• Asian-Australasian Journal of Animal Sciences
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• v.32 no.2
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• pp.233-240
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• 2019

Objective: This study was to investigate the effects of dietary cotton stalk on nitrogen and free gossypol in sheep. Methods: Treatments included 25% cotton stalk (Treat 1), 50% cotton stalk (Treat 2), and a control (no cotton stalk). Six Xinjiang daolang wethers were cannulated at the rumen and duodenum and fed one of these diets. The effects of these diets on nitrogen and free gossypol absorption and metabolism were determined. Fifteen healthy Xinjiang daolang wethers were assessed for daily gain, tissue lesions, and free gossypol accumulation. Results: Dry matter intake decreased with increasing dietary cotton stalk. Total tract dry matter digestibility did not significantly differ among treatments. Dietary cotton stalk significantly decreased volatile fatty acids and increased ammonium nitrogen in the rumen. Nitrogen intake was significantly higher in Treat 2 than in the control or Treat 1. Nitrogen retention and free gossypol intake increased with dietary cotton stalk. Duodenal free gossypol flow did not increase, and free gossypol almost disappeared from the rumen. The free gossypol content of plasma and tissue was increased with dietary cotton stalk with liver free gossypol>muscle free gossypol>kidney free gossypol. Elevated dietary free gossypol decreased platelets, hemoglobin, and serum iron. Aspartate aminotransferase and ${\gamma}$-glutamyltransferase increased in Treat 2. With high long-term dietary cotton stalk intake, liver cells were swollen, and their nuclei dissolved. Renal cells were necrotic and the interstitia were enlarged. Conclusion: With short-term cotton stalk administration, only a small amount of free gossypol is retained in the body. In response to long-term or high free gossypol cotton stalk feeding, however, free gossypol accumulates in, and damages the liver and kidneys.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.3
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• pp.356-359
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• 2000

The study was conducted to evaluate the nutritional value of seeds and various seed fractions of different varieties of cotton (MNH 147, CIM 240, NIAB 78, FH 87, CIM 109, MNH 93, FH 682, GOHAR 87, SLS I and B 557). Linter, hull, kernel and meal were obtained from cottonseed by physical and chemical methods. Free gossypol and available lysine contents of seed and it's fractions were determined. In vitro protein digestibility of cottonseed meal was also determined. Free gossypol and available lysine contents ranged between 0.22-2.26% and 0.64-1.32% in seed, and 0.03-0.29% and 1.38-2.36% in meal, respectively. FH 87 was highest both in free gossypol and available lysine content, and NIAB 78 was lowest in free gossypol content and FH 862 was lowest in available lysine content. In vitro protein digestibility of cottonseed meal ranged between 66,02-79,96%. Statistical analysis revealed significant (p<0.05) varietal differences in free gossypol, available lyslne and in vitro protein digestibility of cottonseed and derived products.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.1
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• pp.63-70
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• 2018

Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1314-1321
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• 2006

The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at $30^{\circ}C$ for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.9
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• pp.1291-1296
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• 2003

Effects of feeding of 9.95 mg free gossypol/kg live weight through cottonseed meal (CSM) were studied in 20 intact male calves fed barley or sorghum as source of cereal during the experimental duration of 210 days. Serum concentration of total protein, albumin, globulin and their ratio did not vary because of protein (gossypol) or cereal sources. Serum level of cholesterol and urea were lower (p<0.05) in sorghum than barley fed calves. Feeding of gossypol through CSM enhanced (p<0.05) serum cholesterol. An interaction between protein and period was observed with respect to serum concentrations of urea, creatinine and alanine transferase. The levels of serum creatinine and alanine transferase increased (p<0.05) following 120 days of experimental feeding in calves fed CSM diets compared to the control animals fed groundnut meal diets. No effect of feeding gossypol was, however, evident on the serum enzymes viz. alanine and aspartate transferases and alkaline phosphatase. Moreover, the source of cereal and protein did not appear to influence the metabolic profile of the calves. Humoral immune response, measured through antibody titre against Brucella abortus S99 innoculation, revealed a delayed and depressed seroreactivity indicative of immunocompromisation because of the phytotoxin gossypol. In conclusion, the feeding of gossypol at the designated levels, although had no deleterious clinico-biochemical manifestations, affected the humoral immune response of the calves.

