N-乙酰半胱氨酸对衰老小鼠回肠结构和抗氧化机能的影响

研究旨在探讨N-乙酰半胱氨酸(NAC)对衰老小鼠回肠结构和抗氧化机能的影响。试验利用D-半乳糖构造衰老模型小鼠,并通过饮水给小鼠服NAC,观察小鼠行为、体质量和回肠结构,并测定血液、肝和回肠组织中过氧化氢酶(CAT)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)等抗氧化指标,结果表明:NAC使衰老小鼠毛发和皮肤得到改善,使衰老小鼠体质量显著增加(P0.05)。NAC使衰老小鼠绒毛高度显著升高(P0.05),绒毛宽度和隐窝深度显著降低(P0.05)。NAC使衰老小鼠血液、肝和回肠组织中CAT和GSH-Px含量显著升高(P0.05),MDA含量显著降低(P0.05)。因此,NAC具有抗氧化、清除自由基和延缓衰老的作用。     

牦牛脾脏非特异性转移因子对小鼠抗衰老作用的影响

构建小鼠亚急性衰老模型,注射牦牛脾脏非特异性转移因子(n TF),采用试剂盒检测衰老小鼠组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性和丙二醛(MDA)含量,以研究n TF的抗氧化作用。结果:药物组小鼠的心、肝、肺、肾组织中SOD酶活性与阳性对照组相比有增加的趋势,但各组间差异不显著(P0.05);小鼠心、脾、肾组织中MDA含量显著低于阳性对照组,差异显著(P0.05);药物组小鼠心、肝、脾、肾组织中GSH-Px酶活性显著高于阳性对照组,差异显著(P0.05)。试验表明,注射n TF衰老小鼠组织中GSH-Px酶活性显著增加,同时MDA含量显著降低,表明n TF能有效增强机体抗氧化的作用及降低机体受自由基攻击的程度。     

苜蓿皂苷对断奶仔猪生长性能、肠道菌群、组织抗氧化能力及相关酶mRNA表达的影响

本试验旨在研究苜蓿皂苷对断奶仔猪生长性能、肠道菌群、组织抗氧化能力及相关酶mRNA表达的影响。选取24头平均体重为8 kg的大×长杂交断奶仔猪,随机分为2组,每组3个重复,每个重复4头猪。对照组饲喂基础饲粮,苜蓿皂苷组饲喂在基础饲粮中添加0.25%苜蓿皂苷的饲粮。预试期10 d,正试期30 d。结果表明:1)与对照组相比,饲粮中添加苜蓿皂苷可显著提高断奶仔猪平均日增重(P0.05),并显著降低料重比(P0.05)。2)与对照组相比,饲粮中添加苜蓿皂苷可显著降低仔猪十二指肠和盲肠的pH(P0.05),显著提高十二指肠、空肠和回肠中乳酸菌数量(P0.05)。3)与对照组相比,饲粮中添加苜蓿皂苷可显著提高仔猪肝脏和肾脏中谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)活性(P0.05),并显著提高仔猪肝脏和空肠中GSH-Px mRNA表达量及十二指肠和回肠中CAT mRNA表达量(P0.05)。综上,苜蓿皂苷可以提高断奶仔猪生长性能,增强其组织抗氧化能力并有效改善其肠道菌群。     

