Transglutaminase 2: a novel autoantigen in canine idiopathic central nervous system inflammatory diseases

Necrotizing meningoencephalitis (NME), necrotizing leukoencephalitis (NLE) and granulomatous meningoencephalomyelitis (GME) are common idiopathic inflammatory central nervous system (CNS) diseases with unknown etiology in dogs. We previously showed that IgG autoantibodies in the cerebrospinal fluid (CSF) of NME cases reacted to unknown brain proteins as well as to glial fibrillary acidic protein (GFAP). In the present report, we evaluated the autoantibodies against transglutaminase2 (TG2) in the canine CNS diseases. CSF samples obtained from dogs with NME (n=19), NLE (n=7), GME (n=11) and miscellaneous CNS diseases (n=12) were subjected. CSFs from 20 healthy dogs were used as controls. Indirect fluorescent antibody test on the canine cerebrum revealed astrocyte-binding IgG in the CSF of NME. After absorption of the CSF with bovine GFAP, the CSF still possessed the reactivity to astrocytes. Double-color staining showed clear colocalization of the autoantibodies and anti-human TG2 rabbit polyclonal IgG. An immunoblot assay against human recombinant TG2 revealed anti-TG2 IgG in the CSF from dogs with NME, NLE and GME. The CSF of canine idiopathic encephalitis cases, notably of NME, tended to show high ELISA OD values against human recombinant TG2 compared to healthy controls. The presence of anti-TG2 autoantibodies in the CSF may contribute to the elucidation of the etiology of canine NME, NLE and GME.     

Rumen development and growth of Balouchi lambs offered alfalfa hay pre- and post-weaning

The consumption of solid feed is essential for successful transition from a pre-ruminant to a functional digestive tract. Lambs fed starter rations containing highly fermentable carbohydrates often experience dramatic changes in concentrations of rumen and blood metabolites. The optimal amount of roughage required in the diet of pre-ruminant animals is still unclear. The objective of this study was to determine the effect of feeding alfalfa hay on performance and rumen development in young Balouchi lambs. In a completely randomized design, 30 lambs were fed one of three experimental diets consisting of a control, without alfalfa hay (C), a diet containing 7.5% alfalfa hay (A1), and a diet containing 15% alfalfa hay (A2). Lambs fed A1 and A2 diets had lower dry matter intake during the pre-weaning period (P?P?=?0.02), but feed conversion ratio and average daily gain were not affected by feeding alfalfa hay. Concentration of beta-hydroxybutyric acid was higher in C compared with the A1 and A2 groups (P?P?=?0.04) and increased thickness of muscular layer (P?=?0.05). We concluded that including 15% alfalfa hay in the starter diet could reduce thickness of the keratinization layer and increase muscularity of rumen wall without adverse effects on growth and performance of newborn lambs.     

Improvement in the diagnosis of <Emphasis Type="Italic">Brucella abortus</Emphasis> infections in naturally infected water buffaloes (<Emphasis Type="Italic">Bubalus bubalis</Emphasis>) using an ELISA with a Protein-G-based indicator system

Brucellosis caused by Brucella abortus in domestic water buffaloes (Bubalus bubalis) raised under the traditional system of husbandry in northern India was diagnosed using an enzyme-linked immunosorbent assays (ELISA) with a Protein-G-based indicator system (Protein-G ELISA). A total of 1,551 animals that are positive (N = 61), negative (N = 243), and suspected (N = 1,247) for brucellosis were examined. Rose bengal test (RBT) was used to predict the disease, and accordingly, animals were dichotomized in positive and negative population for receiver operating characteristic (ROC) curve analysis to determine the sensitivity, the specificity, and the performance index of Protein-G ELISA. Taking all animals (N=1551) into account, the ROC curve analysis revealed cut off value of 29.6% positivity (%P) with 98.40% and 94.94%, sensitivity and specificity, respectively. The results were compared with ELISA in which anti-bovine conjugate was used. The cut off in ELISA was 37.9%P and sensitivity and specificity were 96.26% and 97.07%, respectively. The performance indexes of both the assays were almost equal and were 193.34 for Protein-G ELISA and 193.33 for ELISA. The cut off values of both the tests changed, if only known positive (N = 61) and known negative (N=243) animals were used for ROC curve analysis, and accordingly, changes in sensitivity and specificity were observed with significant decrease of performance indexes of both the tests. The high optical density (P<0.0001) background signal with negative serum control and high %P (P<0.0001) in sera from negative population were noticed in ELISA in comparison to Protein-G ELISA.     

