Nosocomial Pneumocystis jirovecii infections

Airborne transmission of Pneumocystis sp. from host to host has been demonstrated in rodent models and several observations suggest that interindividual transmission occurs in humans. Moreover, it is accepted that the Pneumocystis organisms infecting each mammalian species are host specific and that the hypothesis of an animal reservoir for Pneumocystis jirovecii (P. jirovecii), the human-specific Pneumocystis species, can be excluded. An exosaprophytic form of the fungus cannot be strictly ruled out. However, these data point toward the potential for the specific host to serve as its own reservoir and for Pneumocystis infection in humans as an anthroponosis with humans as a reservoir for P. jirovecii. This review highlights the main data on host-to-host transmission of Pneumocystis in rodent models and in humans by the airborne route and provides a rationale for considering the occurrence of nosocomial infections and measures for their prevention     

实验鼠群的健康监测管理

虽然实验鼠种群的总体健康水平得到了显著的提高,但还是有很多重要的传染性病原体在实验鼠群中流行.实验鼠群的健康状况对于动物福利、科学研究甚或人类健康都很重要.所以必须要对实验鼠群进行健康监测.笔者依据自身的实践和经验,对鼠群健康监测的基本原理进行了分析,包括以下的内容：鼠群健康监测的必要性、健康检测规程的建立、哨兵鼠、常用的监测力法、样本采集原则、对检测结果的解读和应对.相信读者在了解这些基本的原理后,能设定出符合自身设施实际的鼠群健康监测计划并逐步在实践中完善.     

Pneumocystis carinii--animal production perspective.

Pneumocystis carinii is an important cause of pneumonia in immunocompromised human patients. The organism is also found as a saprophyte in the lungs of many species of animals. Animal models have been used as a source of P. carinii organisms for study of the disease. The rat model has been especially useful. Initially, the infection was latent in most colonies, and P. carinii pneumonia readily developed when animals were immunosuppressed. Today, many barrier raised rodent colonies are free of adventitious viruses, bacteria, Mycoplasma sp., and parasites, including P. carinii. Variability is now seen in the rat model. The use of cultured organisms to experimentally infect rats and mice prior to immunosuppression has met the need for some investigators, however, latent-infected, barrier-raised and isolator-raised rodents are still required. Colonies specifically infected with P. carinii can provide latent-infected animals and are better protected from potentially interfering organisms than barrier-raised animals. The development of these colonies is feasible as investigators and animal producers work together to define and develop this resource.     

Molecular phylogeny of Pneumocystis based on 5.8S rRNA gene and the internal transcribed spacers of rRNA gene sequences

To clarify the phylogenetic relationships and species status of Pneumocystis, the 5.8S rRNA gene and the internal transcribed spacers (ITS, 1 and 2) of Pneumocystis rRNA derived from rat, gerbil and human were amplified, cloned and sequenced. The genetic distance matrix of six Pneumocystis species compared with other fungi like Taphrina and Saccharomyces indicated that the Pneumocystis genus contained multiple species including Pneumocystis from gerbil. The phylogenetic tree also showed that Pneumocystis from human and monkey formed one group and four rodent Pneumocystis formed another group. Among the four members, Pneumocystis wakefieldiae was most closely related to Pneumocystis murina and Pneumocystis carinii, and was least related to gerbil Pneumocystis.     

Detection, eradication, and research implications of Helicobacter infections in laboratory rodents

Researchers first isolated and characterized Helicobacter hepaticus in 1994 as a cause of hepatitis that progressed to hepatocellular carcinoma in A/JCr mice. During the past decade, isolation and characterization of additional novel helicobacters from rodents has continued. In addition to causing overt disease, rodent helicobacter infections are important because intercurrent disease in select models will confound research data. Emerging evidence suggests that inflammatory responses to enterohepatic helicobacter infections may alter host responses to other experimental stimuli in unanticipated ways. Additionally, scientists have experimentally infected a variety of inbred mouse strains and genetically engineered mice with a variety of Helicobacter spp. isolated from rodents, birds, and higher mammals (including humans) to develop animal models of gastrointestinal diseases as well as idiopathic human disease syndromes. This review highlights current information about helicobacter infections in laboratory rodents and provides recommendations for the detection and eradication of these infections. The authors discuss the impact of subclinical and clinical disease and offer recommendations for managing helicobacter-free rodent colonies.     

