the parameters considered in this selection. Moreover, HNT-GABA H1 remained stable for 3-month storage period and showed higher cell viability values than GABA. Rats were used in in vivo studies and potential of anticonvulsant effect of GABA was determined in the pentylenetetrazole model of seizure. HNT-GABA H1 was found to increase latency of seizure, decrease ending time of the convulsion, duration of severe convulsion and mortality rate significantly compared to pure GABA. After administration of HNT-GABA H1, GABA concentration in Stratum corsatum measured by enzyme immune assay showed that it was not significantly higher than GABA administered alone. These findings suggest that GABA loaded HNTs reduces the duration of all phases of convulsion indicating efficient delivery of GABA to all brain areas to interfere with epileptic mechanism., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Halloysite is a clay mineral with chemical similarity to kaolin, a pharmaceutical ingredient. It consists of mainly aluminosilicate nanotubular particles in the size range of ∼ 200-1000 nm. Many studies have tried to empirically explore this novel clay for its potential in drug delivery systems but no work has yet studied its cytotoxicity from the perspective of oral drug delivery system. In this study, the halloysite nanotubes (HNTs) were subjected to size distribution analyses, which reveal more than 50% of nanotubes in the size range of 500 nm and rest mainly in the sub micrometer range. HNTs were then evaluated for in-vitro cytotoxicity against HCT116 (colorectal carcinoma) and HepG2 (hepatocellular carcinoma) cells which represent the earliest entry point and the first accumulating organ, respectively, for nanoparticles en-route to systemic circulation after oral delivery. Moreover, HNTs were tested for their cytogenetic toxicity against human peripheral blood lymphocytes. Both these results collectively indicated that HNTs are generally safe at practical concentrations of excipients for oral dosage forms., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Twenty 10-month-old Japanese Black steers were used to evaluate the effects of clay on fat necrosis and carcass characteristics. Ten steers (Clay group) were fed the clay (50 g/day) during 10-30 months of age. The other 10 steers (Control group) were not fed it. There was no significant difference in body weight or average daily gain between the two groups (P > 0.05). The occurrence of fat necrotic mass in the Clay group (30%) was lower (P < 0.05) than that in the Control group (90%) at slaughter. The size of necrotic masses in the Clay group was smaller (P < 0.05) than that in the Control group. There was no significant difference in the marbling score, beef color, Longissimus muscle area or subcutaneous fat thickness between the two groups. These results suggest that the clay prevented the occurrence of fat necrosis and did not affect the carcass characteristics in Japanese Black steers., (© 2015 Japanese Society of Animal Science.)

Introduction: Quick-Set (Primus Consulting, Bradenton, FL) is a calcium aluminosilicate cement that is a potential alternative to mineral trioxide aggregate (MTA) with greater acid resistance and faster setting. The purpose of this study was to compare the effects of Quick-Set and MTA on pulpal tissues in response to pulpotomy procedures., Methods: The pulp chambers of 42 maxillary teeth in 7 beagle dogs were accessed, and the coronal pulpal tissue was removed. Pulpotomy procedures were performed, placing the experimental materials directly over the radicular pulp tissues. The dogs were sacrificed at 70 days, and the teeth and surrounding tissues were removed and prepared for histologic analysis. The sections of the pulpotomy areas were scored for inflammation, pulp tissue organization, reactionary dentin formation, and quality of dentinogenesis., Results: The Quick-Set group exhibited significantly more pulpal inflammation (P = .002) and significantly less pulp tissue organization (P = .004). No significant difference was noted for reactionary dentin formation (P = .526) and quality of dentinogenesis (P = .436)., Conclusions: Compared with ProRoot White MTA (Dentsply Tulsa Dental Specialties, Tulsa, OK), Quick-Set exhibited more pulpal inflammation and decreased pulp tissue organization. No significant differences were noted for reactionary dentin formation and quality of dentinogenesis., (Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

Biotite and bentonite are phyllosilicate minerals that were originally used in industrial applications. Several beneficial activities of them have recently been reported, especially regulation of the immune system and antimicrobial effects. Therefore, we investigated the immune-enhancing and bacterial clearance effects of a biotite and bentonite mixture (BBM) on experimental infection of Salmonella enterica serovar Typhimurium (S. Typhimurium) to determine whether the BBM could be used as an alternative antibiotic. We administered 1% or 2% BBM as a feed supplement. We then evaluated the bacterial clearance effects of the BBM against S. Typhimurium. We also evaluated the immune-enhancing effect of the BBM through several immunological experiments that included examination of the lysozyme activity, CD4(+)/CD8(+) T lymphocyte ratio and the T-helper type 1 (Th 1) cytokine profile. The clinical signs of S. Typhimurium and the number of viable bacteria in feces and tissues were significantly decreased in both BBM groups, especially in the 2% BBM group. The BBM also markedly enhanced the lysozyme activity, CD4(+)/CD8(+) T lymphocyte ratio and expression levels of IFN-γ and IL-12 in S. Typhimurium-challenged pigs. Therefore, the BBM could be a good candidate as an alternative antibiotic that improves Th 1-specific immune responses and the bacterial clearance effect.

