In this study, we investigated the aortic arch (AA) branching pattern in the Eurasian otter (Lutra lutra). We performed arterial silicone casting of the AA of 18 Eurasian otters (8 males and 10 females). We analyzed the AA branching pattern at three levels: the AA, brachiocephalic trunk (BCT), and subclavian artery (SB), using different classification methods at each level. We introduced new criteria for classifying the SB branching pattern applicable for Eurasian otter and other carnivores based on the sequence of the four main branches: vertebral artery (VT), internal thoracic artery (IT), costocervical artery (CCT), and superficial cervical artery (SC). In all Eurasian otters, two major branches emerged directly from the AA, i.e., the BCT and left SB. The BCT branched off the left common carotid artery and terminated in the right common carotid artery and right SB in 17 of 18 Eurasian otters; the BCT formed a bicarotid artery in the remaining case. The SBs showed various branching patterns, with the main branching pattern involving branching to the VT and IT at the same position, followed by the CCT and SC. The SB branching pattern in the Eurasian otter differed from that in dogs in that the two first branching arteries were VT and IT, rather than VT and CCT. Here, we present the anatomical characteristics of the AA branching patterns in the Eurasian otter and new analysis methods applicable for comparative studies of other carnivores.

The expression of sex determining region of the Y chromosome (Sry) in the fetal gonads is important for male development. In a mouse model of disorders of sex development (C57BL/6 (B6)-XY^POS), the gonadal phenotype and the timing of Sry expression differ due to differences among B6 substrains as the genetic background. Since differences in Sry expression among B6 substrains have been speculated, the present study examined Sry expression in B6J, B6JJmsSlc, and B6NCrl mice. These substrains differed in the number of Sry-expressing cells in the gonads of embryonic mice at each developmental stage, with B6NCrl having more than the other strains. The substrains differed also in the number of Sry-expressing cells between the left and right gonads, with B6J and B6NCrl, but not B6JJmsSlc, showing left gonad-dominant Sry expression. Substrain differences existed also in the distribution of Sry-expressing cells in the medial and lateral directions of gonads. In addition, in the left gonad-dominant Sry-expressing substrains B6J and B6NCrl, the medial and central regions of the left gonad had more Sry-expressing cells than those of the right gonad. Substrains of B6 mice have not always been considered in sex differentiation studies. In the present study, however, we observed substrain differences in the number of Sry-expressing cells, left–right distribution, and medial/lateral distribution during the early stages of gonadal development in B6 mice. Therefore, future studies on sex differentiation in B6 mice should consider substrain differences.

Infectious diseases are an important issue in the poultry industry, requiring early diagnosis and countermeasures. To address this, we present a system based on TaqMan real-time PCR to detect pathogen genome in specimens collected from chickens. We designed 12 primer–probe sets for pathogens causing respiratory or systemic symptoms. In field samples, we detected three viruses, including DNA and RNA viruses, and three bacteria. The chicken anemia virus and Avibacterium paragallinarum were detected only in young and laying hens, respectively. Bacteria were detected only in throat swabs, and gallid alphaherpesvirus 2 was detected in different specimens at each developmental stage. Our novel TaqMan real-time PCR system effectively detects pathogen’s gene in chickens, while taking age into account.

A 3-year-old spayed female domestic short-haired cat presented with a head turning to the left, circling to the right, seizures, and opisthotonos for approximately one month. Neurological examination revealed a deficit in the postural reaction of the left limbs and visual abnormalities. Forensic computed tomography revealed a hyperattenuating round mass of 1.3 cm diameter with a hypoattenuating center in the right hemisphere. Histopathology showed multifocal granuloma lesions with the major mass mostly affecting the right basal ganglia. Cryptococcus neoformans variety grubii molecular type VNI/ST31 was isolated from the cryptococcal granulomas. This report highlights the epidemiological distribution and differential diagnosis of a feline central nervous system cryptococcosis caused by C. neoformans that occurred in an Asian country.