Geographic variation in rodent-flea relationships in the presence of black-tailed prairie dog colonies

We characterized the relationship between fleas and their rodent hosts in the presence of prairie dog colonies and compared them to adjacent assemblages away from colonies. We evaluated the rodent-flea relationship by quantifying prevalence, probability of infestation, flea load, and intensity of fleas on rodents. As prairie dog burrows provide refugia for fleas, we hypothesized that prevalence, flea load, and intensity would be higher for rodents that are associated with black-tailed prairie dog colonies. Rodents were trapped at off- and on-colony grids, resulting in the collection of 4,509 fleas from 1,430 rodents in six study areas. The rodent community composition varied between these study areas. Flea species richness was not different between prairie dog colonies and the surrounding grasslands (p = 0.883) but was positively correlated with rodent species richness (p = 0.055). Prairie dog colonies did not increase the prevalence of fleas (p > 0.10). Flea loads on rodents did not vary between off- and on-colony grids at three of the study areas (p > 0.10). Based on the prevalence, infestation rates, and flea loads, we identified Peromyscus maniculatus, Onychomys leucogaster, and two Neotoma species as important rodent hosts for fleas and Aetheca wagneri, Orchopeus leucopus, Peromyscopsylla hesperomys, Pleochaetis exilis, and Thrassisfotus as the most important fleas associated with these rodents. Prairie dog colonies did not seem to facilitate transmission of fleas between rodent hosts, and the few rodent-flea associations exhibited significant differences between off- and on-colony grids.     

A serological survey of accredited breeding colonies in the United Kingdom for common rodent viruses.

This paper reports the results of a general survey, the first in the United Kingdom, carried out on accredited breeding colonies of mice, rats and guinea-pigs over a period of a year. While the results show the potential usefulness of a viral accreditation grading scheme, they also show that contamination of breeding colonies with inapparent viral infections is widespread. This situation can only be improved by the continuous monitoring of animal stocks for rodent viruses, with the aim of improving the standard of animals available for research and for pharmacological, toxicological and routine diagnostic procedures.     

Prevalence of naturally occurring viral infections, Mycoplasma pulmonis and Clostridium piliforme in laboratory rodents in Western Europe screened from 2000 to 2003

In this report prevalence rates of rodent viruses in laboratory animals are presented based on routine serological screening of mouse and rat colonies from European institutes. The prevalences found during the period 2000-2003 are compared with those reported for 1981-1984 and 1990-1993. It is shown that some infections were eliminated from laboratory animal colonies (e.g. K-virus and polyomavirus) by taking preventative measures whereas other infections such as mouse hepatitis virus and parvoviruses remained at a high rate. Further decreases in prevalence rates in the last 10 years were found for infections such as pneumonia virus of mice, reovirus type 3, Sendai virus, sialodacryoadenitis/rat coronavirus and Mycoplasma pulmonis. The introduction of new detection methods showed that mouse parvovirus and rat parvovirus, both members of the Parvoviridae family, remain a major threat to laboratory mice and rats. Guinea pig cytomegalovirus and para-influenza virus appeared to be the most prevalent agents among laboratory guinea pigs. The importance of a standardized, up-to-date screening programme is discussed.     

Factors influencing Pneumocystis infection in the immunocompromised rat.

A compromised immune system is the primary predisposing condition for Pneumocystis infection. Factors that contribute to this underlying state of immunosuppression are poorly understood. The presence of common rodent viruses and the role of anti-Pneumocystis antibodies on the progression of natural infection in the corticosteroid-treated rat model of Pneumocystis pneumonia were evaluated. The development and intensity of infection were not affected by the presence or absence of antibodies to these viruses or to major Pneumocystis antigens. A significant increase in survival of Pneumocystis-infected viral antibody-positive rats was observed when these rats were housed under barrier conditions.     

New building, old parasite: Mesostigmatid mites--an ever-present threat to barrier facilities

Mesostigmatid mites are blood-sucking parasitic mites found in wild rodent populations. Periodically they can also become a problem for laboratory rodent colonies, particularly when building construction or renovations disturb colonies of commensal (building) rodents that had been acting as hosts. Mesostigmatid mites infest both rats and mice and, unlike the more common rodent fur mites (Myobia, Myocoptes, and Radfordia sp.), can survive for long periods in the environment and travel considerable distances in search of new hosts. They easily penetrate barrier caging systems, including individually ventilated cages, thus circumventing the usual precautions to protect rodents from infection. The two mites reported in laboratory rodent colonies, Ornithonyssus bacoti and Laelaps echidnina, also bite humans and have the potential to transmit zoonotic diseases. Once the mites gain access to a colony, eradication requires elimination of commensal rodent reservoirs in addition to insecticide treatment of both the laboratory rodents and the environment. In view of the undesirability of insecticide use in the animal facility, it is advisable to investigate the effectiveness of preventive treatments, such as environmental application of insect growth regulators or silica-based products. This article summarizes available information on mesostigmatid mites and their laboratory incursions, and provides suggestions for diagnosis, treatment, and control based on the authors experience with several outbreaks at a large academic institution..     

A comparison of modified methenamine silver and toluidine blue stains for the detection of Pneumocystis carinii in bronchoalveolar lavage specimens from immunosuppressed patients.

