Claudia Giannetto

Analysis of serum proteins in clinically healthy goats (Capra hircus) using agarose gel electrophoresis

BACKGROUND Electrophoretic patterns of serum proteins provide useful information on pathological conditions in ruminants. Their reference values, however, are dissimilar to those of other species. Reference values for goats using agarose gel as the supporting matrix have not been reported. OBJECTIVE The aim of this study was to evaluate serum concentrations of total protein and protein fractions (albumin and globulins) by means of agarose gel electrophoresis (AGE) in goats in order to establish electrophoretic reference intervals and to evaluate potential changes associated with aging. METHODS Blood was collected from 105 clinically healthy Girgentana goats by means of jugular venipuncture. Serum protein concentrations were assessed by AGE. Three age groups were compared: 1-1.5 years, 2-4 years, and 5-12 years. RESULTS Values (mean ± SD) were determined for concentrations of total protein (72.26 ± 6.40 g/L), albumin (31.80 ± 4.00 g/L), α-globulins (6.40 ± 1.23 g/L), β(1) -globulins (10.50 ± 2.58 g/L), β(2) -globulins (5.18 ± 1.60 g/L), and γ-globulins (18.65 ± 5.90 g/L) and for albumin/globulin (A/G) ratio (0.82 ± 0.20). One-way ANOVA showed statistically significant age-related differences for total protein and α-globulin concentrations and A/G ratios. Age influenced protein concentrations with the 5-12-year-old group having higher total protein and α-globulin concentrations and lower albumin concentration and A/G ratios than the 2-4-year-old group. CONCLUSIONS This study provides reference values for total protein concentrations and protein fractions obtained by AGE in goats. Some values vary with age. Age-specific reference intervals are reported in order to provide clinicians with an additional diagnostic aid.

Reference intervals for total protein concentration, serum protein fractions, and albumin/globulin ratios in clinically healthy dairy cows.

The aim of the current study was to evaluate total serum protein concentration measured by the biuret reaction as well as albumin and globulin protein fractions determined by agarose gel electrophoresis. These data were used to establish reference intervals in dairy cows of different ages. Blood was collected from 111 clinically healthy Modicana dairy cows by means of jugular venipuncture. Reference intervals (mean ± standard deviation) were determined for total protein (67.54 ± 11.53 g/l), albumin (31.86 ± 4.60 g/l), α1-globulin (5.77 ± 2.20 g/l), α2-globulin (5.84 ± 1.90 g/l), β-globulin (7.46 ± 1.94 g/l), and γ-globulin (16.73 ± 4.54 g/l) concentrations as well as for albumin/globulin (A/G) ratio (0.88 ± 0.43). Values from 2-, 3-, 4-, 5-, and 6-year-old cows were compared statistically. One-way analysis of variance showed age-related differences for α-globulin and β-globulin fractions only. The results of the current study provide reference intervals for total protein concentration as well as albumin and globulin protein fractions in 2- to 6-year-old dairy cows.

Daytime profile of the intraocular pressure and tear production in normal dog

OBJECTIVE To evaluate the circadian rhythms of intraocular pressure (IOP) and tear production in dog exposed to a natural photoperiod. ANIMALS STUDIED We used 12 clinically healthy Beagles dog housed under natural photoperiod at indoor temperature and humidity. PROCEDURE Intraocular pressure and Schirmer tear test (STT) I were measured every 4 h over a 48-h period in both eyes in each animal. Statistical analysis of the data was performed by one-way repeated-measures anova, Students t-test, and single cosinor method. RESULTS On each day, there was a highly significant effect of time on both parameters. A statistically significant difference of STT I values was observed comparing left and right eyes (P < 0.0001). Robust daily rhythms were observed for both parameters, IOP values showed diurnal acrophase (left eye: 09:33 +/- 00:50 h; right eye: 09:25 +/- 00:22 h), while STT I values showed nocturnal acrophase (left eye: 20:27 +/- 00:46 h; right eye: 20:00 +/- 00:05 h). CONCLUSION This study has demonstrated circadian rhythms in both IOP and STT I.

