Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive interstitial lung disease characterized by excessive collagen deposition and irreversible scarring. Reliable animal models are essential for elucidating IPF pathogenesis and evaluating antifibrotic therapies. However, conventional bleomycin (BLM) models generally induce only transient and moderate fibrosis. Considering the profibrotic role of eosinophils, we developed a novel IPF model using the spontaneous eosinophilic mutant Yama mice strain to achieve persistent and severe fibrosis. Male Yama and C57BL/6 mice (6 weeks old, n = 24) received eight intranasal administrations of BLM (2 mg/kg) at two-week intervals, while controls received saline. Peripheral eosinophil counts were monitored, and tissues were analyzed histologically and biochemically. Yama mice exhibited a sharp eosinophil peak after the first BLM exposure, followed by a gradual decline. Histological analysis revealed markedly enhanced fibrosis, greater collagen deposition, and higher Ashcroft scores in Yama mice than in C57BL/6 mice. Alcian blue-PAS-positive fibroblast foci, a characteristic feature of human IPF, were abundant in Yama mice. Immunohistochemistry demonstrated increased expression of eosinophil cationic protein (ECP). Moreover, mRNA expression of Th2 cytokines (IL-4, IL-5, IL-13) and profibrotic mediators (IL-6, Col1a1) was significantly higher in Yama mice. However, the pronounced fibrotic changes, TGF-beta expression exhibited only a modest increase in Yama mice, suggesting that fibrosis might develop via pathways not totally dependent on conventional TGF-beta signaling. This repeated BLM administration model successfully reproduces the key pathological features of severe IPF and emphasizes the pivotal contribution of eosinophils to fibrogenesis.
Behavioral differences during and before/after dispersal were identified in two male raccoons (Procyon lotor) fitted using global positioning system (GPS) collars in Oita Prefecture, Japan. One male raccoon dispersed in a yearling spring, whereas the other dispersed in a yearling winter. The timing and direction of dispersal might reflect raccoon density in terms of differences in harvest pressure between municipalities in Oita. In both individuals, the average travel speed increased, and subsequently, the average travel distance per day rose during dispersal compared with periods before/after dispersal. During dispersal, forest use decreased, whereas artificial and agricultural land use increased in one individual, suggesting that this individual preferred anthropogenic areas for dispersal.
Pentosan polysulfate sodium (PPS) is a semi-synthetic sulfated polysaccharide with a heparin-like structure, known for its anticoagulant and anti-inflammatory properties. This study investigated the effect of PPS on adhesion molecules involved in neutrophil-endothelial interactions using an in vitro canine model. Canine aortic endothelial cells (CnAOEC) and neutrophils were pretreated with various concentrations of PPS and stimulated with recombinant canine tumor necrosis factor-alpha (rh-cTNF alpha). The mRNA expression of E-selectin, intercellular adhesion molecule-1 (ICAM-1), P-selectin, C-X-C motif chemokine receptor 2 (CXCR2), P-selectin glycoprotein ligand-1 (PSGL-1), and L-selectin was measured through real-time polymerase chain reaction (qPCR). C-X-C motif chemokine ligand 1 (CXCL1) secretion was quantified by enzyme-linked immunosorbent assay (ELISA). Lymphocyte function-associated antigen-1 (LFA-1/CD11a) expression was examined using flow cytometry. A static adhesion assay was used to evaluate the effect of PPS on neutrophil adhesion to endothelial cells. In conclusion, PPS displayed no cytotoxicity toward CnAOEC, and endothelial P-selectin expression was significantly reduced. Other adhesion molecules showed no significant differences compared with controls. CXCL1 secretion and neutrophil gene expression (CXCR2, PSGL-1, and L-selectin) varied slightly, although without statistical significance. PPS did not affect CD11a expression or neutrophil adhesion to endothelial cells. These results indicate that PPS is non-cytotoxic and may weakly modulate selectin-mediated endothelial activation without affecting integrin- or chemokine-dependent adhesion.