N-乙酰半胱氨酸对肉鸭肝脏能量代谢状况及组织抗氧化指标的影响

为考查N-乙酰半胱氨酸(NAC)是否可以改善肉鸭肝脏的能量代谢状况及组织抗氧化能力,试验选用90只1 d樱桃谷肉鸭,随机分成3个处理组:对照组(基础日粮)、500 mg/kg NAC组(基础日粮+500 mg/kg NAC)、1 000 mg/kg NAC组(基础日粮+1 000 mg/kg NAC),每组3个重复,每个重复10只鸭。试验期为3周(1~21日龄)。结果表明:与对照组相比,日粮添加500 mg/kg NAC显著提高了血浆、肝脏的过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的活力及肝脏谷胱甘肽过氧化物酶(GSH-Px)的酶活力、三磷酸腺苷(ATP)、二磷酸腺苷(ADP)的含量和能荷(EC)值(P0.05),显著降低了血浆丙二醛(MDA)和肝脏过氧化氢(H2O2)的含量及肝脏AMP/ATP值和肝脏低氧诱导因子1(HIF-1)、黄嘌呤氧化还原酶(XOR)的m RNA水平(P0.05);1 000 mg/kg NAC组显著提高了血浆和肝脏CAT、SOD的活力及肝脏GSH-Px的酶活力、ATP的含量、EC值和AMPK-α1的m RNA水平(P0.05),显著降低了血浆和肝脏H2O2的含量及肝脏MDA的含量、HIF-1、XOR的m RNA水平。表明NAC可以提高肉鸭的组织抗氧化能力、改善肝脏的能量代谢,这可能与NAC对AMPK-α1、HIF-1及XOR的基因表达调控有关。     

母鼠围产期铅暴露对仔鼠的氧化损伤及乙酰半胱氨酸的保护效应

为探讨发育期铅暴露所致仔鼠氧化损伤及N-乙酰半胱氨酸(NAC)的保护效应,从母鼠怀孕开始通过饮水染铅至仔鼠21日龄断奶建立动物模型,测定断奶仔鼠脑组织和血清谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、乙酰胆碱酯酶(AchE)活性和丙二醛(MDA)含量,并观察海马的组织结构变化。结果显示,染铅使出生后21d仔鼠脑组织和血清MDA含量极显著升高(P0.01),GSH-Px,SOD,AchE活性极显著降低(P0.01),血清CAT活性极显著降低(P0.01);NAC拮抗组与染毒组比较,血清和脑组织MDA含量极显著或显著降低(P0.01或P0.05),脑组织SOD活性显著升高(P0.05),血清SOD,GSH-Px,CAT,AchE活性极显著升高(P0.01),其余指标均差异不显著(P0.05)。组织学观察表明,海马神经细胞形态正常,组间未见明显病理组织学变化,超微结构可见染毒组部分神经细胞异常。结果表明,母鼠铅暴露可致仔鼠脑组织脂质过氧化损伤,NAC可提高仔鼠脑组织的抗氧化能力,对铅损伤具有一定的保护作用。     

黄花蒿多糖对肉仔鸡生长性能及组织抗氧化指标的影响北大核心

试验旨在探究添加不同剂量的黄花蒿多糖(AAP)对肉仔鸡生长性能及组织抗氧化指标的影响。选用1日龄AA肉仔鸡240只,随机分为5组,每组6个重复,每个重复8只鸡。各组分别在基础日粮中添加0、250、500、750和1 000 mg/kg的AAP。试验期42 d。结果显示:22~42日龄时,与对照组相比,添加750 mg/kg的AAP肉仔鸡的平均日增重(ADG)极显著升高(P<0.01);添加750 mg/kg和1 000 mg/kg的AAP肉仔鸡的料重比(F/G)显著降低(P<0.05)。42日龄时,日粮中添加750 mg/kg的AAP肉仔鸡的末重和ADG显著提高(P<0.05)。21日龄时,添加250 mg/kg、500 mg/kg AAP肉仔鸡脾脏中总超氧化物歧化酶(SOD)活性极显著提高(P<0.01);添加750 mg/kg AAP肉仔鸡十二指肠和回肠中过氧化氢酶(CAT)的活性以及空肠中谷胱甘肽过氧化物酶(GSH-Px)、SOD的活性和总抗氧化能力(T-AOC)显著提高(P<0.05);添加750 mg/kg和1 000 mg/kg AAP肉仔鸡脾脏中GSH-Px活性以及十二指肠中SOD、GSH-Px活性和回肠中T-AOC极显著提高,肝脏、脾脏、空肠和回肠中丙二醛(MDA)含量极显著降低(P<0.01),肝脏中T-AOC和SOD活性显著提高(P<0.05)。42日龄时,添加750 mg/kg AAP肉仔鸡肝脏中T-AOC以及空肠中GSH-Px的活性和回肠中SOD、CAT活性均显著提高(P<0.05);添加500 mg/kg和750 mg/kg AAP肉仔鸡肝脏和空肠中SOD活性、脾脏中CAT活性、十二指肠中GSH-Px活性、回肠中GSH-Px活性、T-AOC均极显著提高(P<0.01),回肠中MDA含量极显著降低(P<0.01);添加1 000 mg/kg AAP肉仔鸡十二指肠中CAT活性和空肠中T-AOC显著提高(P<0.05),十二指肠中MDA含量显著降低(P<0.05)。研究表明,日粮中添加AAP可提高肉仔鸡的生长性能及抗氧化功能,且适宜的添加量为750 mg/kg。     

