Isolation and characterization of alpha 1-acid glycoprotein from horses, and its evaluation as an acute-phase reactive protein in horses.

Equine alpha 1-acid glycoprotein (alpha 1AG) was isolated from equine serum by successive ammonium precipitation, anion- and cation-exchange chromatographies, and gel filtration. Purified equine alpha 1AG had a molecular weight of 46,000 +/- 1,000, and contained 31.4% carbohydrate. Gel isoelectric focusing revealed an isoelectric point range of 2.8 to 3.7. With immunoelectrophoresis, it was found that alpha 1AG migrated to the alpha 1-globulin region. Single radial immunodiffusion was used for quantitative measurement of alpha 1AG in equine serum. In clinically normal foals, serum alpha 1AG was undetectable (less than or equal to 20 micrograms/ml) in less than or equal to 7-day-old foals, but was detected by 14 days. The alpha 1AG concentration (mean +/- SD) increased to reach mean adult values of 99.23 +/- 26.90 micrograms/ml by 1 year of age. The alpha 1AG concentration in pregnant mares decreased at 2 to 3 months before parturition, then gradually increased until 1 day after parturition, when a brief decrease was observed. The concentration increased again at 2 weeks after foaling, then a decrease was observed, after which the alpha 1AG concentration increased again by 2 to 4 months after parturition. The concentration of serum alpha 1AG quickly rose to peak values 2 to 3 days after castration and jejunojejunostomy in adult horses, returning to baseline values by 14 to 28 days after surgery. The alpha 1AG was concluded to be an acute-phase reactive protein in horses.     

Serological evidence of akabane virus infection in northern Israel in 2001

In February 2002 the first cases of a "blind newborn calves" syndrome with hydranencephaly appeared in Israel. Eighty-one serum samples, from 54 animals on farms where the syndrome was recorded and 27 others from unaffected farms were examined by neutralization of Akabane virus (AKAV, strain OBE-1) by the micro-titer method. Forty-seven of the 54 samples from the affected farms contained high serum neutralization titers against AKAV (mean SN titer 79.5 and +/- 44.7, standard deviation), whereas only one of the 27 samples from the unaffected farms was positive (titer of 8). These results suggest that the vector(s) of AKAV was circulating in Israel in August through December, 2001.     

Development of spermatogenic function in the sex maturation process in male cats

The spermatogenic function and plasma testosterone (T) level in the sex maturation process were investigated as to 180 mixed breed cats ranging from 4 months to 2 years in age to be castrated. Testis/epididymis weights reached a peak at 10 and 8 to 9 months of age, respectively. In the testis, sperm appeared at 5 months of age. At 7 months of age, sperm were observed in 96.2% of the cats. In the tail of the epididymis, sperm appeared in 46.9% of the cats at 6 months of age and in all cats at 8 or more months of age. Furthermore, the mean plasma T level rapidly increased at 8 months of age, and reached a peak (2.64 +/- 0.68 (SE) ng/ml) at 10 months of age. Three of 180 cats (1.67%) had unilateral cryptorchidism. These results suggest that the spermatogenic function in male cats becomes mature at 8 to 10 months of age.     

A newly established bovine intestinal epithelial cell line is effective for in vitro screening of potential antiviral immunobiotic microorganisms for cattle

We evaluated whether a bovine intestinal epithelial (BIE) cell line could serve as a useful in vitro model system for studying antiviral immune responses in bovine intestinal epithelial cells (IECs) and for the primary screening of immunobiotic microorganisms with antiviral protective capabilities. Immunofluorescent analyses revealed that toll-like receptor 3 (TLR3) was expressed in BIE cells, and the results of real-time quantitative PCR showed that these cells respond to stimulation with poly(I:C) by up-regulating pro-inflammatory cytokines and type I interferons. In addition, we demonstrated that BIE cells are useful for the primary screening of immunobiotic lactic acid bacteria strains which are able to beneficially modulate antiviral immune responses triggered by TLR3 activation in bovine IECs. The characterization of BIE cells performed in the present study represents an important step towards the establishment of a valuable bovine in vitro system that could be used for the development of immunomodulatory feed for bovine hosts.     

Reliability in somatic cell count measurement of clinical mastitis milk using DeLaval cell counter

Somatic cell counts (SCC) measurements are typically performed using quantitative methods, such as the Breed method (Breed) and the Fossomatic method (FSCC). The DeLaval cell counter (DCC) developed recently is a quantitative somatic cell counter with a low initial cost and superior portability. However, since the DCC was specifically developed for measuring SCC of ≤ 4 × 10⁶ cells/mL milk from bulk tanks or individual cows, its reliability for estimating SCC that exceed this concentration has not yet been clarified. This study therefore examined whether it is possible to accurately measure SCC by diluting milk samples with initial SCC of 4 × 10⁶ cells/mL, as seen in clinical mastitis milk. We collected milk samples from 99 quarters of 99 Holstein cows with clinical mastitis. These milk samples were diluted 10‐fold with saline and thoroughly mixed before performing SCC measurement with the DCC. The correlation coefficients of SCC measured by the FSCC, Breed and DCC methods indicated strong correlations between each pair of methods. The findings showed that DCC can be used to identify bovine clinical mastitis milk and is useful as a quantitative SCC measurement device on farm sites.     

