A review of feline serum protein electrophoresis

A review of the current literature available on feline serum proteins is presented. Early studies concentrated on comparative aspects of species variations in the electrophoretic pattern. The feline electrophoretogram was divided into five basic regions: albumin, alpha-1, alpha-2, beta and gamma. Different subdivisions of these areas were recognized depending on the support medium used. Current papers have compared the relative migration distances of each globulin peak to the migration of albumin. This "Rf" value enables reliable peak identification. To date, no data exists identifying the individual proteins responsible for the peaks in the alpha and beta regions. The only feline globulin to be studied is haptolobin; however its precise location on the electrophoretic strip was not identified.     

Preliminary observations on plasma fibrinogen and plasma protein concentrations and on plasma protein: Fibrinogen ratios in clinically healthy buffaloes

The plasma fibrinogen concentration, the total plasma protein concentration and the plasma protein to fibrinogen ratio (PP:F) were determined in clinically healthy Nili Ravi buffaloes. The plasma fibrinogen concentrations in calves, lactating and non-lactating buffaloes were 513±62, 615±90 and 544±74 mg/dl, respectively, and were statistically different (p<0.05). Total plasma protein concentrations in these animals were 7.15±0.28, 9.32±0.53 and 8.79±0.58 g/dl. PP:F for all animals was between 11 and 19. Fibrinogen levels were positively correlated with plasma protein (r=0.59) and negatively correlated with PP:F (r=–0.59).Abbreviations PP:F plasma protein to fibrinogen ratio     

Proteomic analysis of soluble protein extract of adult Toxocara cati

Toxocara cati is a cat roundworm and the causative agent of toxocariasis as a cosmopolitan zoonotic disease. As no information has been reported so far, identification of T. cati proteins can be useful for the development of new diagnostic strategies. This study was conducted to identify the major proteins in the adult T. cati tegument using bi-dimensional electrophoresis (2-DE) and shotgun proteomics. A total proteins were identified, among them the metabolic enzymes were the largest group, including: Enolase, triose phosphate isomerase, fructose-bisphosphate aldolase, aldehyde dehydrogenase. The other important protein groups recognized in T. cati, belong to the HSP-family, the structure and motor proteins, such as actin. The role of these proteins have been implicated in parasite–host interactions and modulating cellular immune response, immune regulation in evasion mechanisms of the host immune response. Characterizing T. cati adult proteins play a key role not only in host-parasite interactions, but also in the discovery of drug targets, subunit vaccines against toxocariasis, immunodiagnostic kits for toxocariasis and the identification of novel immuno-modulators that can form the next generation of therapeutic possibilities for inflammatory diseases.     

Studies of composition and major protein level in milk and colostrum of mares

The aim of the study was to determine the changes in composition and physicochemical features (pH, density, thermostability and acidity) of mare colostrum and milk, and of protein fraction contribution (serum albumin, β-casein, γ-casein, α-lactalbumin, G class immunoglobulins) depending on lactation stage. The research material was colostrum and milk samples from 12 Arabian mares. Colostrum samples were collected within 2 h after parturition and milk samples were collected twice, in the 3rd and 6th weeks of lactation. The level of basic milk components decreased significantly (only lactose content increased) as compared to colostrum. Total bacteria count and somatic cell count decreased significantly with an increase in resistance and urea level. The changes observed were connected to differentiated contribution of particular protein fractions and their relative proportions. Lower levels of γ-casein (P ≤ 0.05), β-casein, serum albumin as well as α-lactalbumin were observed in colostrum as compared to those in milk. Any relationship between lactation stage and β-casein content was observed. Serum albumin and α-lactalbumin content increased in subsequent milkings. The level of G class immunoglobulins decreased significantly and its highest level was noted in colostrum. Any significant differences between the 3rd and 6th lactation weeks were obtained.     