镉对大鼠大脑皮质神经细胞的氧化损伤及N-乙酰半胱氨酸的保护效应

为研究镉对新生大鼠原代大脑皮质神经细胞的脂质过氧化损伤及N-乙酰半胱氨酸(N-acetyl cysteine,NAC)的保护效应,对神经细胞用不同浓度(0,5、10、20 μmol/L)醋酸镉染毒和NAC(100 μmol/L)进行保护,检测细胞内活性氧(ROS)水平、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)、过氧化氩酶(CAT)活性及丙二醛(MDA)含量.结果表明,与时照组比较,染毒组细胞内ROS水平、MDA含量和CAT活性极显著升高(P＜0.01),GSH-Px活性降低,20μmol/L组出现显著差异(P＜0.05);NAC保护组与相应染毒组比较,ROS水平、MDA含量和CAT活性呈不同程度降低,部分组间出现显著差异(P＜0.05);说明镉有促进神经细胞脂质过氧化的作用,NAC可提高神经细胞的抗氧化能力,对镉暴露所致的脂质过氧化损伤有一定的保护作用.     

N-乙酰半胱氨酸和冷应激对肉鸡肠道组织形态、能量代谢和抗氧化能力的影响

本试验旨在研究N-乙酰半胱氨酸(NAC)和冷应激对肉鸡肠道组织形态、能量状况和抗氧化能力的影响。试验将200只1日龄健康雄性科宝肉鸡随机分为4个处理组:对照组(基础日粮)、NAC组(基础日粮+0.1%NAC)、冷应激组(基础日粮)与冷应激+NAC组(基础日粮+0.1%NAC),每组5个重复,每个重复10只鸡。试验第1~2周按常规饲养方法控制温度,试验第3~6周冷应激组和冷应激+NAC组的温度控制在(10±2)℃,而对照组与NAC组维持在(26±2)℃。结果表明:冷应激处理使肉鸡平均日采食量、平均日增重、空肠低氧诱导因子1(HIF-1)m RNA水平显著下降(P0.05),饲料转化率、回肠一磷酸腺苷(AMP)含量和空肠隐窝深度极显著升高(P0.01);日粮添加0.1%NAC显著提高了冷应激肉鸡肠道绒毛高度(VH)、十二指肠与空肠绒毛高度与隐窝深度比值(VH/CD)、空肠三磷酸腺苷(ATP)和腺苷酸池(TAN)含量、回肠ATP和二磷酸腺苷(ADP)及能荷(AEC)含量、十二指肠超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活力、回肠谷胱甘肽过氧化物酶(GSH-Px)活力、空肠血红素加氧酶(HMOX)m RNA水平(P0.05)。因此,日粮中添加0.1%的N-乙酰半胱氨酸可改善肠道组织形态和能量状况,提高冷应激肉鸡肠道抗氧化能力。     

