Biochemical and hematologic reference intervals for free-ranging desert bighorn sheep

Over 200 clinically normal desert bighorn sheep (Ovis canadensis) from multiple geographic areas were sampled utilizing a uniform protocol. The goals of this study were to develop comprehensive reference intervals for hematologic and biochemical analytes using central 90th percentile nonparametric analyses. Adult female sheep had greater erythrocyte mass (hemoglobin and hematocrit) compared with adult male sheep. Young animals < or = 1-yr-old had greater erythrocyte mass (hemoglobin, hematocrit and red blood cell count), higher alkaline phosphatase activity, and lower serum protein and globulin concentrations compared with adult animals. Because of the large sample size, wide geographic range, and uniform sample and handling protocol in this study, these reference intervals should be robust and applicable to other free-ranging desert bighorn sheep populations.     

嗜麦芽寡养单胞菌脂多糖对斑点叉尾免疫保护作用

嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)是近年来引起斑点叉尾高致死性、传染性疾病的主要病原之一。为了研究嗜麦芽寡养单胞菌脂多糖对斑点叉尾的免疫保护作用,实验选用了400尾健康斑点叉尾,随机分成Ⅰ、Ⅱ、Ⅲ、Ⅳ四个组,每组用鱼100尾,每组下设两个重复,每个重复用鱼50尾,以腹腔注射的方式分别向Ⅰ、Ⅱ、Ⅲ、Ⅳ四个组的斑点叉尾注射嗜麦芽寡养单胞菌脂多糖、无花果多糖加脂多糖、全菌灭活苗和生理盐水,在试验第0、第28天时分别进行首次免疫和加强免疫,试验期间每隔7d,对试验鱼的血液白细胞杀菌活性、补体C3含量、IgM含量和血清凝集效价滴度进行测定;试验第49天时进行活菌攻毒。结果表明,首免后,接种脂多糖、无花果多糖加脂多糖的试验鱼血清凝集效价滴度明显升高,白细胞杀菌活性、补体C3含量、IgM含量也明显增加;加强免疫后,脂多糖、无花果多糖加脂多糖的试验鱼血清凝集效价滴度峰值分别为1∶256和1∶512,白细胞杀菌活性峰值分别为0.565和0.511,补体C3含量峰值分别0.194和0.180mg/mL,IgM含量峰值分别1.415和1.464mg/mL,免疫保护率分别为70.0%和65%。嗜麦芽寡养单胞菌脂多糖和脂多糖+无花果多糖受免鱼的上述指标均显著(P<0.05)或极显著(P<0.01)地高于生理盐水注射组,这两者的免疫保护效果优越于全细胞灭菌苗。     

Phagocytosis by catfish neutrophils

Channel catfish peripheral blood leucocytes were separated on a Percoll gradient to establish the phagocytic function of the neutrophils. Four fractions of leucocytes were formed on the Percoll gradient, including a fraction that contained 50–80% neutrophils at a density of 1.08–1.09 g ml⁻¹ and a fraction that contained 10% monocytes at a density of 1.071–1.074 g ml⁻¹. Phagocytic assays, using ³H-uridine, showed that the two fractions had similar phagocytic indices, although neutrophils were less phagocytic than monocytes. Neutrophils were confirmed to be phagocytic when examined with transmission electron microscopy. Staining with 3,3-diaminobenzidine-tetrahydrochloride demonstrated peroxidase-positive granules in the cytoplasm of actively phagocytic cells as well as peroxidase reaction products in a number of phagosomes containing bacteria. Phagocytosis of bacteria by channel catfish neutrophils was further confirmed by differential staining of external bacteria and cell surfaces with ruthenium red during the fixation process.     

