Pathology of female mice experimentally infected with an in vitro cultured strain of Trypanosoma equiperdum

Dourine, caused by infection with Trypanosoma equiperdum, is one of the trypanosomiasis in equids. The clinical course of dourine is long-term, ranging from 1–2 months to several years. Since the pathogenesis of dourine has not yet been elucidated, experimental studies using mouse infection models are needed. Although mice are not susceptible to most T. equiperdum strains, some strains can infect mice. Even in such strains, infected mice develop rapidly transient parasitemia and die within 2–8 days. Therefore, mice experimentally infected with these T. equiperdum strains are not suitable for mouse infection models to analysis the pathogenesis of dourine. A sequential method of isolating parasites from dourine-affected horses and adapting them to in vitro cultures using soft agarose media was recently developed. Various T. equiperdum strains adapted to in vitro conditions have been established using this technique. We used one of these strains, the T. equiperdum IVM-t2 strain. In the present study, T. equiperdum IVM-t2 strain inoculated mice developed periodic parasitemia during the experimental period of 60 days. Histopathologically, vaginitis and dermatitis were observed. These findings were comparable to those of dourine-affected horses. Therefore, mice infected with T. equiperdum IVM-t2 strain may be a valuable tool for pathological, immunological, and parasitological in vivo research, and will contribute to investigations on the mechanisms underlying the disease process and the host-parasite relationship.     

Muscle growth in full-cycle cultured Pacific bluefin tuna Thunnus orientalis from early larval to juvenile stage: histochemical,immunohistochemical and ultrastructural observation

With the development of successful aquaculture techniques, the Pacific bluefin tuna (PBT) has become the most important fish species in Japan. Early muscle growth and cellularity of muscle fibers have a profound effect on ultimate body size and meat quality of fish. Larvae and juveniles of full-cycle cultured PBT were sampled at 3, 15, 29, 41, 70, 128 and 218 days post hatch (dph). Transverse body sections until 41 dph and muscle sections from 41 dph thereafter from dorso-cranial or caudal areas were stained for muscle fiber morphological and morphometric analysis and muscle sections from the dorso-cranial region were used for ultra-structural observation. Red muscle fibers appeared by 15 dph at the horizontal septum both in epaxial and hypaxial regions. Other than a single layer of superficial red fibers and those at the horizontal septum, existence of red muscle fibers was apparent in the PBT myotome. The muscle fiber diameters varied in their size even in adjacent myotomes of PBT. Overall, growth throughout the various stages was a result of both hyperplasia and hypertrophy of muscle fibers. Muscle fiber diameters were greater in the dorso-cranial than in dorso-caudal region at 41, 70 and 128 dph. These results provide additional characterization of PBT growth, which may assist with managing fish production efficiency, harvest age and eating quality.     

Fluctuations in serum steroid hormone concentrations and body mass during growth and sexual maturation in captive northern fur seals (Callorhinus ursinus)

Northern fur seals (Callorhinus ursinus) have a distinct life history pattern comprising annual terrestrial breeding and oceanic migration, and the physiological changes associated with these patterns are of particular interest for understanding their environmental adaptations. However, owing to their oceanic distribution, limited information is available on the reproductive physiology of wild individuals during the immature stage and the winter migration period. This study aimed to determine the relationships among the seasonal hormone profiles, body growth, age, and pregnancy using monthly serum samples collected over 3–5 years from two male and two female captive individuals during pubescence and sexual maturation. Small increases in the serum testosterone signaled puberty in males aged 3 and 4 years. Thereafter, males showed considerable increases in testosterone during breeding seasons, indicating sexual maturity. Immature female serum progesterone was maintained at low levels, but after pubescence, females showed an increase in serum progesterone in August, the month next to the peak of delivery, followed by a decrease. In non-pregnant females, progesterone did not increase significantly until the next breeding season, but in pregnant females, they increased again from February to March and then gradually decreased. Immature males increased body mass constantly and reached puberty when their body mass exceeded 20 kg, and they showed seasonal weight fluctuations after puberty. These results provide fundamental information for determining sexual maturity and pregnancy in this species based on sex steroid hormones and body mass measurements.     

