Timing of Puberty and Its Relationship with Body Growth and Season in Male Raccoons (Procyon lotor) in Hokkaido

The raccoon (Procyon lotor), indigenous to North America, has naturalized in Japan as an invasive alien species, having been introduced into the country in the 1970s. In Hokkaido, the northernmost island of Japan, feral raccoons have been increasing in number and spreading throughout the island. The age at the onset of puberty for raccoons is important for estimating individual lifetime reproductive success and population growth. The present study investigated the timing of and potential factors affecting the onset of puberty in male raccoons in Hokkaido. External characteristics and histology of testes were studied in 151 male feral raccoons and in 1 captive juvenile. For the majority of feral yearling raccoons, prepubertal development began in May, and spermatozoa production began in October prior to their second mating season. However, some larger juveniles attained puberty during the juvenile period. The captive juvenile, which was fed throughout the winter, attained puberty only 11 months after birth. These results suggest that if male raccoons can achieve enough body growth before the first mating season, puberty can be attained early. In both juveniles and yearlings, spermatozoa production was only observed after autumn. This timing coincided with the recrudescence of seasonally active spermatogenesis in adult males. Therefore, attaining puberty in male raccoons appears to require both adequate body nutrient development and several environmental factors that control seasonal testicular changes.     

Self-EcoTILLING to identify single-nucleotide mutations in multigene family

TILLING (Targeting Induced Local Lesions IN Genomes) is a low-cost, high-throughput reverse genetic technique that employs a mismatch-specific endonuclease CEL-1 to discover induced point mutations in the genes of interest. The use of the TILLING technique to survey natural variation in genes is called EcoTILLING. Here, we report a modified EcoTILLING method for the discovery of mutations in multigene family, which we coin “Self-EcoTILLING” by using an allotetraploid Monochoria vaginalis ALS multigene family as an example. The mutations could be detected by TILLING of PCR products resulting from the primers specific to both Als1 and Als3 without involving the experimental step of mixture of reference and query DNA. Either of the two co-amplified loci could serve as reference DNA to the other. We demonstrate with this example that Self-EcoTILLING is a fast, reliable and economical technique of detecting single-nucleotide mutations in polyploid plants containing multigene family.     

Physicochemical and morphological properties of termite (Macrotermes bellicosus) mounds and surrounding pedons on a toposequence of an inland valley in the southern Guinea savanna zone of Nigeria

Termites play a significant role in soil-forming processes of the tropics. The influence of termites on pedogenesis as affected by the toposequence, however, has rarely been explored. We investigated the soil physicochemical and morphological characteristics of epigeal mounds constructed by Macrotermes bellicosus (Smethman) compared with those of surrounding pedons along a toposequence (bottom, fringe and upland sites) of an inland valley in central Nigeria. The physicochemical and morphological properties of the mound soils varied according to structural units but were generally different from those of the adjacent pedons. The differences included finer texture, higher electrical conductivity, total N, exchangeable bases (Ca, Mg and K) and effective cation exchange capacity and lower C/N ratio and exchange acidity in the mound than the pedon at each toposequence position. This tendency to modify the soil properties was more prominent in the nest body where the termites actually live, that is, in the hives, royal cell and base-plate, than in the soils below the nest and the other mound parts, that is, the external wall, internal wall and pillars. We found this trend to a greater or lesser degree at all toposequence positions. Our findings suggest that: (1) M. bellicosus can manipulate the mound soils according to functional applications of structure units or environmental requirements for its livelihood, regardless of local soils; (2) M. bellicosus makes ecological patches (hot spots) at all toposequence positions in the same measure; (3) the influence of M. bellicosus on the pedogenesis is reduced in the lowlands compared with the uplands because the number and volume of the mounds were substantially lower in the bottom and fringe sites compared with the upland site.     

Pharmacokinetics of fentanyl after single intravenous injection and constant rate infusion in dogs