红腹锦鸡、石鸡和雉鸡的部分血液生理生化指标

为了解红腹锦鸡、石鸡和雉鸡的血象、血液生理生化参数正常值及其特点,为人工饲养繁殖与健康检测提供参考资料,采用改良纽巴氏法、沙利氏比色法、离心法、瑞氏染色等方法和全自动血液生物化学分析仪对红腹锦鸡、石鸡和雉鸡的血象和16项生化指标进行了测定,通过SDS-PAGE分析了血浆蛋白质。结果表明:红腹锦鸡的平均红细胞体积、淋巴细胞、嗜酸性粒细胞、嗜中性粒细胞、血浆总蛋白、血浆白蛋白、血浆钠和血浆氯在性别之间有显著差异,所测其它生理生化指标在性别之间无显著差异;红腹锦鸡的红细胞、平均红细胞体积、凝血细胞、尿素氮和肌肝与石鸡和雉鸡存在显著差异,红腹锦鸡与石鸡的白细胞存在显著差异,所测其它生理生化指标在三者之间无显著差异。种属不同可能是导致3种雉科鸟类血液生理常值差别的主要原因,而采样时的机体状况、营养状况及测试操作方法也有一定的影响。     

圈养毛冠鹿血液生理生化指标测定

本文采集了8只不同年龄段毛冠鹿的14个血液样品,并检测和分析了样品中的血细胞、血蛋白、电解质、生化酶类等44项指标的含量。初步建立了圈养毛冠鹿的血液学生理生化指标参考值,为今后加强毛冠鹿的人工饲养及繁育研究提供了参考依据。     

豚鹿血液细胞和血液化学指标测定

应用Sysmex F-820血球计数仪和Olympus AU400全自动生化分析仪,对成都动物园圈养的16只豚鹿的46个血液样本进行血液细胞及血液化学43项指标检测,并对成体、亚成体及幼体进行统计分析,建立了一个参考指标.这对豚鹿的疾病诊断和繁育研究具有重要的意义.     

Hematological and serum biochemical indices in healthy bonnet macaques (Macaca radiata)

Background Blood reference values for bonnet macaques (Macaca radiata) are limited. The goal of this study was to determine reference ranges for hematological and serum biochemical indices in healthy, socially housed bonnet macaques for males and females over a range of ages. Methods Blood hematological and serum biochemical values were obtained from 50 healthy bonnet macaques of both sexes and aged 10–234 months. Results Age and sex differences were present in a number of measures. Globulins, total protein, and creatinine (CREAT) values were highest among older subjects, while alkaline phophatase, albumin, and phosphorus values were higher in juveniles. Sex differences were present in concentrations of red blood cells and CREAT, with higher values in males. Conclusion The blood parameter data reported here as age‐specific reference values for laboratory‐housed, healthy bonnet macaques may be used to inform clinical care and laboratory primate research.     

Haematological and biochemical blood reference values for Canary Island camels (Camelus dromedarius), an endangered dromedary species

The purpose of this research was to develop reference values for haematological and biochemical variables in the Canary camel breed (Camelus dromedarius). 114 clinically healthy dromedary camels were assessed. Age, sex, and pregnancy status was also recorded. The reference range for red blood cells (RBCs) was 8.45 – 13.65 X10⁶/µL, haemoglobin (HGB) was 10.61 – 15.29 g/dL, packed cell volume (PCV) was 19.93 – 32.51 %, and white blood cells (WBCs) 7.35 – 18.36 X10³/µL. A correlation was established between the haemoglobin concentration (HGB) (g/dl) and packed cell volume (PCV) obtaining a linear regression (HGB = 0.31 PCV + 4.67). Young animals had higher RBC and WBC values than adult animals. Additionally, blood urea nitrogen (BUN), phosphorus, calcium, albumin/globulin (A/G) ratio, alkaline phosphatase, cholesterol, and lipase were higher in young animals compared with adults. Female dromedary camels showed higher values for the three main variables: RBC, HGB and PCV, but no differences between sexes were detected in the biochemical variables results. The WBC count was higher in non-pregnant females than in pregnant animals. These results provide references values for the Canary camel breed and may contribute to the understanding of differences in 18 haematological and biochemical parameters in dromedary camels with a potential impact in health and welfare for this species.     