This study measured serum or plasma levels of reactive oxygen metabolites (d-ROMs), biological antioxidant potential (BAP), and oxidative stress index (OSI) in dogs. The subjects were classified into four groups according to sex and gonadal status: intact males, castrated males, intact females, and spayed females, both off-therapy and during therapy. No significant differences were observed in d-ROMs, BAP, or OSI between the off-therapy and during therapy groups. Among off-therapy dogs, intact males showed significantly higher mean d-ROMs and BAP values compared with females and neutered dogs. In contrast, among during therapy dogs, no significant differences were found in d-ROMs among the groups; however, intact males had significantly higher mean BAP values than castrated males, and the OSI was significantly higher in castrated males than in intact males. These results suggest that gonadectomy in male dogs may influence the balance between oxidative and antioxidative processes through factors such as sex hormones. The findings highlight the importance of considering the effects of spaying and neutering when establishing reference values for oxidative stress markers in dogs.
Bovine mastitis has a high morbidity rate and causes significant economic losses due to milk wastage and antimicrobial use. Mastitis is treated with antimicrobial agents, but criteria for disk diffusion tests for pathogens that cause bovine mastitis in Japan are lacking. In this study, we used a simplified agar disk diffusion (ADD) method that is easy to implement in clinical practice to develop breakpoints for bovine mastitis-associated organisms and generate a corresponding antibiogram. We established new breakpoints for a simplified ADD method for Staphylococcus aureus, coagulase-negative Staphylococcus (CNS), Streptococcus uberis, Streptococcus spp. (except for S. uberis), and Klebsiella spp. We also revalidated previously reported breakpoints against Streptococcus spp. and E. coli. The antibiogram showed that the susceptibility rates of Streptococcus spp. (excluding S. uberis) to marbofloxacin in eastern Japan were significantly lower than those in Hokkaido and western Japan (P < 0.01). The susceptibility rate of E. coli to oxytetracycline (OTC) in eastern Japan was significantly lower than that in Hokkaido (P < 0.01). Low susceptibility rates were observed for ampicillin (ABPC) and erythromycin among CNS strains, OTC among S. uberis strains, and cefazolin, cefuroxime, and ABPC among Klebsiella spp. strains. The use of breakpoints in the simplified ADD method in clinical practice should promote the prudent use of antimicrobial agents. In cases of acute mastitis with systemic symptoms in which antimicrobial agents must be administered prior to susceptibility testing, the antibiogram can be used as a criterion for determining the drug of choice.
Classical swine fever (CSF) is an important viral disease of pigs caused by classical swine fever virus (CSFV), with major impacts on animal health and the livestock industry. This study aimed to estimate the seroprevalence of CSFV antibodies in pigs in Bali, assess potential risk factors associated with selopositivity, and describe pathological changes in clinically suspected cases confirmed by rRT-PCR. From September to December 2023, 47blood samples, from pigs with possilable vaccination, racords were collected across Bali and tested using C-ELISA. In addition, 15 pigs showing CSF signs were necropsied for pathological and rRT-PCR evaluation. Six were confirmed CSFV-positive. The overall seroprevalence was 40.8% (192/470, 95% CI: 36.4-45.4). The highest prevalence occurred in Klungkung regency (65.4%, 17/26). Pigs from urban areas were 1.76 times more likely to be seropositive (OR: 1.76, 95% CI: 1.18-2.64) than those from rural areas. By age, pigs under two years were 1.21 times more likely to be seropositive (OR: 1.21, 95% CI: 0.65-2.23), while crossbred pigs were 1.27 times more likely (OR: 1.27, 95% CI: 0.85-1.88) compared to purebreds; however, these differences were not statistically significant. Only location (urban vs. rural) showed a significant association. Pathological changes were observed in the skin, brain, trachea, lungs, kidney, spleen, and intestines. This study demonstrates a relatively high seroprevalence of CSFV in Bali. The limited vaccination records suggests minimal vaccine coverage, indicating that the high seroprevalence most likely reflects natural infection.
