Histopathology of incidental findings in cynomolgus monkeys ( macaca fascicularis ) used in toxicity studies

The purpose of our publication is to widely communicate pictures of spontaneous findings occurring in cynomolgus monkeys. Focal lymphoplasmacytic infiltration is commonly seen in the general organs. The frequency and severity of these lesions may be influenced by the administration of drugs with an effect on the immune system. Lymphoplasmacytic infiltration in the lamina propria of the stomach is also frequently seen in cynomolgus monkeys, and it is caused mainly by a Helicobacter pylori infection. Various degrees of brown pigments are observed in various organs, and it is possible to distinguish the material of the pigments by its morphological features and site. A focal/segmental glomerular lesion is occasionally seen in a section of the kidney, and the minimal lesion has no influence on the urinalysis. We showed the common glomerular lesions in HE-stained sections, as well as in PAM- or PAS-stained sections, for understanding the details. Young and pubertal monkeys are usually used in toxicity studies; therefore, understanding various maturation stages of the genital system is important. In particular, the female genital system needs to be understood in the morphology, because their cyclic changes are different from other laboratory animals. Thus, we present the normal features of the cyclic changes of the female genital organs. Furthermore, we provide more information on spontaneous findings in cynomolgus monkeys for exact diagnoses in toxicity studies.     

Population structure of pigs determined by single nucleotide polymorphisms observed in assembled expressed sequence tags

We have collected more than 190 000 porcine expressed sequence tags (ESTs) from full‐length complementary DNA (cDNA) libraries and identified more than 2800 single nucleotide polymorphisms (SNPs). In this study, we tentatively chose 222 SNPs observed in assembled ESTs to study pigs of different breeds; 104 were selected by comparing the cDNA sequences of a Meishan pig and samples of three‐way cross pigs (Landrace, Large White, and Duroc: LWD), and 118 were selected from LWD samples. To evaluate the genetic variation between the chosen SNPs from pig breeds, we determined the genotypes for 192 pig samples (11 pig groups) from our DNA reference panel with matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry. Of the 222 reference SNPs, 186 were successfully genotyped. A neighbor‐joining tree showed that the pig groups were classified into two large clusters, namely, Euro‐American and East Asian pig populations. F‐statistics and the analysis of molecular variance of Euro‐American pig groups revealed that approximately 25% of the genetic variations occurred because of intergroup differences. As the F_IS values were less than the F_ST values_, the clustering, based on the Bayesian inference, implied that there was strong genetic differentiation among pig groups and less divergence within the groups in our samples.     

Commensal bacteria coated by secretory immunoglobulin A and immunoglobulin G in the gastrointestinal tract of pigs and calves

A large amount of secretory immunoglobulin A (S‐IgA) is secreted in the alimentary tract of mammals. It has been reported that S‐IgA coats a portion of commensal intestinal bacteria in human and mouse. However, S‐IgA‐coated bacteria have not been studied in pigs and calves. In this study, we evaluated the distribution of S‐IgA‐coated commensal intestinal bacteria in each portion of the gastrointestinal tracts of pigs and calves. Immunoglobulin G (IgG)‐coated bacteria were also analyzed because a considerable amount of IgG is secreted in the gastrointestinal tracts of pigs, and in particular, calves. S‐IgA‐ or IgG‐coated bacteria were detected in all the segments of the gastrointestinal tracts of pigs and calves. The proportion of S‐IgA‐coated bacteria to total bacteria (i.e. S‐IgA coating ratio) varied in the segments of the gastrointestinal tract in pigs, whereas those of calves were nearly the same throughout the gastrointestinal tract. The S‐IgA and IgG coating ratios were higher in pigs than in calves for all segments of the gastrointestinal tract.     

Carbon balance in a cool–temperate deciduous forest in northern Japan: seasonal and interannual variations, and environmental controls of its annual balance

We monitored variation in seasonal and annual net ecosystem production (NEP), gross primary production (GPP), and ecosystem respiration (R _E) based on 7-year eddy covariance measurements above a cool?Ctemperate deciduous broad-leaved forest (Japanese beech forest). The 7-year means (±SD) of annual NEP, GPP, and R _E were 312?±?64, 1250?±?62, and 938?±?36?g?C?m^?2?year^?1, respectively. Variation in NEP was much larger than variation in GPP and R _E. During the growing season, the main factor controlling carbon balance was air temperature; variation in seasonal integrated NEP was regulated by accumulated air temperature (degree-day) with a significant negative correlation, whereas the seasonal ratio of R _E to GPP was correlated positively with accumulated air temperature. Because the deviation of seasonal NEP was also significantly correlated with seasonal R _E/GPP, NEP was controlled by R _E/GPP, depending on air temperature during the growing season. Seasonal R _E in the defoliation and snow seasons was also important for evaluating the annual carbon balance, because the total number of days in the two seasons was quite large owing to a long snowy winter. In the defoliation and snow seasons, we found defoliation season length was a major factor determining seasonal integrated R _E, illustrating the positive correlation between R _E and defoliation season length. The major factors controlling interannual variations in forest carbon balance are discussed.     

