The improvement of veterinary care has prolonged the lifespan of rabbits, and the number of rabbits suffering from age-related, male genital disorders may increase in the near future. This could result in increased opportunities for male genital surgery, requiring knowledge of their arterial anatomy, which, however, has not been sufficiently studied. Therefore, the arteries supplying the genitals were observed in 20 male New Zealand White rabbits. The testis was supplied by the testicular artery originating from the abdominal aorta. The right testicular artery usually emerged at a more cranial level than the left artery (65%). The testicular artery encircled the testis in the sagittal plane and bifurcated (95%) or trifurcated (5%) at the caudal extremity of the testis before entering the parenchyma. The epididymis was supplied by the epididymal branches, either from only the testicular artery (75% of the right and 80% of the left halves) or from both the testicular artery and aorta. The deferent duct was supplied in all halves by the dorsal and ventral branches of the deferential artery, which usually arose from the umbilical artery. The accessory genital glands were supplied by the dorsal branch of the deferential artery and the prostatic artery. The latter, which emerged from the internal iliac artery, exhibited 3 branching types. The most frequent type (55% of the right and 45% of the left halves) had 3 branches supplying the accessory genital glands. These findings will help improve rabbit genital surgery.

Four concentrations of potassium peroxymonosulfate (PPMS) were evaluated for bactericidal activities and indicated that the concentration is less than the manufacturing-recommended concentrations, must extend the exposure time for bacterial inactivation. However, even with and without of organic material contamination, did not show marked inactivation difference. In addition, all concentrations were inactivated on all carrier surfaces within 30 sec, except on rubber where inactivation occurred within 1 min. However, quaternary ammonium compounds were inactivated on stainless steel and plastic within 1 min and 30 sec, respectively, but not inactivated within 5 min on rubber surfaces. Conclusion, PPMS inactivated bacteria under optimal concentration, organic material conditions, exposure timing and on carrier surfaces which can useful as an alternative disinfectant for biosecurity enhancement.

Paenibacillus larvae and Melissococcus plutonius are bacterial pathogens of honey bee brood. As decontamination of beekeeping equipment, including combs, is essential to control these pathogens, we evaluated the disinfecting effects of slightly acidic hypochlorous acid water (SAHAW) and weakly acidified chlorous acid water (WACAW) on the pathogens. Both disinfectants exhibited strong disinfecting effects in suspension tests under no organic matter conditions and reduced both pathogens by >5 log₁₀ CFU/ml. Although the microbicidal activity of SAHAW with an available chlorine concentration (ACC) of 10–30 ppm was decreased by organic matter, it reduced viable P. larvae spores in combs more efficiently than H₂O when the comb was not as dirty. However, its efficacy on combs decreased at 4°C and when overused or highly contaminated combs were tested. WACAW with an ACC of ≥600 ppm had a higher disinfecting capacity than SAHAW, and efficiently removed P. larvae spores from combs even under organic matter-rich and low-temperature conditions. However, even by WACAW, the amount of viable spores in combs was not markedly reduced depending on contamination levels and P. larvae genotypes. These results suggest the usefulness of both disinfectants for decontaminating beekeeping equipment depending on the situations expected.

Among domestic animals, melioidosis is one of the most common diseases reported in goat, sheep, and swine. To evaluate the specific antibodies in goats with melioidosis, we developed a serology test using recombinant outer membrane protein A (OmpA) and flagellin (FliC) of Burkholderia pseudomallei as antigens. DNA corresponding to each antigen was cloned into a pET32a vector and expressed in Escherichia coli. Essentially, the recombinant OmpA and FliC were expressed in a soluble form that could be isolated with 95% homogeneity. Both recombinants could be recognized by rabbit antibodies prepared against heat-inactivated B. pseudomallei (1:1,000) on a Western blot. Subsequently, we demonstrated that both recombinants could capture the antibodies present in goat with naturally occurring melioidosis (optimized titer 1:40) while not cross-reacting with the serum samples of goats naturally infected by Corynebacterium pseudotuberculosis or Staphylococcus aureus. Finally, an enzyme-linked immunosorbent assay (ELISA) using 20 goat serum samples without melioidosis and 10 goat serum samples with melioidosis demonstrated that the infected group has significantly higher antibody titer levels than the normal group (P<0.001) when using either OmpA or FliC as an antigen. However, the sensitivity (100%) of the assay using OmpA was superior to that (90%) from using FliC. Serological tests that are commonly used often rely on antigens from crude cell extracts, which pose risks for laboratory-acquired infections and inconsistency in their preparation; however, use of recombinant OmpA is safe; it can potentially be used as a reagent in testing for goat melioidosis.