As Johne's disease progresses, clumps of Mycobacterium avium subsp. paratuberculosis (MAP) are identified in the feces. MAP shows resistance to disinfectants, and effective disinfection methods for MAP cells excreted in feces have not been verified. Furthermore, given that biofilm-like vesicles and spore-like morphotypes were observed in feces from infected cattle, we aimed to compare reduction rates between cultured and naturally infected MAP cells after disinfectant treatment. Although the logarithmic reduction value for all cultured MAP cells was > 3.0, four of 10 infected MAP cells showed values < 2.0 after dichloroisocyanuric acid treatment. Our findings suggest that materials contaminated with infected cattle feces should be exposed to higher concentrations of disinfectants than those reported to be effective based on cultured cells
Using serological tools for bat-and shrew-borne hantaviruses, we investigated the seroprevalence of these viruses in 336 human sera from Sri Lanka, consisting of 168 healthy individuals and 168 febrile patients. Twelve and 31 sera were positive for Mix antigen-bat (recombinant N antigens of Brno, Dakrong, Quezon, Robina, and Xuan Son viruses) and Mix antigen-shrew (recombinant N antigens of Altai, Asama, Cao Bang, Nova, Seewis, and Thottapalayam viruses), respectively. 2 of 168 control sera and 39 of 168 febrile patient sera were positive for Mix antigen-bat or Mix antigen-shrew. Individual antigen reactions were positive for Quezon, Robina, Altai, Asama, Cao Bang, Seewis, and Thottapalayam viruses. These findings suggest that bat-and shrew-derived hantaviruses may infect humans in Sri Lanka.
African pygmy hedgehog adenovirus 1 (AhAdV-1) was isolated from a hedgehog exhibiting respiratory symptoms in Japan. In this study, we evaluated the cellular tropism of AhAdV-1 using various mammalian cell lines and investigated its genetic stability during long-term serial passage in MDCK cells. AhAdV-1 exhibited high replication efficiency in MDCK, MDBK, CPK, and Vero cells but showed limited growth in BHK, HeLa, and HC11 cells. No replication was detected in RAW 264.7 and 3LL cells. Since MDCK cells demonstrated the highest viral production, serial passage was performed up to 110 passages. Whole-genome analysis revealed only four mutations, suggesting that AhAdV-1 is a genetically stable virus. These findings contribute to the understanding of the host range and evolutionary stability of AhAdV-1.
Aichivirus D (AiV-D) is a newly emerging diarrhea-associated pathogen in cattle and sheep, and comprised two officially recognized genotypes (AiV-D1 and AiV-D2) and two potential novel genotypes (AiV-D3 and AiV-D4). This study aimed to establish a fluorescent RT-PCR assay for detecting AiV-D and to investigate its prevalence in yak diarrheic samples. A SYBR Green fluorescent RT-PCR assay was successfully established by targeting the conserved region of viral 3D genes, enabling detection of all known AiV-D genotypes. The established assay was specific for AiV-D, exhibiting no cross-reactivity with other common diarrhea-causing pathogens in ruminants, with coefficients of variation for reproducibility ranging from 0.23% to 2.16% and detection limits for the positive plasmids of AiV-D1 to AiV-D4 at 2.21, 4.78, 4.10, and 5.94 copies/mu L, respectively. Of the 166 diarrheic samples collected from calf yaks across nine farms in Sichuan province, China, between May and August 2023, 22.3% (37/166) tested positive for AiV-D, suggesting its substantial prevalence within the sampled region. Furthermore, seven complete 3D gene sequences were cloned from positive samples, and subsequent phylogenetic analysis revealed distinct evolutionary patterns among these isolates. In conclusion, this newly established assay represents a reliable diagnostic tool for AiV-D detection, while our findings provide significant insights into the epidemiological distribution and evolutionary dynamics of AiV-D in the yak population.
Bovine respiratory disease (BRD) significantly impacts the beef and dairy industries. The acute phase proteins procalcitonin (PCT) and haptoglobin (Hp) have been proposed as useful biomarkers for BRD. Here, we examined the time course of these two biomarkers in BRD-affected calves and their relationship to prognosis. Hp and PCT were elevated on the 1st day of examination (day 1). The PCT value on day 1 showed a positive correlation with the BRD score on day 7. Therefore, serum PCT concentration on day 1 may be used to predict whether symptoms are present on day 7. This study provides useful insights into the use of PCT and Hp in the diagnosis of BRD, particularly the use of PCT to determine prognosis.