Spontaneous Malignant T Cell Lymphoma in a Young Male Common Marmoset (Callithrix jacchus)

We histopathologically and immunohistochemically investigated a case of malignant lymphoma that spontaneously developed in a male common marmoset at two years of age. Beginning at two years four months of age, the animal had an enlargement of the submandibular and inguinal lymph nodes, small subcutaneous nodules near the right breast and an approximately fivefold increase in peripheral lymphocyte count compared with the previous examination value. The postmortem findings at two years eight months of age showed lymphadenopathy with enlargement of the thymus and spleen. Small- to intermediate-sized neoplastic lymphocytes had diffusely proliferated in the enlarged nodes. The neoplastic cells were pleomorphic and had irregularly shaped nuclei. The nuclear chromatin staining revealed hyperchromatism in the small-sized cells, and the intermediate-sized cells exhibited vesicular staining. An immunohistochemical examination indicated that the neoplastic lymphocytes were positive for CD3 and negative for CD20, thus suggesting that they had originated from T cells. In addition, the proliferation of high endothelial venules and reactive epithelioid histiocytes was observed. Scattered tingible body-laden macrophages were infrequently detected. Neoplastic lymphocytes were also observed in the thymus, spleen, heart, lungs, liver, kidneys, adrenal glands and femoral and sternal bone marrow. This malignant lymphoma in a young male common marmoset was considered to fit the category of “peripheral T-cell lymphoma, not otherwise specified (PTCL-NOS)” according to the new WHO system of classification.     

Analysis of Phytosterol,Fatty Acid,and Carotenoid Composition of 19 Microalgae and 6 Bivalve Species

ABSTRACT

In this study, the functional components of 19 microalgae and 6 bivalve species were investigated in the context of the application in bivalve feeding and human health food. Principal component analysis was performed to detect any association between the functional components and individual microalgal species or taxonomic group. The proportions of the functional components differed depending on the taxonomic group or species of microalga. The genera Cheatoceros, Thalassiosira, and Isochrysis contained high concentrations of fucosterol and fucoxanthin, which are present in large amounts in brown algae. Diatoms, haptophytes, and eustigmatophytes, which are used as feed for bivalves, were rich in fucosterol and eicosapentenoic acid (EPA); further, haptophytes were rich in docosahexaenoic acid (DHA). In addition, the microalgae associated with red tide, i.e., the raphidophytes, were found to be rich in fucoxanthin, β-sitosterol, and EPA, whereas dinoflagellates were rich in DHA. Seven bivalve species also contained high concentrations of fucosterol, EPA, and DHA, as did microalgae, which were used to feed by bivalves. These results are useful in selecting microalgae effectively as feed of the bivalves.     

Characteristics of two populations of Thai river sprat Clupeichthys aesarnensis from man-made reservoirs in Thailand and Laos,with aspects of gonad development

Fisheries Science - Growth and reproduction of the Thai river sprat Clupeichthys aesarnensis (Teleostei: Clupeidae) in Sirindhorn Reservoir, Thailand, and Nam Ngum Reservoir, Laos, were...     

Liquid-chromatographic hybrid triple–quadrupole linear-ion-trap MS/MS analysis of fatty-acid esters of dinophysistoxin-1 in bivalves and toxic dinoflagellates in Japan

Fatty-acid esters of dinophysistoxin-1 (DTX1) in scallops Patinopecten yessoensis, mussels Mytilus coruscus, and toxic dinoflagellate Dinophysis species, collected from Japanese seawater, were analyzed by liquid-chromatography mass spectrometry (LC–MS). Precursor ion monitoring, multiple reaction monitoring for 18 fatty-acid esters of DTX1, and full-scan MS/MS spectra obtained with a hybrid triple–quadrupole linear-ion-trap mass spectrometer showed that 14:0, 16:0, and 16:1 esters were the most abundant 7-O-acyl-DTX1 analogues in bivalves. Fatty-acids esters formed by conjugations at hydroxyl positions other than the 7-position of DTX1 were not detected in the bivalves. DTX1 and okadaic acid-16:0 fatty-acid esters have been reported as the most abundant ester in bivalves in several previous studies; however, we found that 7-O-16:1-DTX1 was the most abundant ester in some mussels in which 16:1 was more dominant than 16:0 in the free fatty-acid profile. Comparison between 7-O-acyl-DTX1 and free fatty-acid profiles in the same bivalve samples suggests that polyunsaturated fatty acids are selectively excluded in enzymatic acylation of DTX1. No 7-O-acyl-DTX1 was detected in any single-cell isolates of D. fortii, D. acuminata, D. mitra, D. norvegica, D. tripos, D. infundibulus, and D. rotundata.