The aim of this study was to determine the susceptibility of Lactobacillus bacteria to selected coccidiostats. Seventy-five Lactobacillus isolates obtained from chickens were classified by MALDI-TOF mass spectrometry and 16S rDNA restriction analysis into seven species, among which L. salivarius (33%) and L. johnsonii (24%) were dominant. Susceptibility of lactobacilli to coccidiostats was determined by broth microdilution method. The ranges of minimum inhibitory concentrations (MICs) were 0.5–≥128 µg/ml for monensin, 0.125–8 µg/ml for salinomycin, ≤0.03–2 µg/ml for lasalocid A, and 4–16 µg/ml for robenidine. Coccidiostats in low concentrations inhibited in vitro growth of most lactobacilli and therefore there is a high probability that administration of this drugs to chickens would reduce the number of lactobacilli in the gut.

Undesirable behaviors exhibited by cats are regarded as feline behavioral problems. Like canine behavioral problems, feline behavioral problems are commonly reported by owners. Thus, in order to advise cat owners appropriately, veterinarians, particularly general veterinary practitioners, are encouraged to have sufficient knowledge of these problems. However, in ordinary Japanese homes, only a small number of feline behavioral problems have been investigated in terms of prevalence and relevant factors. Comprehensive survey data may improve understanding of feline behavioral problems. In the present study, we distributed an online survey including general information and questions about 17 feline behaviors which can be problematic for owners, to investigate: (1) the prevalence of feline behavioral problems, (2) the most common behavioral problems, and (3) relevant factors for each behavior. We collected 1,376 valid responses. Among all cats, 75.7% exhibited at least one behavioral problem. “Pica” and “Showing fear while on the examination table” were the most frequently reported behavioral problems. To identify factors related to the expression of each behavior, we conducted multiple logistic regression analyses, with the presence of behaviors as objective variables and cats’ general information as explanatory variables. Age, sex, breed, number of cats in the household, and type of residence were each associated with more than one behavior. The present study revealed the prevalence of 17 feline behavioral problems in Japanese homes and relevant factors of each behavior. These findings will help veterinarians understand and address feline behavioral problems.

Fluid responsiveness, defined as the response of stroke volume to fluid loading, is a tool to individualize fluid administration in order to avoid the deleterious effects of hypovolemia or hypervolemia in hospitalized patients. To evaluate the accuracy of two ultrasound indices, the caudal vena cava to abdominal aorta ratio (CVC/Ao) and the respiratory collapsibility of the caudal vena cava (cCVC), as independent predictors of fluid responsiveness in a heterogeneous population of spontaneously breathing, conscious, hospitalized dogs. A prospective, multicenter, observational, cross-sectional study was designed in twenty-five dogs. The accuracy of CVC/Ao and cCVC in predicting fluid responsiveness was evaluated by the area under the receiver operating characteristic curve (AUROC) in a group of hospitalized dogs after receiving a mini-fluid bolus of 4 ml/kg of Hartmann’s solution. Dogs with an increased aortic velocity time integral >15% were classified as fluid responders. Twenty-two dogs were finally included. Ten were classified as responders and 12 as non-responders. The AUROC curves were 0.88 for the CVC/Ao ratio (95% confidence interval, CI, 0.67–0.98; P=0.0001) and 0.54 for cCVC (95% CI 0.32–0.75; P=0.75). The CVC/Ao threshold optimized for best sensitivity (SE) and specificity (SP) values was 0.83 (SE 100%; SP 75%). In spontaneously breathing hospitalized dogs, the CVC/Ao measurement predicted stroke volume increase after a fluid bolus, while the respiratory variations in the cCVC did not discriminate between fluid responders and non-responders.

In dogs, reports on thoracic lymph nodes are lacking compared to abdominal lymph nodes. This report analyzed the position, number, size, shape, and homogeneity of thoracic lymph nodes (cranial sternal, cranial mediastinal, tracheobronchial, aortic thoracic, and pulmonary lymph nodes) using thoracic computed tomography (CT) images of 100 dogs without any lesions in the dominated areas of thoracic lymph nodes. The position and number of intrathoracic lymph nodes could be observed in CT, consistent with macroscopic anatomical studies. It was difficult to set a clinical index associated with size using CT scans. Image findings that indicated abnormalities, such as circular shapes and non-uniform, may be routinely found in dogs and may not be considered abnormal on CT scans.