In vitro inhibitory effect of trichostatin A on canine grade 3 mast cell tumor

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect skin and soft tissue in dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. Trichostatin A (TSA), an antifungal antibiotic, has shown inhibitory effects on the proliferation and induction of apoptosis in various types of cancer cells. In order to evaluate the potential of trichostatin A as a therapeutic drug, cells of grade 3 MCT were cultured and treated with concentrations of 1 nM to 400 nM of TSA. MTT assay and trypan blue exclusion assays were performed to estimate cell growth and cell viability, and cell cycle analysis was evaluated. TSA treatment showed a reduction in numbers of viable cells and an increase of cell death by apoptosis. The cell cycle analysis showed an increase of hypodiploid cells and a reduction of G0/G1 and G2/M –phases. According to these results, trichostatin A may be an interesting potential chemotherapeutic agent for the treatment of canine MCT.     

Characterization of Doayo and Kapsiki taurine cattle breeds of Cameroon in their natural environment

Data were collected on live weight (LW), heart girth (HG), height-at-withers (HW), trunk length (TL), age, sex, and coat color of 207 taurines cattle—122 of the Doayo (Namchi) breed and 85 of the Kapsiki (Kirdi) breed. The animals, aged 1 to 20 years, were selected from 60 herds randomly selected from villages of Poli of Faro and Mokolo of Tsanga, divisions of the North and Far North Regions of Cameroon. The data were analyzed using the SAS program with a linear model, applying standard tests. Results indicated no breed effect (P > 0.05) in the growth trends of LW, HG, HW, and TL. HG and TL were highly significantly (P < 0.0001) related to LW. The growth pattern for the two breeds was the same since the linear contrast of least square means for the traits at various age groups did not differ (P > 0.05) significantly. The breeds attained maturity as from 4 years. In the absence of breed effect (P > 0.05), a single regression equation was established for the estimation of live weight as thus LW = - 244.42 ( ±22.57 ) kg + 2.49 ( ±0.23 ) HG + 1.04 ( ±0.25 ) TL {\hbox{LW}} = - {244}.{42 }\left( {\pm {22}.{57}} \right){\hbox{ kg}} + {2}.{49 }\left( {\pm 0.{23}} \right){\hbox{ HG}} + {1}.0{4 }\left( {\pm 0.{25}} \right){\hbox{ TL}} , with HG contributing up to 70% of total variation and TL, 2%. This equation could be used to develop a measuring band useful in the rural environment for commercial and clinical veterinary purposes.     

Establishment of a large-scale purification procedure for purified recombinant bovine interferon-tau produced by a silkworm-baculovirus gene expression system

We developed a procedure for the large-scale purification of bovine interferon-tau (boIFN-tau) by means of a silkworm-baculovirus gene expression system. Recombinant boIFN-tau (rboIFN-tau) was efficiently produced in the silkworm infected with boIFN-tau cDNA recombinant baculovirus and accumulated in the haemolymph. To establish a purification method suitable for mass production, we tried three crude purification methods, namely, an acidification and neutralization treatment (ANT), silica gel column chromatography (SGCC), and Blue sepharose column chromatography (BSCC) with a combination of Q-sepharose (QSC) and chelating sepharose column chromatographies (CSCC). As a result, the acidification and neutralization treatment was found to be the most efficient and cost effective. With this combination, we obtained 91% pure products. To confirm the applicability of the procedure for mass production, we inoculated 100 silkworms with the recombinant virus, and recovered about 4.55 mg (1.26 x 10(8)U/mg) of 91% pure rboIFN-tau by means of a combination of the ANT, followed by QSC and CSCC.     