The poultry industry is always at a risk due to high feed prices and frequent disease outbreaks. A global trend of using Antibiotic Growth Promoters (AGPs) for prophylaxis, improvement production, and enhanced growth have entered the poultry industry. Being banned in European countries, the AGPs are still extensively being used in South Asian's poultry industry which has higher bacterial infections as compared to the European countries. Zinc bacitracin, an AGP, at sub-therapeutic concentration has shown advantageous results in poultry. In this study zinc bacitracin was made in CASVAB University of Balochistan labs and applied for the trial. In this study birds were distributed in three groups, group 1 used feed with no feed additive, group 2 used feed with 20 mg zinc bacitracin/kg and group 3 used 40 mg zinc bacitracin/kg in feed as feed additive. Body weight gain was observed as 513.7 grams (g) as compare to control group of birds which was 424.3 (g), which is highly significant and significant with group 2. Final body weight was observed 1250.1 (g) in group 3 significant with group 2 observed 1200.0 (g) and highly significant with group 3 which was 1100.2 (g). Mortality rate in group 3 was observed 1.8 which is highly significant P <= 0.05 with group 3 and significant with group 2. We also observed significant result in group 3 as compare to group 2 and 1, regarding to blood leukogram, hemogram and various serum chemistry attributes.
In domestic cats, feline coronavirus (FCoV) infection is widespread, and its outcomes range from no signs to severe or potentially fatal disease. This study investigated the prevalence of infection and associated factors in a population of 277 owned cats in Thailand. The overall proportion of FCoV infections was 45.8%, with positive detections observed for all sample types. The positivity rates were 60.6% for rectal swabs, 51.3% for body fluids, and 11.1% for EDTA blood samples. Although the positive rate was slightly higher in males and more common in younger cats, neither difference was statistically significant. Mathematical modeling revealed a significant association between FCoV infection and the sampling period, with the highest positivity observed in April and June. Through S gene sequencing, positive samples from rectal swabs and body fluids [feline infectious peritonitis (FIP)-cats] were identified as FCoV genotype I. All rectal swab samples had a methionine codon at position 1058 of the S gene, and 62.5% of body fluid samples exhibited a notable substitution (M1058L). Additionally, analysis of the S1/S2 cleavage motif revealed that most FCoV-positive samples contained a polybasic R-R-S/A-R-R-S recognition motif. Notably, 37.5% of rectal swabs exhibited a mutation at the P1 residue (arginine to serine) and 14.3% of FIP-suspected cats exhibited a mutation at the P4 residue (arginine to glycine). These findings contribute to our understanding of the epidemiology of FCoV and its behavior in infected cats. Such insights are crucial for developing effective diagnostics, understanding the FCoV transmission dynamics, and informing potential control strategies.
A 3-year-old castrated male cat was presented with lethargy, anorexia, and melena. A complete blood count revealed regenerative anemia. Abdominal ultrasonography revealed a layer-disrupted intestinal mass at the ileocecal junction. Fine needle aspiration performed on the mass revealed large lymphocytes, suggesting lymphoma. The patient was treated with CHOP-based chemotherapy but admitted for emergency care with vomiting and depression during the CHOP protocol. Abdominal ultrasonography suggested annular intestinal stricture at the site of lymphoma. The patient underwent ileocecectomy, which resolved the small intestinal obstruction. Histopathologic examination of the resected specimen demonstrated transmural enterotyphilitis with prominent fibrosis. The patient remained stable without relapse. To our knowledge, this is the first case of intestinal stricture occurring during chemotherapy for feline high-grade intestinal lymphoma.
The present study was designed to investigate the oral pharmacokinetics and bioavailability of roxithromycin, a macrolide class antimicrobial drug, in broiler chickens at a single dose of 20 mg/kg body weight. Plasma concentrations of roxithromycin were measured by High Performance Liquid Chromatography (HPLC) method using UV (ultraviolet) detector. The pharmacokinetic (PK) parameters were calculated from plasma concentrations versus time data using 'PK Solver 2.0' software. Following single intravenous administration, the volume of distribution at steady state (Vss) and total body clearance (Cl) were 3.04 L/kg and 0.45 L/hr/kg, respectively. Following oral administration of roxithromycin at the dose rate of 20 mg/kg body weight in broilers, a characteristic short lag phase of 15 minutes in absorption was observed and thereafter mean maximal plasma concentration (Cmax: 3.60 mu g/ml) was achieved at 2 hr. The mean values of elimination half-life (t1/2 beta) and mean resident time (MRT) were 8.30 and 9.57 hr, respectively. Calculated oral bioavailability (F) ranged from 51.95 to 62.57% with a mean value of 56.86%. Based on pharmacokinetic-pharmacodynamic (PK-PD) integration efficacy predictor, oral dose at 20 mg/kg body weight of roxithromycin twice a day, is predicted to be effective for treating the susceptible bacterial infections in the broiler chickens with a MIC value <= 1.0 mu g/ml.