水产饲料中大豆蛋白替代鱼粉的应用研究

凭借充足的供应源和低廉的价格,大豆蛋白成为我国最为常用且最具前途的鱼粉替代物。研究表明,豆类蛋白可部分或全部代替鱼粉而不影响鱼类的生长和发育,但也存在各种各样的限制因素,使得鱼类对其利用仍然不够理想。文章介绍大豆蛋白在水产饲料上的应用情况,探讨限制大豆蛋白利用的抑制因子及其相应的解决措施,以期为大豆蛋白的广泛研究和应用提供参考依据。     

Identification of sperm motility markers in bovine transition protein genes

Transition proteins (TNPs) are essential in chromatin condensation during spermiogenesis, and hence, they are the candidate genes for identifying sperm motility markers. Coding and in silico predicted promoter regions of these genes were investigated in crossbred and purebred cattle, and also, their mRNA quantification was done to explore its use as a diagnostic tool of infertility. PCR‐SSCP analysis revealed two band patterns in fragment III of TNP1 and fragment II of TNP2 gene. Sequence analysis revealed a deletion of “G” nucleotide in 3′UTR region of TNP1 and C>T SNP in intronic region of TNP2 gene. Least square analysis of variance did not reveal any significant influence of nucleotide deletion on any sperm motility parameters in both crossbred and purebred cattle. However, C>T SNP had a significant effect on initial progressive motility (p < 0.05) in purebred cattle and post‐thaw motility in overall cattle population. RT‐qPCR analysis did not reveal any significant variation in TNP1 and TNP2 gene expression among poorly motile and good quality spermatozoa of Vrindavani bulls.     

Evidence for a functional bone morphogenetic protein (BMP) system in the porcine ovary

Bone morphogenetic proteins (BMPs) play important roles in controlling fertility and ovulation rate. There is however, little information on the BMP system in the ovary of a large polyovular species. The aims of the present study were to investigate BMP-2 and -6 protein expression in the porcine ovary, their effects on granulosa cells in culture and their mechanism of action. Cells and oocytes were recovered from healthy antral follicles 2–6 mm in diameter. When assessed by Western blotting, oocytes and follicular fluid contained BMP-2 and -6. In addition, BMP-2 and -6 were observed in granulosa cells and BMP-2 was also found in theca cells. Granulosa cells were cultured in a serum-free system for 144 h in the presence of increasing doses (0, 3, 30 and 100 ng/ml) of BMP-2 or BMP-6. Both BMPs suppressed progesterone production in a dose-dependent manner after 48 h (P < 0.001) and 144 h (P < 0.05). Only BMP-6 stimulated cell proliferation at 100 ng/ml (P < 0.05). Investigation into the mechanism of action found that BMP-2 and -6 decreased cyclic adenosine monophosphate (cAMP) production (P < 0.01), expression of 3β-hydroxysteroid dehydrogenase (3β-HSD) protein (P < 0.001) and steroidogenic acute regulatory protein (StAR) (BMP-6 only; P < 0.05). This supports the hypothesis that BMP-2 and -6 act as luteinization inhibitors. In conclusion, these findings provide evidence for the presence of a complex signalling mechanism in the porcine ovary and suggest that both BMP-2 and -6 may act in a paracrine manner to control granulosa cell function in this large polyovulatory species.     

两型饲料的能量和新型蛋白质替代原料

两型饲料是指资源节约型和环境友好型饲料。目前,市场上猪、肉鸡和蛋鸡的日粮以玉米豆粕型为主,需要新的解决方案来改善饲料生产可持续性和解决工农争粮问题。有许多原料可作为替代能量和蛋白质来源正受到人们的关注,其中包括高粱、大麦、昆虫源性原料、藻源性蛋白和单细胞蛋白等。文章主要论述了两型饲料中纤维的重要性以及几种能量和新型蛋白质替代猪只饲料原料的应用前景。     

Analysis of serum protein concentrations after severe thermal injury in a dog

Serial serum electrophoreses and routine serum protein assays were used to assess changes in serum protein concentrations after severe thermal injury in a dog. Electrophoretic patterns during the month of evaluation were consistent with protein-losing dermatopathy (thermal burn) and inflammatory dysproteinemias.     