Network simulation modeling of equine infectious anemia in the non-racehorse population in Japan

An equine infectious anemia (EIA) transmission model was developed by constructing a network structure of horse movement patterns in a non-racehorse population. This model was then used to evaluate the effectiveness and efficiency of several EIA surveillance strategies. Because EIA had not been detected in Japan since 1993, it was appropriate to review the current surveillance strategy, which aims to eradicate EIA by intensive testing, and to consider alternative strategies suitable for the current EIA status in Japan.The non-racehorse population was divided into four sectors based on horse usage: the equestrian sector, private owner sector, exhibition sector, and fattening sector. To evaluate the risk of disease spread within and between sectors accompanied by horse movements, a stochastic individual-based network model was developed based on a previous survey of horse movement patterns. Surveillance parameters such as targeting sectors and frequency of testing were added into the model to compare surveillance strategies.The disease spread heterogeneously among sectors. Infection occurred mainly in the equestrian sector; the infection was less disseminated in other sectors. Therefore, we considered that the equestrian sector posed a higher risk of disease dissemination within and between sectors through horse movements. However, surveillance strategies targeting only the equestrian sector were not effective enough for early detection of the disease. Alternatively, targeting horses that moved permanently and those in the private owner sector in addition to the equestrian sector is recommended to achieve effectiveness equivalent to that of the current surveillance. In terms of surveillance efficacy, by increasing the testing interval (once yearly to once every 3 years), this testing scheme could reduce the number of tested horses to 44% of the current surveillance, while maintaining almost equivalent effectiveness. Intensive strategies targeting high-risk populations are considered to enhance effectiveness and efficiency of surveillance. The approach in this study may be helpful in the decision-making process that is involved in setting up strategies for risk-based surveillance.     

Genetic detection of Babesia bigemina from Mongolian cattle using apical membrane antigen-1 gene-based PCR assay

We developed a new nested PCR (nPCR) assay based on the Babesia bigemina apical membrane antigen-1 (AMA-1) gene sequence for parasite-specific detection. The primers were designed to amplify 738-bp and 211-bp fragments of the AMA-1 gene by primary and nested PCRs, respectively. The assay was proven to be specific for the B. bigemina, whereas the previously established SpeI-AvaI nPCR assay amplified not only the target fragment of B. bigemina but also a homologous one from Babesia ovata. The AMA-1 nPCR assay was also evaluated using field DNA samples extracted from 266 bovine blood samples collected from Mongolia in 2010. In a comparative evaluation, 90 (33.8%) and 25 (9.4%) of the blood samples showed positive reactions for B. bigemina by the SpeI-AvaI nPCR and AMA-1 nPCR assays, respectively. The sequencing analysis of the nPCR products confirmed that the AMA-1 nPCR method had specifically detected the target B. bigemina DNA. However, 4 different kinds of sequences were determined among the SpeI-AvaI nPCR amplicons. Two of them were derived from B. bigemina and B. ovata, while the origins of the others were unknown. In the current study, the presence of B. bigemina was clearly demonstrated among Mongolian cattle populations by the current nPCR assay for the first time. Furthermore, our findings also indicate that the AMA-1 nPCR assay may be a useful diagnostic tool for the specific detection of B. bigemina.     

Plasmid-mediated florfenicol resistance in Mannheimia haemolytica isolated from cattle

The aim of this study was to analyse a florfenicol-resistant Mannheimia haemolytica isolated from a calf to determine the genetic basis of its florfenicol-resistance. The antimicrobial susceptibility and plasmid content of the isolate were determined. A florfenicol resistant plasmid carrying the floR gene was identified by PCR and transformed into Escherichia coli JM109 and HB101 strains. The plasmid was then mapped and sequenced completely. The isolate was resistant to chloramphenicol, florfenicol, oxytetracycline, kanamycin, dihydrostreptomycin, nalidixic acid, ampicillin, and amoxicillin; it carried a floR plasmid of 7.7kb, designated pMH1405. The mobilisation and replication genes of pMH1405 showed extensive similarity to the 5.1-kb pDN1 plasmid from Dichelobacter nodosus and the 10.8-kb pCCK381 plasmid from Pasteurella multocida. An adjacent 2.4-kb segment was highly homologous to the TnfloR region of the E. coli BN10660 plasmid. A plasmid-mediated floR gene was responsible for florfenicol resistance in the bovine respiratory tract pathogen M. haemolytica. The pMH1405 plasmid is the smallest floR-carrying plasmid reported to date. To the best of our knowledge, this is the first report of a florfenicol-resistant gene in M. haemolytica.     

Essential thrombocythemia in a dog

A two-year old male Welsh Corgi was referred for persistent thrombocytosis and occasional seizure. Hematological findings indicated marked thrombocytosis, eosinophilia, basophilia and moderate anemia. Bone marrow examination revealed marked megakaryocytic hyperplasia with morphologic abnormality. A diagnosis of essential thrombocythemia was made and the treatment was initiated with combination chemotherapy and maintained by prednisolone and busulfan. The dog successfully achieved complete remission on 100 days after initial presentation and has been good in health without chemotherapy since then.