Introduction: Quick-Set (Avalon Biomed Inc, Bradenton, FL) is a calcium aluminosilicate cement that is a potential alternative to mineral trioxide aggregate (MTA) with greater acid resistance and faster setting. The purpose of this study was to compare the regeneration of apical tissues after root-end surgery when the apical tissues were exposed to Quick-Set or White ProRoot MTA (Dentsply Tulsa Dental Specialties, Tulsa, OK) by root-end resection., Methods: The root canals of 42 mandibular premolars in 7 beagle dogs were accessed, cleaned and shaped, and obturated with Quick-Set or white MTA. Osteotomies and root-end resections were performed immediately. The dogs were sacrificed at 90 days, and the teeth and surrounding tissues were removed and prepared for histologic analysis. The sections of the apical areas were scored for inflammation, new cementum formation, periodontal ligament formation, and bone quality., Results: At 90 days, both materials supported some degree of cementum formation on the surface of the material, periodontal ligament regeneration, and excellent bone quality. The only significant difference was greater inflammation found in the Quick-Set group., Conclusions: Quick-Set and White ProRoot MTA had a similar effect on bone quality, cementum formation, and periodontal ligament formation after root-end surgery in dogs. Quick-Set was associated with greater inflammation., (Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.)

The aims of this study were, first, to investigate the toxicity of zearalenone (ZEN) through the analysis of biochemical parameters, oxidative stress, pathological changes and inflammatory response in the kidney of gestation sows and offspring; and, second, to evaluate the efficacy of modified halloysite nanotubes (MHNTs) for the alleviation to the adverse effects induced by ZEN. This study focused on the period of organogenesis between days 35 and 70 of gestation, and treatments included (1) a control diet; (2) contaminated grain (50% control corn and 50% mouldy corn); and (3) contaminated grain (50% control corn and 50% mouldy corn) + 1% MHNTs. ZEN treatment significantly increased most of the biochemical parameters and inflammatory cytokines and degenerative changes in the kidney and induced oxidative damage in plasma, whereas the addition of MHNTs in combination with ZEN induced a re-establishment of the biochemical parameters, the plasma oxidative stress enzyme activities and the normal histology of the kidney. Thus, the data strongly suggest that the deleterious effects of ZEN can be significantly diminished by MHNTs.

Since the outbreaks of foot-and-mouth disease (FMD) in South Korea in 2010-2011, a trivalent vaccine has been used as a routine vaccination. Despite the high efficacy of the trivalent vaccine, low antibody formation was reported in the pig industry and there is considerable concern about the ability of the vaccine to protect against the Andong strain responsible for recent outbreaks in South Korea. To overcome these problems, immunostimulators have been widely used to improve vaccine efficacy in South Korea, although without any scientific evidence. Based on the current situation, the aim of this study was to investigate the effects of germanium biotite, a feed supplement used to enhance the immune system, on the immune responses to FMD vaccination through the Andong strain challenge experiment in trivalent vaccinated pigs. Following the challenge, the germanium biotite-fed pigs showed high levels of IL-8 in serum, and increased cellular immune responses to stimulation with the Andong strain antigen compared to nonsupplemented pigs. In addition, higher FMD virus (FMDV) neutralizing antibody titers were detected in the germanium biotite-fed group than in the nonsupplemented group before the challenge. The findings of this study indicate that germanium biotite supplement might enhance immune responses to the FMD vaccine in pigs.