Effects of aging and delivery order on the breeding capacity of F344/N rats

Reproductive performance affects the efficiency of the production of animals. Therefore, knowing the reproductive properties of each species or strain of animal is important for proper management of breeding stocks. To elucidate the reproductive properties of female F344/N rats, frequently used for longevity and gerontology research, we monitored the breeding duration, number of deliveries, litter size and weaning rate of their pups. The first delivery was observed at 2.9 ± 0.1 (mean ± standard deviation) months of age (M) and the last delivery was at 15.1 ± 1.8 M. The number of deliveries was 10.4 ± 2.8 (range of 7–16) within the life span of the mother (24.7 ± 5.6 M). The litter sizes at the third (10.0 ± 3.0) or fourth (10.3 ± 2.7) deliveries were higher than for other deliveries. The breeding outcome declined after the fifth delivery yielding reduced litter size or weaning rate, number of delivered mother. These results suggests that though female F344/N rats are able to deliver more than 10 times, the reproductive performance lowered after fifth delivery.     

Effect of progesterone on the in vitro response of peripheral blood mononuclear cells stimulated by Escherichia coli in mares

Escherichia coli(E. coli) isolated from the uterus of a Thoroughbred mare with bacterial endometritis was used to evaluate the effect of progesterone (P(4)) on the immune response of mares. Peripheral blood mononuclear cells (PBMCs) were collected from 10 nonpregnant clinically healthy adult mares (range, 4-12 years) during diestrus, four Thoroughbreds and six Hokkaido native horses. Cell proliferation and expression of cytokine mRNA, including interferon (IFN)-γ, tumor necrosis factor (TNF)-α and interleukin (IL)-10, of PBMCs stimulated with E. coli and P(4) were examined in vitro. P(4) was shown to have significantly inhibited E. coli induced proliferation and expression of IFN-γ in PBMCs. These results indicate that P(4) inhibits the immune response to E. coli in mares.     

Anti-Müllerian hormone profiles as a novel biomarker to diagnose granulosa-theca cell tumors in cattle

This study was carried out to evaluate the blood profile and tissue expression of Anti-Müllerian hormone (AMH) as a biomarker for granulosa-theca cell tumors (GTCTs) in cattle. Five cases with unilateral ovarian GTCTs (GTCT group) were investigated in comparison to other groups of Japanese Black cows, which had either cystic ovarian disease (COD group, n=5), a functional corpus luteum on Days 9 to 11 of the estrous cycle (Day 0=estrus; CL group, n=13) or received superovulation treatment (SOT group, n=13). We used transrectal ultrasonography and measured plasma AMH, estradiol-17β (E(2)), progesterone (P(4)) and testosterone (T) levels. Moreover, GTCT tissues were collected and examined by immunohistochemical staining (IHC) for AMH. In the GTCT group, ultrasound images of GTCTs were variable and not definitive. However, the AMH level in the GTCT group (n=3, 58.1 ± 66.3 ng/ml) was significantly higher than in the COD, CL and SOT groups (0.1 ± 0.1 ng/ml for GTCT vs. COD, P<0.05; 0.2 ± 0.1 and 0.3 ± 0.2 ng/ml, respectively for GTCT vs. CL and SOT, P<0.01). The other hormonal levels in the GTCT group had no significant differences compared with the COD or SOT group. Neoplastic granulosa cells labeled with AMH antibody clearly demonstrated a variety of tissue patterns in all cases by IHC. To the best of our knowledge, this is the first study to investigate the blood profile and IHC of AMH in bovine GTCTs. Our findings indicate that AMH may be a novel biomarker to diagnose GTCTs in cattle.     

Immunomodulatory effects of Alliums and Ipomoea batata extracts on lymphocytes and macrophages functions in White Leghorn chickens: In vitro study

We previously described that supplementary garlic, onion and purple sweet potato (PSP) enhance humoral immune response in White Leghorn chickens. In the present in vitro study, we investigated the effects of garlic (GE), onion (OE) and PSP (PSPE) extracts on proliferation, interleukin (IL)‐2 and interferon (INF)‐γ gene expression of stimulated lymphocytes. The effects on microbicidal activity, reactive oxygen species (ROS) and nitric oxide (NO) productions of stimulated peritoneal macrophages were studied as well. The results showed that GE augmented Concanavalin A (ConA)‐induced splenocytes (4, 8 and 16 µg/mL) and thymocytes (2, 4 and 8 µg/mL) proliferations, and gene expression of IL‐2 (8 and 16 µg/mL) and INF‐γ (16 µg/mL). None of the examined extracts had mitogenic effect nor stimulated bursacytes response to phorbol 12‐myristate 13‐acetate (PMA). Macrophages exhibited superior microbicidal activity and ROS production with GE at 4 and 8 µg/mL and with OE at 25.6 µg/mL. None of the extracts showed stimulatory effects on NO production. The extracts showed concentration‐dependent inhibitory effects on all measured parameters at higher concentrations. Taken together, it is likely that garlic has direct stimulatory effects on immune cell functions, whereas the in vitro inhibitory effects of onion and PSP were likely attributed to high flavonoid contents.     