The serum concentration and activity of diamine oxidase (DAO) in calves with or without diarrhea during the first month of life were examined. The serum DAO concentration was highest on day 0 after birth, and gradually decreased. No differences in serum DAO concentrations were observed between calves with diarrhea and control calves. In contrast, serum DAO activity was lowest on day 0 after birth, and gradually increased. Serum DAO activity in calves with diarrhea was lower than that in the control calves from day 20 after birth. These findings suggest that serum DAO activity and concentration are not correlated, and serum DAO activity is useful for assessing damage to the small intestinal mucosa in calves.

A 2-month-old female mixed cat was emaciated due to dysphagia, and inspection of the mouth revealed a 2 cm pedunculated mass elongated from the palate, which occupied the oral cavity. The mass was surgically removed, and histopathological examination revealed that the tumor was composed of three germ cell layers: ectodermal (skin and skin appendages), mesodermal (cartilaginous and osseous structures), and endodermal (glandular and respiratory mucosa) tissues. An immature teratoma was diagnosed because of the presence of immature neuroectodermal tissues, and the presence of nephroblastic components was a characteristic finding in this case. This is the first report of an oropharyngeal teratoma in cats and the first case of an immature teratoma with nephroblastic components in a domestic species.

Understanding the antimicrobial resistance of Campylobacter jejuni and Salmonella spp. isolated from patients with enteritis will aid in therapeutic decision-making. This study aimed to characterize C. jejuni and Salmonella spp. isolates from patients with enteritis. For C. jejuni, the resistance rates against ampicillin, tetracycline, and ciprofloxacin were 17.2%, 23.8%, and 46.4%, respectively. All the C. jejuni isolates were susceptible to erythromycin, which is recommended as a first-choice antimicrobial if Campylobacter enteritis is strongly suspected. C. jejuni was classified into 64 sequence types (STs), and the five major STs were ST22, ST354, ST21, ST918, and ST50. The ciprofloxacin-resistance rate of ST22 was 85.7%. For Salmonella, the resistance rates against ampicillin, cefotaxime, streptomycin, kanamycin, tetracycline, and nalidixic acid were 14.7%, 2.0%, 57.8%, 10.8%, 16.7%, and 11.8%, respectively. All the Salmonella spp. isolates were susceptible to ciprofloxacin. Therefore, fluoroquinolones are the recommended antimicrobials against Salmonella enteritis. S. Thompson, S. Enteritidis, and S. Schwarzengrund were the three most prevalent serotypes. The two cefotaxime-resistant isolates were serotyped as S. Typhimurium and were found to harbor bla_CMY-2. The results of this study would help select antimicrobials for treating patients with Campylobacter and Salmonella enteritis.

Bupivacaine, levobupivacaine and ropivacaine are potent, long acting, amide-type local anesthetics that have several clinical applications including intra-articular administration. The objectives of this study were to evaluate their in vitro effects on cell viability and caspase activity to elucidate whether they activate the extrinsic or intrinsic pathways of apoptosis in canine articular chondrocytes. Chondrocytes in monolayer culture were treated with culture medium as the control, or with 0.062% (0.62 mg/mL) bupivacaine, 0.062% levobupivacaine, and 0.062% ropivacaine for 24 hr. Cell viability was evaluated using the live/dead, 3-(4,5-dimehylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT), and Cell Counting Kit-8 (CCK-8) assays. Evaluation of caspase-3, caspase-8, and caspase-9 activity was performed using colorimetric assays. The MTT and CCK-8 assays were used to evaluate the effect of caspase inhibitors on local anesthetic chondrotoxicity. All three local anesthetics decreased chondrocyte viability after 24 hr (P<0.001). Apoptosis was induced through both the extrinsic and intrinsic pathways. Bupivacaine increased caspase-3, caspase-8, and caspase-9 activity (P<0.001). Levobupivacaine increased caspase-3 (P=0.03) while ropivacaine did not significantly upregulate activity for all three caspases. Caspase inhibition did not suppress bupivacaine chondrotoxicity whereas inhibition of caspase-8 and caspase-9 decreased ropivacaine chondrotoxicity and mildly attenuated levobupivacaine chondrotoxicity. In summary, the level of chondrotoxicity, the type of caspase activated, the level of caspase activation, and the response to caspase inhibitors was dependent on the type of local anesthetic. Therefore, ropivacaine may be a safer choice for intra-articular administration compared to levobupivacaine and bupivacaine.