Serum protein reference values in foals during the first year of life: comparison of chemical and electrophoretic methods

Normal reference values for serum proteins of foals from birth to 1 year of age have been established. Chemical and electrophoretic/refractometric methods for total protein, albumin, total globulin and Albumin/Globulin (A/G ratio) have also been compared. The biuret total protein method and Bromcresol Green (BCG) method on the Technicon SMA 12/60 autoanalyzer were used and compared with total protein determined via refractometry and albumin determined by Cellulose Acetate (CA) electrophoresis/densitometry. Globulin and A/G ratios were calculated from the chemical method data and compared with that obtained electrophoretically. Total protein, albumin, total globulins and A/G ratios all were in agreement at all sample times studied. Data on the subfractionation of serum globulins via CA electrophoresis is also presented. Wide variations in alpha and beta globulin levels were noted among the foal sera early in life. As a result, two distinct populations of foals with respect to both globulin content and A/G ratio were identified. One of these populations (Group A) appeared to have obtained passive immunity more slowly than the other (Group B) animals. Comparison of these data with clinical cases of foals in which failure of passive transfer was a part suggests that the A/G ratio may be useful in assessing adequate colostral antibody levels in the newborn foal.     

Acute phase protein levels in pregnancy and oestrus cycle in bitches

The purpose of this study was to evaluate the acute phase protein (serum amyloid A, C-reactive protein, haptoglobin, ceruloplasmin and fibrinogen) profiles in pregnant dogs as well as bitches at various stages of the estrus cycle and to verify possible relationship(s) among the APPs profile and hormonal status. A total of 60 bitches of various breeds were enrolled. The animals were divided into six groups. B-mode ultrasonographic scanner (concept M/C) was used for pregnancy diagnosis, involving the first and second halves of pregnancy. Proestrus, estrus, diestrus and anestrus stages of sexual cycles were differentiated by use of vaginal cytology and blood progesterone and estradiol 17-β levels. Serum C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin, ceruloplasmin and fibrinogen levels were determined. No statistically significant differences were observed between the groups with regard to the serum SAA and CRP concentrations. Haptoglobin, ceruloplasmin and fibrinogen concentrations were elevated in pregnant dogs compared to non-pregnant groups’, independently from the pregnancy stage. No statistically significant differences in acute phase protein concentrations in dogs at various stages of estrus could be detected. Results of this study indicated that ceruloplasmin and fibrinogen levels may have potential importance in the diagnosis of pregnancy, and acute phase proteins may be used for monitoring the health status of the pregnant dogs.     

Serum C-reactive protein concentrations in healthy Miniature Schnauzer dogs

Background: C‐reactive protein (CRP) is a sensitive marker for inflammation in people and dogs. In people, an association between CRP concentration and atherosclerosis has been reported. Atherosclerosis is rare in dogs, but the Miniature Schnauzer breed may be at increased risk for developing this vascular disease. It is not known if CRP concentrations in Miniature Schnauzer dogs differ from those in other dog breeds. Objectives: Our objectives were to validate an automated human CRP assay for measuring CRP in dogs and compare CRP concentrations in healthy Miniature Schnauzer dogs with those in non‐Miniature Schnauzer breeds. Methods: Sera from 37 non‐Miniature Schnauzer dogs with inflammatory disease were pooled and used to validate a human CRP immunoturbidimetric assay for measuring canine CRP. Blood was collected from 20 healthy Miniature Schnauzer dogs and 41 healthy dogs of other breeds. Median serum CRP concentration of healthy Miniature Schnauzer dogs was compared with that of healthy non‐Miniature Schnauzer dogs. Results: The human CRP assay measured CRP reliably with linearity between 0 and 20 mg/L. CRP concentration for healthy Miniature Schnauzer dogs (median 4.0 mg/L, minimum–maximum 0–18.2 mg/L) was significantly higher than for the healthy non‐Miniature Schnauzer dogs (median 0.1 mg/L, minimum–maximum 0–10.7 mg/L); 17 of the 20 Miniature Schnauzer dogs had values that overlapped with those of the non‐Miniature Schnauzer dogs. Conclusions: Median CRP concentration of Miniature Schnauzer dogs was slightly higher than that of other breeds of dogs. A relationship between higher CRP concentration in Miniature Schnauzer dogs and idiopathic hyperlipidemia, pancreatitis, and possible increased risk for atherosclerosis remains to be determined.     