Ferric citrate is an oral iron-based phosphate binder, being known to affect iron status and improve iron deficiency anemia (IDA) in chronic kidney disease (CKD) patients. We examined whether oral administration of ferric citrate could change iron status and improve anemia without affecting phosphorus metabolism in iron deficiency anemia rats. In Normal rat study, normal rats were fed a diet containing 0.3 or 3% ferric citrate for 11 days for setting the dose and administration period of ferric citrate. The effects of ferric citrate on iron status- and phosphorus metabolism-related parameters were evaluated using blood and urine samples. Next, an iron deficiency anemia was induced by feeding iron-depleted diet in rats. After 7 days of starting the iron-depleted diet, 0.3% ferric citrate was administered for 7 days by dietary admixture. Iron status- and phosphorus metabolism-related parameters were evaluated with blood and urine samples. In Normal rat study, 3% ferric citrate treatment increased serum iron level and transferrin saturation (TSAT), and decreased serum phosphorus level, intact fibroblast growth factor 23 (iFGF23) level, and urinary phosphorus excretion, but 0.3% ferric citrate treatment showed no effects. On the other hand, in Iron deficiency anemia rat study, 0.3% ferric citrate treatment increased iron status-related parameters and improved anemia, but did not show any apparent changes in phosphorus metabolism-related parameters. In conclusion, ferric citrate could have hematopoietic effects without affecting phosphorus metabolism, and could be a potential option for the treatment of IDA in patients without CKD.

In Sudan, donkeys are important animals, providing transportation and income possibilities. However, the prevalence of parasites in donkeys in Sudan has not been thoroughly characterized. Accordingly, in this study, we aimed to detect selected hemoprotozoan parasites in donkeys in West Omdurman, Khartoum State, Sudan, wherein people depend mainly on donkeys for their daily life. In total, 198 blood samples collected from donkeys in a local market in West Omdurman, were screened using serological and molecular diagnostic techniques. Serologically, 52 (26.3%), 56 (28.3%), and 19 (9.6%) samples were positive for trypanosomosis using Card Agglutination Test for Trypanosoma evansi, Trypanosoma evansi crude antigen -based enzyme-linked immuno sorbent assay (ELISA) and recombinant Trypanosoma evansi GM6-4r-based ELISA, respectively. ELISA for equine piroplasmosis revealed 156 (78.8%) and 10 (5.1%)Theileria equi- and Babesia caballi-positive samples, respectively. PCR detected Trypanosoma congolense, subgenus Trypanozoon, Theileria equi, and Babesia caballi in 18 (9.1%), 77 (38.9%), 18 (9.1%), and 8 (4%) samples, respectively. Of the 77 Trypanozoon-positive samples, 35 (45.5%) were confirmed as Trypanosoma evansi type A. To our knowledge, this is the first report of detection of Trypanosoma congolense in donkeys outside of tsetse-infested areas in Sudan.

In chicken, peritoneal cystic lesions have not been clearly categorized. In this study, diffuse peritoneal multiple cysts were observed in two layer hens. The cysts in the serosa were lined with single layers of squamous or cuboidal cells. The papillary proliferations of columnar cells were also observed in one case. The smooth muscle layer or mass were observed around the cysts in both cases. The cystic lining cells were positive for pan-cytokeratin, vimentin, S100 and Wilms tumor 1. Ultrastructurally, they had sparsely microvilli on the luminal surface. The histological results indicated the present cases were multicystic mesothelioma, but also had characteristics of Mullerian epithelium. This is the first report describing the detailed pathological feature of unique multicystic tumor in chicken.

Small extracellular vesicles (sEV) contain various molecules and mediate cell-to-cell communication under both physiological and pathological conditions. We have recently reported that sEV isolated from plasma of normotensive Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) regulate systemic blood pressure. The initiation and development of hypertension partly rely on proliferation and migration of vascular smooth muscle cells (SMCs) followed by the structural remodeling of vascular wall. In the present study, we examined the effects of plasma sEV in WKY and SHR on the proliferative and migratory functions of primary rat aortic SMCs. There was no difference in the concentration and size distribution of plasma sEV between WKY and SHR, while the protein expression of CD81 in plasma sEV from SHR was lower than that from WKY. Both plasma sEV from WKY and SHR were internalized into SMCs and stimulated the migration and proliferation with a similar potency. In summary, we, for the first time, demonstrated that plasma sEV in WKY and SHR are physiologically active in terms of proliferative and migratory functions, however, these effects do not seem to be related to the pathogenesis of hypertension development.