Daily rhythm of tear production in normal horse

OBJECTIVE This study was designed to assess the daily rhythm of tear production in clinically healthy horses, to compare left eye and right eye, and to evaluate the influence of time and gender. ANIMAL STUDIED For our study 18 horses (nine female and nine male) were subjected to three different light/dark (L/D) schedules: 12/12 L/D cycles, constant light (24/0 L/D) and constant darkness (0/24 L/D). PROCEDURES In all subjects Schirmer tear tests were performed at 4-h intervals over a 24-h period. RESULTS A circadian rhythm of tear production was found during the 12/12 L/D period and during constant darkness. Also our results showed a statistical significant difference comparing left eye and right eye, and between gender. CONCLUSIONS Our findings would suggest that the time of tear evaluation is significant in the diagnosis of tear film disorders, and that the difference between the two eyes and the two genders should be taken into consideration during the ophthalmic exam.

Daily rhythms of activity in horses housed in different stabling conditions

Abstract Total locomotor activity was studied in 10 Thoroughbreds housed under a natural 12/12 light/dark cycle. Five horses were housed in individual boxes, and five were housed in individual boxes with a paddock. In order to record locomotor activity, on each horse was placed an Actiwatch-Mini® (Cambridge Neurotechnology Ltd, UK), actigraphy-based data loggers that record a digitally integrated measure of motor activity. Locomotor activity in the different experimental conditions was evaluated by visual inspection. Average amount of activity (bout of activity/hour) during light and dark phase and cosine Peak (time of peak activity) were calculate using Actiwatch Activity Analysis 5.06. Students t-test was used to determine significant differences. The results from this study underline the influence of stabling conditions on activity rhythms in horses; furthermore, we clearly established that in horses, the activity rhythm reaches its peak in the middle of the day.

Seasonal variations in daily rhythms of activity in athletic horses

Circadian rhythms reflect extensive programming of biological activity that meets and exploits the challenges and opportunities offered by the periodic nature of the environment. In the present investigation, we recorded the total activity of athletic horses kept at four different times of the year (vernal equinox, summer solstice, autumn equinox and winter solstice), to evaluate the presence of seasonal variations of daily activity rhythms. Athletic Thoroughbred horses were kept in individual boxes with paddock. Digitally integrated measure of total activity of each mare was continuously recorded by actigraphy-based data loggers. Horse total activities were not evenly distributed over the day, but they were mainly diurnal during the year. Daily activity rhythms showed clear seasonal variations, with the highest daily amount of activity during the vernal equinox and the lowest during the winter solstice. Interestingly, the amount of activity during either photophase or scotophase changed significantly throughout the year. Circadian analysis of horse activities showed that the acrophase, the estimated time at which the peak of the rhythm occurs, did not change during the year, it always occurred in the middle of the photoperiod. Analysing the time structure of long-term and continuously measured activity and feeding could be a useful method to critically evaluate athletic horse management systems in which spontaneous locomotor activity and feeding are severely limited. Circadian rhythms are present in several elements of sensory motor and psychomotor functions and these would be taken into consideration to plan the training schedules and competitions in athletic horses.

Influence of transportation on serum concentrations of acute phase proteins in horse

The modifications of Haptoglobin (Hp), Serum Amyloid A (SAA), Fibrinogen (Fbg) and White Blood Cells (WBCs) were evaluated in 15 Saddle Italian horses. Ten horses were transported covering a distance of about 320 km within 4 h with an average speed of 80 km/h (experimental group) and five horses were not subject to transportation (control group). Blood was collected via jugular venipuncture before the transportation (T0), immediately after the transportation (T1), 12 (T12), 24 (T24) and 48 (T48)hours after the transportation in experimental group and at the same time point in control group. For each parameter statistical analysis of different groups and sampling time was performed using a two-way analysis of covariance, with the data before the transportation (T0) as the covariate, by the GLM procedure of SAS. For all parameters the interaction (Group × Time) was tested and it was resulted no significant. The application of statistical analysis showed significant differences between the control group and horses subjected to transportation (P<0.01), and the influence of sampling time (P<0.05) on Hp, SAA and WBCs. These modifications appeared to be innovative showing that equine Hp, generally considered as moderate acute phase protein, increases more rapidly than the SAA after transportation-induced stress.