Prevalence of the virulence plasmid in Salmonella Typhimurium isolates from pigs

To determine the prevalence of the virulence plasmid in Salmonella Typhimurium isolates from pigs in Japan, a total of 106 porcine isolates were subjected to PCR amplification for the detection of the virulence plasmid. Out of the isolates of S. Typhimurium, 38 (35.8%) harbored the virulence plasmid. The presence of the virulence plasmid was widely observed in the isolates from systemically infected pigs (92.0%, 23/25), compared with diarrheic (18.8%, 12/64) and apparently healthy pigs (17.6%, 3/17) (P<0.01).     

Determination of S-methyl-, S-propyl-, and S-propenyl-L-cysteine sulfoxides by gas chromatography-mass spectrometry after tert-butyldimethylsilylation

A gas chromatographic-mass spectrometric method for the determination of S-methyl-L-cysteine sulfoxide (1), S-propyl-L-cysteine sulfoxide (2), and S-propenyl-L-cysteine sulfoxide (3), specific marker compounds in the genus Allium, is described. The target amino acids were converted to the tert-butyldimethylsilyl derivatives. The products were silylated on the amino and carboxyl groups and on an additional oxygen atom and were separated on a nonpolar capillary column. That incorporation of three tert-butyldimethylsilyl groups had occurred was verified by mass spectrometry, which gave an m/z 302 fragment as base peak (amino acid side chain eliminated ion) and m/z 436 (1), 464 (2), or 462 (3) as major peaks (tert-butyl function eliminated ion), by electron impact ionization. The detection limits for 1 and 2 under selected ion monitoring at m/z 436 (1) and m/z 464 (2), respectively, were determined to be 0.3 and 1.8 ng per injection. To clean up the analytes from the solvent extract of onion, as a representative food material, onion, the sample solution was subjected to combined solid phase extraction. The eluate from a Sep-Pak C(18) cartridge was applied to a Bond Elut SCX cartridge (H(+) form), followed by washing with 0.1 M hydrochloric acid and elution with 0.5 M ammonia. From a simulated matrix solution containing 5% sucrose, 1 and 2 were extracted quantitatively, and the detection yield was approximately 75%. The contents of 1, 2, and 3 in commercial onion were estimated to be 0.3, 3.1, and 3.0 mg, respectively, per gram of fresh weight.     

The mineralization rate of black soil carbon in the deep layers of Japanese volcanic ash soil may be easily accelerated by labile carbon supply

ABSTRACT

The stability of black soil carbon in the deep layers of Japanese volcanic ash soil (i.e., buried A horizons) is often explained by its unique chemical (molecular structure) and physical (associated with short-range-order minerals) recalcitrance. However, the stability of black soil C in buried A horizons may be changed by labile C supply for soil microbes. Here, we hypothesized that the mineralization of black soil C in buried A horizons of Japanese volcanic ash soil could be easily accelerated by a supply of labile C (i.e., a priming effect; PE). To test our hypothesis, we investigated the direction and magnitude of the PE with a buried A horizon in Japan using ¹³C-labeled glucose (2.188 atom %) in a short-term (21 days) incubation study. We also investigated the effect of mineral nitrogen (N), which could contribute to microbial activity in this incubation study. We found that a positive PE occurred by glucose supply with (182%) or without (181%) mineral N input over the 21-day incubation, and its values were very similar to the PE ratios previously reported in other deep soils. The estimated mean residence time (MRT) of black soil C considering PE was clearly accelerated by glucose supply, regardless of mineral N input, compared with the initial soil MRT. These results strongly support our hypothesis that the mineralization rate of black soil C in buried A horizons is easily accelerated by a labile C supply, and it also demonstrates important implications for the effects of global warming on buried A horizons (e.g., increased root exudation, fine root biomass supply, and N deposition) in Japanese volcanic ash soils.