• Korean Journal of Animal Reproduction
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• v.17 no.1
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• pp.21-25
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• 1993

We have previously shown that gossypol injection into the stroma of testis was found to effectively inhibit the process of spermatogenesis. In this study, properties and chemistry of blood were investigated weekly in ICR mouse after the injection of gossypol into the testicular stroma(5, 10 or 15mg per kg body weight). There were no significant differences in red blood cell(RBC), hematocrit, white blood cell(WBC), basophils, eosinophils and monocytes during first 4 weeks after injection of gossypol between treatments, but neutrophils increased and lymphocytes decreased, respectively(P<0.05). Total content of protein, albumin and globulin were not different, compared with control. However, the concentration of glucose after injection of gossypol increased significantly(P<0.05). In conculsion, the results of this study indicated that injection of gossypol into stroma of testis might affect both properties and chemistry of blood in mice.

• 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.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.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.

• Fisheries and Aquatic Sciences
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• v.13 no.4
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• pp.284-293
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• 2010

To reduce anti-nutritional factors in plant protein sources for fish meal replacement in fish feeds, cottonseed and soybean meal (CS) were fermented with Aspergillus oryzae. A feeding trial was conducted to verify the effects of fermented CS (FCS) with phytase supplementation on gossypol detoxification, phosphorus digestibility, antioxidant activity, and growth performance of juvenile olive flounder over 10 weeks. Four diets were formulated to replace 0, 30, or 40% fish meal protein with CS or FCS (designated as CS0, CS30, FCS30P, and FCS40P). Phytase (1,000 FTU/kg) was added to FCS30P and FCS40P. The microbial fermentation significantly increased dietary total polyphenols and consequently led to higher DPPH radical-scavenging activities in fish feed and fish tissue. Dietary and liver gossypol concentrations were dramatically decreased by the fermentation process. Phosphorus digestibility was significantly increased in fish fed the FCS40P diet. However, growth performance decreased in fish fed FCS diets. This study demonstrates that the fermentation process and phytase supplementation can improve the phosphorus availability of plant protein sources in fish. The fermentation of CS by A. oryzae could increase antioxidant activities in feed and fish and effectively degrade toxic gossypol in cottonseed meal.

• 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.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1106-1109
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• 2001

This experiment was conducted with 30 adult wool producing Angora rabbits of either sex, to evaluate the effect of replacing soyflakes (SF), on equal protein basis, with low gossypol containing cottonseed meal (CSM) either alone or in combination with lysine and methionine; on the biological performance, total wool yield, wool yield per shearing, wool characteristics and mortality. Three experimental mash diets were prepared by incorporating SF (6%) as standard / control protein source $(T_1)$ and CSM (9%) as test protein source ($T_2$ and $T_3$). In $T_3$, amino acids-lysine and methionine (0.1 % each) were added. Animals were given the experimental diets about $150g{\cdot}day^{-1}{\cdot}head^{-1}$, for a period of 225 d or three shearing, and ad libitum Kudzu-vines. No significant effect of $T_2$ or $T_3$, on the body weight gain, total wool yield, wool yield per shearing and wool characteristics, was observed compared to $T_1$. However, the digestibility of dry matter, crude fibers, ether extract, acid detergent fibers, neutral detergent fibers, cellulose and hemicellulose was significantly (p<0.05) depressed in CSM based diets. Mortality of about 20% was recorded in $T_2$ and $T_3$, but not related to the addition of CSM or gossypol toxicity. More studies are needed to standardize the safe level of CSM, duration of safe feeding of CSM, and level of amino acids supplementation in CSM based diets.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.3
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• pp.400-409
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• 2017

Objective: This study was conducted to evaluate the effects of dietary protein sources (soybean meal, SBM; low-gossypol cottonseed meal, LCSM; double-zero rapeseed meal, DRM) on laying performance, egg quality, and plasma parameters of laying hens. Methods: A total of 432 32-wk-old laying hens were randomly divided into 6 treatments with 6 replicates of 12 birds each. The birds were fed diets containing SBM, $LCSM_{100}$, or $DRM_{100}$ individually or in combination with an equal amount of crude protein (CP) ($LCSM_{50}$, $DRM_{50}$, and $LCSM_{50}-DRM_{50}$). The experimental diets, which were isocaloric (metabolizable energy, 11.11 MJ/kg) and isonitrogenous (CP, 16.5%), had similar digestible amino acid profile. The feeding trial lasted 12 weeks. Results: The daily egg mass was decreased in the $LCSM_{100}$ and $LCSM_{50}-DRM_{50}$ groups (p<0.05) in weeks 41 to 44. The $LCSM_{50}$ group did not affect egg production compared to the SBM group in weeks 41 to 44 (p>0.05) and showed increased yolk color at the end of the trial (p<0.05). Compared to the SBM group, the $LCSM_{100}$ and $LCSM_{50}-DRM_{50}$ groups showed decreased albumen weight (p<0.05), CP weight in the albumen (p<0.05) and CP weight in the whole egg (p<0.05) at 44 weeks. Plasma total protein (TP) levels were lower in the $LCSM_{100}$ group than in the SBM group at 44 weeks (p<0.05); however, TP, albumin, and globulin levels were not significantly different between the $LCSM_{50}$ group and the SBM group or between the $DRM_{50}$ group and the SBM group (p>0.05). Conclusion: Together, our results suggest that the $LCSM_{100}$ or $DRM_{100}$ diets may produce the adverse effects on laying performance and egg quality after feeding for 8 more weeks. The 100.0 g/kg LCSM diet or the 148.7 g/kg DRM diet has no adverse effects on laying performance and egg quality.

• Animal Bioscience
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• v.36 no.4
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• pp.619-628
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• 2023

Objective: This study aimed to determine and compare the apparent ileal digestibility (AID) and the standardized ileal digestibility (SID) of amino acids (AA) in soybean meal (SBM), cottonseed meal (CSM), and low-gossypol cottonseed meal (LCSM) fed to broiler chickens and laying hens. Methods: Three semi-purified diets containing the identical crude protein concentration at 20% were formulated to contain SBM, CSM, or LCSM as the sole source of N. A N-free diet was also formulated to estimate the basal ileal endogenous losses of AA for broilers and hens. A total of 300 male Ross 308 chicks at one-day-old and 144 Hy-Line Brown laying hens at 30-week-old with initial egg production rate of 88.3%±1.0% were randomly allocated into 1 of 4 dietary treatments, respectively. Results: CSM and LCSM showed more Arg and Cys+Met while less Lys, Ile, Leu, and Thr relative to SBM. Significant interactions existed between species and experimental diets for AID (except for Arg, Asp, Glu, Gly, and Pro) and SID (except for Arg, His, and Phe) of most AA. Most AA in diets showed higher AID (except for Lys) and SID (except for Lys, Met, and Ser) in broilers relative to laying hens. The AID and SID of all AA were significantly different between the three diets. In broilers, the AID and SID of most indispensable AA except for Arg in SBM and LCSM was higher than CSM. In laying hens, the AID and SID of most indispensable AA except for Arg, Met+Cys, and Phe in SBM was higher than CSM and LCSM. Conclusion: The accurate determination of AID and SID of AA in CSM and LCSM for broilers and layers benefits the application of CSM and LCSM in chicken diets. The cottonseed by-products CSM or LCSM showed the species-specific AA digestibility values for broilers and layers.