Background: Ge-imogolites are short aluminogermanate tubular nanomaterials with attractive prospected industrial applications. In view of their nano-scale dimensions and high aspect ratio, they should be examined for their potential to cause respiratory toxicity. Here, we evaluated the respiratory biopersistence and lung toxicity of 2 samples of nanometer-long Ge-imogolites., Methods: Rats were intra-tracheally instilled with single wall (SW, 70 nm length) or double wall (DW, 62 nm length) Ge-imogolites (0.02-2 mg/rat), as well as with crocidolite and the hard metal particles WC-Co, as positive controls. The biopersistence of Ge-imogolites and their localization in the lung were assessed by ICP-MS, X-ray fluorescence, absorption spectroscopy and computed micro-tomography. Acute inflammation and genotoxicity (micronuclei in isolated type II pneumocytes) was assessed 3 d post-exposure; chronic inflammation and fibrosis after 2 m., Results: Cytotoxic and inflammatory responses were shown in bronchoalveolar lavage 3 d after instillation with Ge-imogolites. Sixty days after exposure, a persistent dose-dependent inflammation was still observed. Total lung collagen, reflected by hydroxyproline lung content, was increased after SW and DW Ge-imogolites. Histology revealed lung fibre reorganization and accumulation in granulomas with epithelioid cells and foamy macrophages and thickening of the alveolar walls. Overall, the inflammatory and fibrotic responses induced by SW and DW Ge-imogolites were more severe (on a mass dose basis) than those induced by crocidolite. A persistent fraction of Ge-imogolites (15% of initial dose) was mostly detected as intact structures in rat lungs 2 m after instillation and was localized in fibrotic alveolar areas. In vivo induction of micronuclei was significantly increased 3 d after SW and DW Ge-imogolite instillation at non-inflammatory doses, indicating the contribution of primary genotoxicity., Conclusions: We showed that nm-long Ge-imogolites persist in the lung and promote genotoxicity, sustained inflammation and fibrosis, indicating that short high aspect ratio nanomaterials should not be considered as innocuous materials. Our data also suggest that Ge-imogolite structure and external surface determine their toxic activity.

Aqueous leachates prepared from natural antibacterial clays, arbitrarily designated CB-L, release metal ions into suspension, have a low pH (3.4-5), generate reactive oxygen species (ROS) and H2O2, and have a high oxidation-reduction potential. To isolate the role of pH in the antibacterial activity of CB clay mixtures, we exposed three different strains of Escherichia coli O157:H7 to 10% clay suspensions. The clay suspension completely killed acid-sensitive and acid-tolerant E. coli O157:H7 strains, whereas incubation in a low-pH buffer resulted in a minimal decrease in viability, demonstrating that low pH alone does not mediate antibacterial activity. The prevailing hypothesis is that metal ions participate in redox cycling and produce ROS, leading to oxidative damage to macromolecules and resulting in cellular death. However, E. coli cells showed no increase in DNA or protein oxidative lesions and a slight increase in lipid peroxidation following exposure to the antibacterial leachate. Further, supplementation with numerous ROS scavengers eliminated lipid peroxidation, but did not rescue the cells from CB-L-mediated killing. In contrast, supplementing CB-L with EDTA, a broad-spectrum metal chelator, reduced killing. Finally, CB-L was equally lethal to cells in an anoxic environment as compared to the aerobic environment. Thus, ROS were not required for lethal activity and did not contribute to toxicity of CB-L. We conclude that clay-mediated killing was not due to oxidative damage, but rather, was due to toxicity associated directly with released metal ions.

Background: Infectious diseases in dairy cows often follow a time of nutritional or physiological stress and the subsequent altered immune system function. This study aimed to determine if the immunomodulatory effects of a feed additive previously observed in experimental animals and housed cattle fed total mixed rations could be reproduced in pasture-fed dairy cattle under Australian conditions., Methods: The study included 34 pasture-fed dairy cattle given the treatment (n = 17) or placebo (bentonite, n = 17) for an acclimation period of 15 days followed by 60 days of supplementation. Blood tests were taken pre-trial and then 30, 60 and 90 days after acclimation. Blood samples were extracted and preserved in Trizol and analysed for immune markers., Results: Pasture-fed dairy cows in the treatment group had significantly higher levels of the immune markers interleukin-8R and L-selectin in comparison with placebo-fed cows at 60 days after the start of supplementation., Conclusion: The immunomodulatory effects of the additive observed in the current study and the associated enhanced neutrophil function demonstrated by other studies suggest a role in decreasing the rates of mastitis and other infectious diseases of dairy cattle, particularly during times of nutritional or physiological stress., (© 2014 Australian Veterinary Association.)