• Animal Bioscience
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• v.35 no.4
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• pp.556-566
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• 2022

Objective: This study aimed to determine the appropriate supplementation level of lactic acid bacteria (LAB; Lactobacillus plantarum and Bacillus clausii), yeast (Saccharomyces cariocanus and Wickerhamomyces anomalus) for degrading free gossypol and glucosinolate in the fermented total mixed ration (TMR) containing cottonseed meal (CSM) or rapeseed meal (RSM), to improve the utilization efficiency of these protein sources. Methods: For LAB, L. plantarum or B. clausii was inoculated at 1.0×10⁸, 1.0×10⁹, 1.0×10¹⁰, and 1.0×10¹¹ colony-forming unit (CFU)/kg dry matter (DM), respectively. For yeast, S. cariocanus or W. anomalus was inoculated at 5×10⁶, 5×10⁷, 5×10⁸, and 5×10⁹ CFU/kg DM, respectively. The TMR had 50% moisture and was incubated at 30℃ for 48 h. After fermentation, the chemical compositions, and the contents of free gossypol and glucosinolate were determined. Results: The results showed that the concentration of free gossypol content was reduced (p<0.05), while that of the crude protein content was increased (p<0.05) in the TMR containing CSM inoculated by B. clausii (1×10⁹ CFU/kg DM) or S. cariocanus (5×10⁹ CFU/kg DM). Similarly, the content of glucosinolate was lowered (p<0.05) and the crude protein content was increased (p<0.05) in TMR containing RSM inoculated with B. clausii (1×10¹⁰ CFU/kg DM) or S. cariocanus (5×10⁹ CFU/g DM). Conclusion: This study confirmed that inclusion of B. clausii with 1.0×10⁹ or 1.0×10¹⁰ CFU/kg DM, or S. cariocanus (5×10⁹ CFU/kg DM) to TMR containing CSM/RSM improved the nutritional value and decreased the contents of anti-nutritional factors.

• Journal of Animal Science and Technology
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• v.55 no.2
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• pp.123-130
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• 2013

Whole cotton seed (WCS) has become one of the major feed ingredients in TMR for dairy cattle in Korea, and WCS for feed use is mostly imported from abroad. Since this genetically modified oil seed is usually fed to the animal in raw state, its germination ability, if last long, often causes concerns about ecological disturbances. In the process of looking for effective conditions to remove germination ability of WCS this study had the objectives to evaluate the nutritional effects of gamma irradiation at doses of 8, 10 and 12 kGy on changes in nutrient contents, anti-nutritional factors, in vitro digestibility and ruminal degradability. No significant differences were found in proximate analysis of nutrients between raw WCS and gamma irradiated one. Glycine and threonine contents significantly increased when the WCS was exposed to gamma ray as compared to untreated WCS (p<0.05). As for fatty acid composition, no significant differences were observed with the irradiation treatment. Free gossypol in WCS was decreased (p<0.05) by gamma irradiation treatment. Of the 3 different levels of gamma irradiation, a dose of 12 kGy was found to be the most effective in reducing free gossypol concentration. Results obtained from in situ experiment indicated that gamma irradiation at a dose of 10 kGy significantly (p<0.05) lowered rumen degradability of both dry matter and crude protein as compared with raw WCS. However, there were no significant differences in rapidly degradable and potentially degradable fractions of crude protein due to 10 kGy gamma irradiation. Overall, this study show that gamma irradiation at a dose of 10 kGy is the optimum condition for removing germination ability of WCS, and could improve nutritive value for the ruminant with respect to the decrease in both ruminal protein degradability and gossypol content of WCS.

• Korean Journal of Food Science and Technology
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• v.9 no.3
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• pp.211-219
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• 1977

Dehulled and defatted Korean cottonseed flour was extracted with alkaline solution for 30 minutes and had precipitated the crude portein by adjusting pH $1{\sim}12$. The general composition and the amino acid composition of cottonseed protein were analyzed. Crude protein was purified with sephadex G-100 and G-200, and its component had been identified by disc electrophoresis. Toxic gossypol was removed by n-hexane, acetone and other solvents. The results were as follows. (1) pH 5, pH 7 and pH 4 were the best condition of precipitation of curde protein at single, two step and water extraction, respectively. (2) The cottonseed flour which was dehulled and defatted, contained 61.3% of crude protein. (3) The protein which was isolated from cottonseed flour, contained 20% of glutamic acid, and comparatively high levels of essential amino acids. (4) Dehulled cottonseed flour contained 0.97% of total gossypol and could be romoved 70% of total gossypol by extraction with n-hexane. (5) 10-13 bands of water soluble protein were found in disc electrophoresis, and 10-12 bands in protein were isolated by single and two step procedures. (6) The cottonseed protein could be purified by sephadex G-100 and G-200. (7) 10-20% of gossypol-free cottonseed fluor could be used for animal and human comsumption.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.57-62
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• 2003