OBJECTIVE: To determine the plasma concentration and define the pharmacokinetic characteristics of fentanyl (10 microg kg(-1)) administered as a single intravenous (IV) injection followed by: (a) no further drug; or (b) a constant rate infusion (CRI) of fentanyl 10 microg kg(-1) hour(-1) lasting 1, 3 or 4 hours in dogs. Animals Fourteen healthy adult beagles (seven males and seven females). EXPERIMENTAL DESIGN: Randomized cross-over design. MATERIALS AND METHODS: Dogs were randomly assigned to four treatment groups. Drugs were administered to each dog in a randomized cross-over design with at least a 14-day washout interval between experiments. All dogs received an IV loading dose of fentanyl (10 microg kg(-1)). One group received no further fentanyl. In others, the loading dose was followed by a CRI of fentanyl (10 microg kg(-1) hour(-1)) for 1, 3 or 4 hours. Blood samples were collected and plasma fentanyl concentrations determined using high-performance liquid chromatography-mass spectrometry. Plasma pharmacokinetic estimates were obtained by plotting plasma concentrations versus time data and by fitting the change in concentration to a pharmacokinetic model, using a purpose-built program written by the Graduate School of Pharmaceutical Sciences (Kyoto University) in Visual Basic (VBA) on Excel (Microsoft Corporation). RESULTS: Plasma fentanyl concentration decreased rapidly after single IV injection: the plasma concentration-time curve best fitted a two-compartment model. Pharmacokinetic variables for IV injection were characterized by a short distribution half-time (t1/2alpha was 4.5 minutes), a relatively long elimination half time (t1/2beta was 45.7 minutes), a large volume of distribution (approximately 5 L kg(-1)) and high total body clearance (77.9 mL minute(-1) kg(-1)). Stable plasma fentanyl levels were obtained in all CRI groups although pharmacokinetic variables were influenced by the duration of administration. CONCLUSIONS AND CLINICAL RELEVANCE: While this study clarified the pharmacokinetic features of rapid IV fentanyl injection and CRI in dogs, the plasma concentration achieving analgesia was not and so further research is needed. Further studies on the effects of other sedatives and/or anaesthetics on fentanyl's disposition are also required as the drug is commonly used with other agents.     

Maturation-associated changes in internal distribution and intra-ovarian microdistribution of tetrodotoxin in the pufferfish <Emphasis Type="Italic">Takifugu pardalis</Emphasis>

To clarify the maturation-associated accumulation profile of tetrodotoxin (TTX) in the female pufferfish Takifugu pardalis, specimens collected from Omura Bay, Nagasaki Prefecture, Japan, from November through December, 2014, were separated based on the histology of ovarian sections into four groups: yolk vesicle stage (Yv), yolk globule stage (Yg)-1, Yg-2, and Yg-3. TTX quantification in each tissue in each group by liquid chromatography–mass spectrometry revealed that the TTX concentration [mouse unit (MU)/g] in the ovary increased from Yv to Yg-1, and the TTX amount (MU/individual) in the ovary increased from Yg-1 through Yg-3. In Yv, TTX in the skin accounted for 86% of the total TTX in the body, whereas from Yg-1 through Yg-3, ovarian TTX accounted for 53–76% of the total. In female specimens collected from the same bay from February to May 2015, the TTX amount in the ovary markedly decreased after spawning, and the TTX amount in the liver increased. Immunohistochemical observation of the ovarian sections indicated that TTX localized mainly in the nucleus of immature oocytes, but then distributes to the yolk vesicles and yolk globules as maturation advances, and is partly transferred to the egg membrane as spawning approaches.     

Genetic characterization of chicken anemia viruses newly isolated from diseased chicks in Japan in 2020

In this study, a total of nine chicken samples obtained from two broiler flocks in Oita and Tottori prefectures in 2020 were examined for Chicken anemia virus (CAV) infection. The samples were collected from clinically suspected flocks and diseased chickens. The CAV genome was detected in all nine samples tested by real-time PCR. Phylogenetic analyses and sequence comparisons of the full-length VP1 gene sequences indicated that all the Japanese CAV strains obtained in this study formed a similar cluster of genotype III and shared high nucleotide (99.62–100%) identity. The current Japanese CAV strains were closely related to Chinese CAV strains but not related to vaccine strains. One positive selection site of VP1 was detected among the Japanese CAV strains.     

Effect of drug-eluting bead transarterial chemoembolization loaded with cisplatin on normal dogs

Transcatheter arterial embolization (TAE) and transcatheter arterial chemoembolization (TACE) are standard treatments for advanced hepatocellular carcinoma (HCC) and particularly for unresectable tumors or liver metastases in humans. However, reports on TACE used in veterinary medicine are few. This study aimed to evaluate the feasibility and safety of drug-eluting bead transarterial chemoembolization (DEB-TACE). We performed DEB-TACE in four clinically normal dogs and pharmacokinetically compared the results against hepatic arterial infusion (HAI) of cisplatin in two dogs. Drug-eluting beads (DEB) loaded with cisplatin were injected through a microcatheter for selective embolization of the left hepatic artery. After embolization, computed tomography (CT) images and histological examination findings were obtained during a 4-week observation period. Serum platinum concentrations were measured to evaluate cisplatin after each procedure. Biochemical analysis was performed during a 12-week observation period. Embolization was successful in all dogs, and there were no clinically apparent abnormalities. Embolization was confirmed up to 4 weeks after DEB-TACE in two of the four dogs and up to 1 week in the other two dogs using postoperative CT images. Cisplatin was not detected in peripheral veins in all dogs after DEB-TACE, but it was detected in trace amounts after HAI. DEB-TACE using cisplatin was safe and well tolerated by normal dogs. DEB-TACE may be useful in terms of determining systemic toxicity and drug concentration within tumors.