Voluntary running in an activity wheel establishes aversion to paired taste in rats. A proposed mechanism underlying this taste aversion learning is gastrointestinal discomfort caused by running. We tested this hypothesis by measuring the pica behavior (kaolin clay intake) of rats, because it is known that rats engage in pica behavior after various nausea-inducing treatments including irradiation, motion sickness, and injection of emetic drugs such as lithium chloride (LiCl). Following a demonstration of the already-known phenomenon of LiCl-based pica in Experiment 1, we successfully showed running-based pica behavior in Experiment 2 where the running treatment was compared with a non-running control treatment (i.e., confinement in a locked wheel). These results suggest that not only LiCl but also running induces nausea in rats, supporting the gastrointestinal discomfort hypothesis of running-based taste aversion learning., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We discuss new trends in Layer-by-Layer (LbL) encapsulation of spherical and tubular cores of 50-150 nm diameter and loaded with drugs. This core size decrease (from few micrometers to a hundred of nanometers) for LbL encapsulation required development of sonication assistant non-washing technique and shell PEGylation to reach high colloidal stability of drug nanocarriers at 2-3mg/mL concentration in isotonic buffers and serum. For 120-170 nm spherical LbL nanocapsules of low soluble anticancer drugs, polyelectrolyte shell thickness controls drug dissolution. As for nanotube carriers, we concentrated on natural halloysite clay nanotubes as cores for LbL encapsulation that allows high drug loading and sustains its release over tens and hundreds hours. Further drug release prolongation was reached with formation of the tube-end stoppers., (Copyright © 2014. Published by Elsevier Ltd.)

Aim: To evaluate the effect of muscovite in preventing small bowel injury induced by nonsteroidal anti-inflammatory drugs (NSAIDs)., Methods: We recruited and screened thirty-two healthy volunteers who were randomly allocated equally into two groups: an NSAID control group, who received 75 mg slow-release diclofenac, twice daily for 14 d; and an NSAID-muscovite group, who received 3 g of muscovite in addition to the 75 mg of slow-release diclofenac, twice daily for 14 d. For gastroprotection, both groups were administered 20 mg/d of the proton pump inhibitor omeprazole. All eligible subjects underwent video capsule endoscopy (CE) prior to and 14 d after treatment., Results: Thirty subjects (NSAID-muscovite group, n =16; NSAID control group, n =14) finally completed the whole trail. At the baseline CE examination, no statistically significant differences between the two groups have been observed. However, after 14 d of drug treatment, a significant difference was observed in the percentage of subjects with mucosal breaks when comparing the NSAID-muscovite group with the NSAID control group. While 71.4% (10/14) of subjects in the NSAID control group had at least one mucosal break, co-administration of muscovite in the NSAID-muscovite group reduced the rate to 31.3% (5/16) (P = 0.028). Moreover, higher number of mucosal breaks was found in the NSAID control group vs that in the NSAID-muscovite group (P < 0.05)., Conclusion: Muscovite co-therapy reduced the incidence of small intestinal injury after 14 d of diclofenac administration.

In vivo and in vitro experiments were conducted to test for beneficial effects of dietary clays on broiler chicks challenged with Salmonella enterica serovar Typhimurium and to explore potential mechanisms. First, two hundred forty 1-d-old male broilers (initial BW: 41.6 ± 0.4 g) were allotted in a 2 × 4 factorial arrangement in a randomized complete block design. There were 2 infection treatments (with or without Salmonella) and 4 diets: basal (BAS), 0.3% smectite A (SMA), 0.3% smectite B, and 0.3% zeolite. The Salmonella reduced (P < 0.05) the growth rate of chicks fed the BAS, and feeding clay largely restored it (challenge × diet interaction, P < 0.05). Goblet cell number and size were increased (P < 0.05) by Salmonella in chicks fed the BAS and were reduced (P < 0.05) in Salmonella-challenged chicks by feeding SMA. Villus height was reduced by the Salmonella challenge in the chicks fed dietary clays (P < 0.01) but not in chicks fed the BAS (interaction P < 0.05). A human adenocarcinoma cell line (LS174T) was cultured in vitro in 3 separate experiments in the absence or presence of 3 concentrations (0.05, 0.10, and 0.50%) of SMA. Expression of mucin 2 (MUC2), resistin-like molecule β (RELMß), and trefoil factor 3 (TFF3) were determined by real-time reverse-transcription PCR. The expression of RELMβ was increased and expression of MUC2 was reduced (P < 0.05) by 0.10% SMA. Also, LS174T cells were cultured without or with SMA (0.05 and 0.10%) and the medium and cell lysate were analyzed for RELMβ using an immunoblot assay. Protein expression of RELMß in the cell lysate was reduced (P < 0.05) by SMA addition but increased in the medium, indicating that SMA increased secretion of RELMß, thus depleting the cell and concentrating this protein in the medium. In conclusion, the dietary clays restored the growth depression caused by Salmonella, and changes in goblet cell function may contribute to the benefits of one of the clays, specifically SMA.