Pneumonia due to Pneumocystis carinii is an increasingly frequent occurrence; a prompt, accurate diagnosis is important to successfully manage this infection. Methenamine silver and toluidine blue stainings of material recovered by bronchoalveolar lavage (BAL) have been the most widely employed techniques for detecting Pneumocystis organisms. The value of these two staining techniques for the detection of Pneumocystis organisms was prospectively compared in 220 BAL specimens obtained from 186 immunosuppressed individuals. The patients included those with disease-related and therapy-related immunosuppression; half of the BAL specimens came from organ transplant recipients. The results indicate that neither method is superior to the other and that both will correctly identify almost all Pneumocystis infections if a sufficient aliquot of BAL material is examined systematically by trained individuals. The toluidine blue method is somewhat simpler to perform, however. The reasons for discrepant results between the two staining methods were analyzed.     

The characteristics of the course of Leishmania and Pneumocystis opportunistic infections in immunosuppressed laboratory animals

Specific features of the course of Leishmania and Pneumocystis infections were studied on experimental models. Laboratory animals were subjected to medicinal suppression with Tricort-40. Leishmania infection in naturally susceptible animals was considerably aggravated (in comparison with the controls) in the presence of even short-term immunosuppression. Pneumocystis infection developed in experimental animals under the effect of double suppression. The process of the reactivation of Pneumocystis infection was influenced by Leishmania infection, rapidly developing in the presence of prolonged immunodeficiency. The results thus obtained make it possible to regard Leishmania infection in animals with reactivated Pneumocystis infection as co-infection having the properties of a biological, immunosuppressant. Such situation may be extrapolated on HIV/visceral leishmaniasis co-infection.     

两个封闭群SPF级昆明小鼠遗传背景调查

目的：比较两个封闭群SPF级昆明（ KM）小鼠的遗传差异,调查引进的SPF级KM小鼠封闭繁殖6年后,其遗传构成是否发生变化。方法应用微卫星DNA标记方法对18个位点在两个群体中的遗传差异进行分析,主要包括观察等位基因数（ Na）、有效等位基因数（ Ne）、观察杂合度（ Ho）、期望杂合度（ He）、多态信息含量（PIC ）、Shan-non信息指数、遗传分化系数（ F st）、遗传距离等遗传参数。结果两个封闭群SPF 级KM小鼠在18个微卫星位点共发现67个等位基因,Na为2～8个,平均3．7222个;Ne为1．9459～6．5442,平均2．7966个;Ho为0．4225～1．0000,平均0．8823;He为0．4892～0．8527,平均0．6162;Shannon信息指数0．6792～1．9526,平均1．0598;PIC为0．3680～0．8301,平均0．5317;Fst平均值为0．0159,表明群体间的遗传差异仅1．59％,二者间的遗传距离（DA）为0．0499。结论两个封闭群SPF级KM小鼠遗传结构相似度极高,它们与原引进群体的分化差异极小。     

Opportunistic infections of mice and rats: Jacoby and Lindsey revisited

Adventitious infections among rodents used in biomedical research and teaching continue to be problematic even with improved housing and disease-deterrent methodologies. In addition to well-documented viral diseases (e.g., mouse hepatitis virus and rodent parvoviruses) and parasites (mites and pinworms), new pathogens such as murine norovirus have emerged in recent years. Infectious agents can enter colonies via incoming rodent shipments, in unscreened biological materials, on people (especially husbandry or investigative staff) who move from a location where animals have a lower health status to an area where health status is higher and operational procedures are more stringent, or by introduction of contaminated food, bedding material, or other fomites. These factors, coupled with the very high volume of movement of rodents within and between institutions, increase the risk of spreading infectious agents. The challenge to the laboratory animal community is to implement control measures that halt the passage of these organisms from one location to another while still enabling collaborative scientific discovery to proceed with minimal disruption. It is therefore critical to make appropriate decisions about identifying outbreaks in a timely fashion and controlling the spread of infection once identified. Such efforts should be practical, reproducible, and cost-effective.     

The piglet as a potential model of Pneumocystis carinii pneumonia.

Spontaneous Pneumocystis carinii infections occur in piglets. In this report we describe the symptoms, pathology and predisposing conditions of P. carinii pneumonia in the pig. We also discuss the advantages and disadvantages of the pig as an experimental system to study P. carinii pneumonia.     

Multilocus Genotyping of Pneumocystis jirovecii in Patients Developing Diverse Forms of Parasitism: Implication for a Wide Human Reservoir for the Fungus

ABSTRACT. Pneumocystis jirovecii ITS and DHPS genotypes were identified in 3 patient groups developing diverse forms of P. jirovecii infections: 13 patients with Pneumocystis pneumonia, 8 patients merely colonized by the fungus, and 19 immunocompetent infants with bronchiolitis developing mild P. jirovecii infection. Common P. jirovecii genotypes were found in the 3 patient groups, suggesting that common sources of P. jirovecii were involved in the fungus acquisition, and that transmission cycles of P. jirovecii infections in these patient groups are not independent. Parasitized patients, whatever the form of parasitism they present, may he part of a common reservoir for P. jirovecii.     