Growth,feed efficiency and blood profile of buffalo calves consuming high levels of fluoride

Twelve male buffalo calves of 10 to 12 months of age were divided into 3 groups of four each. They were fed wheat straw+concentrate mixture +3 Kg greens. The chemical composition of the diet was same in all the three groups except fluoride which was added (as NaF) in concentrate mixture of group B and C to make the final fluoride concentration 30 ppm and 60 ppm respectively. The animals were kept on scheduled diet for a period of 90 days. Body weights were recorded at the start of the experiment and at fortnightly interval thereafter. Analysis of data revealed that the dry matter intake decreased non significantly in group B and C as compared to control group. A significant decrease in serum calcium and a significant increase in phosphorus concentration were observed in group C animals. A significant increase was observed in alkaline phosphatase activity in group C animals. A non significant decrease was observed in T₄ values in group C animals. On the basis of these results it could be concluded that fluoride in the diet of buffalo calves @ 30 ppm is a safe level whereas 60 ppm has affected the blood metabolites.     

Prematurational Culture with 3-Isobutyl-1-methylxanthine Synchronizes Meiotic Progression of the Germinal Vesicle Stage and Improves Nuclear Maturation and Embryonic Development in In Vitro-grown Bovine Oocytes

The objective of this study was to clarify the effects of prematurational culture (pre-IVM) supplemented with 3-isobutyl-1-methylxanthine (IBMX) on nuclear and cytoplasmic maturation of in vitro-grown bovine oocytes. In experiment 1, oocytes (95 μm in diameter) derived from early antral follicles (0.5–1 mm in diameter) were cultured for 12 days for in vitro growth (IVG). IVG oocytes with a normal appearance were subjected to examinations of diameter and chromatin structure in the germinal vesicle (GV) before IVM. In addition, percentages of metaphase II (M II) were examined after IVM. Regardless of pre-IVM, the mean diameters of IVG oocytes were about 115 μm. The proportions of GV3 (50.0%) and M II stages (80.1%) of IVG oocytes with pre-IVM were higher than those without pre-IVM (28.0 and 49.4%, respectively). In experiment 2, the fertilizability and developmental competence of IVG oocytes were examined. Regardless of pre-IVM, the normal fertilization rates of IVG oocytes were similar (around 70%) but were lower than that of in vivo-grown oocytes (88.0%). Cleavage and blastocyst rates of IVG oocytes with pre-IVM (63.0 and 26.1%, respectively) were higher than those without pre-IVM (45.8 and 12.7%, respectively). The blastocyst rate based on cleaved IVG oocytes with pre-IVM (41.7%) was similar to that of in vivo-grown oocytes (48.7%), although the cleavage rate of IVG oocytes with pre-IVM was lower than that of in vivo-grown oocytes. In conclusion, pre-IVM with IBMX improved the maturational and developmental competences of IVG oocytes, probably due to promotion of their chromatin transition and synchronization of meiotic progression.     

Increase in plasma total antioxidant capacity of grazing Japanese Black heifers and cows in forestland in Japan

Blood total antioxidant capacity (TAC) has become a key bio‐marker for animal health. Forest‐grazing cattle are known to forage various native plants that have high TAC. This study evaluated differences of plasma TAC between forest‐grazing (FG) and pasture‐grazing cattle (PG). Experiment 1 monitored the plasma TAC levels of 32 Japanese Black cattle. The level in PG did not change throughout the grazing period. However, that in FG, which increased from summer, was significantly higher than that in PG through fall (P < 0.05). In experiment 2, we used nine Japanese Black heifers and investigated their blood antioxidant parameters and the TAC in plants that the cattle consumed in late June and September. The plasma TAC levels in FG were significantly higher than those in PG in both periods (P < 0.05). Plasma levels of lipid peroxidation in FG tended to be lower than that in PG (P = 0.098). Furthermore, the TAC levels in various species of shrubs and trees consumed by FG were higher than those in pasture grasses. Results of this study show that plasma TAC of grazing Japanese Black cattle in forestland increase from summer through fall.