Although clays are wildly used in a range of applications, the toxicity assessment of these new materials is still scarce. In the present study, oxidative stress induced by Clay 1, a novel clay, was determined in rats after 90 d of oral exposure. The activities of antioxidant enzymes, namely, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST), were examined. In addition, genetic expressions of SOD and CAT and relative protein abundance of CAT were also determined. Data showed that most of the biomarkers assayed remained unaltered. Only CAT activity, as well as its genetic and protein expressions, appeared enhanced in the kidney. Therefore, further studies are needed to clarify the relevance and consequences of these findings to ensure the safety of this clay.

A total of 1,280 1-d-old ducks were used in a study to investigate the effects of increasing aflatoxin B1 (AFB1) concentrations from naturally contaminated corn on young ducklings, and the effectiveness of a clay adsorbent (CA) to protect against those effects. Ducks were randomly allotted to 8 treatments (TRT) in a 4 × 2 factorial arrangement with 4 levels of AFB1 (0, 25, 50, and 100 μg/kg) and 2 levels of CA (0 and 0.1%) with 8 pens per TRT and 20 ducks per pen. All ducks were allowed ad libitum access to feed and water during the 21-d experiment. The ADG, ADFI, feed conversion rate, mortality, bill color, and CV of BW of each replicate were measured at the end of the study. Blood and tissue samples from 8 ducks per TRT were obtained on d 21 of the experiment to determine the serum immunoglobulin and protein concentrations, relative organ weights, and intestinal morphology. Average daily gain and relative weights of the liver, spleen, thymus, and bursa of Fabricius decreased linearly (P < 0.05) as dietary AFB1 increased. Serum proteins and intestinal villi heights and villus/crypt ratio followed the same pattern. Bill decolorization ratio, CV of BW, and mortality increased linearly (P < 0.05) as dietary AFB1 increased. Adding 0.1% CA to the diet improved (P < 0.05) the relative weights of the small intestine, spleen, and thymus, and the villus height and villus/crypt ratio of the duodenum and jejunum, as well as the serum IgG and IgM concentrations. Adding CA also reduced (P < 0.05) bill decolorization ratio, CV of BW, mortality, and serum IgA concentration. Therefore, duck performance was negatively affected by increasing AFB1 concentrations in diets. But the addition of 0.1% CA can protect against the detrimental effects caused by AFB1-contaminated corn in diets for ducks.

Germanium biotite (GB) is an aluminosilicate mineral containing 36 ppm germanium. The present study was conducted to better understand the effects of GB on immune responses in a mouse model, and to demonstrate the clearance effects of this mineral against Porcine reproductive and respiratory syndrome virus (PRRSV) in experimentally infected pigs as an initial step towards the development of a feed supplement that would promote immune activity and help prevent diseases. In the mouse model, dietary supplementation with GB enhanced concanavalin A (ConA)-induced lymphocyte proliferation and increased the percentage of CD3+CD8+ T lymphocytes. In pigs experimentally infected with PRRSV, viral titers in lungs and lymphoid tissues from the GB-fed group were significantly decreased compared to those of the control group 12 days post-infection. Corresponding histopathological analyses demonstrated that GB-fed pigs displayed less severe pathological changes associated with PRRSV infection compared to the control group, indicating that GB promotes PRRSV clearance. These antiviral effects in pigs may be related to the ability of GB to increase CD3+CD8+ T lymphocyte production observed in the mice. Hence, this mineral may be an effective feed supplement for increasing immune activity and preventing disease.

This paper deals with the synthesis of a biocompatible chitosan ammonium salt N-(2-hydroxy) propyl-3-trimethylammonium chitosan chloride (HTCC) and using it in montmorillonite ion-exchange process. HTCC-modified montmorillonite (Mt) with different chemical ratios was successfully synthesized, and their characteristics have been verified by XRD and FTIR analyses. Produced samples have been evaluated in terms of antibacterial efficiency and biocompatibility (cell culture test). Antibacterial efficiency of synthesized HTCC/Mt samples has been confirmed against both gram negative bacteria (Escherichia coli) and gram positive bacteria (Staphylococcus aureus). The results disclosed that the antibacterial efficiency of HTCC-modified montmorillonite was unexpectedly even more than HTCC. This excellent synergistic effect has been referred to entrapping bacteria between the intercalated structures of HTCC-modified montmorillonite. Then HTCC on clay layers can seriously attack and damage the entrapped bacteria. An extraordinary biocompatibility, cell attachment, and cell growth even more than tissue culture polystyrene (TCPS) have been recorded in the case of this novel kind of modified clay. Due to existing concerns about serious and chronic infections after implant placement, this natural-based bioactive and antibacterial modified clay can be used in electrospun nanofibers and other polymeric implants with promising mechanical properties for tissue engineering applications.