Hematological and serum biochemical reference intervals of rainbow trout,Oncorhynchus mykiss cultured in Himalayan aquaculture: Morphology,morphometrics and quantification of peripheral blood cells

De novo reference intervals (RIs) for a total of thirty two hematological and serum biochemical attributes were established for rainbow trout (Oncorhynchus mykiss) cultured in Himalayan aquaculture system. For this purpose, long term assessment of hemato-biochemical parameters was carried over a period of one year from March 2019 to February 2020 and a total of 444 blood samples were analysed. Blood examination results were recorded systematically and reference intervals were established, notably for erythrocyte parameters: hematocrit (Hct) 29–40%, hemoglobin 8.32–12.28 g/dL, red blood cell (RBC) count 1.01–2.04 (×10⁶/mm3); leukocyte parameters (x 10³/mm3): total leukocytes 31.32–90.60, neutrophils 4.21–18.85, total lymphocytes 20.55–63.63, small lymphocytes 14.86–46.50, large lymphocytes 6.35–22.34 and monocytes 1.22–7.56; thrombocyte count 23.00–68.00 (×10⁶/mm3). RIs were also established for red blood cell indices, vital serum constituents involved in carbohydrate, protein, lipid and nitrogen metabolism including the less known, diagnostically important, serum enzymes and electrolyte concentrations. Principal component analysis revealed that certain serum components were more efficient at distinguishing between the life stages (juvenile, adult) of fish by explaining about 92.7% of variation in the whole dataset compared to the principal hematological components which explained only about 80% of the variation. Significant (P < 0.05) differences were noted for RBC count, total leukocyte count (TLC), total protein, total cholesterol and uric acid with respect to the sex of fish. Moreover, clearly differentiable morphometric and morphological attributes were also noticed among erythrocytes, leukocytes (lymphocytes, neutrophils and monocytes) and thrombocytes. To our knowledge, the present study is the first of its kind that elucidates blood chemistry of cultured rainbow trout, O. mykiss in accordance to the guidelines framed by the American society of veterinary clinical pathologists (ASVCP). RIs reported here can help monitor the fish health status by improving the use of non-lethal diagnostics in piscine medicine.     

Hematologic and biochemical profiles in Standardbred mares during peripartum

The purposes of this study were to determine physiological changes occurring in hematologic and biochemical parameters in mares between the last month of gestation and the first week after parturition. If a significant change was observed with respect to the reference interval of an adult horse, a further aim of the study was to establish different reference intervals. Blood samples were collected from 62 healthy pregnant Standardbred mares. Seventeen nonpregnant and nonlactating mares were used as a control group. In pregnant mares, blood sampling was conducted every three days from 1 month before the expected foaling date (335 days after the last mating), at parturition, and 7 days after foaling. The barren mares in the control group were sampled once. Results from samples collected 20 and 10 days before parturition, at parturition, and 7 days after were considered in the statistical analysis. A parametric method for all the parameters studied was used to establish reference intervals. Results were compared by repeated measures ANOVA. When significant differences were observed in relation to sampling time, a post hoc analysis was performed (Tukey test). The one-way ANOVA test followed by Dunnett's test was performed to evaluate the presence of a significant difference between each sampling time and the control group. Any significant difference in the blood count parameters at different sampling times was observed by repeated measure ANOVA. Hemoglobin (P < 0.01) and hematocrit (P < 0.01) 7 days after parturition and white blood cell count (P < 0.01) at parturition were significantly different from the control group. Erythrocyte indices and platelet count were within the normal reference intervals as established in the control group. In the biochemical panel, gamma-glutamyltransferase, creatinine, glucose, biliar acids, total protein, albumin-to-globulin ratio, and calcium were significantly different at different sampling times. Moreover, serum concentration of creatine kinase, aspartate aminotransferase, creatinine, blood urea nitrogen, glucose, lactate, total protein, albumin, albumin-to-globulin ratio, calcium, magnesium, sodium, chloride, potassium, and total, direct, and indirect bilirubin was different from that of the control group. Remarkable changes were not observed in alkaline phosphatase, triglyceride, and fibrinogen concentrations. Temporal changes in the hematologic and biochemical parameters observed in the present study in the peripartum and the differences with reference intervals made up for nonpregnant and nonlactating mares could be used to better evaluate the conditions of periparturient mares.     

