Salmonella enterica serovar Kentucky recovered from human clinical cases in Maryland,USA (2011–2015)

Salmonella Kentucky is among the most frequently isolated S. enterica serovars from food animals in the United States. Recent research on isolates recovered from these animals suggests there may be geographic and host specificity signatures associated with S. Kentucky strains. However, the sources and genomic features of human clinical S. Kentucky isolated in the United States remain poorly described. To investigate the characteristics of clinical S. Kentucky and the possible sources of these infections, the genomes of all S. Kentucky isolates recovered from human clinical cases in the State of Maryland between 2011 and 2015 (n = 12) were sequenced and compared to a database of 525 previously sequenced S. Kentucky genomes representing 12 sequence types (ST) collected from multiple sources on several continents. Of the 12 human clinical S. Kentucky isolates from Maryland, nine were ST198, two were ST152, and one was ST314. Forty‐one per cent of isolates were recovered from patients reporting recent international travel and 58% of isolates encoded genomic characteristics similar to those originating outside of the United States. Of the five isolates not associated with international travel, three encoded antibiotic resistance genes conferring resistance to tetracycline or aminoglycosides, while two others only encoded the cryptic aac(6′)‐Iaa gene. Five isolates recovered from individuals with international travel histories (ST198) and two for which travel was not recorded (ST198) encoded genes conferring resistance to between 4 and 7 classes of antibiotics. Seven ST198 genomes encoded the Salmonella Genomic Island 1 and substitutions in the gyrA and parC genes known to confer resistance to ciprofloxacin. Case report data on food consumption and travel were, for the most part, consistent with the inferred S. Kentucky phylogeny. Results of this study indicate that the majority of S. Kentucky infections in Maryland are caused by ST198 which may originate outside of North America.     

Kaempferol alleviates the reduction of developmental competence during aging of porcine oocytes

Kaempferol (KAE) is a natural flavonoid present in different plant species and exhibits anti‐inflammatory, antioxidant, and anticancer therapeutic properties. In the present study, we investigated the influence and underlying mechanisms of KAE supplementation on porcine oocytes during in vitro aging. The results show that KAE treatment can alleviate the aging‐related reduction of developmental competence. We observed that the blastocyst production rate in aged oocytes treated with 0.1 μM KAE was significantly higher than in untreated aging oocytes (36.78 ± 0.86% vs. 27.55 ± 2.60%, respectively, p < .05). The KAE‐treated aging oocytes had significantly reduced levels of reactive oxygen species (p < .05). Furthermore, the mRNA levels of the embryonic pluripotency‐related genes Oct4, NANOG, and ITGA5 were significantly increased in blastocysts derived from KAE‐treated oocytes (p < .05). During excessive oocyte culture, KAE treatment maintained the mitochondrial membrane potential and reduced apoptosis; however, this was not observed in untreated aging oocytes. In conclusion, our results suggest that KAE treatment can alleviate the aging of porcine oocytes by reducing oxidative stress and improving mitochondrial function.     

Micro-computed tomography evaluation and pathological analyses of female rats with collagen-induced arthritis

Imaging techniques have been introduced to assess the efficacy and toxicity of developing pharmaceuticals. The purpose of this study was to perform a comprehensive characterization of collagen-induced arthritis (CIA) in rats using micro-computed tomography (micro-CT) and to compare the results with data from conventional pathological examination. Arthritis was induced by collagen in 24 female Wistar rats. Micro-CT and pathological analyses were performed to assess arthritis progression. Micro-CT analysis showed marked joint destruction occurring in a time-dependent manner following collagen administration. Bone volume was significantly decreased in the tibia at weeks 3 and 4 compared to week 0 (p < 0.05 and p < 0.01, respectively). Additionally, percent bone volume was significantly reduced in the tibia at week 4 compared to week 0 (p < 0.05). In contrast, bone surface/bone volume and trabecular separation were significantly increased in the tibia of the animals at week 4 compared to week 0 (p < 0.05). Severe joint destruction with extensive inflammation, erosion of cartilage and bone, and infiltration of inflammatory cells were observed in the knee joints of the collagen-treated rats. Taken together, micro-CT made it possible to quantify CIA lesions and should be performed with pathological examination in rats.     

Cytotoxicity of gamma-ray in rat immature hippocampal neurons

This in vitro study evaluated the detrimental effect of acute gamma (γ)-irradiation on rat immature hippocampal neurons. Rat immature hippocampal neurons (0.5 day in vitro) were irradiated with 0~4 Gy γ-rays. Cytotoxicity was analyzed using a lactate dehydrogenase release assay at 24 h after γ-irradiation. Radiation-induced cytotoxicity in immature hippocampal neurons increased in a dose-dependent manner. Pre-treatments of pro-apoptotic caspase inhibitors and anti-oxidative substances significantly blocked γ-irradiation-induced cytotoxicity in immature hippocampal neurons. The results suggest that the caspase-dependent cytotoxicity of γ-rays in immature hippocampal cultured neurons may be caused by oxidative stress.     