The purpose of the current study was to provide a mechanistic basis for in vitro and in vivo performance differences between lipid-based formulations solidified by adsorption onto a high surface area material and their respective liquid (i.e., nonadsorbed) counterparts. Two self-emulsifying formulations (based on either medium-chain or long-chain lipids) of the poorly water-soluble drug danazol were solidified by adsorption onto Neusilin US2. Liquid and adsorbed lipid-based formulations were subjected to in vitro dispersion-digestion tests, and additional in vitro experiments were performed to elucidate the cause of performance differences. The bioavailability of danazol after oral administration to rats was also assessed. The percentage of the dose solubilized in the aqueous phase during in vitro dispersion-digesting was ∼35% lower for the adsorbed formulations when compared to their liquid counterparts. This trend was also reflected in vivo, where the bioavailability of danazol after administration of the adsorbed formulations was ∼50% lower than that obtained after administration of the equivalent liquid formulation. Incomplete desorption of the microemulsion preconcentrate from the carrier on dispersion-digestion was identified as the main contributor to the reduced pharmaceutical performance of the adsorbed formulations. The results of the current study indicate that solidification of lipid-based formulations through adsorption onto a high surface area carrier may limit formulation (and drug) release in vivo and thereby reduce oral bioavailability.

The presence of circulating tumor cells (CTCs) is believed to lead to the formation of secondary tumors via an adhesion cascade involving interaction between adhesion receptors of endothelial cells and ligands on CTCs. Many CTCs express sialylated carbohydrate ligands on their surfaces that adhere to selectin protein found on inflamed endothelial cells. We have investigated the feasibility of using immobilized selectin proteins as a targeting mechanism for CTCs under flow. Herein, targeted liposomal doxorubicin (L-DXR) was functionalized with recombinant human E-selectin (ES) and polyethylene glycol (PEG) to target and kill cancer cells under shear flow, both when immobilized along a microtube device or sheared in a cone-and-plate viscometer in a dilute suspension. Healthy circulating cells such as red blood cells were not targeted by this mechanism and were left to freely circulate, and minimal leukocyte death was observed. Halloysite nanotube (HNT)-coated microtube devices immobilized with nanoscale liposomes significantly enhanced the targeting, capture, and killing of cancer cells. This work demonstrates that E-selectin functionalized L-DXR, sheared in suspension or immobilized onto microtube devices, provides a novel approach to selectively target and deliver chemotherapeutics to CTCs in the bloodstream., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Hydroxypropyl methylcellulose (HPMC) tablets containing nicotine-magnesium aluminum silicate (NCT-MAS) complex particles and pH modifiers, namely, sodium chloride, citric acid, and magnesium hydroxide, were prepared using the direct compression method. The effects of HPMC viscosity grades and pH modifiers on NCT release and permeation of the matrix tablets were examined. The results showed that the higher the viscosity grade of HPMC that was used in the tablets, the lower was the unidirectional NCT release rate found. The unidirectional NCT permeation was not affected by the viscosity grade of HPMC because the NCT diffusion through the mucosal membrane was the rate-limiting step of the permeation. Incorporation of magnesium hydroxide could retard NCT release, whereas the enhancement of unidirectional NCT release was found in the tablets containing citric acid. Citric acid could inhibit NCT permeation due to the formation of protonated NCT in the swollen tablets at an acidic pH. Conversely, the NCT permeation rate increased with the use of magnesium hydroxide as a result of the neutral NCT that formed at a basic microenvironmental pH. The swollen HPMC tablets, with or without pH modifiers, gave sufficient adhesion to the mucosal membrane. Furthermore, the addition of magnesium hydroxide to the matrix tablets was the major factor in controlling buccal delivery of NCT. This study suggests that the NCT-MAS complex-loaded HPMC tablets, which contained magnesium hydroxide, are potential buccal delivery systems of NCT.

Background: Clay is often used in cosmetic treatments, although little is known about its action., Aim: To evaluate the effect of topical clay application on the histoarchitecture of collagen fibres in rat skin., Methods: Animals received a daily application of clay and retinoic acid (RA) 0.025% to the dorsal skin over 7 and 14 days, under vaporization at 37 °C for 40 min. Control skin was not vaporized. Samples from each region were excised, and stained with picrosirius red for collagen evaluation., Results: Seven days after clay treatment, an increase in the number of collagen fibres was observed in treated skin compared with control skin (51.74 ± 1.28 vs. 43.39 ± 1.79%, respectively, P < 0.01), whereas RA did not alter the collagen level (45.66 ± 1.10%). Clay application over 14 days did not induce a further increase in skin collagen, whereas treatment with RA did (58.07 ± 1.59%; P = 0.001 vs. control)., Conclusion: Clay application promotes an increase in the number of collagen fibres, which may account for its beneficial effects., (© The Author(s). CED © 2012 British Association of Dermatologists.)