Reference values of blood biochemical indices in Russian cosmonauts

In the period from 2000 to 2012, the values of 40 diagnostically significant biochemical indices in venous blood were determined during the preflight clinical and physiological examination of 28 Russian cosmonauts aged 35 to 54, members of the main and backup crews of missions on board the International Space Station. The examination was conducted 45-30 days prior to the launch. Since most cosmonauts performed several flights during this period and they were repeatedly included in backup crews, each of the cosmonauts participated in preflight examinations one to five times. The reference values were calculated for each of the studied indices. It was found that reference interval boundaries for 15 indices were narrower as compared to the generally accepted ranges. The upper boundaries of the reference ranges of the activities of a number of enzymes, as well as of the concentrations of some metabolites of energy and plasticity metabolism, were higher than the population’s mean values. Thus, it was found that Russian cosmonauts, being a specific professional team, had reference values of a number of blood biochemical indices that were different from the population’s mean values. This was due to the characteristics of selection, physical training, and psychic and emotional states of crew members during the preflight period.     

Gender and age-specific reference intervals of common biochemical analytes in Chinese population: Derivation using real laboratory data

BackgroundIndirect sampling methods are not only inexpensive but also efficient for establishing reference intervals (RIs) using clinical data. This study was conducted to select fully normal records to establish ageand gender-specific RIs for common biochemical analytes by laboratory data mining.MethodsIn total, 280,206 records from 2014 to 2018 were obtained from Peking Union Medical College Hospital. Common biochemical analytes total protein, albumin, total bilirubin (TBil), direct bilirubin (DBil), alanine aminotransferase (ALT), glutamyltranspeptidase (GGT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), potassium, sodium, chlorine, calcium, urea, glucose, uric acid (UA), inorganic phosphorus, creatinine (Cr), total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol] were measured using an automatic analyzer. Sources of variation were identified by multiple regression analysis. The 2.5th and 97.5th percentiles were calculated as the lower and upper limits of the RIs, respectivelyResultsGender was the major source of variation among the 13 common biochemical analytes with an rp > 0.15. In contrast to the value listed in the WS/T 404, nearly all RIs established in this study were significantly narrower. Furthermore, age-specific RIs should be determined for DBil, LDH, and urea, whereas gender-specific RIs are suggested for GGT, LDH, and urea.ConclusionsWe recommend that gender-specific RIs should be established for ALT, AST, GGT, DBil, TBil, UA, and Cr as well as genderand age-specific RIs for urea and ALP. Through indirect sampling, ageand gender-specific RIs for common biochemical analytes were established and analyzed.     

南方鲇血型的初步鉴定

本研究旨在通过观察南方鲇血清与其红细胞的交叉反应以鉴定南方鲇的血型.实验结果表明:南方鲇的血清与同种其他个体的红细胞进行交叉反应时均未出现凝集现象,这表明南方鲇可能不存在血型或南方鲇具备血型但血清中相应的凝集素含量不足.以南方鲇的红细胞为抗原免疫日本种大耳白兔制备的抗血清与南方鲇的红细胞进行交叉反应,出现了不同程度的凝集反应,这表明南方鲇存在血型.据上述两个实验结果可以推断,南方鲇可能存在4种血型,分别命名为NA、NB、NAB和NO型;同时也证实,在鉴定南方鲇血型的研究中,通过制备抗血清与红细胞进行交叉反应的方法更为可靠.     

Phylogeny of lymphocyte heterogeneity: the cellular requirements for in vitro antibody responses of channel catfish leukocytes

Three functionally distinct leukocyte subpopulations were isolated from the peripheral blood of channel catfish. Surface immunoglobulin-positive (sIg+) and sIg- lymphocytes were isolated by an indirect "planning" procedure employing monoclonal antibodies specific for channel catfish Ig. A third population composed of macrophages was isolated by adherence to baby hamster kidney cell microexudate-coated surfaces. Functional features of these three cell types were established by assessing their role(s) in primary in vitro anti-hapten PFC responses to known murine thymus-dependent (TD) and thymus-independent (TI) antigens. The results indicated that anti-hapten PFC responses to a TI antigen required the presence of sIg+ lymphocytes and macrophages. In contrast, all three cell types were required for responses to TD antigens. Furthermore, the results of studies involving the depletion of antigen-reactive lymphocytes demonstrated that both hapten-specific sIg+ cells and carrier-specific sIg- cells were required for anti-hapten responses to TD antigens. These studies provide direct evidence that catfish have separable B cells (sIg+ lymphocytes), T helper cells (sIg- lymphocytes), and accessory cells (macrophages) quite similar to those seen in higher animals.     

