Reference intervals and age-related changes for platelet count, mean platelet volume and plateletcrit in healthy pre-weaning piglets in Italy

Platelet count (PLT) mean platelet volume (MPV) and plateletcrit (PCT) were determined for 117 Landrace x Large White piglets aged 3-21 days; counts were performed with an automated blood cell counter (ABX Pentra 120). Reference values were estimated following the guidelines of the International Federation of Clinical Chemistry (IFCC) and the International Committee for Standardization in Haematology (ICSH). The calculated central 95% reference limits for PLT was 49.9-516.2 x 10(9)/1, for MPV 6.71-9.91 fl and for PCT 0.009-0.395%. When observations are divided into three age groups (about 1 week each) there is an increase in mean PLT count and PCT in 2-week piglets, and a decrease in MPV from the first to the third week of life. These reference values provide guidelines for interpreting for experimental and clinical observations, as well as for monitoring of the health status of similar aged piglets determined using automated impedance-light focusing methodology.     

alpha(1)-Acid glycoprotein modulates apoptosis in bovine monocytes

alpha(1)-Acid glycoprotein (AGP, orosomucoid) is a normal constituent of bovine blood. AGP is an immunocalin, a binding protein that can also exert several immunomodulatory functions. In this paper we investigated the effect of bovine alpha(1)-acid glycoprotein (boAGP) on spontaneous and staurosporine-induced apoptosis of blood derived monocytes purified using magnetic cell sorting techniques. Bovine AGP was purified from blood following a chromatographic protocol. The homogeneous protein was used to stimulate the cells as well to raise a polyclonal antibody, that was used throughout all the experiments. When monocytes were incubated with high concentrations of boAGP (0.9 mg/ml), similar to those found in bovine plasma during systemic reaction to inflammation, their spontaneous apoptosis rate was suppressed, as determined by caspase-3/7 enzymatic activity assay. Similar results were obtained when apoptosis was induced by adding staurosporine, a potent protein kinase inhibitor. The apoptosis-modulating activity of boAGP was dependent on its concentration, since physiological concentrations of boAGP (0.3 mg/ml) did not exhibit a statistically significative anti-apoptotic activity. We also investigated whether this apoptosis-modulating activity was dependent on the terminal sialic acid residues exposed on the surface of the protein. Enzymatic treatment with neuraminidase, that cleaves terminal sialic acid residues, completely abolished boAGP's anti-apoptotic activity. These results suggest that the protective effect of AGP is likely mediated by its sialic acid terminal groups.     

Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes

MicroRNAs (miRNAs) are a family of regulatory molecules involved in many physiological processes, including activation of cells of the immune system. This study investigated the effect of Escherichia coli lipopolysaccharide (LPS) and Staphylococcus aureus enterotoxin B (SEB) on the expression of five miRNAs involved in the inflammatory response, including miR-9, miR-125 b, miR-155, miR-146 a and miR-223, in bovine CD14(+) cells (monocytes). Incubation of monocytes with SEB induced down-regulation of miR-155, miR-223 and miR-125 b, but not the anti-inflammatory miRNA miR-146 a. Conversely, incubation with LPS upregulated both miR-155 and miR-146 a. In vitro incubation of isolated CD14(+) bovine monocytes with LPS and SEB elicited different and opposite expression of miRNAs reportedly involved in inflammatory reactions.     

Milk hygiene and udder health in the periurban area of Hamdallaye, Niger

The prevalence of intra-mammary infections in dairy herds was studied in Hamdallaye, Niger. A total of 956 milk samples were collected in 2007 from 239 lactating cows of four local breeds in eight traditional herds; the first sampling was undertaken in the dry season at morning milking, and the second in the rainy season at evening milking. Staphylococcus aureus, Coagulase-Negative Staphylococci (CNS) and environmental microorganisms were detected in significantly (p < 0.05) more samples in the rainy season, 55.2%, than in the dry season, 27.1%. Statistically significant (P < 0.05) differences in prevalence were observed among herds and according to lactation number. Infections were assigned to four classes, according to the major pathogen, and the respective mean somatic cell counts during the dry season were: S. aureus, 775 × 10³ cells/ml; CNS, 447 × 10³ cells/ml; environmental microorganisms, 407 × 10³ cells/ml; and non-infected, 262 × 10³ cells/ml. Most of the tested strains were sensitive to antibiotics, and selected strains of S. aureus (n = 15) were negative to the multiplex PCR tests for production of enterotoxins.     

Post-operative analgesic effects,after orthopaedic surgery in the dog,of loco-regional ropivacaine and bupivacaine blockade using the nerve locator technique: 159 cases

Veterinary Research Communications -     

Bovine respiratory syncytial virus seroprevalence and risk factors in endemic dairy cattle herds

The herd seroprevalence of bovine respiratory syncytial virus (BRSV) was studied in 59 dairy cattle herds using serology on random selected animals stratified by two age classes (heifers, cows). Risk factors for primary infections in heifers were investigated using a questionnaire on management conditions and data on bovine viral diarrhoea (BVD) status. At least one seropositive cow was present in all the herds. In 25% of the herds all individual were seropositive and 22% of herds had all heifers seronegative. Analysis of the influence of risk factors retained summer pasture and BVD status. In particular, absence of summer pasture and the BVD positive status of heifers were associated with an increased risk of BRSV infection in heifers group.     

Alpha(1)-acid glycoprotein is contained in bovine neutrophil granules and released after activation

The present study was designed to investigate the capability of bovine neutrophil granulocytes to produce the minor acute phase protein alpha(1)-acid glycoprotein (AGP, Orososmucoid). Bovine neutrophils contain a high MW (50-60kDa) AGP isoform (PMN-AGP), as determined by Western blotting and confirmed by fluorescence microscopy. The presence of AGP in bovine neutrophils has been confirmed by fluorescence immunocytometry. In addition, bovine neutrophils contain also a 42-45kDa isoform, which has the same MW as plasma-, liver-delivered, AGP. cDNA sequence of plasma- and PMN-AGP revealed that (i) the two proteins are products of the same gene; (ii) the differences in molecular weight are due do different post-translational modifications. This result was confirmed by deglycosylation of the two glycoforms. Exocytosis studies showed that isolated neutrophils exposed to several challengers, including Zymosan activated serum (ZAS) and phorbol 12-myristate 13-acetate (PMA), which mimic the inflammatory activation, released PMN-AGP as early as 15min. AGP's mRNA is physiologically expressed by mature resting neutrophils. Real-time PCR on LPS, ZAS and PMA challenged cells revealed that the level of expression apparently does not increase after inflammatory activation. Collectively, the findings reported in this paper proved that PMN-AGP: (i) is a hyperglycosylated glycoform of plasma AGP, (ii) is stored in granules, and (iii) is released by neutrophils in response to activation. Due to its anti-inflammatory activity, PMN-AGP may work as a fine tuning of the neutrophils functions in the inflammatory focus, i.e. it can reduce the damages caused by an excess of inflammatory response.