Increase in intoxication products, such as medium size peptides, indole and myoglobin, in urine was observed in Wistar rats after exposure of their chest to gamma-radiation at a dose of 6 Gy (dose rate 4 Gy/min). The rats exhibited moderate ischemic ECG. Administration of enterosorbents, such as Smekta and Clay of Kaluga deposit, to the irradiated rats resulted in the decrease of the toxicant content in the animals and the recovery of the cardiac function on the 28th day. These sorbents had practically a similar efficacy.

Natural halloysite nanotubes with a 15-nm internal lumen and a 50 nm outer diameter were investigated as a nanocontainer for the loading and extended release of glycerol for cosmetic applications. Cytotoxicity testing of the halloysite was conducted on 3T3 and MCF-7 cells, and the tubules showed no toxic effect on the cells for over 48 h. The capability of halloysite for loading glycerol was higher with the USA halloysite than with the New Zealand's, being approximately 20% and 2.3% by weight, respectively. The total elapsed time for releasing glycerol from the nanotubes exceeded 20 h. To further retard the glycerol release rate, the halloysite samples filled with glycerol were coated with several alternate layers of polyethyleneimine and polyacrylic acid. The release rate remained at the same level, however, probably due to the low molecular weight of the polyelectrolytes and the high solubility of glycerol in water.

Chondrokola Rosh (CKR) is a traditional metallic Ayurvedic preparation widely used by the rural and ethnic people of Bangladesh in dysuria. It is a preparation of various roasted metals (Hg and Cu), non-metal (sulphur and Mica) and medicinal herbs. Considering the controversy over the risk of toxic heavy metals in Ayurvedic herbo-mineral preparations, toxicological parameters on liver functions were investigated. A single dose of 100mg/kg body weight of the preparation was administered orally to the rats of both sexes for ninety days. In this evaluation a statistically significant (p<0.001) increase of serum albumin levels in male (17%) and female (15%) rat groups were observed. On the other hand, the plasma bilirubin level was decreased 50% and 28% respectively in both rats groups. But no remarkable differences were observed in plasma protein, sGPT, sGOT and ALP activities from their corresponding control values. This study showed that CKR had no remarkable toxic effect on liver of the animals despite the presence of traces of transformed heavy metals.

Background and Aims: The aim of this study was to investigate whether rectal administration of muscovite can ameliorate colonic inflammation in a rat model of experimental colitis, and its possible mechanism., Methods: Female Sprague-Dawley (SD) rats with trinitrobenzene sulfonic acid (TNBS)-induced colitis were treated with rectal administration of muscovite or 5-aminosalicylic acid (5-ASA) daily for 14 days. The changes in body weight, macroscopic damage and histologic scores were subsequently evaluated. Gene expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), mucin2 (MUC2) and trefoil factor 3 (TFF3) in the colonic tissues was assessed by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) while protein levels of TNF-alpha and IL-1beta were detected by ELISA. Mucin2 expression in colonic mucosa was detected by immunohistochemistry. The capacity of muscovite to adsorb cytokines in vitro was determined by the changes in the amount of TNF-alpha, IL-1beta secreted by lipopolysaccharide (LPS)-stimulated THP-1 cells and IL-8 secreted by LPS-stimulated HT-29 cells., Results: Rectal administration of muscovite improved the loss of body weight, macroscopic and histologic scores of TNBS-induced colitis in a dose-dependent manner. Trinitrobenzene sulfonic acid-induced expression of TNF-alpha and IL-1beta was reduced by muscovite and 5-ASA treatment. Reduction of MUC2 expression in colitis rats was reversed by muscovite and 5-ASA treatment. However, the expression of TFF3 mRNA in colonic mucosa was not affected. In addition, we found muscovite inhibited the expression of TNF-alpha, IL-1beta secreted by THP-1 and IL-8 secreted by HT-29 cells in a dose-dependent manner., Conclusions: Our study demonstrated for the first time that rectal administration of muscovite can ameliorate colonic inflammation of TNBS-induced colitis. Further confirmatory studies are needed to prove that muscovite might be a potential therapeutic agent for the treatment of ulcerative colitis.

