Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein

Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.     

Methods of evaluating the spermatogenic ability of male raccoons (Procyon lotor)

Feral raccoons (Procyon lotor) have been growing in number in Japan, and they are becoming a problematic invasive species. Consequently, they are commonly captured and killed in pest control programs. For effective population control of feral raccoons, it is necessary to understand their reproductive physiology and ecology. Although the reproductive traits of female raccoons are well known, those of the males are not well understood because specialized knowledge and facilities are required to study them. In this study, we first used a simple evaluation method to assess spermatogenesis and presence of spermatozoa in the tail of the epididymis of feral male raccoons by histologically examining the testis and epididymis. We then evaluated the possibility of using 7 variables—body weight, body length, body mass index, testicular weight, epididymal weight, testicular size and gonadosomatic index (GSI)—to estimate spermatogenesis and presence of spermatozoa in the tail of the epididymis. GSI and body weight were chosen as criteria for spermatogenesis, and GSI was chosen as the criterion for presence of spermatozoa in the tail of the epididymis. Because GSI is calculated from body weight and testicular weight, this model should be able to be used to estimate the reproductive state of male raccoons regardless of season and age when just these two parameters are known. In this study, GSI was demonstrated to be an index of reproductive state in male raccoons. To our knowledge, this is the first report of such a use for GSI in a member of the Carnivora.     

Pharmacokinetic analysis of plasma concentration and urinary excretion of sulphamonomethoxine and its metabolite in Goettingen minipigs

The plasma concentration and renal excretion after a bolus intravenous injection of a low (10 mg/kg) or high (100 mg/kg) dose of sulphamonomethoxine (SMM) were studied in five Goettingen minipigs. The time data of plasma concentration after a low dose decreased rapidly and appeared to be linear on semilog graph paper. On the other hand, a decrease in the plasma concentration after a high dose was slow at first, gradually accelerated, then became rapid, showing that SMM disposition after a high dose (100 mg/kg) seemed to be non-linear with capacity-limited elimination. A large amount of the acetyl derivative of SMM (AcSMM), which was determined to be the main excretory product of SMM in urine, was detected in the plasma after SMM injection. As the ratios of the area under plasma concentration-time curve of AcSMM to that of SMM were not significantly different at either dose, the acetylation of SMM may be unsaturable by injection of 100 mg/kg of SMM. Immediately after the injection of a low dose, a rapid hyperbolic increase of the fraction of the cumulative amount of the excretory products in urine was observed. On the other hand, the fraction curve at the high dose rose slowly at first, then rapidly and hyperbolically. These results suggested that the non-linear drug disposition after a high dose (100 mg/kg) of SMM in pigs may be the result of a limited capacity for renal excretion of SMM and excretory products, especially the acetyl derivative.     

Evaluation of the transmission risk of foot-and-mouth disease in Japan

The transmission risk of foot-and-mouth disease (FMD) in Japan was evaluated using a mathematical FMD transmission model. The distance-based transmission rate between farms, which was parameterized using the FMD epidemic data in 2010 in Japan, was used to calculate the local-level reproduction numbers—expected numbers of secondary infections caused by one infected farm—for all cattle and pig farms in the country, which were then visualized as a risk map. The risk map demonstrated the spatial heterogeneity of transmission risk in the country and identified risk areas with higher possibility of disease spread. This result suggests that, particularly in high-risk areas, it is important to prepare for the smooth and efficient implementation of control measures against FMD outbreaks.     

The effects of migration on the immunity of Black-Headed Gulls (Chroicocephalus ridibundus: Laridae)

In order to elucidate the relationship between migration period and immunity related to susceptibility, we conducted research on Black-headed gulls (Chroicocephalus ridibundus). We captured 260 gulls and collected their peripheral blood. Their leukocyte (WBC) count, percentages of heterophils (Het) and lymphocytes (Lym), heterophil and lymphocyte ratio (H/L ratio), and CD4 and CD8α expression levels (CD4 and CD8α, respectively) were quantitatively analyzed over three migration periods (Autumn migration, Wintering, Spring migration). In Adult gulls, WBC counts and CD4 levels significantly increased. Moreover, the Het and H/L ratio decreased from the Autumn migration to Wintering. Conversely, only WBC counts and CD4 levels measurements significantly decreased from Wintering to Spring migration (P<0.05). The tested parameters of the Tokyo-bay population show a greater significant difference than the measurements of immunity of the Mikawa-bay population. This study suggests that the migratory period has a negative effect on an aspect of the immune system. Including the period-difference in the immune systems in the local population, it is necessary to investigate the relationship between the ecology of migratory birds and their immunity.     

