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Development of a new quantification method of <i>Sarcocystis cruzi</i> through detection of the acetyl-CoA synthetase gene

Sarcocystis cruzi is a member of the genus Sarcocystis, infecting bovine animals such as cattle and bison as intermediate hosts, and canids such as dogs and raccoon dogs as definitive hosts. Acute sarcocystosis of S. cruzi causes occasional symptoms in cattle, including weight loss, reduced milk production, abortions, and death, and similar to other Sarcocystis species can potentially cause food poisoning in humans when raw or undercooked infected cattle meat is consumed. Despite these issues, genetic information on S. cruzi is scarce, and there is no specific quantitative method for the detection and quantification of the parasite in infected cattle. In this study, we aimed to develop a method based on high-throughput sequencing of S. cruzi genome and transcriptome that specifically and quantitatively detects the S. cruzi acetyl-CoA synthetase gene (ScACS). Cardiac muscles were collected from slaughterhouses in Saitama Prefecture to obtain sarcocysts from which DNA and RNA were extracted for the high-throughput sequencing. Using the sequences, we developed a specific quantitative PCR assay which could distinguish S. cruzi ACS from that of Toxoplasma gondii by taking advantage of the differences in their exon/intron organizations and validated the assay with the microscopic counting of the S. cruzi bradyzoites. Thus, this assay will be useful for future studies of S. cruzi pathogenesis in cattle and for the surveillance of infected animals, thereby easing public health concerns.

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Morphology of aortic arch branching patterns in the Eurasian Otter (<i>Lutra lutra</i>, Linnaeus, 1758)

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.

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Reconstruction of a partial esophageal defect using tunica vaginalis and buccal mucosa autograft: an experimental study in mongrel dogs

In veterinary clinics, esophageal reconstruction is essential in many clinical situations. In this study, two autografts, the tunica vaginalis (TV) and the buccal mucosa (BM), were proposed to reconstruct a semi-circumferential cervical esophageal defect in dogs. This study aimed to verify whether these two grafts could successfully patch esophageal defects. Twelve male mongrel dogs were divided into two groups. Following cervical esophagoplasty, the defective area was patched with either a TV or a BM graft. Comprehensive clinical, serum biochemical, and histological analyses were performed to evaluate the two grafts. Throughout the study (120 days), the dogs survived the procedure well with minor complications. The lumen of the patched areas was covered with mucosa, with slight scar retraction. Compared with that of the natural esophagus, the average relative luminal diameter was not significantly decreased. Importantly, the measured cortisol and inflammatory marker levels returned to the preoperative levels after 14 days. Although histological examination revealed that both grafts repaired the esophageal defect with complete re-epithelialization, the BM graft showed a histological structure similar to that of the natural esophagus. Both grafts effectively repaired the esophageal defect with minor complications; therefore, both are recommended as promising low-cost clinical alternatives for cervical esophagoplasty in dogs.

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Molecular detection and phylogenetic analysis of lymphocytic choriomeningitis virus in ticks in Jilin Province, northeastern China

Lymphocytic choriomeningitis virus (LCMV) is a neglected rodent-borne zoonotic virus in the genus Mammarenavirus and family Arenaviridae, that can cause aseptic meningitis in humans. A recent study identified infectious LCMV in ticks in northeastern (NE) China. To explore the distribution of LCMV, we determined the prevalence and genetically characterized LCMV in ticks in Jilin Province, NE China. Ticks collected in Huadian, Dunhua, and Jiaohe were pooled and LCMV was detected using reverse transcription polymerase chain reaction. The complete genomes of the LCMV-positive pools were amplified and used for phylogenetic analysis. A total of 1679 questing and engorged ticks were collected and divided into 170 pools, including Ixodes persulcatus (5%), Dermacentor silvarum (89%), and Haemaphysalis japonica (6%). Twenty-four pools of D. silvarum (14.9%, 95% CI:9.5-22.3) and three pools of H. japonica (36.3%, 95% CI:9.8-99.5) collected from cattle were LCMV-positive. No I. persulcatus pools were identified as LCMV-positive. Two complete genome sequences (strains JL-DH01 and JL-DH02) were successfully amplified. They had nucleotide identities of 96.4-99.8% with strains JX31, JX14, DH46, and JX4 identified in ticks from Jilin Province. Phylogenetic analysis indicated that JL-DH01 and JL-DH02 clustered with Jilin strains in the same branch and belonged to genotype I. The findings add to the knowledge of the genetic diversity and geographical distribution of LCMV in ticks in NE China.

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Effects of burdock inulin-type fructans exposure on the physiological function of healthy mice and their filial generation

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.

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Antimicrobial resistance profiles of <i>Campylobacter jejuni</i> and <i>Salmonella</i> spp. isolated from enteritis patients in Japan

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 blaCMY-2. The results of this study would help select antimicrobials for treating patients with Campylobacter and Salmonella enteritis.

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Substrain differences in Sry expression at the stage of sex determination in C57BL/6 mouse strains

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)-XYPOS), 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.

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