To prevent aspiration in Japanese White (JW) rabbits, the maximum single volume of medetomidine administered intranasally is 0.3 mL per nostril using a mucosal atomization device (MAD). This study aimed to examine the sedative effect of intranasal administration of medetomidine using MAD in eight healthy female JW rabbits. Each rabbit received intranasal atomization (INA) of saline (Control treatment) along with three doses of 1 mg/mL medetomidine (0.3 mL to one nostril [MED0.3 treatment]; 0.3 mL each to both nostrils [MED0.6 treatment]; 0.3 mL twice to both nostrils [MED1.2 treatment]), with a washout period of at least 7 days between treatments. The actual doses of medetomidine were 82 (75–84) μg/kg (median [25th–75th percentile]), 163 (156–168) μg/kg, and 323 (295–343) μg/kg for the MED0.3, MED0.6, and MED1.2 treatments, respectively. A medetomidine-dose dependent sedative effect was detected, and the loss of righting reflex (LRR) was achieved in one rabbit at 18 min, seven rabbits at 11 (9–18) min, and eight rabbits at 7 (4–18) min after the MED0.3, MED0.6, and MED1.2 treatments, respectively. The LRR was maintained for 63 (29–71) min and 83 (68–101) min after the MED0.6 and MED1.2 treatments, respectively. Additionally, the INA of medetomidine produced a significant dose-dependent cardiorespiratory depression including a decrease in pulse rate, respiratory rate, percutaneous oxygen saturation, and arterial partial pressure of oxygen, and an increase in arterial partial pressure of carbon dioxide in the rabbits.

Periparturient stress can have long-term negative effects on both dairy cows and their calves and may contribute to lower productivity. The purpose of this study was to determine how periparturient stress is related to differences in calving difficulty and health status by measuring hair cortisol concentration in periparturient dairy cows and their calves. Calving environment (calving pen, tie stall, and group paddock), calving difficulty, calving progress, health status, and calf weight were recorded, and tail hair from 25 of the cows and their calves was collected at calving and 1 month after calving to measure hair cortisol concentration. There were no significant correlations between hair cortisol concentration and calving environment, calving difficulty, calf weight gain rate. Hair cortisol concentrations at calving were significantly higher in cows with oversized calves or twin births than in cows with normal-sized singleton calves (4.2 ± 2.2 pg/mg vs. 2.1 ± 1.5 pg/mg, P<0.05). Cows with clinical disease within one month of calving had significantly higher levels of hair cortisol one month after calving compared to healthy cows (3.8 ± 1.1 pg/mg vs. 2.3 ± 1.9 pg/mg, P<0.05). Calves with clinical disease within the first month after birth tended to have higher hair cortisol levels at birth than healthy calves (4.7 ± 2.4 pg/mg vs. 3.2 ± 0.9 pg/mg, P<0.1). These results suggest that calving of oversized calves and twin births and suffering clinical diseases can cause more stress for cows during the periparturient period.

Testicular ultrasound enables the evaluation of changes in the testicular parenchyma. This study aimed to report the occurrence of hypoechogenic testicular alterations and their relationship with semen quality in five breeding buffaloes. Two buffaloes presented with hyperechoic points characteristic of fibrosis and anechoic density content between the parietal and visceral tunica. The two bulls without ultrasonographic changes showed higher average trajectory speed, linear velocity, curvilinear velocity, amplitude of lateral displacement of the spermatic head, total motility, progressive motility, fast speed, and acrosomal membrane values within the normal range. The number of spermatozoa with major and total defects was higher in the group of animals without alterations. The three buffaloes that presented with testicular alterations produced semen within established freezing standards.

Inulin-type fructans (ITFs) have been shown to possess various biological activities. However, studies on their safety and side effects are limited. Hence, we aimed to evaluate the possible effects of burdock ITFs on the physiological indices of healthy mice and their filial generation when fed for six months. Thirty-two C57BL/6J mice were randomly divided into two groups; a normal control (NC) and an ITFs group. The parental generations were kept in one cage with free access to a normal diet and double-distilled water (P-NC group) or burdock ITFs drinking water (P-ITFs group, 2% w/v). The filial generations (F-NC group and F-ITFs group) were kept separately and were fed as their parental generation. Behavior, organ/body weight, serum indices, histopathology, time of production, and number of pup births were observed. There were no significant adverse effects on these indices. Functional indices of the spleen, lung, heart, and pancreas of the ITFs groups were higher than those of the NC groups, respectively. Interestingly, the serum glucose (GLU), total cholesterol (TC), uric acid (UA) and creatine kinase (CK) levels of the ITFs groups were lower than those of the NC groups. Meanwhile, the pregnancy number and pup birth number of the P-ITFs group were more than those of P-NC group. Therefore, long-term consumption of burdock ITFs has no obvious adverse effects on the health of parental mice and their offspring, but may contribute to reproductive capacity, fatigue reduction, and risk reduction of renal disease.

Diamide insecticides activate ryanodine receptors expressed in lepidopteran skeletal muscle and promote Ca²⁺ release in the sarcoplasmic reticulum, causing abnormal contractions and paralysis, leading to death of the pest. Although they had been thought not to act on nontarget organisms, including mammals, adverse effects on vertebrates were recently reported, raising concerns about their safety in humans. We investigated the neurotoxicity of the acute no-observed-adverse-effect level of chlorantraniliprole (CAP), a diamide insecticide, in mice using clothianidin (CLO), a neonicotinoid insecticide, as a positive control. The CLO-administered group showed decreased locomotor activities, increased anxiety-like behaviors, and abnormal human-audible vocalizations, while the CAP-administered group showed anxiety-like behaviors but no change in locomotor activities. The CAP-administered group had greater numbers of c-fos–immunoreactive cells in the hippocampal dentate gyrus, and similar to the results in a CLO-administered group in our previous study. Blood corticosterone levels increased in the CLO-administered group but did not change in the CAP-administered group. Additionally, CAP was found to decreased 3-Methoxytyramine and histamine in mice at the time to maximum concentration. These results suggest that CAP-administered mice are less vulnerable to stress than CLO-administered mice, and the first evidence that CAP exposure increases neuronal activity and induces anxiety-like behavior as well as neurotransmitter disturbances in mammals.

Neonicotinoid pesticides (NN) were recently reported to exhibit adverse effects in higher vertebrates. Moreover, NNs are routinely transferred from mother to offspring, raising concerns about their effects on future generations. The fetal and neonatal periods are the most critical to the formation of neural circuits in the brain through neurogenesis and differentiation, neuronal migration, axon guidance, and synaptogenesis. NN exposure throughout the fetal and neonatal periods was found to affect the neurobehavior of the offspring, but the stage-specific neurobehavioral effects are unclear. We exposed fetal and neonatal mice to a no-observed-adverse-effect level (NOAEL) of clothianidin (CLO) for 4 days during each of four developmental stages: neurite proliferation and differentiation (fetal days 9–12, CLO-1), neurite outgrowth (fetal days 15–18, CLO-2), synapse formation and astrocyte differentiation (days 1–4 after birth, CLO-3), and synapse remodeling (days 11–14 after birth, CLO-4). CLO’s neurobehavioral effects were evaluated in juveniles and adults, revealing that CLO-1 and CLO-2 caused behavioral abnormalities in adult mice. CLO-3 significantly increased locomotor activity and decreased juvenile neurons in the hippocampal dentate gyrus in adulthood. Comprehensive gene analysis of CLO-3 revealed high expression of genes related to neurite outgrowth and axonal branching in the hippocampus in juveniles and adults. These results revealed developmental stage-specific effects of a NOAEL of CLO in the fetal and neonatal periods, suggesting that the susceptibility of the fetus and neonate to CLO varies by developmental stage.

Recently, the effects of exposure to clothianidin (CLO) on the thymus and gut microbiota have become clear, but no report has examined its next-generation impacts. Pregnant C57BL/6N mice were administered a no-observed-adverse-effect-level dose of CLO until weaning. We examined CLO’s effects on the gut microbiota and immune organs of dams and their 3- and 10-week-old male offspring. CLO administration led to several alterations of the top 30 bacterial genera in the gut microbiota in dams and 3-week-old mice. Compared to controls, 10-week-old mice had more thymic Hassall’s corpuscles, and both dams and 10-week-old mice had fewer macrophages. These results suggest that fetal and lactational CLO exposure may affect the immune system and gut microbiota of the next generation.