D-Amino acids exert various physiological functions and are widely present in animals. However, they are absorbed to a lesser extent than L-amino acids. Little is known about D-arginine (D-Arg); however, its isomer L-Arg serves as a substrate for several metabolites and exhibits various functions including promotion of growth hormone secretion. Milk is the only nutrient source for infants; it plays an important role during their initial growth and brain development. No studies have evaluated the availability of D-Arg in the brain and milk in mammals. Here, we have studied the differential availability of orally administered D- and L-Arg in the brain and milk using ICR mice. Our results revealed that without D-Arg administration, D-Arg was undetectable in both plasma and brain samples. However, the plasma D-Arg was about twice the concentration of L-Arg post administration of the same. In the cerebral cortex and hypothalamus, L-Arg concentration remained almost constant for over period of 90 min after L-Arg treatment. Nevertheless, the L-Arg concentration decreased after D-Arg administration with time compared to the case post L-Arg administration. Contrastingly, D-Arg level sharply increased at both the brain regions with time after D-Arg treatment. Furthermore, L-Arg concentration in the milk hardly increased after L-Arg administration. Interestingly, oral administration of D-Arg showed efficient enrichment of D-Arg in milk, compared with L-Arg. Thus, our results imply that D-Arg may be available for brain development and infant nourishment through milk as an oral drug and/or nutrient supplement.

The emergence and prevalence of antimicrobial-resistant bacteria in wild animals are a great concern for public health. A total of 963 Escherichia coli isolates from 475 wild mammals (242 sika deers, 112 wild boars, 113 small mammals, 4 Japanese badger, 2 Tokara cows, and 2 Amani rabbits), collected between 2013 and 2017, were examined for antimicrobial susceptibility. Resistance to at least one antimicrobial was observed in 92 of 963 isolates (9.3%). No isolates exhibited resistance to carbapenem (meropenem). Resistance to third-generation cephalosporin (cefotaxime) and fluoroquinolone (ciprofloxacin) was observed in less than 1% of the isolates. Thus, low prevalence of bacterial antimicrobial resistance was observed in wild mammals between 2013 and 2017 in Japan.

Dinotefuran (DIN) belongs to the neonicotinoids (NNs), a class of globally applied pesticides originally developed to exhibit selective toxicity in insects. However, several reports have suggested that NNs also exert neurotoxic effects in mammals. We previously demonstrated neurobehavioral effects of DIN on mice under non-stressful conditions. For further toxicity assessments in the present study, we investigated the effects of DIN on mice exposed to stressful conditions. After subacutely administering a no-observed-effect-level (NOEL) dose of DIN and/or chronic unpredictable mild stress (CUMS) to mice, we conducted three behavioral tests (i.e., open field test [OFT], tail suspension test [TST] and forced swimming test [FST]). In addition, serotonin (5-HT) and tryptophan hydroxylase 2 (TPH2) of the dorsal raphe nuclei (DRN) and median raphe nuclei (MRN) and tyrosine hydroxylase (TH) of the ventral tegmental area and substantia nigra (SN) were evaluated immunohistochemically. A NOEL dose of DIN or CUMS alone increased of the total distance in OFT, decreased or increased the immobility time in TST or FST, respectively, and increased the positive intensity of 5-HT and TPH2 in the DRN/MRN, and TH in the SN. These changes were suppressed under the conditions of combined exposure to DIN and CUMS, though the blood corticosterone level was increased depending on the blood DIN values and the presence of CUMS. The present study suggests the multifaceted toxicity of the neurotoxin DIN.

Neonicotinoid pesticides (NNs) act as agonists on nicotinic acetylcholine receptors (nAChRs) of insects, and there have been concerns about the effects of NNs on the health of mammals. Since nAChRs are expressed in immune cells, it is possible that NNs disturb the immune system. However, few reports have examined the immunotoxicity of clothianidin (CLO), a widely-used NN. Here, we report the effects of CLO on immune organs and type IV allergic reactions in ear auricles. We orally administered CLO at 0, 30 and 300 mg/kg/day (CLO-0, 30 and 300) to Sprague-Dawley rats for 28 days. The effects were evaluated by organ and body weights, histopathology, and immunohistochemistry (TCRαβ, CD4, CD8, CD11b, CD68, CD103). In addition, some cecal contents were subjected to preliminary gut microbiota analysis, because microbiota contribute to host homeostasis, including the immunity. Our results showed loose stool, suppression of body weight gain, significant changes in organ weights (thymus: decreased; liver: increased) and changes of the gut microbiota in the CLO-300 group. There were no obvious histopathological changes in immune organs. Granulomas of the ear auricles were found in one rat of each of the CLO-30 and 300 groups, but CLO had no apparent effect on the thickness or immunohistochemistry in the ear auricles. We present new evidence that CLO affects the thymus and intestine, and might enhance the local inflammatory response. These findings should contribute to the appropriate evaluation of the safety of NNs in the future.