An effort was made to assess comparative production performance in broiler chickens fed diets containing solvent extracted cottonseed meal (CSM) processed from BT and Parental Non-BT lines. Processed meal of national check and commercial produce cottonseeds were also used for comparison. The free gossypol contents were 0.02, 0.02, 0.44 and 0.03% in meals of BT, Parental Non-BT, national check and commercial produce cottonseeds, respectively. Day-old broiler chicks (n=243) were divided to 27 groups of 9 each. Nine dietary treatments (iso-nitrogenous, 23% CP and iso-caloric, 2,800 kcal ME/kg) were formulated viz., D1 (control, soybean meal-SBM based), D2 and D3 (BT CSM at 10% of diet with and without additional iron), D4 and D5 (non-BT CSM with and without additional iron), D6 and D7 (national check CSM with or without additional iron), and D8 and D9 (commercial produce CSM with or without additional iron at 2 ppm for every 1 ppm of free gossypol, respectively). Each dietary treatment was offered to three replicated groups up to 6 weeks of age. At the end of 6 weeks of age, 10 birds were taken out randomly from each treatment and were sacrificed to study carcass traits, organs' yield and histo-pathological changes in vital organs. The broiler chickens received CSM processed from BT (D2, 1,753 g and D3, 1,638 g) and Parental Non-BT (D4, 1,653 g and D5, 1,687 g) with or without additional Fe grew at same rate as observed in soybean meal (solvent ext.) based diet (D1, 1,676 g). The feed intake and feed conversion efficiency (feed: gain) in these dietary treatments (BT, non-BT line based diets) also did not differ significantly (p>0.05) from control diet. Similar observation was also observed in dietary treatments (D8 and D9) containing solvent extracted cottonseed meal of commercial produce origin. However, a decrease (p<0.05) in body weight gain and feed intake was observed in D6 containing national check CSM with high gossypol content. Addition of Fe in the diet (D7) improved (p<0.05) feed intake and weight gain but not to the extent as observed in diets containing BT, parental non-BT, and commercial produce CSM or control. However, any type of CSM did not affect feed conversion efficiency when fed with or without additional iron. The carcass characteristics in terms of dressing percentage, liver weight and heart weight was not significantly (p>0.05) different between the treatments. The eviscerated yields emanated from diets containing either BT, non-BT or commercial produce were statistically similar to control. However, eviscerated yield of broilers fed national check CSM with or without iron supplementation was lower (p<0.05) than BT cotton with Fe supplementation and commercial produce CSM. The study envisaged that BT, parental non-BT and commercial produce solvent extracted cottonseed meal can be included at 10% in soybean meal based broiler diet replacing soybean meal and rice bran without additional iron.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.1
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• pp.29-35
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• 2003

In order to find the feasibility of feeding raw or processed cotton (Gossypium) seed meal (CSM), 30 male crossbred lambs were randomly assigned to 5 dietary treatments with 6 animals each. Lambs were fed each of the isonitrogenous and isocaloric concentrate mixtures containing 30% deoiled groundnut cake (DGNC) (reference diet), 40% raw, 45 minute cooked, 1% $Ca(OH)_2$ or iron (1 part free gossypol:0.3 parts iron) treated CSM (replacing about 50% nitrogen moiety of reference concentrate mixture). The concentrate mixture was fed to meet 80% of CP requirements (NRC, 1985) along with ad libitum maize (Zea mays) hay for 180 days. Incorporation of raw or processed CSM did not affect the palatability of diets as evidenced by higher (p<0.01) or comparable overall daily intakes of DM and protein (CP and Digestible CP) per kg $W^{0.75}$ by lambs in comparison to reference group. Intakes of DE and ME increased (p<0.01) due to inclusion $Ca(OH)_2$ treated CSM in diets, while the intakes were lower (p<0.01) on iron treated CSM based diet when compared with reference diet. The intakes of DM, CP, TDN, DE and ME by reference and experimental lambs were higher than the requirements stipulated by NRC (1985). The growth rate was highest (p<0.01) in lambs fed on diets with cooked CSM followed by those fed raw or $Ca(OH)_2$ treated CSM based diets. The growth of lambs fed on iron treated CSM diets was depressed (p<0.01) in comparison to that of other experimental lambs probably due to increased iron intake (889 mg/kg DM of diet) which was much higher than the toxic levels of 500 ppm (NRC, 1980). The lambs on various diets utilized DM with similar efficiency while the utilization of energy (TDN, DE and ME) was more efficient (p<0.01) when the diets contained raw or cooked CSM in comparison to that of reference diet. The lambs fed raw and $Ca(OH)_2$ treated CSM incorporated diets utilized DCP with higher (p<0.01) efficiency than the lambs fed reference diet. The feed cost per kg weight gain was lower (p<0.01) on raw, cooked and $Ca(OH)_2$ treated CSM based diets in comparison to reference diet. Cooking of CSM for 45 minutes further reduced the feed cost of weight gain.