Three compartment model for pyrimethamine disposition in the pig

Plasma concentration and urine excretion of pyrimethamine (Py) after intravenous (i.v.) administration (10 mg/kg) were determined in four pigs, using NP-FID gas chromatography. A three-compartment model system adequately fitted the observed plasma data. Py may undergo an extensive extravascular distribution, because of the ratio of total volume of distribution to volume of the central compartment was extremely large (about 10). From the calculation of theoretical distribution, only 30% of the dose appears to remain in the central compartment 15 min after administration. Within 2 h of administration the proportion of drug within the two identified peripheral compartments exceeded the proportion remaining in the central compartment. Calculation of cumulative drug excretion showed that up to 90% of an administered dose is excreted within 24 h of administration, although only about 3% of the dose could be detected in urine in an unchanged form during the 24 h period.     

Experimental production of porcine gastroesophageal ulcers by betazole and reserpine

The ulcerogenic and secretogenic properties of betazole, (an H₂ agonist), and/or reserpine on pig stomachs were studied. (1) Betazole (50 mg/body) and reserpine (0.02 mg/kg) were successively injected (i.m.) for 5–10 days into starved animals. Gastroesophageal ulcers, pathologically similar to naturally occurring ulcers were observed in all pigs (10/10), whether a nutrient solution (5% glucose, 1 litre/day) was supplied or not. In the starved pigs given either betazole or reserpine, only a small ulcer or erosion was observed. (2) In four pigs which were given commercial mash at a rate of 3% of body weight per day and killed on the 6th or 10th day of successive injections of both drugs, the incidence of ulcers was 2/4. (3) The gastric secretogenic effect of betazole and/or reserpine was confirmed in pigs with Heidenhain gastric pouches. (4) From these data, a new method for the experimental induction of pig gastroesophageal ulcers, by means of simultaneous injections of betazole and reserpine for 5 days under the nutrient (glucose solution) drinking condition, is proposed. The authors consider that this method may be superior to the repository histamine method, with respect to the absence of adverse effects, practical convenience and pathological and etiological similarities with naturally occurring ulcers.     

Reproductive characteristics of Japanese monkeys on Shimokita Peninsula,Japan, the northernmost habitat of wild primates in the world

The Shimokita Peninsula in Aomori Prefecture, Japan, which is inhabited by Japanese monkeys (Macaca fuscata), is the northernmost habitat for wild primates in the world. This study was the first to determine the conception dates of specific individuals and estimate the pregnancy rate of wild populations in this region. The pregnancy rate of animals aged 5 years or more at delivery was estimated to be 40.9% (27/66). Conception dates of each fetus were also estimated using a regression line of Pig-tail monkeys (Macaca nemestrina), which are taxonomically related to Japanese monkeys and have a similar physique. The conception dates were distributed across 90 days between September 24th and December 23rd, with a mean conception date of November 4th (SD=22.3 days, n=53). Using these findings, the mean birth date was estimated as April 25th, more than two weeks earlier than the mean birth date in previous research determined using direct observations carried out over the past 20 years ago. Global warming due to climate change is thought to be one of the main causes of this difference.     

The role of neighboring infected cattle in bovine leukemia virus transmission risk

A cohort study was conducted to evaluate the risk of bovine leukemia virus (BLV) transmission to uninfected cattle by adjacent infected cattle in 6 dairy farms. Animals were initially tested in 2010–2011 using a commercial ELISA kit. Uninfected cattle were repeatedly tested every 4 to 6 months until fall of 2012. The Cox proportional hazard model with frailty showed that uninfected cattle neighboring to infected cattle (n=53) had a significant higher risk of seroconversion than those without any infected neighbors (n=81) (hazard ratio: 12.4, P=0.001), implying that neighboring infected cattle were a significant risk factor for BLV transmission. This finding provides scientific support for animal health authorities and farmers to segregate infected cattle on farms to prevent spread of BLV.     

Potential impact of species and livestock density on the epidemic size and effectiveness of control measures for foot-and-mouth disease in Japan

The characteristics of a livestock area, including farm density and animal species, influence the spread of foot-and-mouth disease (FMD). In this study, the impact of livestock area on FMD epidemics was examined using an FMD transmission model. For this simulation, three major livestock areas were selected: the 2010 FMD epidemic area in Japan as the baseline area (BS), a cattle and pig mixed production area (CP) and a cattle production area (C). Simulation results demonstrated that under the 24-hr culling policy, only 12% of epidemics among 1,000 simulations were abated within 100 days in the CP area, whereas 90% of the epidemics ceased in the BS area. In the C area, all epidemics were successfully contained within 100 days. Evaluation of additional control measures in the CP area showed that the 0.5-km pre-emptive culling, even when only targeting pig farms, raised the potential for successful containment to 94%. A 10-km vaccination on day 7 or 14 after initial detection was also effective in halting the epidemics (80%), but accompanied a large number of culled or vaccinated farms. The combined strategy of 10-km vaccination and 0.5-km pre-emptive culling targeting pig farms succeeded in containing all epidemics within 100 days. The present study suggests the importance of preparedness for the 24-hr culling policy and additional control measures when an FMD outbreak occurs in a densely populated area. Considering the characteristics of the livestock area is important in planning FMD control strategies.