Histopathological effects of cadmium on the gills of the freshwater fish, Macropsobrycon uruguayanae Eigenmann (Pisces, Atherinidae)

Evaluations of histopathological lesions in gill tissue were carried out in the freshwater fish Macropsobrycon uruguayanae following 30 and 60 days of exposure to 1.5 mg 1^-1 of cadmium. The study was conducted on both fed and starved animals in order to determine the influence of feeding condition on cadmium toxicity. The main lesions observed and quantified were: (1) hyperplasia of primary lamellar epithelium; (2) hyperplasia of secondary lamellar epithelium; (3) separation of respiratory epithelium; (4) shortening of secondary lamellae; (5) epithelial necrosis; (6) fusion of adjacent secondary lamellae; (7) hypertrophy of respiratory epithelium; (8) lamellar telangiectasis; (9) hyperplasia of chloride cells; (10) mucinous metaplasia; and (11) inflammatory infiltration. Lesions 6, 8, 9 and 11 were only induced by exposure to cadmium, while lesion 4 could be produced only by starvation. Starved fish also showed a reduction in total body weight and length. Lesion 2 was shown to be non-specific, and produced by either cadmium, starvation or even exposure time. The possible mode of action of the experimental factors are discussed in relation to the observed pathologies.     

Use of touch-down polymerase chain reaction to enhance the sensitivity of Mycobacterium bovis detection.

The confirmatory diagnosis of Mycobacterium bovis (M. bovis) in animal samples is carried out by culture in Stonebrink media. However, culture is very slow because of the extremely long duplication time of the bacillus and difficult because of the scarcity of bacilli in diagnostic samples. This study describes the development of a single-tube touch-down polymerase chain reaction (PCR) protocol for the detection of M. bovis using primers that target the IS6110 element. Spiked water and milk as well as routine diagnostic samples (milk and nasal swabs) from M. bovis-positive cattle were tested. This protocol allows the rapid and sensitive detection of M. bovis in bovine samples by enhancing the sensitivity of standard PCR amplification.     

Plasma and urine electrolyte and mineral concentrations in Thoroughbred horses with recurrent exertional rhabdomyolysis after consumption of diets varying in cation-anion balance

OBJECTIVE: To determine whether plasma, urine, and fecal electrolyte and mineral concentrations differ between clinically normal horses and Thoroughbreds with recurrent exertional rhabdomyolysis (RER) after consumption of diets varying in cation-anion balance. ANIMALS: 5 Thoroughbred mares with RER and 6 clinically normal mixed-breed mares. PROCEDURE: Each of 3 isocaloric diets designated as low, medium, and high on the basis of dietary cation-anion balance (DCAB) values of 85, 190, and 380, respectively, were fed to horses for 14 days. During the last 72 hours, 3 horses with RER and 3 control horses had daily urine and fecal samples obtained by total 24-hour collection. Remaining horses had urine samples collected daily by single catheterization. RESULTS: For each diet, no differences existed between horses with RER and control horses in plasma pH, electrolyte concentrations, and creatine kinase activity or in urine pH and renal fractional excretion (FE) values. Plasma pH, strong ion difference, bicarbonate and total carbon dioxide concentrations, and base excess decreased and plasma chloride and ionized calcium concentrations increased with decreasing DCAB. Urine pH decreased with decreasing DCAB. The FE of chloride and phosphorus were greatest for horses fed the low diet. The FE values for all electrolytes exept magnesium did not differ between urine samples obtained by single catheterization and total 24-hour collection. Daily balance of calcium, phosphorus, sodium, chloride, and potassium did not differ significantly among horses fed the various diets. CONCLUSIONS: In clinically normal horses and in horses with RER, the DCAB strongly affects plasma and urine pH and the FE of sodium, potassium, chloride, and phosphorus.     

Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition

Impaired polymorphonuclear neutrophil leukocyte (PMN) function around parturition has been associated with increased clinical mastitis in dairy cows. Rolling and attachment of PMN to the endothelium is the first step in the recruitment process and is accomplished by interaction between L-selectin on PMN and its ligand on endothelial cells. Furthermore, tyrosine phosphorylation is involved in the initiation of many PMN functions. The objective of this work was to determine changes in expression of L-selectin and tyrosine phosphorylation in the perinatal period. Eight clinically healthy Holstein cows were used as PMN donors at d-21, -14, -7,0 (calving), +1, +2, +7, +14, +28. Evaluation of L-selectin expression was carried out on activated and resting PMN. Anti-bovine L-selectin monoclonal antibody (MAB) and flow cytometric analysis were used to measure the percentage of PMN fluorescing and receptor expression (log mean fluorescent channel, LMFC). Activated and resting PMN showed similar trends in % PMN fluorescence and LM FC. The percentage of PMN fluorescing tended to decrease at parturition, followed by a significant increase at d +14 and +28 (P < 0.02). For LMFC a decrease was observed on d +1 followed by an increase through d +28 (P < 0.01). Protein tyrosine phosphorylation of lysates prepared from PMN isolated throughout the study was detected by electrophoresis and western blotting using anti-phosphotyrosine MAB. Several protein bands were tyrosine phosphorylated. Two of these bands (42-44 kDa and 90 kDa) varied in intensity over time. The intensity of the 42-44 kDa band gradually increased from d -7, peaked at d +7 (P < 0.03), and steadily decreased to d +28 (P < 0.02). Antibody to activated mitogen protein kinase reacted with the 42-44 kDa band. Reduced PMN function during the periparturient period could be related to reduced L-selectin adhesion molecules on the cell surface, and to modulation in the phosphorylation of functionally important molecules.     