Comparison of methods for detection of pinworms in mice and rats

Though pinworm infestation remains common in laboratory rodent colonies, there is little information regarding current practices for pinworm detection and their relative efficacy. The authors surveyed research institutions to find out the prevalence of pinworm infestations and the detection methods they used. They also tested mice and rats from colonies that were known to be infested with Syphacia sp. and compared the following detection methods: perianal tape test, fecal flotation, fecal concentration, cecal content examination, cecal flotation and histological examination. Though the different methods yielded comparable efficacy overall, the authors recommend using more than one type of test to increase detection potential.     

Studies of gene transfer and reversion to mitomycin C resistance in Fanconi anemia cells

As a first step to the cloning of the Fanconi anemia (FA) gene, we have attempted to correct the sensitivity of FA cells to DNA crosslinking agents by the introduction of wild-type DNA. The protocol involved the introduction of both genomic and pRSVneo DNA, selection for G418-resistant colonies and the subsequent selection of mitomycin C-resistant cells from the latter. Preliminary experiments indicated that untransformed FA cells were not suitable recipients for the introduction of foreign DNA, so all experiments were performed with an SV40-transformed FA cell line. Approximately 40,000 G418-resistant colonies were obtained in 5 separate experiments at an overall frequency of about 5 X 10(-4). These were then selected in mitomycin C and 15 colonies were recovered. Colonies were obtained with wild-type DNA (both human and rodent) and with FA DNA at about the same frequency of 2 X 10(-7). Colonies were isolated and shown to have a stable, partial (from 25 to 90% of wild-type) resistance to mitomycin C. One colony was also shown to be partially resistant to two other DNA crosslinking agents, diepoxybutane and nitrogen mustard. This clone also had an intermediate level of spontaneous and MMC-induced chromosome aberrations. pRSVneo, but not rodent, DNA could be demonstrated in the high molecular weight fraction of several colonies. Thus, it is likely that these colonies represent partial revertants rather than transfectants. These mitomycin C-resistant FA cells should be useful for the biochemical analysis of the FA mutation.     

Identification and Characterization of an Endo/exonuclease in Pneumocystis carinii that is Inhibited by Dicationic Diarylfurans with Efficacy Against Pneumocystis Pneumonia

ABSTRACT Dicationic diarylfurans and dicationic carbazoles are under development as therapeutic agents against opportunistic infections. While their ability to bind to the minor groove of DNA has been established, the complete mechanism of action has not. We demonstrate here that an effective diarylfuran, 2,5-bis[4-(N-isopropylguanyl)phenyl]furan. inhibits an endo/exonuclease activity present in Pneumocystis carinii, Cryptococcus neoformans, Candida albicans , and Saccharomyces cerevisiae. This activity was purified from the particulate fraction of P. carinii. The enzyme requires Mg⁺⁺ or Mn⁺⁺, and shows preferences for single- over double stranded DNA and for AT-rich over GC-rich domains. A panel of 12 dicationic diarylfurans and eight dicationic carbazoles, previously synthesized, were evaluated for inhibition of the purified nuclease and for efficacy against Pneumocystis pneumonia in rats. Among the diarylfurans, potency of nuclease inhibition, in vivo antimicrobial activity, and DNA binding strength were all strongly correlated (p < 0.001). These findings suggest that one target for antimicrobial action of the diarylfurans may be a nucleolytic or other event requiring unpairing of DNA strands. Dicationic carbazoles which were strong nuclease inhibitors all displayed anti- Pneumocystis activity in vivo, but there were also noninhibitory carbazoles with in vivo efficacy.     

Evaluation of the association of zoonotic Ljungan virus with perinatal deaths and fetal malformation

More and more epidemiologic and experimental data support the notion that Ljungan virus (LV), originally isolated from some rodent populations in Sweden, Denmark, and the United States, plays an important role in stillbirth and fetal malformation. Mouse dams infected with LV may result in uterine resorption and perinatal deaths that may cross generations, and their offspring may suffer high rates of malformations including cranial, brain, and limb malformations. In humans, researches founded that LV infection is related to malformation, intrauterine fetal death, and even central nervous system malformation. Although molecularly characterized, little is known about the biophysical nature of LV. Consequently, the role of LV infections in sudden infant death syndrome is still confusing, and the mechanism of how LV infections cause diseases is not clear. More research is clearly necessary to explore the mechanisms of LV infection in human and animal diseases to bring improvement to the clinical outcomes. Birth Defects Research (Part C) 105:81–85, 2015. © 2015 Wiley Periodicals, Inc.