It can be judged that if the detection frequency of prevalent pathogenic viruses decreases, biosecurity has been enhanced. To monitor bovine farm biosecurity levels, one-step multiplex reverse transcription polymerase chain reaction (RT-PCR) for the simultaneous detection of group A rotavirus (RVA), bovine torovirus (BToV), bovine enterovirus (BEV), and bovine coronavirus (BCV) was designed, with the aim of configuring candidates for “viral pathogen indicators”. A total of 322 bovine fecal samples were collected from calves aged less than three months at 48 bovine farms in Ibaraki and Chiba prefectures. At farm A, 20 calves were selected and sampled weekly for 12 weeks (184 samples); at farm B, 10 calves were selected and sampled for five weeks (50 samples); and at the rest of the 46 farms, 88 calves were sampled once. The screening on the 358 field samples proved positive for 27 RVA, 4 BToV, 55 BEV, and 52 BCV. In the successive sampling, RVA was detected once but not continuously, whereas BEV and BCV were detected in succession for up to five weeks. The results revealed that RVA was the primary agent among the positive samples obtained from calves aged three weeks or less, while BEV was the primary among those from the older than three weeks old. They can be employed as useful viral pathogen indicators for soundly evaluating biosecurity at bovine farms.

Adenosine 5’-triphosphate (ATP), the major energy currency of the cell, is involved in many cellular processes, including the viral life cycle, and can be used as an indicator of early signs of cytopathic effect (CPE). In this study, we demonstrated that CPE can be analyzed using an FRET-based ATP probe named ATP indicator based on Epsilon subunit for Analytical Measurements (ATeam). The results revealed that as early as 3 hr, the virus infected cells showed a significantly different Venus/cyan fluorescent protein (CFP) ratio compared to the mock-infected cells. The ATeam technology is therefore useful to determine the early signs of ATP-based CPE as early as 3 hr without morphology-based CPE by light microscopy, and enables high throughput determination of the presence of microorganisms in neglected samples stored in laboratories.

Infectious pancreatic necrosis virus (IPNV) causes economic losses in Mexican rainbow trout industry. In this study, virulence and genetic fingerprints of Mexican IPNV isolates was investigated for the first time. Two Mexican IPNV isolates were analyzed in rainbow trout fry and the Sp strain was included as high virulence. One of the Mexican IPNV isolate was obtained from diseased fish and the other from fish without clinical signs. The infection was performed using a standardized immersion. Clinical signs were observed at 4 days post infection in fry group infected with strain Sp, two days earlier than in trout infected with IPNV isolates Mexican. Severe lesions were found in 100% of the individuals of Sp group, but only in 25% of each isolated Mexican group. Results suggest that Mexican IPNV isolates are pathogenic, but less virulent than strain Sp. The amino acid motif residues of both Mexican isolates, corresponded to a subclinical disease. Nevertheless, the accumulated motility observed in the field, suggest that other factors play a role in the virulence of the disease.

Autopsy imaging (Ai) was performed for a King Penguin. Ai-computed tomography (CT) revealed air sac membrane thickening, multiple nodules in the cranial air sac, suspected abscess, lung infiltration, and air sac contraction. Based on these findings, respiratory disorder was concerned. Aspergillosis, which is the highly observed in penguins, was considered as the primary differential diagnosis. The cultured sample showed characteristic conidial head of Aspergillus spp., the DNA of which was 100% identical to that of A. fumigatus. The cause of death was determined to respiratory failure due to aspergillosis. Ai-CT findings facilitated the dissection workflow and alerted the pathologist to potential hazards during the autopsy. Ai is useful to determine the cause of death and for readiness and safe pathological dissection.

Osteoporosis is a skeletal disease that occurs in many mammals. Our report describes osteoporosis in an Asian small-clawed otter (Aonyx cinereus). Gross, histological, and radiographic observations showed that all of the bones had numerous pockmarks on their surfaces. Histologically, the pockmarks were filled with fibrous tissue without inflammation. However, the spongy bone was normal according to the histological and radiographic results. Overall, the results showed that this was a case of osteoporosis that mainly involved external rather than internal surfaces.