Follicular Dynamics in Heifers during Pre-pubertal and Pubertal Period Kept under Two Levels of Dietary Energy Intake

The objective of this study was to characterize follicular dynamics in pre-pubertal, pubertal and post-pubertal periods, as well as the effect of high-energy intake on follicular development and age at puberty in heifers. Thirty-one Nelore (Bos indicus) heifers, 6 months old, were randomly assigned to receive two different diets: one of low (GI) and other of high dietary energy intake (GII). Animals were evaluated in relation to body weight gain by being weighed every 21 days. Heifers were evaluated every other day by real-time linear ultrasonography to characterize ovarian structures development from weaning to post-pubertal period. Blood samples were collected to determine plasmatic concentrations of progesterone by RIA method. The ovulation was determined when progesterone concentrations were >1 ng/mL in three consecutive samples, and by ultrasound images of corpus luteum; and oestrous behaviour in some animals. Age at puberty differed among heifers of GII (17.00 +/- 0.46 months) compared with heifers of GI (19.87 +/- 0.47 months; p < or = 0.05). Maximum size of the dominant follicles at pre-pubertal period was greater in GII heifers than in GI (10.52 +/- 0.33 and 9.76 +/- 0.15 mm, respectively; p < or = 0.05). As heifers approached first ovulation time, size of dominant follicle increased (11.75 +/- 0.37 mm for GI and 12.52 +/- 0.91 mm for GII; p < or = 0.05). Body weight at puberty was not different in both groups (302.33 +/- 27.31 kg for GI and 326.19 +/- 27.78 kg for GII heifers; p > 0.05). We conclude that animals receiving high dietary energy intake attained the puberty earlier and the development of follicles were different than in low dietary energy intake.     

A new hemolytic assay for bovine serum complement and its application during experimental bovine anaplasmosis

A new hemolytic assay for bovine complement is presented. Using this assay we found a significant reduction in bovine serum complement activity during the acute phase of anaplasmosis, and an increase in the sensitivity of the red blood cells (RBC) to bovine complement lysis in vitro. The new hemolytic test is performed with bovine RBC, rabbit anti-bovine RBC serum and bovine serum complement. An isotonic sucrose Tris-buffered saline solution of ionic strength 0.094 and pH 7.2 was found to be adequate for this test. The titres obtained with this new assay, which uses autologous RBC, are comparable with those obtained using the guinea pig RBC assay. The finding of a reduction in bovine serum complement during anaplasmosis may be suggestive of a mechanism responsible for the pathology of this disease.     

Phenological investigations of different winter-deciduous species growing under Mediterranean conditions

Phenological stages are the result of biorhythms and environmental factors, these last are probably the same ones that caused, during evolution, adjustments of the species to different climate. The present study was carried out in a Phenological Garden located in central Italy (Perugia, Umbria Region) which contains indicator species, common to all International Phenological Gardens. The aim of this study was to determine and analyse the average trends of development of eight plant species and their phenological adjustment to the Mediterranean environment, over a nine-year period (1997–2005). The results of the statistical analyses show a strong relationship between the temperature trends and vegetative seasonal evolutions interpreted by phenological data for all the species considered. Moreover, it was demonstrated that the plants studied may approach or close completely the timing gaps eventually created during the first phenological phases, adjusting thus the beginning of subsequent phenophases.     

Virus interference between H7N2 low pathogenic avian influenza virus and lentogenic Newcastle disease virus in experimental co-infections in chickens and turkeys

Low pathogenicity avian influenza virus (LPAIV) and lentogenic Newcastle disease virus (lNDV) are commonly reported causes of respiratory disease in poultry worldwide with similar clinical and pathobiological presentation. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. In this study chickens and turkeys were infected with a lNDV vaccine strain (LaSota) and a H7N2 LPAIV (A/turkey/VA/SEP-67/2002) simultaneously or sequentially three days apart. No clinical signs were observed in chickens co-infected with the lNDV and LPAIV or in chickens infected with the viruses individually. However, the pattern of virus shed was different with co-infected chickens, which excreted lower titers of lNDV and LPAIV at 2 and 3 days post inoculation (dpi) and higher titers at subsequent time points. All turkeys inoculated with the LPAIV, whether or not they were exposed to lNDV, presented mild clinical signs. Co-infection effects were more pronounced in turkeys than in chickens with reduction in the number of birds shedding virus and in virus titers, especially when LPAIV was followed by lNDV. In conclusion, co-infection of chickens or turkeys with lNDV and LPAIV affected the replication dynamics of these viruses but did not affect clinical signs. The effect on virus replication was different depending on the species and on the time of infection. These results suggest that infection with a heterologous virus may result in temporary competition for cell receptors or competent cells for replication, most likely interferon-mediated, which decreases with time.