1. A study was conducted to evaluate the possible protective effect of a feed additive containing aluminosilicate and phytogenic substances against the adverse effects of aflatoxins in turkey poults. 2. Dietary treatments (6) were given to turkey poults from d 1 to d 42 of age. From d 1 to 21 the dietary treatments were as follows: 1, negative control, no aflatoxins or feed additive added; 2, feed additive control, 1 kg/t feed additive, no aflatoxins; 3, 250 ppb (microg/kg) aflatoxins, no feed additive; 4, 250 ppb aflatoxins + 1 kg/t feed additive; 5, 500 ppb aflatoxins, no feed additive; and 6, 500 ppb aflatoxins + 1 kg/t feed additive. From d 22 to 42, the dietary concentration of the feed additive was increased from 1 to 2 kg/t in all treatment groups receiving the feed additive (2, 4 and 6), while keeping constant the dietary concentrations of aflatoxins. 3. Aflatoxins at 250 ppb did not cause adverse effects on performance but affected certain toxicopathological parameters. At 500 ppb, adverse effects on performance and several toxicological parameters were observed. 4. Some of the adverse affects were partially or completely overcome by supplementation with the feed additive, including amelioration of the performance parameters, suppression of mortality and correction of the immunological alterations induced by the exposure to the aflatoxins.

1. To investigate the growth performance and histological intestinal alterations of Aigamo ducks fed on dietary combinations of zeolite, plant extract and vermiculite (ZEM, 14-d-old Aigamo ducks were divided into 4 groups, with 3 replicates of 3 male and 3 female ducks. They were fed ad libitum on a basal commercial duck mash diet with 0, 0.1, 0.5 and 1.0 g/kg dietary ZEM for 63 d. 2. Body weight gain tended to be higher for the 0.1 and 0.5 g/kg ZEM groups than for the control group at 9 weeks. 3. In light microscopic observation, most values of the intestinal villus height, villus area, cell area and cell mitosis numbers were higher in the ZEM group than those of the control in all intestinal segments, and the duodenal villus height, cell area and cell mitosis of the 0.5 g/kg ZEM group, as well as jejunal cell mitosis in the 0.1 g/kg ZEM group, increased (P < 0.05). In the scanning electron microscope results, all ZEM groups showed protuberant epithelial cells and cell clusters on the villus apical surface of the duodenum and ileum. In the jejunum, villus gyri were frequently observed in the 0.1 g/kg ZEM group. These histological intestinal alterations suggest that intestinal villi and epithelial cellular functions might have been activated. 4. From the present results, dietary ZEM showed hypertrophied functions of intestinal villi and epithelial cells at the duodenum and ileum, and the 0.1 and 0.5 g/kg levels improved body weight gain. These suggest that the ZEM can be supplemented until a level of 1.0 g/kg.

The aim of this study was to investigate the ability of liquid loadable tablets (LLT) to be loaded with a self-microemulsifying drug delivery system (SMEDDS) containing cyclosporine (CyA). LLT were prepared by direct compression of the porous carrier magnesium aluminometasilicate and subsequently loaded with SMEDDS by a simple absorption method. SMEDDS was evaluated regarding visual appearance and droplet size distribution after dispersion in aqueous media. The developed SMEDDS was found to be similar to Neoral. LLT were characterized before and after loading regarding weight variation, tablet hardness, disintegration time, and in vitro drug release. It was found that LLT with high porosities suitable for liquid loading and further processing could be prepared. Adding a tablet disintegrant was found to improve in vitro drug release. Additionally, the volume-based loading capacity of LLT was evaluated and found to be comparable to soft gelatin and hard two-piece capsules. Furthermore, the pharmacokinetic performance of CyA from loaded LLT was tested in two PK-studies in dogs. Absorption of CyA from SMEDDS loaded into LLT was found in the first study to be significantly lower than the absorption of CyA from SMEDDS filled into a capsule. However, addition of a superdisintegrant improved the absorption markedly. The bioavailability of CyA from SMEDDS loaded into disintegrating LLT was found in the second study to be at the same level as from capsule formulation. In conclusion, the LLT technology is therefore seen as a promising alternative way of achieving a solid dosage form from liquid drug delivery systems.

Studies with rats and white mice demonstrated that combined radiation injury (CRI) comes to reducing protease activity in small intestine tissues. In liver tissue the activity of cathepsine D increased under the action of new enterosorbent--clay of Kaluga deposit ("CKD"). In blood serum of the damaged animals the amount of histogenic toxins--middle mass molecules (MMM) and oligopeptides decreased after "CKD" administration. An effective fixation and withdrawal of bacteria from the bowl of the affected animals took place. The survival level of mice in 30 days after CRI comprised 20% as compared with 60% in the case of "CK D" administration. It is supposed that the increase of survival after "CKD" administration is the result of enhancement of macrophage detoxification in liver, determined on the increase of activity of cathepsine D (lysosomal protease), elimination of MMM and normalization of gut microflora.