Gene and protein expression in the reproductive tract of Brazilian Somalis rams

Brazilian Somalis is a locally‐adapted breed of rams raised in tropical climate and native pastures. The present study was conducted to evaluate gene expression and proteome of the reproductive tract of such rams. Samples were collected from testes, epididymides, seminal vesicles and bulbourethral glands of four rams. Expression of clusterin (CLU), osteopontin (OPN) and prostaglandin D2 synthase (PGDS) genes were evaluated in all samples by real‐time PCR. Shotgun proteomic analysis was performed using samples from the head, corpus and cauda epididymides and from all other structures as well. Gene ontology terms and protein interactions were obtained from UniProtKB databases and MetaCore v.6.8 platform. CLU trasncripts were detected in the testes, epididymides, seminal vesicles and bulbourethral glands of the Somalis rams. The initial region and body of the epididymis had the greatest CLU expression. OPN mRNA was localized in all tissues of the ram reproductive tract. PGDS mRNA was detected in the testes and epididymides. Lable‐free mass spectrometry allowed the identification of 137 proteins in all samples. Proteins of the epididymis head mainly participate in cellular processes and response to stimulus, participating in catalityc activity and binding. Proteins of epididymis body acted as regulatory proteins and in cellular processes, with binding and catalytic activity. Cauda epididymis molecules were associated with cellular processes and regulation, with binding function and catalytic activity as well. Testis proteins were mainly linked to cell processes and response to stimuli, and had catalytic function. Seminal vesicle proteins were involved in regulation and mainly with binding functions. Most bulbourethral gland proteins participated in cellular processes. The present study is the first to evaluate the proteome and gene expressions in the reproductive tract of Brazilian Somalis rams. Such pieces of information bring significant cointribution for the understanding of the reproductive physiology of locally‐adapted livestock.     

Characterisation of the pig acute phase protein response to road transport

The acute phase protein (APP) response was evaluated after prolonged transportation of pigs under commercial conditions. Elevated serum APP concentrations were observed in two groups of boars immediately after their arrival at a destination farm compared with within-animal control samples obtained one month later. The effect was more pronounced in the first group of pigs conveyed under average transport conditions (Transport 1, 24 h), although the second group was transported for a longer time period (Transport 2, 48 h) but in superior transport conditions. In a second trial, pigs were sampled before transport, on arrival at an abattoir (following 12 h transport), and at the slaughter-line (after 6 h lairage). Significant increases in major acute phase protein (Pig-MAP), haptoglobin, serum amyloid A, C-reactive protein, and a decrease in apolipoprotein A-I, were observed at slaughter. The results demonstrate that shipment of pigs by road can result in an APP response that is probably related to the stress of transport.     

The influence of anticoagulants on the measurement of total protein concentration in equine peritoneal fluid

The aim of this study was to evaluate the influence of two commonly used anticoagulants (K3EDTA and lithium heparin) on refractometric and spectrophotometric measurement of total protein (TP) concentration in equine peritoneal fluid samples. The influence of a commercial solution of K3EDTA, a solution of K3EDTA in distilled water and lithium heparin on the refractometric and spectrophotometric (biuret) quantification of TP content in peritoneal fluid samples was assessed. Total protein concentration measured by refractometry was consistently overestimated in samples with commercial K3EDTA. The solution of K3EDTA in distilled water only caused TP overestimation at high K3EDTA concentrations (>5 micromol/ml). By contrast, lithium heparin did not influence the refractometric values of TP. Neither anticoagulant modified TP values when measured by the biuret method. In conclusion, the use of K3EDTA as anticoagulant may result in a significant overestimation of TP values of peritoneal fluid samples measured by refractometry.     