Hematologic and biochemical values for Spanish ibex (Capra pyrenaica) captured via drive-net and box-trap

Between October 2002 and September 2004, 70 free-ranging Spanish ibex (Capra pyrenaica) were captured in Catalonia, northeastern Spain, using two different physical methods, drive-net (n=26) and box-trap (n=44). Blood samples were taken to determine 20 hematologic and 23 biochemical variables. Values obtained fell within already published reference intervals, with the following exceptions: higher values for red blood cells (RBC), packed cell volume (PCV), white blood cells (WBC), eosinophil count, triglyceride concentration, creatine kinase (CK; in box-trap), chloride, sodium, alpha-1, alpha-2, and gamma electrophoretic fractions of serum proteins; and lower values for hemoglobin, mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), urea concentration, CK (in drive-net), albumin, and albumin:globulins ratio (A:G). Published values for aspartate aminotransferase (AST) concentration are both higher and lower than observed in this study. In our study, monocyte and eosinophil counts, as well as triglyceride and potassium concentrations, were lower in animals captured via box-trap than those captured via drive-net. Conversely, MCHC, neutrophil count, total bilirubin concentration, urea, and AST were higher in animals captured via box-trap. Hematologic and biochemical values obtained from Spanish ibexes show that the drive-net is a newer, less-stressful method of capture than the box-trap.     

Isolation, culture and characterization of a primary fibroblast cell line from channel catfish

A primary cell line (designated as CCf) derived from caudal fin tissue of channel catfish, Ictalurus punctatus, was developed using explant techniques. The cell line grew fastest in media supplied with FBS and channel catfish serum. The duplication time of the cell line under optimal conditions was ∼56 h at a plating density of 1.1 × 10⁵ cells/ml. The cell line has been propagated continuously for 25 passages (1:4 dilution per passage), cryopreserved, and recovered successfully at different passages. The cultured cells had fibroblastic morphology, and synthesized fibronectin and Type I and III collagens in the cytoplasm. The cell line maintained the normal diploid chromosome number (58) of channel catfish throughout the experiment. Nucleolus organizer regions were located on the short arms of a pair of medium-sized submetacentrics, which is typical for channel catfish. This study provides a method for acquiring a cell line from juvenile catfish without sacrifice, and is especially useful for early screening of valuable fishes. This revised version was published online in August 2006 with corrections to the Cover Date.     

Thioredoxin acts as a B cell growth factor in channel catfish

To identify differentially expressed genes from channel catfish macrophages, a cDNA library from LPS-stimulated catfish macrophages was screened by subtractive hybridization. This screening yielded a 552-bp cDNA coding for catfish thioredoxin (CF-TRX). The deduced amino acid sequence revealed that CF-TRX contains 107 amino acids and is 59% homologous to human adult T cell leukemia-derived factor/TRX, originally described as an IL-2R alpha-inducing factor. Northern blot analyses showed that CF-TRX is expressed in catfish T and macrophage cell lines, but weakly in B cell lines. Similar results were also observed in Western blot analyses using a mAb specific for recombinant CF-TRX (rTRX). The use of rTRX in functional studies demonstrated that rTRX induces in vitro proliferative responses of catfish PBL that were synergistically enhanced by the addition of culture supernatants from catfish T cell lines. In addition, cell separation studies and flow cytometric analyses revealed that the cells proliferating in rTRX-stimulated cultures were mostly B cells. These results suggest that CF-TRX may have an important role in the activation and proliferation of channel catfish B cells.     