A 5-year-old spayed female domestic short haired cat presented with chronic nasal obstruction. The cat was diagnosed with B-cell nasal lymphoma based on histological examination of biopsy specimens and intracranial invasion was suspected. We initiated treatment with nimustine (ACNU), known to pass the blood-brain barrier, as the primary protocol with prednisolone, with the owner's consent. The ACNU dose was 20-25 mg/m2 (4-5-week intervals, four or six administrations, two courses). The cat remained oligosymptomatic until day 962 after the first dose and no serious adverse events were reported. The cat lived until day 1089. To our knowledge, this is the first report of long-term survival achieved using ACNU as the primary treatment in a cat with nasal lymphoma.
Oligoadenylate synthetase (OAS) plays a role in intracellular viral defense. Mouse Oas1b is responsible for the susceptibility to flaviviruses. The flavivirus replication inhibitory effect of Oas1b is independent of its enzymatic activity; however, its mechanism remains unclear, though mouse Oas1b possibly targets the 3 '-stem-loops (SLs). We measured the effect of Oas1b by removing and overexpressing SLs in the 3 '-untranslated regions (UTRs) of the West Nile virus (WNV) replicon. Both SL2 deletion and SL2 RNA overexpression in the 3 '-UTR of the WNV replicon resulted in the loss of Oas1b-mediated replication inhibition. These results revealed that WNV SL2 is essential for Oas1b-mediated replication inhibition.
A new transvaginal artificial insemination (AI) procedure by frozen-thawed semen using a Foley catheter in ewes was performed in the present study. A 30-ml ballon was fixed in the vagina and frozenthawed semen expelled into the vagina was retained near the cervical os for approximately 30-60 min. In the first trial, a single AI was performed on eight ewes after estrus synchronization and one ewe became pregnant (12.5%). In the second trial, AI was performed for two consecutive days on the seven ewes that did not become pregnant in the first trial. Three of the seven (42.9%) ewes become pregnant. These results suggest the usefulness of transvaginal AI by frozen-thawed semen using a Foley catheter for two consecutive days.
Present study was done to delineate the immune-regulated interplay following the toxicological deterring effect of ethion. The study was conducted in chicks and they were equally and randomly divided into four groups (n=6) with group I served as control, group II, III and IV were administered with ethion at 2.5, 5.0 and 7.5 mg/kg body weight per os route respectively for 30 days. On 30th day the blood and tissue samples were collected to assess the immunotoxic potential of ethion. The study revealed significant (P < 0.05) reduction in total protein, albumin and globulin, splenic cell cellularity count, lymphocytic count whereas significant (P < 0.05) increase was observed in dose dependent manner on apoptotic cell count via FACS. The non significant changes were observed in HI titre, DTH response and lymphocyte proliferation assay following 30 days administration of toxicant to chicks. Histopathological examination of spleen and skin tissue revealed more pronounced alteration in the normalcy of spleen and skin in group III and IV as compared to control. The study therefore concluded that ethion intoxication in poultry birds produced significant (P < 0.05) apoptosis in splenocytes, heterophilia and lymphocytopenia as a part of defensive mechanism albeit at given experimental doses direct immunotoxic effect was not advocated. Further downstream apoptotic signaling studies are warranted.
Rectal swabs containing Salmonella enterica may be exposed to desiccation stress while awaiting laboratory isolation. Growth of injured S. enterica is inhibited on selective media but the extent to which this varies across different media remains unclear. After storage at 25 degrees C, we compared growth recovery using four enrichment media and revealed that log 2.7 CFU and > log 4.0 CFU was required to confirm the growth of monophasic S. enterica Typhimurium K13-4-49 in buffered peptone water and selective enrichment media (Hajna tetrathionate broth, tetrathionate broth, and modified semisolid Rappaport agar for stab culture), respectively. Since < log 1.7 CFU was required for all four media, we recommend sample storage at <= 4 degrees C or the application of transport medium.