Immunohistochemical and histomorphometric evaluation of vascular distribution in intact canine cranial cruciate ligament

Objectives: To (1) describe vascular distribution in the grossly intact canine cranial cruciate ligament (CCL) using immunohistochemical techniques specific to 2 components of blood vessels (factor VIII for endothelial cells, laminin for basement membrane); and (2) compare the vascularity in different areas of interest (craniomedial versus caudolateral bands; core versus epiligamentous regions; and proximal versus middle versus distal portions) in the intact normal canine CCL. Study Design: In vitro study. Animals: Large, mature dogs (n=7) of breeds prone to CCL disease that were euthanatized for nonorthopedic conditions. Methods: Intact CCL were collected from fresh canine cadavers free from stifle pathology. CCL tissue was processed for immunohistochemistry and stained for factor VIII and laminin. Vascular density was determined by histomorphometric analysis. Results: Specific vascular staining was sparsely identified throughout the CCL; however, the proximal portion of the CCL appears to have a greater number of vessels than the middle or distal portion of the ligament. Conclusions: The CCL is a hypovascular tissue and its vascular distribution is not homogeneous.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

Kinematic gait analysis of the hind limb after tibial plateau levelling osteotomy and cranial tibial wedge osteotomy in ten dogs

This study identifies and compares the kinematic gait changes occurring in tibial plateau levelling osteotomy (TPLO) and cranial tibial wedge osteotomy (CTWO) limbs after transection of the cranial cruciate ligament (CrCL). Ten, healthy, adult beagle dogs were assigned to TPLO (five dogs) and CTWO (five dogs) groups. Hind limb kinematics were assessed, while dogs were trotted at speeds ranging from 2.0 to 2.3 m/s. The animals were evaluated preoperatively (prior to TPLO and CTWO surgery) and at both 8 and 12 weeks after surgery. Two-dimensional evaluation was synchronized to obtain the three-dimensional coordinates using the APAS motion analysis software. Gait patterns were assessed by measuring stifle, tibiotarsal joint angles and stifle joints angular velocity. Stifle and tibiotarsal joint functions were not affected by TPLO surgery, but stifle and tibiotarsal joint angles were changed, following CTWO surgery, compared with their preoperative values. The angular velocity patterns of CTWO were characterized by increased stifle joint extension velocity from the middle to end swing phase and decrease in the peak velocities (flexion) during swing phase. None of these changes was observed in the stance phase after the CTWO surgery. These kinematic results showed that dogs that underwent a CTWO procedure were more likely to have significantly hyperextended gait patterns of the swing phase postoperatively than the dogs that had a TPLO procedure for repair of a ruptured CrCL.     

Influence of equalizing the gross composition of milk replacer to that of whole milk on the performance of Holstein calves

This study compared the performance of female Holstein calves fed either whole milk (WM) or milk replacer (MR) having similar gross composition to WM. Calves (n = 20) were separated from their mothers within 2 h of birth, weighed, moved into individual pens, and fed colostrum (10% of their BW over 2 feedings daily) for the first 3 d. Calves were alternately assigned to either WM (n = 10) or MR (n = 10) and were fed using mobile plastic bottles. At each feeding, diluted MR was prepared in buckets by mixing 0.143 kg of MR powder (as-is basis) in 1 L of lukewarm water. Calves on both treatments were fed (1.8 L/feeding) for 4 times daily for the first 25 d of age. Feeding frequency was reduced to 3 times daily for next 5 d, then to 2 times daily for next 14 d, and then to once daily during the last 5 d of the preweaning period. Feed intake, growth, and health variables were monitored until calves were 70 d of age. Mean daily consumption of WM and MR was similar in both treatment groups (P = 0.74). Initial BW of calves fed either WM or MR was similar (42.1 +/- 3.3 vs. 41.9 +/- 2.9 kg, respectively; P = 0.83). Calves fed WM or MR daily consumed similar amounts of calf starter (1,019 vs. 1,056 g, respectively; P = 0.32), hay (121.5 vs. 126.3 g, respectively; P = 0.30), and water (5.2 vs. 5.0 kg, respectively; P = 0.54). At weaning (d 49) and postweaning (d 70), BW was greater (P = 0.03 and P = 0.02, respectively) in calves fed WM than in those fed MR. Mean DMI from both solid and liquid feeds was not affected by the treatments during the preweaning (P = 0.21) and postweaning (P = 0.16) periods. Body weight gain efficiency during preweaning and overall was improved (P = 0.001 and P = 0.002, respectively) in calves fed WM than in those fed MR. Health (days scoured, rectal temperature, respiratory score, and general appearance score) and serum chemistry variables (glucose, total protein, urea N, nonesterified fatty acids, and creatinine) in calves were not affected (P >or= 0.12 and P >or= 0.12, respectively) by the treatments. Even though gross composition of the MR and WM was similar, growth was greater in calves fed WM. Calves fed WM consumed similar amounts of DM and were heavier than those fed MR, probably because of better bioavailability (digestion and assimilation) of nutrients and availability of some unknown growth factors from WM.