Construction of Genetic Linkage Maps and Comparative Genome Analysis of Catfish Using Gene-Associated Markers

A genetic linkage map of the channel catfish genome (N = 29) was constructed using EST-based microsatellite and single nucleotide polymorphism (SNP) markers in an interspecific reference family. A total of 413 microsatellites and 125 SNP markers were polymorphic in the reference family. Linkage analysis using JoinMap 4.0 allowed mapping of 331 markers (259 microsatellites and 72 SNPs) to 29 linkage groups. Each linkage group contained 3–18 markers. The largest linkage group contained 18 markers and spanned 131.2 cM, while the smallest linkage group contained 14 markers and spanned only 7.9 cM. The linkage map covered a genetic distance of 1811 cM with an average marker interval of 6.0 cM. Sex-specific maps were also constructed; the recombination rate for females was 1.6 times higher than that for males. Putative conserved syntenies between catfish and zebrafish, medaka, and Tetraodon were established, but the overall levels of genome rearrangements were high among the teleost genomes. This study represents a first-generation linkage map constructed by using EST-derived microsatellites and SNPs, laying a framework for large-scale comparative genome analysis in catfish. The conserved syntenies identified here between the catfish and the three model fish species should facilitate structural genome analysis and evolutionary studies, but more importantly should facilitate functional inference of catfish genes. Given that determination of gene functions is difficult in nonmodel species such as catfish, functional genome analysis will have to rely heavily on the establishment of orthologies from model species.     

Branchial chemoreceptors mediate ventilatory responses to hypercapnic acidosis in channel catfish

The effects of hyperoxic hypercapnia on cardiovascular and ventilatory variables and blood gas and acid/base parameters were examined in conscious and anesthetized spontaneously breathing (ASB) channel catfish, Ictalurus punctatus. These separate experiments were designed to determine: (1) if channel catfish show a ventilatory response to hypercapnic acidosis when blood O(2) content is maintained in conscious animals; and (2) whether branchial receptors innervated by cranial nerves IX and X mediate this response. The combination of high O(2) and CO(2) tensions allowed the cardioventilatory effects of hypercapnic acidosis to be assessed independently of Root or Bohr mediated changes in blood O(2) content. In the absence of significant changes in dorsal or ventral aorta O(2) content, hyperoxic hypercapnia significantly stimulated ventilation, relative to hyperoxic exposure. Hypercapnic acidosis, however, had no significant effects on blood pressure or heart rate. Branchial denervation in ASB fish abolished the ventilatory response to hypercapnic acidosis. The results indicate that hypercapnic acidosis independently stimulates ventilation in channel catfish. This response is mediated by CO(2)/pH-sensitive branchial receptors innervated by cranial nerves IX and X.     

Valoración de la función tiroidea durante la gestación: intervalos de referencia de tirotropina y tiroxina no unida a proteína durante el primer trimestre

Background and objectivesThe physiological changes that occur during pregnancy affect the physiology of the thyroid gland. Consequently, interpretation of thyroid function markers during pregnancy requires trimester-specific reference intervals. The aims of our study were to: 1) establish first-trimester reference intervals for biochemical markers of thyroid function [thyroid-stimulating hormone (TSH) and free thyroxine (T4)] and 2) to establish the prevalence of autoimmune thyroid disease in pregnant women resident in Cartagena (Murcia, Spain).Patients and methodA total of 441 women between weeks 11 and 13 of pregnancy were included in this study. A blood sample was extracted from all women to measure TSH, free T4 and antithyroid antibodies. Reference intervals for TSH and free T4 were determined in 400 pregnant women without autoimmune thyroid disease or known thyroid disease.ResultsAutoimmune thyroid disease was detected in 23 pregnant women (5.2%) who showed TSH levels higher than those in pregnant women without thyroid autoimmunity. First-trimester reference intervals were as follows: TSH: 0.130–3.710 mUI/L; free T4: 0.89–1.50 ng/dL. These reference intervals differed from the non-pregnant reference intervals used in our laboratory.ConclusionsThe reference intervals established are useful to evaluate thyroid function in women between 11 and 13 weeks of pregnancy. Interpretation of thyroid function requires intervals established in a reference population without autoimmune thyroid disease and with the methodology usually used to analyze these markers.