Influence of age on profile of serum proteins in the calf.

The authors studied the course of serum proteins during the first week and the first month of life in the calf in order to obtain useful information for neonatal care. Eight Limousine calves, four males and four females, clinically healthy, were used. From all animals blood samples were collected from the external jugular vein from the day after the birth every day for one week and every five days for thirty days. Blood samples were taken at the same hour (09:00), and the serum concentrations of albumin, globulins (α1-globulins, α2-globulins, β- globulins, γ-globulins) and total proteins were determined by means of automated multiparametric agarose gel electrophoresis system. One-way repeated measures analysis of variance (ANOVA), followed by the Bonferronis test, was used to determine significant differences. Data analysis of variance showed a significant effect of days of life, with p < 0.05, only on albumin, α1-globulins and α2-globulins during the first month of life. These results confirm that despite the fact that newborns must adapt to various environmental factors after birth, including nutrition that can influence the serum protein profile during neonatal period, the evaluation of electrophoretics parameters during the first months of life results useful for the diagnosis and treatment of neonatal diseases.

Annual rhythms of some physiological parameters in Ovis aries and Capra hircus

The aim of this study was to evaluate the annual variation of some physiological parameters (total lipid, NEFA, phospholipids, triglycerides, creatinine, β-idrossibutirrate, albumin, total protein, total cholesterol, urea, uric acid, glucose, rectal temperature and heart rate) in five female goats and five female sheep housed under natural photoperiod and natural ambient temperature in a 12 m2 sound proof box equipped with 50 × 100 cm opening window, that allowed natural ventilation. ANOVA was used to determine significant differences (p values < 0.05 were considered statistically significant). A trigonometric statistical model was applied to describe the periodic phenomenon analytically, by individuating the main rhythmic parameters according to the single cosinor procedure. Serum total lipid, NEFA, phospholipids, triglycerides, rectal temperature and heart rate, both in sheep and goats, showed an annual rhythm with acrophases in autumn. The results show that sheep and goats present similar annual patterns, probably influenced by various environmental factors.

Pattern of serum protein fractions in dairy cows during different stages of gestation and lactation

In dairy cows the period of transition from late gestation to early lactation is recognized as inducing considerable metabolic adaptation. The aim of this study was to analyse modifications in serum protein values occurring during the dry and the transition period and during lactation in a group of five Holstein cows of high average milk production. For all subjects, selected on the basis of their pregnancy status, blood samples were collected at different physiological phases: dry period (-60, -30 d to calving), transition period (almost 7 d to calving, 7 d after calving), and lactation (weeks 2, 5 and 15 after calving), for a total of eight blood samples for each cow. On each blood sample total proteins and electrophoresis analysis were performed. On the data obtained, normally distributed (P<0·05, Kolmogorov-Smirnovs test), one-way Repeated Measure Analysis of Variance (ANOVA), was applied to evaluate the influence of different stages of gestation and lactation on the considered parameters. Results showed a significant effect on total proteins, α1-globulins, β-globulins, γ-globulins and albumin/globulin ratio. Most of the detected modifications were related to the transition from gestation to lactation, indicating that it is a period of great metabolic stress for cows. On the basis of the obtained results we can affirm that the pattern of serum protein fraction rn could give information about dehydration, plasma volume expansion and hepatic function occurring during the peripartum period in dairy cows.