饲粮中添加白藜芦醇对“杜×长×大”猪空肠抗氧化能力及紧密连接蛋白表达的影响

试验旨在研究饲粮中添加白藜芦醇(resveratrol,RES)对"杜×长×大"猪空肠抗氧化酶活性及抗氧化基因指标和紧密连接蛋白相关基因表达水平的影响。试验选用12头体重约为65 kg的育肥猪分为2组,每组6头。对照组育肥猪饲喂基础饲粮,试验组育肥猪饲喂在基础饲粮中添加400 mg/kg的RES的饲粮,试验期41 d。结果表明,与对照组相比,饲粮中添加400 mg/kg的RES显著提高了育肥猪空肠过氧化氢酶(CAT)、总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)的活性及总抗氧化能力(T-AOC)(P<0.05),显著降低丙二醛(MDA)的含量(P<0.05);显著提高育肥猪空肠抗氧化基因铜/锌超氧化物歧化酶(Cu/Zn-SOD)、锰超氧化物歧化酶(Mn-SOD)、谷胱甘肽过氧化物酶-1 (GPx-1)和谷胱甘肽过氧化物酶-4 (GPx-4)活性(P<0.05);显著提高育肥猪空肠抗氧化酶活相关基因CAT、T-SOD和GSH-Px的表达水平(P<0.05),显著降低MDA的表达水平(P<0.05);显著提高育肥猪空肠紧密连接蛋白咬合蛋白(Occludin)、闭合蛋白-1 (Claudin-1)和链接黏附分子A (JAM-A)的表达水平(P<0.05)。研究表明,饲粮添加400 mg/kg的RES能够显著提高育肥猪空肠抗氧化能力及紧密连接蛋白的表达,改善动物肠道健康。     

孕鼠补充高羊毛氨酸硒对新生乳鼠十二指肠和空肠抗氧化功能的影响

为探究在母鼠妊娠期补充高羊毛氨酸硒对新生乳鼠肠道抗氧化功能的影响,18只健康Wistar孕鼠被随机平均分成缺硒组(0.03 mg/kg)、正常硒对照组(0.15 mg/kg)和富硒组(1.5 mg/kg),母鼠分娩后第4天对新生乳鼠进行称重,安乐死后测定新生乳鼠的胸腺指数和脾脏指数,并检测新生乳鼠十二指肠和空肠的谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、总超氧化物歧化酶(T-SOD)的活性,以及总抗氧化能力(T-AOC)和丙二醛(MDA)含量。结果显示:与正常硒对照组相比,缺硒组和富硒组新生乳鼠的体重均无显著变化(P>0.05);缺硒组新生乳鼠脾脏指数、胸腺指数及十二指肠和空肠的T-AOC、CAT、GSH-Px活性均显著降低(P<0.05),MDA含量均极显著升高(P<0.01);富硒组新生乳鼠脾脏指数、胸腺指数及十二指肠和空肠的T-AOC、CAT和GSH-Px活性均显著升高(P<0.05),十二指肠内MDA含量显著降低(P<0.05)。结果表明:母鼠在妊娠期补充高羊毛氨酸硒,能够提高新生乳鼠十二指肠与空肠的抗氧化能力。     

Phase I evaluation of CCNU (lomustine) in tumor-bearing cats

1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.     

Prevalence of gastric lesions in racing Alaskan sled dogs

Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.     

Furosemide continuous rate infusion in the horse: evaluation of enhanced efficacy and reduced side effects

Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.     

Internal Parasites and Haematological Values in Cattle Slaughtered in Buea Subdivision of Cameroon

Tropical Animal Health and Production -     

The Epidemiology of Gastrointestinal Nematodes of Dairy Cattle in Central Kenya

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.     

Diversity of Ectoparasites in Sheep Flocks in Sa~o Paulo,Brazil

The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.     

Chemotaxis of macrophage in inflammation

Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.     

Evaluation of biohydrogenation rate of canola vs. soya bean seeds as unsaturated fatty acids sources for ruminants in situ

An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.     

Serum biochemical values and mineral element contents of tissues in yaks

Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein