Ovariectomized (OVX) mice serve as a key model for studying postmenopausal metabolic changes, particularly obesity, as they mimic the hormonal state of postmenopausal women. However, our understanding remains limited regarding how hormonal and dietary factors affect different adipose tissues. Furthermore, precise documentation of experimental procedures and their effects on specific adipose tissue depots is essential for reproducible translational research. This study investigated depot-specific adiposity development in OVX mice fed a high-fat diet (HFD), focusing on how reduced estrogen levels and dietary intervention affect distinct fat depots. We composed subcutaneous and visceral white adipose tissue (WAT) depots from sham-operated (Sham) and OVX female C57BL/6JJcl mice on a regular diet (RD) and high-fat diet (HFD) for 20 weeks. OVX mice on HFD gained significantly more weight than Sham controls. Adiposity increased in abdominal subcutaneous WAT (sWAT) and perirenal WAT (prWAT) of OVX mice, but not in mesenteric WAT (mWAT). Analysis of adipose tissue morphology revealed that OVX mice exhibited enlarged adipocyte cross-sectional areas under low estrogen (E2) conditions, suggesting enhanced adipogenesis in an estrogen-deficient state. These findings suggest that low estrogen condition accelerated adiposity, in a tissue site-dependent manner.

Apoptosis, an important pathological event associated with kidney disease progression, is expected to be a therapeutic target in chronic kidney disease (CKD). However, its role in naturally occurring CKD in aged cats remains unclear. Therefore, here, we investigated kidney tissues from aged cats (≥10 years) with or without azotemic CKD to evaluate apoptotic events using a terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. The positive TUNEL signals of the renal cells were quantified and statistically analyzed for correlation with the severity of plasma creatinine (pCre) concentration, renal lesions (glomerulosclerosis, interstitial cell infiltration, peritubular capillaries, and interstitial fibrosis), and oxidative damage of the kidney tissue. Oxidative damage was evaluated using immunohistochemistry for 8-hydroxy-2’-deoxyguanosine (OHdG) and 4-hydroxynonenal (HNE). In the TUNEL assay, regardless of azotemia, positive nuclear signals were observed in the tubular epithelial and intraluminal cells, interstitial infiltrating cells, and glomerular cells. Quantitative TUNEL scores showed no significant differences between the azotemic and non-azotemic groups in any compartment of the kidney tissues. In the azotemic group, TUNEL scores did not correlate with pCre or renal lesion severity. However, the scores showed a significant positive correlation with the scores of 8-OHdG and 4-HNE. These findings suggest that apoptosis associated with oxidative damage in renal tissue is an initial pathological event that leads to CKD, rather than a change following CKD progression, in aged cats. Inhibiting apoptosis by antioxidant treatment may be a key strategy to prevent the development of CKD.

In recent years, the importance of using local disinfectants instead of systemic antibiotics for the treatment of infectious skin diseases to prevent the emergence of resistant bacteria has become widely recognized. Chlorhexidine gluconate (CHG) is commonly used in veterinary antibacterial shampoos; however, the daily topical application of diluted CHG solutions has also been adopted. Despite its widespread use, few studies have investigated the effects of CHG on the canine skin barrier. This study examined the skin barrier-damaging effects of CHG in dogs by applying diluted CHG (0.05%, 0.5%, and 4%) daily to six healthy dogs for 2 weeks. At the end of the trial, transepidermal water loss (TEWL), skin surface hydration (SSH), and number of skin bacteria were evaluated. Additionally, the cytotoxicity of CHG to the canine progenitor epidermal keratinocytes (CPEK) was examined using the water-soluble tetrazolium salt (WST-8) assay. Continuous application of 0.5% and 4% CHG to the skin led to a significant reduction in the number of skin bacteria. Additionally, at 4% CHG, a notable increase in TEWL and a decrease in SSH was observed. The WST assay revealed cytotoxicity of CHG at concentrations of 0.05%. In conclusion, although daily topical application of 4% CHG demonstrated the potential to disrupt the skin barrier, 0.5% CHG demonstrated sufficient antimicrobial activity without skin barrier disruption. Additionally, when treating dogs’ skin with a compromised stratum corneum, use of lower concentration of CHG is suitable.

A 24-day-old female Japanese Black calf presented a sudden paraplegia after a history of watery diarrhea. Antemortem magnetic resonance imaging confirmed the suspicion of thrombotic component in the abdominal aorta, without any spinal cord abnormality at the lumbar region. On necropsy, a massive thrombus occupied the lumen from the distal abdominal aorta to the bifurcation of the external iliac arteries. In the thoracic aorta, another mural thrombus developed from the caudal side of the incompletely closed ductus arteriosus orifice, with aortic wall erosion. Both thrombi were mainly composed of platelets. Any microbes were undetected during organ and thrombus incubations. A saddle embolism in the abdominal aorta occurred by an abacterial white thrombus suspiciously originated from the thoracic aortic mural thrombus.

We investigated the distribution and antimicrobial resistance of 120 Staphylococcus felis isolates from feline patients in Japan, mainly from the urinary tract (28.3%), abscesses (23.3%), ears (22.5%), and nasal cavity (10.8%). The distribution of S. felis differed from those of previous studies in Japan and other countries. Antimicrobial susceptibility testing revealed a relatively high resistance to penicillin (PEN, 33.3%), followed by erythromycin (ERY, 15.8%), clindamycin (CLI, 13.3%), and levofloxacin (5.0%). However, oxacillin resistance was not detected. Notably, 11/120 (9.2%)S. felis isolates exhibited multidrug resistance, i.e., resistance to more than three classes of drugs, which mainly consisted of PEN-ERY-CLI resistance phenotypes. This is the first investigation on antimicrobial-resistant S. felis isolates from feline patients in Japan.

A homozygous individual for ITGB7 gene mutation, an autosomal recessive congenital disorder in Holstein cattle, was retrospectively identified by genotyping of 195 stored blood from patients less than 12 months of age. Other 24 patients (12.3%) showed heterozygous. The homozygous individual was a 107-day-old female calf born on March 2017, who presented with chronic diarrhea and severe hypocholesterolemia suggesting hereditary cholesterol deficiency (CD), but genotyping analysis showed negative for CD. The patient showed watery diarrhea, dehydration, and extreme emaciation. Necropsy revealed no apparent cause of chronic diarrhea. Histopathological examination revealed mild mucosal inflammation from the jejunum to the colon. Seven years after the patient’s death, the availability of ITGB7 gene mutation testing revealed the patient to be homozygous.

The three-drug anesthetic mixture (medetomidine, midazolam and butorphanol), developed as an injectable anesthetic for laboratory animals, has been verified from various perspectives and applied to mice and other laboratory animals. However, the effects of its storage conditions and periods on its efficacy have not yet been studied. This study investigated the mixture’s efficacy after storage under various conditions (room temperature, 4°C and −20°C) for 1 and 2 years. Mice in all groups were induced into a stable anesthetic state for at least 15 min after administration. The mice recovered from the anesthetized state 35 min after administration of the antagonist. These findings demonstrate the mixture’s stability under different storage conditions and durations, potentially improving laboratory mouse welfare.

Fasciola-induced fascioliasis is a zoonotic disease with significant health and economic impacts on humans and livestock. Freshwater Lymnaea snails serve as intermediate hosts, contributing to the increasing prevalence of fascioliasis in cattle in coastal areas. The salinity tolerance of Lymnaea snails was investigated along with their distribution and Fasciola infection rates in both snails and grazing cattle in Ben Tre, Tra Vinh, and Soc Trang provinces in Mekong Delta, Vietnam, where seawater reversely enters into the paddy field during the dry season. Lymnaea snails were collected from 53 communes across the three provinces and analyzed for Fasciola larval infections. Additionally, cattle fecal samples were examined for the presence of Fasciola eggs. The salinity levels in the study areas ranged from 0.03% to 1.90%. In total, 1,152 Lymnaea snails including L. viridis and L. rubiginosa were collected. The maximum salinity levels of L. viridis and L. rubiginosa habitats were 0.64% and 0.74%, respectively. The Fasciola infection rates of L. viridis were 2.0% in low-salinity areas and 3.7% in high-salinity areas, whereas no L. rubiginosa infections were detected in either salinity area. The prevalence of cattle fascioliasis in high-salinity areas (35.7%) was similar to low-salinity areas (32.9%). This study is the first to demonstrate that L. viridis can thrive and sustain Fasciola transmission under high-salinity conditions, suggesting the salinity tolerance of these intermediate host snails as a key factor contributing to the expansion of fascioliasis in high-salinity coastal areas.

Lipocalin-2, a siderophore-binding protein, is an antimicrobial, exerting both pro-inflammatory and anti-inflammatory actions. Lipocalin-2 is also involved in glial activation, matrix metalloproteinase stabilization, and cellular iron flux, all of which play roles in autoimmune diseases. The present study aimed to determine the expression of lipocalin-2 in the eyes of Lewis rats with interphotoreceptor-binding protein-induced experimental autoimmune uveitis (EAU). Significantly elevated serum lipocalin-2 levels were also detected in rats by enzyme-linked immunosorbent assay (ELISA). Lipocalin-2 immunostaining was detected predominantly in activated glial cells, including glutamine synthase-positive Müller cells, and ionized calcium-binding adaptor molecule 1-positive microglia and macrophages in EAU rats. Taken together, the results presented herein show that lipocalin-2 is a potential diagnostic marker for uveitis.

It is challenging to isolate Escherichia albertii from clinical specimens. Therefore, a medium that can selectively grow E. albertii and differentiate it from E. coli is earnestly desired. Here, we describe the evaluation of a recently developed selective differential medium, called cefixime-tellurite-phosphate-xylose-rhamnose-MacConkey (CT-PS-XR-MacConkey) medium, which enables the specific growth of E. albertii and differentiation of E. albertii (colorless) from E. coli (red) based on colony color and thus, facilitating the efficient isolation of E. albertii from diarrheal stool. When three E. albertii negative diarrheal stools were inoculated onto CT-PS-XR-MacConkey and xylose-rhamnose-melibiose (XRM) containing MacConkey agars, no colorless colonies were observed on both the media. However, when E. albertii was spiked into these three diarrheal stools, the ratio of colorless colonies to red colonies was higher on CT-PS-XR-MacConkey agar compared to XRM-MacConkey agar in all three samples. Notably, out of 105 Eacdt-gene PCR negative diarrheal stools 56 yielded colorless colonies on MacConkey agar while out of these 56 diarrheal stools, nine yielded colorless colonies on XRM-MacConkey but no colorless colonies were observed on CT-PS-XR-MacConkey agar. Furthermore, evaluation of these two media with five E. albertii positive-stool specimens revealed that the number of red colonies were constantly less, whereas that of colorless colonies were constantly more on CT-PS-XR-MacConkey agar, thus aiding in efficient isolation. Altogether, these results suggest that the CT-PS-XR-MacConkey agar could be a useful selective differential medium for isolation of E. albertii from diarrheal stool specimens.

Chicken meat is a major source of foodborne salmonellosis. In Japan, fluoroquinolones and third-generation cephalosporins are the first- and second-choice treatments for Salmonella gastroenteritis, respectively. We investigated the prevalence and antimicrobial resistance of Salmonella in 154 chicken meat products from Hokkaido (42), Tohoku (45), Kanto (5), and Kyushu (62), Japan. Salmonella was isolated from 133 products (86.4%). High resistance rates were observed for streptomycin (56.5%), tetracycline (50.7%), and kanamycin (47.8%), while all isolates were susceptible to cefazolin, cefotaxime, gentamicin, ciprofloxacin, colistin, and chloramphenicol. The most common serovar, Salmonella Schwarzengrund (83.3%), showed clear regional differences in multidrug resistance: 100% in Kyushu, 41.5% in Tohoku, and 0% in Hokkaido. These findings highlight significant geographical variation in antimicrobial resistance among Salmonella Schwarzengrund isolates.

Two captive-bred lizards, a Western spiny-tailed iguana (Ctenosaura pectinata) and a bearded dragon (Pogona vitticeps), were evaluated for anorexia and absence of feces. The iguana had a recent cloacal prolapse, whereas the dragon had a repaired prolapse 20 days earlier. Exploratory celiotomy under anesthesia revealed a devitalized distal colon in the iguana and stenosis of ductal organs in the pelvic cavity in the dragon, leading to colostomies. Stomal stenosis, managed through dilation, was the main complication. Both gained weight within 1,000 days post-surgery. At 2,152 days, the iguana weighed 166 g; the dragon, which died at 1,792 days, weighed 274 g. Colostomies may benefit lizards with distal colon or cloaca damage that can still urinate, as demonstrated in this report.

A 9-year-old spayed female mixed breed dog weighing 6.8 kg with a history of previous splenectomy for hemangiosarcoma 4 years earlier was referred for a hepatic mass lesion. Although the dog did not have a clinical sign, a computed tomography revealed a solitary mass in the left medial lobe of the liver. Spleen-like tissue was observed on needle core biopsy of the mass, leading to the diagnosis of intrahepatic splenosis. Therefore, hepatic lobectomy of the left medial lobe of the liver was performed. Histopathological evaluation of the excised tumor confirmed the diagnosis of intrahepatic splenosis. No further development of splenosis was detected on CT performed 217 days after the surgery. The dog died of suspected aspiration pneumonia without any event or clinical signs related to intrahepatic splenosis on postoperative 272 days. The cause of the intrahepatic splenosis observed in this dog was unknown; however, hematogeneous dissemination and autoimplantation of splenic tissue via the portal venous flow was speculated. This case shows the utility of needle core biopsy for the preoperative diagnosis of splenosis. The intrahepatic splenosis in this case was considered to have developed as a long-term sequelae after splenectomy, as in humans.

Microsporidia are emerging, opportunistic fungi that infect a diverse population of vertebrates and invertebrates. Mammals of the superorder Xenarthra can harbor and transmit several pathogens, acting as important sources of infection for spreading various zoonoses. Microsporidia have not yet been described in this group of animals, the aim of this study was to analyze the occurrence of the zoonotic microsporidian Encephalitozoon intestinalis, Encephalitozoon cuniculi and Enterocytozoon bieneusi in the feces of giant anteater (Myrmecophaga tridactyla) and armadillo species (Priodontes maximus, Euphractus sexcinctus, Dasypus novemcinctus, Cabassous squamicaudis) monitored by Wild Animal Conservation Institute (ICAS) in Brazil. Fecal samples (n=127) were subjected to DNA extraction with the QIAamp Fast DNA Stool Mini Kit. Amplification by PCR was performed with generic primers and the product generated from this reaction was subjected to nested PCR with specific primers. Eleven samples tested positive for E. intestinalis, two from M. tridactyla (2/56, 3.6%), seven from E. sexcinctus (7/51, 13.7%), two from P. maximus (2/11, 18%) and one from C. squamicaudis (1/3, 33.3%). There was a predominance of positive results in adult animals, of both sexes across and in the Pantanal and Cerrado biomes. In conclusion, the prevalence in Xenarthra was 9.4%, with a higher occurrence in armadillos than in anteaters. Therefore, the species of wild mammals studied here should be considered reservoirs of microsporidian pathogens and have a relevant role in the concept of One Health.

The Pallas’s squirrel (Callosciurus erythraeus) has invaded fragmented woodlands in urban areas of Kanagawa Prefecture, where frequent human contact occurs. We examined 538 squirrels in Hayama-machi, Kanagawa Prefecture, for ectoparasite infestations. A total of 1,164 lice, 877 fleas, and 231 ticks were retrieved from 297 (55.2%), 338 (62.8%), and 135 (25.1%) squirrels, respectively. The identified ectoparasite species were Neohaematopinus callosciuri, Ceratophyllus anisus, Haemaphysalis flava, Haemaphysalis hystricis, and Ixodes turdus. The prevalence of N. callosciuri and C. anisus was significantly higher in adult males (N. callosciuri 63.6%, C. anisus 70.4%) than that in adult females (N. callosciuri 36.1%, C. anisus 49.0%; P<0.0001). Such information on the dynamics of host-ectoparasite relations is crucial for evaluating the risk to public health.

An 11-year-old neutered male Asian small-clawed otter fell down the stairs while sleeping, after which it developed left-sided paralysis. Initial treatment involved once daily administration of prednisolone at 0.5 mg/kg. Despite slight clinical sign improvements by day 10, paralysis persisted. MRI (T2WI) identified a well-defined, hyper-intense lesion on the left side within the spinal cord at the C2-3 intervertebral level. Based on CT and MRI findings, fibrocartilaginous embolus (FCE) was suspected. Prednisolone was then tapered and by day 23 of illness, the otter was able to walk normally, so prednisolone administration was discontinued. No neurological symptom recurrence was observed, and the course remained favorable. One year later, a follow up MRI revealed a diminished lesion compared to initial examination.