The geneLEAD VIII is a fully-automated nucleic acid extraction/quantitative PCR equipment developed by Precision System Science Co., Ltd., (PSS). To take advantage of its capability, we developed a quantitative assay system to measure growth of animal viruses. The system was used to assay one of the Chinese herbal extracts whose anti-malarial activities were previously reported and demonstrated its dose-dependent anti-viral activity against feline infectious peritonitis virus (FIPV), a feline coronavirus causing the fatal diseases in cats, and relatively low cell toxicity. The assay developed in this study is useful to screen antiviral drugs and the anti-FIPV activity of the herbal extract identified have a potential to lead to development of new drugs against FIPV and other coronaviruses, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Peaton virus (PEAV) is a type of arthropod-borne virus similar to Akabane virus (AKAV), belonging to the genus Orthobunyavirus. AKAV infection is common in cattle, but previous reports have suggested that pigs may play a role in transmission cycle. In addition, antibodies against AKAV were detected in wild boars (Sus scrofa). By contrast, PEAV could infect cattle and pigs, but it remains unknown whether PEAV infects wild boars. This study aimed to reveal the possibility of PEAV infection in wild boars by conducting a serological survey. Consequently, the seropositive rate of PEAV was 26.5% in 264 free-living wild boars in Ehime Prefecture, Japan. This is the first study to report the detection of antibodies against PEAV in wild boars.

The Marabou stork (Leptoptilos crumenifer) is a typical scavenging bird and adapted to the Savannah environment, where they show a carnivorous feeding style. However, Marabou stork recently penetrated into the city areas and acclimatized to the urban environment, where they modified their feeding habits to an omnivorous type toward more carbohydrate. To reveal their adaptation to the variable feeding customs, this study compared the gut microbiomes and chemical compositions of feces of Marabou storks inhabiting two different locations in peri urban Kampala: one is a slaughter house floc that predicted their original carnivorous feeding, and the other is a landfill floc that adapted more to the omnivorous feeding. 16S rRNA gene sequencing analysis revealed more diverse gut microbiome, more enriched Lactobacilli, and less abundant Peptostreptococci in the landfill flock comparing to the slaughter house flock. Isolation work and predicted metagenome analysis confirmed more diverse Lactobacilli and more enriched functions for carbohydrate metabolism in the landfill flock. In addition, chemical composition of feces revealed higher ammonia in the former, which is consisting with higher Peptostreptococci and their practice of carnivorous feeding. These results highlighted their adaptation to the variable feeding environment, which presumably protects their health and ensure survival of species.

Aeromonas hydrophila is a facultative anaerobic gram-negative bacterium regarded as an opportunistic pathogen in animals. A 17-year-old female crab-eating macaque (Macaca fascicularis) died after experiencing anorexia and depression for several days. The carcass was severely emaciated, and the sternum was exposed under subcutaneous lesions in the thorax. Many abnormal pathological lesions were found, including tracheal inflammation, pulmonary inflammatory emphysema, yellowish discoloration of the liver, enlargement of the gall bladder, necrosis of the heart, congested bilateral kidneys, and enlargement of the adrenal glands. The stomach was empty, mucosal ulcerations were found, and the duodenum was congested. Giemsa staining revealed rod-shaped organisms in the whole blood smear and major organs, which were identified as A. hydrophila. The animal had experienced stress, and decreased immune system function possibly contributed to the infection.

Elephant endotheliotropic herpesvirus (EEHV) causes a fatal hemorrhagic disease and is a significant cause of mortality in juvenile Asian elephants. A loop-mediated isothermal amplification (LAMP) method was developed to rapidly diagnose EEHV viremia. However, extracting DNA from whole blood samples to perform LAMP hampers diagnosis in a field setting. Here, we established the Direct-LAMP method, using heparinized plasma without extracting the DNA to speed up and simplify the test. EEHV-positive specimens were tested using the Direct-LAMP. The detection limit was calculated to be 10^1.3 copies/μL using the mimetic samples, which was almost identical to the value determined in LAMP in which DNA was extracted. Hence, the Direct-LAMP provided a more rapid diagnosis to save, which could prevent elephant deaths.