Relationship between serum acute phase protein concentrations and lesions in finishing pigs

To establish the relationship between serum levels of three acute phase proteins, haptoglobin (Hp), C-reactive protein (CRP) and serum amyloid A (SAA), and the occurrence and severity of lesions at slaughter, a study was carried out using 70 fattening pigs from a finishing unit. Pigs were divided into three groups: Group 1 (25 pigs with clinical signs of disease), Group 2 (25 apparently healthy pigs with lesions at slaughter) and Group 3 (20 apparently healthy pigs with no lesions at slaughter). Serum levels of CRP, SAA and Hp were significantly higher in pigs with clinical signs of disease than in apparently healthy animals. Additionally, in apparently healthy pigs, serum levels of Hp and CRP were significantly higher in animals with lesions than those without lesions. The extent and severity of lung lesions were related to serum levels of Hp.     

鸡源SUMO标签蛋白的鉴定及在IBDV VP2原核表达中的促溶功能

VP2蛋白是传染性法氏囊病病毒(infectious bursal disease virus,IBDV)的保护性抗原。原核表达中能否获得具有天然构象的可溶性VP2蛋白是研发IBD亚单位疫苗的关键问题。为了解是否存在鸡自体促溶标签蛋白以及外源促溶标签能否在一定程度上促进蛋白的正确折叠,试验对不同物种来源的类泛素蛋白修饰分子(small ubiquitin-like modifier,SUMO)进行比对研究。结果显示,获取到的3种鸡源SUMO蛋白与酵母源SUMO蛋白的氨基酸同源性均高于40%,其中SUMO-1与SUMO-3两种蛋白可以在大肠杆菌中可溶性表达;通过SUMO-1、SUMO-3与VP2蛋白的融合表达,证明了鸡源SUMO蛋白对VP2具有促溶效果,得到的重组菌株P-28A-s1-VP2和P-28A-Os3-VP2琼脂扩散效价均达到1∶16;经过纯化后,融合蛋白SUMO-1-VP2的琼脂扩散效价可达到1∶256;使用SUMO蛋白酶酶切融合蛋白,SDS-PAGE和Western blot结果表明鸡源SUMO蛋白可以被成功切割,说明试验发现的鸡源SUMO蛋白可以作为促溶标签,在亚单位疫苗关键抗原表达中具有广阔的应用前景。     

Effects of sub-clinical post-weaning colibacillosis and dietary protein on acute phase proteins in weaner pigs

There is an increasing body of evidence that plasma acute phase proteins (APP) can be used as a marker to assess pig health. The objective of this study was to assess whether experimentally induced sub-clinical post-weaning colibacillosis (PWC) affects the concentration of two plasma APP, i.e. haptoglobin (Hp) and C-reactive protein (CRP). Since sub-clinical PWC is sensitive to protein supply, we additionally assessed whether these APP responses are sensitive to dietary protein contents. Weaned pigs (6 groups of 9 each) were offered one of the three foods, i.e. H (230 g CP/kg), L (130 g CP/kg) or HA (23% CP with anti-microbial growth promoters). On day 5 post-weaning, pigs were either dosed with 10⁸ colony forming units of enterotoxigenic E. coli or were sham-infected. Three days later and across feeding treatments, infection had reduced feed intake. Feeding and infection treatment significantly interacted for both Hp and CRP. Infection increased Hp concentration in H and HA pigs, but not in L pigs, whilst infection increased CRP concentration to a larger extent in HA pigs than in H and L pigs. The results suggest that sub-clinical PWC leads to changes in APP profiles and that these changes may be sensitive to dietary protein contents. Usefulness of APP to describe health status in pigs may therefore need to be considered in relation to their nutritional environment.